CD3L1 expression in osteosarcoma and chordoma: a retrospective analysis of 62 cases

CD3L1 expression in osteosarcoma and chordoma: a retrospective analysis of 62 cases | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article CD3L1 expression in osteosarcoma and chordoma: a retrospective analysis of 62 cases Yuhang Wang, Jiuhui Xu, Shouyan Deng, Kunkun Sun, Yaqi Wang, Tingting Ren, and 3 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6689428/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Purpose CD3L1 (ITPRIPL1), a natural ligand of CD3ε, is implicated in tumor-immune interactions and is overexpressed in several solid tumors. However, its expression and functional significance in osteosarcoma and chordoma remain uncharacterized. This study aimed to evaluate CD3L1 expression in osteosarcoma and chordoma and assess its association with clinical features and outcomes. Methods Formalin-fixed, paraffin-embedded tissue samples from 42 osteosarcoma and 20 chordoma patients were analyzed for CD3L1 expression using immunohistochemistry. Multiplex immunofluorescence staining was performed to evaluate immune cell infiltration in chordoma. Associations between CD3L1 expression and clinicopathological parameters were examined using chi-square tests, Fisher’s exact test, and Kaplan-Meier survival analysis. Results CD3L1 was detected in 62.9% (39/62) of cases, with positivity rates of 59.6% in osteosarcoma and 70.0% in chordoma. CD3L1 expression was independent of patient age, sex, or tumor location but was significantly associated with chondroid stroma and prior PD-1 immunotherapy in osteosarcoma. In chordoma, higher CD3L1 expression correlated with increased infiltration of CD8 + T cells, B cells, and M2 macrophages. Conclusion CD3L1 is expressed in osteosarcoma and chordoma and may play a pivotal role in regulating tumor stroma and the immune microenvironment, representing a potential target for future therapeutic strategies. CD3L1 (ITPRIPL1) Osteosarcoma Chordoma Immunohistochemistry Prognosis Figures Figure 1 Figure 2 Figure 3 Figure 4 Figure 5 Introduction Osteosarcoma is the most common primary bone malignancy in adolescents and young adults, and it is characterized by extensive genomic and transcriptomic heterogeneity, largely resulting from chromothripsis. This intratumoral heterogeneity makes the disease difficult to target, and combination therapy has become the mainstay of systemic treatment strategies [ 1 ]. Structural alternations, such as genetic amplifications, translocations, single nucleotide insertions or deletions, and loss of heterozygosity, can activate oncogenes or inactivate tumor suppressor genes, contributing to osteosarcoma pathogenesis [ 2 ]. Despite these insights, the precise mechanisms underlying osteosarcoma remain incompletely understood. Large-cohort studies have highlighted pervasive genomic instability and heterogeneity, necessitating treatment approaches that traditionally rely on cytotoxic and cytostatic agents. A typical first-line chemotherapy regimen includes a three-drug combination, which yields an overall 5-year event-free survival (EFS) rate of 65% [ 3 ]. Among patients with non-metastatic osteosarcoma, the 5-EFS is approximately 70%, compared to 30% in those metastatic disease [ 4 ]. Chordoma is a rare malignant bone tumor derived from notochord remnants, accounting for approximately 8.4% of all bone cancer types [ 1 ]. Although typically classified as histologically low-grade, chordomas exhibit a high rate of local recurrence and are intrinsically resistant to both chemotherapy or targeted therapies. This resistance is attributed to their pathological architecture, with infiltrative and lobulated tumor cells separated by fibrous septa. The transcription factor Brachyury, a T-box gene regulating notochord development, is a sensitive and specific immunohistochemical (IHC) marker for chordoma. Brachyury amplification is commonly observed in familial chordoma, while in sporadic cases, it is more often associated with whole-chromosome gains rather than focal duplications [ 5 ]. Chordomas most frequently arise in the sacrum, skull base, and mobile spine. The current standard of care for tumors in the mobile spine and sacrum includes en bloc resection with wide margins, followed by postoperative external-beam radiation therapy [ 6 ]. According to the SEER database, the median overall survival is 6.29 years, with 5, 10, and 20-year survival rates of 67.6%, 39.9%, and 13.1%, respectively [ 5 ]. CD3L1, also known as ITPRIPL1 (inositol 1,4,5-triphosphate receptor-interacting protein-like 1), was recently identified as a natural ligand of CD3ε, and is known to downregulate T cell activation, thereby facilitating immune evasion by tumors [ 7 ]. Its overexpression has been observed in non-small cell lung cancer (NSCLC), where it correlates with advanced disease stages [ 8 ]. However, to date, the expression and functional relevance of CD3L1 in bone tumors such as osteosarcoma and chordoma have not been reported. This study aims to characterize CD3L1 expression in osteosarcoma and chordoma, and to explore its association with clinicopathological features and patient prognosis, thereby providing rationale for future clinical trials. Materials and methods Patients and samples This study was approved by the Institutional Review Board of Peking University People’s Hospital (No. 2024PHB013-001) and was supported by the Beijing Natural Science Foundation (Nos. 2127000356 and QY24058). Clinical data and tissue microarrays were collected from 42 patients with osteosarcoma and 20 patients with chordoma who were treated at the Musculoskeletal Tumor Center of Peking University People’s Hospital (PKUPH) between June 1, 2021 and July 1, 2024. The inclusion criteria were as follows: (i) Histologically confirmed high-grade osteosarcoma or refractory chordoma. (ii) Presence of evaluable lesions defined by the Response Evaluation Criteria in Solid Tumors (RECIST 1.1) [ 9 ], with documented disease progression after standard therapy. Osteosarcoma patients had progressed following first-line MAP/I chemotherapy with target lesions present; chordoma cases had experienced two or more relapses and were deemed unsuitable for further surgical or radiation intervention, and (iii) Availability of sufficient formalin-fixed, paraffin-embedded (FFPE) tissue for analysis. The exclusion criteria were: (i) Incomplete clinical data, (ii) Presence of other malignancies or serious comorbid conditions, and (iii) Lost to follow-up during the observation period. Written informed consent was obtained from all of the patients or their legal guardians. Response evaluations was conducted per RECIST 1.1 guidelines, and clinical follow-up occurred every two months until the patient’s death. Demographic characteristics of enrolled patients are summarized in Tables 1 and 2 . Table 1 Association between CD3L1 expression and clinicopathological features of patients with osteosarcoma Factors Total no. % CD3L1 expression p value Negative % Low % Medium % Sex 42 100 17 40.4 22 52.4 3 7.1 Male 27 64.3 8 29.6 17 63.0 2 7.4 Female 15 35.7 9 60.0 5 33.3 1 6.7 0.171 Age group < 35 years 39 92.9 16 41.0 20 51.3 3 7.7 ≥ 35 years 3 7.1 1 33.3 2 66.7 0 0 1.000 Specimen source From bone 20 47.6 9 45.0 8 40.0 3 15.0 From lung 20 47.6 8 40.0 12 60.0 0 0 From other tissue 2 4.8 0 0 2 100 0 0 1.000 Multinucleated giant cell Yes 15 35.7 6 40.0 9 60.0 0 0 No 27 64.3 11 40.7 13 48.1 3 11.1 0.558 Giant cell tumor Yes 17 40.5 7 41.2 10 58.8 0 0 No 25 59.5 10 40.0 12 48.0 3 12.0 0.503 Chondroid stroma Yes 10 23.8 1 10.0 6 60.0 3 30.0 No 32 76.2 16 50.0 16 50.0 0 0 0.003 After PD-1 treatment Yes 15 35.7 4 26.7 8 53.3 3 20.0 No 27 64.3 13 48.1 14 51.9 0 0 0.049 Radiotherapy Yes 10 23.8 4 40.0 6 60.0 0 0 No 32 76.2 13 40.6 16 50.0 3 9.4 0.872 Therapy line 1 10 23.8 4 40.0 5 50.0 1 10.0 2 24 57.1 10 41.7 12 50.0 2 8.33 ≥ 3 8 19.1 3 37.5 5 62.5 0 0 1.000 Table 2 Association between CD3L1 expression and clinicopathological features of patients with chordoma Characteristics Total no. % CD3L1 expression p value Negative % Low % Medium % Gender 20 100 6 30 11 55 3 15 Male 12 60 3 25 7 58.3 2 16.7 Female 8 40 3 37.5 4 50 1 12.5 0.835 Age < 56 years 11 55 2 18.2 7 63.6 2 18.2 ≥ 56 years 9 45 4 44.4 4 44.4 1 11.1 0.4695 Treatment before surgery Yes 3 15 0 0 2 66.7 1 33.3 No 17 85 6 35.3 9 52.9 2 11.8 0.5368 Invasion Yes 12 60 5 41.7 6 50 1 8.3 No 8 40 1 12.5 5 62.5 2 25 0.408 State Primary 13 65 5 38.5 6 46.2 2 15.4 Recurrence 7 35 1 14.3 5 71.4 1 14.3 0.6807 Mucus matrix Yes 5 25 3 60 2 40 0 0 No 15 75 3 20 9 60 3 20 0.3615 Immunohistochemistry IHC was performed on FFPE tumor sections from patients with osteosarcoma or chordoma ( n = 62). Tissue sections (5 µm thick) were prepared from the blocks and mounted on glass slides. Slides were deparaffinized in xylene and rehydrated through a graded ethanol series. Endogenous peroxidase activity was quenched using 3% hydrogen peroxide (H 2 O 2 ) for 15 minutes, followed by phosphate-buffered saline (PBS) washes. Antigen retrieval was carried out by heating slides in citrate buffer using a microwave for 15 minutes, after which the slides were cooled to room temperature and rinsed in PBS. Tissue sections were incubated overnight at 4°C with anti-CD3L1 primary antibody (BioTroy, Cat. #BT02-CDX-01; 1:100 dilution in 1% Bovine Serum Albumin). The following day, sections were washed and incubated with a secondary antibody (1:100 in 1% BSA) for 30 minutes at room temperature. Detection was performed using the DAB chromogenic method, and slides were counterstained with hematoxylin for 3 minutes, dehydrated, and mounted for microscopy. IHC scoring A board-certified pathologist (SKK), experienced in bone tumor pathology, assessment assessed CD3L1 expression. Tumors were classified as negative (0 ≤ 1% tumor cells stained) or positive (> 1%). Among positive samples, staining intensity was further graded as weak (1–25%), moderate (26–50%), and strong (≥ 50%) [ 10 ]. Multiplex immunofluorescence (mIF) staining To investigate the immune microenvironment of chordoma, Multiplex immunofluorescence (mIF) staining was conducted using a PANO 7-plex IHC kit (Panovue, Cat. #10004100100, Beijing, China) according to the manufacturer’s instructions, as previously described [ 11 ]. Sides were deparaffinized overnight at 65°C, rehydrated with xylene and ethanol, and subjected to antigen retrieval via microwave treatment. The primary antibodies were grouped into two panels: Panel 1 included CD3, CD8A, CD20, PD-L1, and CD56, and Panel 2 included CD68, CD163, CD206, and IRF8 (Supplementary Table S1). Antibodies were incubated for 1 hour at room temperature, followed by 10-minute incubation with HRP-conjugated secondary antibodies. Nuclei were counterstained using DAPI (Sigma-Aldrich). H&E staining and tumor-stroma ratio assessment Slides were deparaffinized and rehydrated following standard protocols, then stained using hematoxylin and eosin. The parenchyma-to-stroma ratio in chordoma samples was interpreted by a dedicated pathologist. Statistical analysis Statistical analyses were conducted using SPSS Statistics 26.0 software (IBM Corp., Armonk, NY, USA) and GraphPad Prism 10. Continuous variables were expressed as mean ± standard deviation (SD), and categorical as frequencies and percentages. Association between CD3L1 expression and clinicopathological parameters were evaluated using chi-square or Fisher’s exact tests. Progression-free survival (PFS) was assessed using Kaplan-Meier survival analysis, and differences between groups were compared using the log-rank test. Statistical significance was set at p < 0.05. Results CD3L1 protein expression in tumor tissues A total of 42 patients with osteosarcoma and 22 with chordoma were enrolled for CD3L1 IHC analysis. Two chordoma samples were excluded due to tissue loss, resulting in 62 valid cases. Among the osteosarcoma patients, the median age was 16 years old (range: 12–59). Of the 42 osteosarcoma samples, 47.6% (20/42) originated from bone, 47.6% (20/42) from pulmonary metastases, and 4.8% (2/42) from lymph node or skin lesions. In the chordoma group, the median age was 56 years old (range: 10–74), and all of the samples were derived from the sacrococcygeal region. Among them, 5% (1/20) were classified as dedifferentiated chordoma and 95% (19/20) as conventional chordoma. Additionally, 65% (13/20) were primary and 35% (7/20) were recurrent cases (Tables 1 and 2 ). CD3L1 protein was detected via brown cytoplasmic staining, with occasional nuclear localization, in IHC (Fig. 1 ). Of the 62 evaluable samples, 39 (62.9%) showed positive expression. CD3L1 was expressed in 70.0% (14/20) of chordoma samples and 59.6% (25/42) of osteosarcoma samples (Table 3 ). Specimens were categorized as negative, low-positive, or moderately positive, with no cases exhibiting high-intensity staining (Fig. 1 ). Table 3 The positive expression of CD3L1 in osteosarcoma and chordoma Cases Positivity Staining intensity Low Moderate High All sarcoma 62 39 (62.9%) 33 (53.2%) 6 (9.7%) 0 Osteosarcoma 42 25 (59.6%) 22 (52.4%) 3 (7.2%) 0 Chordomas 20 14 (70.0%) 11 (55.5%) 3 (15.0%) 0 These results demonstrate that CD3L1 is broadly expressed in both osteosarcoma and chordoma, suggesting its potential as a disease marker. Assessment stability and repeatability of the CD3L1-targeting antibody in chordoma tissue microarrays The CD3L1 humanized antibody, developed and provided by the Shanghai Key Laboratory of Medical Epigenetics [ 7 , 8 ], was tested for stability and reproducibility. Two sequential tissue microarray (TMA) slides from 20 chordoma specimens were stained in parallel using the anti-CD3L1 antibody. The staining patterns between the two slides exhibited high consistency (Supplementary Fig. S1), confirming the excellent stability of the antibody. Association between CD3L1 expression and clinicopathologic characteristics of osteosarcoma and chordoma To assess the clinical relevance of CD3L1, the association between its expression and patient characteristics were analyzed (Tables 1 and 2 ). In osteosarcoma, CD3L1 expression was significantly correlated with the presence of chondroid stroma (Fig. 2 ), a histological hallmark of chondroblastic osteosarcoma. Additionally, a positive association was observed between CD3L1 expression and prior PD-1 immunotherapy ( p < 0.05), suggesting a potential role in modulating immune responses. No significant associations were found with other clinical variables, such as age, sex, or tumor site. In the chordoma cohort, CD3L1 expression did not significantly correlate with gender, age, preoperative treatment, local invasion, disease state, or presence of mucus matrix (Table 2 ). Relationship between CD3L1 expression and immune cells infiltration in chordoma Given the role of CD3L1 in regulating the tumor immune microenvironment [ 12 ], we assessed immune cell infiltration in chordoma using mIF staining (Fig. 3 A). Increased CD3L1 expression was associated with higher infiltration of CD20 + B cells, CD8A + T cells, and CD68 + macrophages. Specifically, CD3L1 expression positively correlated with the presence of CD206 + and CD163 + M2-type macrophages infiltration, suggesting a potential role in macrophage polarization and tumor-associated immune modulation (Fig. 3 B and 3 C). To further assess the clinical relevance of immune infiltration, we assessed its relationship with prognosis. Notably, recurrent chordoma cases exhibited higher levels of CD206 + macrophages, supporting our previous findings that M2 macrophages promote tumor progression in chordoma [ 11 ] (Fig. 4 ). Prognostic significance of CD3L1 expression in osteosarcoma and chordoma To further validate whether CD3L1 expression impacted clinical outcomes, we performed Kaplan–Meier survival analysis using follow-up data from all 62 patients. CD3L1 expression was not significantly associated with PFS in osteosarcoma or chordoma. No differences in PFS were observed between patients with moderate or low CD3L1 expression, or between those with positive or negative expression (Supplementary Fig. S2A and S2B). Discussion CD3L1 is a single-pass type I membrane protein that functions as a ligand for CD3ε and plays a regulatory role in tumor-immune interactions. Previous studies have reported CD3L1 overexpression in several solid tumors, including NSCLC, breast cancer, rectal cancer, and thyroid cancer, compared to corresponding normal tissues [ 12 ]. This overexpression may contribute impaired T cell activation and immune evasion. Analysis of public databases has also shown that CD3L1 mRNA levels are associated with reduced overall survival in patients with sarcoma [ 13 ]. However, given the substantial histological diversity across more than 100 sarcoma subtypes [ 14 ], the role of CD3L1 in specific bone tumors, such as osteosarcoma and chordoma, remains largely unexplored. To address this knowledge gap, our study provides the first comprehensive assessment of CD3L1 protein expression in osteosarcoma and chordoma using IHC. We also examined its clinicopathological relevance and potential prognostic significance. Our findings revealed that CD3L1 was positivity expressed in 59.6% osteosarcoma and 70.0% of chordoma cases, levels comparable to or exceeding those of established immune checkpoints such as B7-H3 and CD47, and markedly higher than PD-L1 [ 10 ]. These data suggest that CD3L1 may serve as a novel biomarker or therapeutic target in these malignancies. Although the function of CD3L1 in sarcoma remains poorly understood, investigating its association with pathological risk factors could provide mechanistic insights. We found no significant correlation between CD3L1 expression and demographic variables such as age, sex, or tumor site. However, CD3L1 expression was significantly associated with the presence of chondroid stroma in osteosarcoma ( p = 0.003). Chondroid stroma is a defining feature of chondroblastic osteosarcoma, a subtype linked to increased chemoresistance, metastasis, and poorer prognosis, as demonstrated in long-term follow-up studies such as EURAMOS-1 [ 13 , 15 , 16 ]. Some reports also suggest that this subtype has a higher risk of venous tumor thrombus (VTTs), contributing to pulmonary metastases and worse outcomes [ 17 ]. In our study, we observed tumor thrombus formation in samples with chondroid stroma (Fig. 5 ), supporting these findings. Therefore, CD3L1 may be involved in mesenchymal remodeling and the pathogenesis of aggressive osteosarcoma variants, warranting further investigation. Previous studies have demonstrated an inverse relationship between CD3L1 and PD-L1 expression in tumor cells [ 7 ]. Our data further show that CD3L1 expression was positively associated with prior PD-1 immunotherapy in osteosarcoma, suggesting a compensatory immune escape mechanism. It is possible that anti-PD-1 therapy selectively eradicates PD-L1-expressing tumor cells, leaving behind cells with upregulated CD3L1 expression. Deng et al. [ 7 ] reported the anti-tumor efficacy of CD3L1 inhibition in osteosarcoma mouse models, highlighting the therapeutic potential. Given the limited efficacy of anti-PD-1 monotherapy in osteosarcoma, it may be worthwhile to explore combination strategies involving CD3L1 inhibition and PD-1/PD-L1 blockade. Chordoma, being a rare malignancy, has an understudied immune microenvironment. Understanding immune regulatory mechanisms in chordoma is crucial for therapeutic development. Currently, chordomas typically exhibit an immune-excluded or immunosuppressive phenotype, characterized by infiltration of T cells and macrophages. In our study, high CD3L1 expression was significantly associated with increased infiltration of CD8 + T cells, B cells, and macrophages, particularly M2-polarized macrophages (CD163 + and CD206 + ) [ 18 , 19 ]. While B cells and CD8 + T cells are generally associated with anti-tumor responses in solid tumors, M2 macrophages have been implicated in tumor promotion, including in chordoma [ 20 ]. These findings suggest that CD3L1 plays a dual role in immune modulation, promoting both immune activation and immune suppression, depending on the cellular context. Despite the novel insights provided by this study, several limitations must be acknowledged. Kaplan–Meier survival analysis did not reveal a significant association between CD3L1 expression and PFS. This may be attributed to limited sample size, patient heterogeneity, or confounding clinical factors. Furthermore, the complex biology of osteosarcoma and chordoma suggest that CD3L1 alone may not be a sufficient prognostic prognosis. Nonetheless, our findings underscore the potential relevance of CD3L1 in these tumors and support its further evaluation in larger, more homogenous cohorts, ideally with multi-omics integration and immune profiling. Declarations Author Contribution Project Administration: XDT, TTR, JX, LX. Funding Acquisition: XDT, JX, LX. Methodology: YHW, JHX, SYD. Data Curation: YHW, JHX, KKS. Software: YQW. Writing-Original Draft Preparation: YHW, JHX. Writing–Review and Editing: SYD, LX, JX. Accountable for all aspects of the work: YHW, JHX, SYD, KKS, YQW, TTR, XDT, JX, LX. All authors have read and approved the manuscript. Data Availability We promised to maintain patient data confidentially. However, patient data will be made disguisedly available (including data dictionaries) for approved data sharing requests. Individual data that underlie the results reported in this article will be shared after de-identification and normalization of information (text, tables, figures, and appendices). 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Cancer Med 7(10):4903–4913. https://doi.org/10.1002/cam4.1739 Lopez DC, Robbins YL, Kowalczyk JT, Lassoued W, Gulley JL, Miettinen MM, Gallia GL, Allen CT, Hodge JW, London NR, Jr. (2022) Multi-spectral immunofluorescence evaluation of the myeloid, T cell, and natural killer cell tumor immune microenvironment in chordoma may guide immunotherapeutic strategies. Front Oncol 12:1012058. https://doi.org/10.3389/fonc.2022.1012058 Duan W, Zhang B, Li X, Chen W, Jia S, Xin Z, Jian Q, Jian F, Chou D, Chen Z (2022) Single-cell transcriptome profiling reveals intra-tumoral heterogeneity in human chordomas. Cancer Immunol Immunother 71(9):2185–2195. https://doi.org/10.1007/s00262-022-03152-1 Huo X, Ma S, Wang C, Song L, Yao B, Zhu S, Li P, Wang L, Wu Z, Wang K (2023) Unravelling the role of immune cells and FN1 in the recurrence and therapeutic process of skull base chordoma. Clin Transl Med 13(10):e1429. https://doi.org/10.1002/ctm2.1429 Supplementary Table and Figure Supplementary Table and Figure are not available with this version. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. Our growing team is made up of researchers and industry professionals working together to solve the most critical problems facing scientific publishing. Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6689428","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":458125478,"identity":"b422084f-8a74-47eb-ab18-41e4f551c030","order_by":0,"name":"Yuhang Wang","email":"","orcid":"","institution":"Department of Musculoskeletal Tumor, Peking University People’s Hospital","correspondingAuthor":false,"prefix":"","firstName":"Yuhang","middleName":"","lastName":"Wang","suffix":""},{"id":458125479,"identity":"b5e666df-a47c-4a20-a332-0d75b20cb446","order_by":1,"name":"Jiuhui Xu","email":"","orcid":"","institution":"Department of Musculoskeletal Tumor, Peking University People’s Hospital","correspondingAuthor":false,"prefix":"","firstName":"Jiuhui","middleName":"","lastName":"Xu","suffix":""},{"id":458125480,"identity":"e77fce9d-e16b-43fa-81a4-978b56bba840","order_by":2,"name":"Shouyan Deng","email":"","orcid":"","institution":"Institutes of Biomedical Sciences, Shanghai Key Laboratory of Medical Epigenetics, International Co-laboratory of Medical Epigenetics and Metabolism, Zhongshan-Xuhui Hospital, Fudan University","correspondingAuthor":false,"prefix":"","firstName":"Shouyan","middleName":"","lastName":"Deng","suffix":""},{"id":458125481,"identity":"0390bb8f-cfd2-4c09-88a9-a3a5cd9bbccc","order_by":3,"name":"Kunkun Sun","email":"","orcid":"","institution":"Department of Pathology, Peking University People’s Hospital","correspondingAuthor":false,"prefix":"","firstName":"Kunkun","middleName":"","lastName":"Sun","suffix":""},{"id":458125482,"identity":"3f9ea9eb-ce90-4822-99f8-d6223d39cad1","order_by":4,"name":"Yaqi Wang","email":"","orcid":"","institution":"Department of Biomedical Engineering, College of Future Technology, Peking University","correspondingAuthor":false,"prefix":"","firstName":"Yaqi","middleName":"","lastName":"Wang","suffix":""},{"id":458125483,"identity":"55e2a3af-f9fa-41c4-bf73-4ffdf3aeec17","order_by":5,"name":"Tingting Ren","email":"","orcid":"","institution":"Department of Musculoskeletal Tumor, Peking University People’s Hospital","correspondingAuthor":false,"prefix":"","firstName":"Tingting","middleName":"","lastName":"Ren","suffix":""},{"id":458125484,"identity":"94c3f2fd-8958-4b66-b040-6fa73d154e13","order_by":6,"name":"Xiaodong Tang","email":"","orcid":"","institution":"Department of Musculoskeletal Tumor, Peking University People’s Hospital","correspondingAuthor":false,"prefix":"","firstName":"Xiaodong","middleName":"","lastName":"Tang","suffix":""},{"id":458125485,"identity":"782f0d90-38cd-4a92-af53-89faa144acec","order_by":7,"name":"Jie Xu","email":"","orcid":"","institution":"Institutes of Biomedical Sciences, Shanghai Key Laboratory of Medical Epigenetics, International Co-laboratory of Medical Epigenetics and Metabolism, Zhongshan-Xuhui Hospital, Fudan University","correspondingAuthor":false,"prefix":"","firstName":"Jie","middleName":"","lastName":"Xu","suffix":""},{"id":458125486,"identity":"a74b15ca-22c8-4324-bd49-b473ee75b9bb","order_by":8,"name":"Lu Xie","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAz0lEQVRIie3RoQvCQBTH8d84WNqxeiv+DU8Ei/4xaxbzMBgEwaRmQfHPMN94oGXMKnhBy5Jh0SJ4DKPMrRnuGx5c+PB4HOBy/WMCnsYEBAT25Tcj0MhaEZv2Fm0InYRmuTc9uq40yoQRbmf1JJr7MctD0SeTx94mZyij60koArKEh3QZk5ALBqm4nvgiLFnuPuTVhNgtYDnjfkW8JsTeQunuWPQik1G6zEeBuvwgdOb77TE13fV12b09k0En3PwgnzSgqln9Tgvicrlcrq+9AVRFRRx8cyiFAAAAAElFTkSuQmCC","orcid":"","institution":"Department of Musculoskeletal Tumor, Peking University People’s Hospital","correspondingAuthor":true,"prefix":"","firstName":"Lu","middleName":"","lastName":"Xie","suffix":""}],"badges":[],"createdAt":"2025-05-18 02:38:12","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6689428/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6689428/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":83292729,"identity":"e437c6d1-7165-41fa-a5b1-27525a156dc8","added_by":"auto","created_at":"2025-05-22 13:20:35","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":2149296,"visible":true,"origin":"","legend":"\u003cp\u003eCD3L1 expression in osteosarcoma and chordoma. Representative images for different immunohistochemistry staining of CD3L1 in osteosarcoma and chordoma. The IHC score was performed by the pathological specialist. Medium expression defined in chordoma shows slightly lower intensity than that of osteosarcoma, due to differential expression between the two diseases. Scale bars indicate 1 mm in the left images, and scale bars indicate 250 μm in the right magnified images\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-6689428/v1/a15a0f49697e29136e5970ac.png"},{"id":83294052,"identity":"b35baa5a-50b0-486f-a8c2-dfa3d505fdf5","added_by":"auto","created_at":"2025-05-22 13:28:35","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":378087,"visible":true,"origin":"","legend":"\u003cp\u003eCD3L1-positive expression was associated with chondroid stroma in osteosarcoma. Representative images showing CD3L1-positive expression with chondroid stroma in patients with osteosarcoma. The expression of CD3L1 was significantly associated with the chondroid stroma in osteosarcoma. Scale bars indicate 100 μm\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-6689428/v1/fb88ace6176bb98005713264.png"},{"id":83294053,"identity":"57b2d5b9-9963-43ed-9dcd-1e3d25919af3","added_by":"auto","created_at":"2025-05-22 13:28:35","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":1893725,"visible":true,"origin":"","legend":"\u003cp\u003eCD3L1 expression was related to more immune cells infiltration in chordoma. \u003cstrong\u003eA\u003c/strong\u003e Multiplex immunofluorescence staining showing immune cells infiltration in chordoma. Chordoma represents an immune-excluded microenvironment, immune cells were infiltrated into the tumor stroma. Panel 1: CD68 (pan-macrophage, green), CD163 (M2 macrophage, red), CD206 (M2 macrophage, yellow), and IRF (M1 macrophage, pink). Panel 2: CD3 (T cell, blue), CD8 (cytotoxic T cell, pink), CD56 (NK cell, green), CD20 (B cell, red), and PD-L1 (yellow). Scale bars indicate 200 μm. \u003cstrong\u003eB\u003c/strong\u003eImmune cells proportions in different CD3L1 expression groups. \u003cstrong\u003eC\u003c/strong\u003e Representative images showing CD3L1-positive expression is associated with more macrophages, especially the M2 type macrophages infiltration in chordoma. Scale bars indicate 100 μm. *\u003cem\u003ep\u003c/em\u003e\u0026lt; 0.05, ** \u003cem\u003ep\u003c/em\u003e \u0026lt; 0.01, *** \u003cem\u003ep\u003c/em\u003e \u0026lt; 0.001, **** \u003cem\u003ep\u003c/em\u003e \u0026lt; 0.0001\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-6689428/v1/6cff2af3e73fa2f096fb5533.png"},{"id":83292732,"identity":"baf71bb0-8175-4eb0-bfff-49209b9c927c","added_by":"auto","created_at":"2025-05-22 13:20:35","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":871505,"visible":true,"origin":"","legend":"\u003cp\u003eMore CD206 positive macrophage was infiltrated in the recurrent chordoma. \u003cstrong\u003eA\u003c/strong\u003e Statistical analysis of each immune cells proportion in primary and recurrent chordoma. CD206-positive macrophages are related to recurrent chordoma. \u003cstrong\u003eB\u003c/strong\u003e Representative images showed the CD206-positive macrophages in primary and recurrent chordoma patients. CD206 (yellow). Scale bars indicate 50 μm. *\u003cem\u003ep\u003c/em\u003e \u0026lt; 0.05\u003c/p\u003e","description":"","filename":"floatimage4.png","url":"https://assets-eu.researchsquare.com/files/rs-6689428/v1/f8e0716e3b4f3776d21ce24c.png"},{"id":83292734,"identity":"1b24cd92-877e-4810-b93b-060608d39347","added_by":"auto","created_at":"2025-05-22 13:20:35","extension":"png","order_by":5,"title":"Figure 5","display":"","copyAsset":false,"role":"figure","size":1328138,"visible":true,"origin":"","legend":"\u003cp\u003eThe image showing the presence of tumor thrombus and chondroid stroma in tissues of osteosarcoma. Tumor thrombus was observed in tissues with CD3L1-Low positive expression and chondroid stroma in osteosarcoma. Scale bars indicate 1 mm in the left images, and scale bars indicate 250 μm in the right magnified images\u003c/p\u003e","description":"","filename":"floatimage5.png","url":"https://assets-eu.researchsquare.com/files/rs-6689428/v1/81fb667e460fde0ad1ba79cf.png"},{"id":83536472,"identity":"b3269973-82fd-4ba6-87a3-656da92ef260","added_by":"auto","created_at":"2025-05-28 06:47:12","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":6329228,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6689428/v1/703b11f3-4c12-4c73-a5a0-3ce1e20802e5.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"CD3L1 expression in osteosarcoma and chordoma: a retrospective analysis of 62 cases","fulltext":[{"header":"Introduction","content":"\u003cp\u003eOsteosarcoma is the most common primary bone malignancy in adolescents and young adults, and it is characterized by extensive genomic and transcriptomic heterogeneity, largely resulting from chromothripsis. This intratumoral heterogeneity makes the disease difficult to target, and combination therapy has become the mainstay of systemic treatment strategies [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Structural alternations, such as genetic amplifications, translocations, single nucleotide insertions or deletions, and loss of heterozygosity, can activate oncogenes or inactivate tumor suppressor genes, contributing to osteosarcoma pathogenesis [\u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2\u003c/span\u003e]. Despite these insights, the precise mechanisms underlying osteosarcoma remain incompletely understood. Large-cohort studies have highlighted pervasive genomic instability and heterogeneity, necessitating treatment approaches that traditionally rely on cytotoxic and cytostatic agents. A typical first-line chemotherapy regimen includes a three-drug combination, which yields an overall 5-year event-free survival (EFS) rate of 65% [\u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e3\u003c/span\u003e]. Among patients with non-metastatic osteosarcoma, the 5-EFS is approximately 70%, compared to 30% in those metastatic disease [\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eChordoma is a rare malignant bone tumor derived from notochord remnants, accounting for approximately 8.4% of all bone cancer types [\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e]. Although typically classified as histologically low-grade, chordomas exhibit a high rate of local recurrence and are intrinsically resistant to both chemotherapy or targeted therapies. This resistance is attributed to their pathological architecture, with infiltrative and lobulated tumor cells separated by fibrous septa. The transcription factor Brachyury, a T-box gene regulating notochord development, is a sensitive and specific immunohistochemical (IHC) marker for chordoma. Brachyury amplification is commonly observed in familial chordoma, while in sporadic cases, it is more often associated with whole-chromosome gains rather than focal duplications [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e]. Chordomas most frequently arise in the sacrum, skull base, and mobile spine. The current standard of care for tumors in the mobile spine and sacrum includes en bloc resection with wide margins, followed by postoperative external-beam radiation therapy [\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e]. According to the SEER database, the median overall survival is 6.29 years, with 5, 10, and 20-year survival rates of 67.6%, 39.9%, and 13.1%, respectively [\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eCD3L1, also known as ITPRIPL1 (inositol 1,4,5-triphosphate receptor-interacting protein-like 1), was recently identified as a natural ligand of CD3ε, and is known to downregulate T cell activation, thereby facilitating immune evasion by tumors [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Its overexpression has been observed in non-small cell lung cancer (NSCLC), where it correlates with advanced disease stages [\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e]. However, to date, the expression and functional relevance of CD3L1 in bone tumors such as osteosarcoma and chordoma have not been reported. This study aims to characterize CD3L1 expression in osteosarcoma and chordoma, and to explore its association with clinicopathological features and patient prognosis, thereby providing rationale for future clinical trials.\u003c/p\u003e"},{"header":"Materials and methods","content":"\u003cp\u003ePatients and samples\u003c/p\u003e \u003cp\u003e This study was approved by the Institutional Review Board of Peking University People\u0026rsquo;s Hospital (No. 2024PHB013-001) and was supported by the Beijing Natural Science Foundation (Nos. 2127000356 and QY24058). Clinical data and tissue microarrays were collected from 42 patients with osteosarcoma and 20 patients with chordoma who were treated at the Musculoskeletal Tumor Center of Peking University People\u0026rsquo;s Hospital (PKUPH) between June 1, 2021 and July 1, 2024.\u003c/p\u003e \u003cp\u003eThe inclusion criteria were as follows: (i) Histologically confirmed high-grade osteosarcoma or refractory chordoma. (ii) Presence of evaluable lesions defined by the Response Evaluation Criteria in Solid Tumors (RECIST 1.1) [\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e], with documented disease progression after standard therapy. Osteosarcoma patients had progressed following first-line MAP/I chemotherapy with target lesions present; chordoma cases had experienced two or more relapses and were deemed unsuitable for further surgical or radiation intervention, and (iii) Availability of sufficient formalin-fixed, paraffin-embedded (FFPE) tissue for analysis. The exclusion criteria were: (i) Incomplete clinical data, (ii) Presence of other malignancies or serious comorbid conditions, and (iii) Lost to follow-up during the observation period.\u003c/p\u003e \u003cp\u003e Written informed consent was obtained from all of the patients or their legal guardians. Response evaluations was conducted per RECIST 1.1 guidelines, and clinical follow-up occurred every two months until the patient\u0026rsquo;s death. Demographic characteristics of enrolled patients are summarized in Tables\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e and \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAssociation between CD3L1 expression and clinicopathological features of patients with osteosarcoma\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"10\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eFactors\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTotal no.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"6\" nameend=\"c9\" namest=\"c4\"\u003e \u003cp\u003eCD3L1 expression\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNegative\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eLow\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eMedium\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSex\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e40.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e22\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e52.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e7.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e64.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e29.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e63.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e7.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e35.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e60.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e33.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e6.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e0.171\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAge group\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;35 years\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e39\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e92.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e41.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e51.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e7.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026ge;\u0026thinsp;35 years\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e33.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e66.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e1.000\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eSpecimen source\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFrom bone\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e45.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e40.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e15.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFrom lung\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e47.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e40.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e60.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFrom other tissue\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e1.000\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMultinucleated giant cell\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e35.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e40.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e60.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e64.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e40.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e48.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e11.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e0.558\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGiant cell tumor\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e40.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e41.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e58.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e59.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e40.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e48.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e12.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e0.503\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eChondroid stroma\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e23.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e10.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e60.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e30.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e76.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e50.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e50.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e0.003\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAfter PD-1 treatment\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e35.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e26.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e53.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e20.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e27\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e64.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e48.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e14\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e51.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e0.049\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRadiotherapy\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e23.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e40.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e60.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e32\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e76.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e40.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e16\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e50.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e9.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e0.872\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTherapy line\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e23.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e40.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e50.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e10.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e24\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e57.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e10\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e41.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e50.0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e8.33\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026ge;\u0026thinsp;3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e19.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e37.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e1.000\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eAssociation between CD3L1 expression and clinicopathological features of patients with chordoma\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"10\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c10\" colnum=\"10\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eCharacteristics\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTotal no.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"6\" nameend=\"c9\" namest=\"c4\"\u003e \u003cp\u003eCD3L1 expression\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c10\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003e\u003cem\u003ep\u003c/em\u003e value\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eNegative\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eLow\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003eMedium\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003e%\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eGender\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e100\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e30\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e58.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e16.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eFemale\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e37.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e12.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e0.835\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAge\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026lt;\u0026thinsp;56 years\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e11\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e55\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e18.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e63.6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e18.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u0026ge;\u0026thinsp;56 years\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e45\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e44.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e44.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e11.1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e0.4695\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eTreatment before surgery\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e66.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e33.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e17\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e85\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e35.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e52.9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e11.8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e0.5368\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eInvasion\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e12\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e41.7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e50\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e8.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e8\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e12.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e62.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e0.408\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eState\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePrimary\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e13\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e65\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e38.5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e6\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e46.2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e15.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eRecurrence\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e7\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e35\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e14.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e71.4\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e1\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e14.3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e0.6807\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMucus matrix\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c8\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eYes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e5\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e25\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e2\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e40\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c10\"\u003e\u0026nbsp;\u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eNo\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e15\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e75\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e9\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c7\"\u003e \u003cp\u003e60\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e3\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c9\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c10\"\u003e \u003cp\u003e0.3615\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eImmunohistochemistry\u003c/p\u003e \u003cp\u003eIHC was performed on FFPE tumor sections from patients with osteosarcoma or chordoma (\u003cem\u003en\u003c/em\u003e\u0026thinsp;=\u0026thinsp;62). Tissue sections (5 \u0026micro;m thick) were prepared from the blocks and mounted on glass slides. Slides were deparaffinized in xylene and rehydrated through a graded ethanol series. Endogenous peroxidase activity was quenched using 3% hydrogen peroxide (H\u003csub\u003e2\u003c/sub\u003eO\u003csub\u003e2\u003c/sub\u003e) for 15 minutes, followed by phosphate-buffered saline (PBS) washes. Antigen retrieval was carried out by heating slides in citrate buffer using a microwave for 15 minutes, after which the slides were cooled to room temperature and rinsed in PBS.\u003c/p\u003e \u003cp\u003eTissue sections were incubated overnight at 4\u0026deg;C with anti-CD3L1 primary antibody (BioTroy, Cat. #BT02-CDX-01; 1:100 dilution in 1% Bovine Serum Albumin). The following day, sections were washed and incubated with a secondary antibody (1:100 in 1% BSA) for 30 minutes at room temperature. Detection was performed using the DAB chromogenic method, and slides were counterstained with hematoxylin for 3 minutes, dehydrated, and mounted for microscopy.\u003c/p\u003e \u003cp\u003eIHC scoring\u003c/p\u003e \u003cp\u003eA board-certified pathologist (SKK), experienced in bone tumor pathology, assessment assessed CD3L1 expression. Tumors were classified as negative (0\u0026thinsp;\u0026le;\u0026thinsp;1% tumor cells stained) or positive (\u0026gt;\u0026thinsp;1%). Among positive samples, staining intensity was further graded as weak (1\u0026ndash;25%), moderate (26\u0026ndash;50%), and strong (\u0026ge;\u0026thinsp;50%) [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e].\u003c/p\u003e \u003cp\u003eMultiplex immunofluorescence (mIF) staining\u003c/p\u003e \u003cp\u003eTo investigate the immune microenvironment of chordoma, Multiplex immunofluorescence (mIF) staining was conducted using a PANO 7-plex IHC kit (Panovue, Cat. #10004100100, Beijing, China) according to the manufacturer\u0026rsquo;s instructions, as previously described [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e]. Sides were deparaffinized overnight at 65\u0026deg;C, rehydrated with xylene and ethanol, and subjected to antigen retrieval via microwave treatment. The primary antibodies were grouped into two panels: Panel 1 included CD3, CD8A, CD20, PD-L1, and CD56, and Panel 2 included CD68, CD163, CD206, and IRF8 (Supplementary Table S1). Antibodies were incubated for 1 hour at room temperature, followed by 10-minute incubation with HRP-conjugated secondary antibodies. Nuclei were counterstained using DAPI (Sigma-Aldrich).\u003c/p\u003e \u003cp\u003eH\u0026amp;E staining and tumor-stroma ratio assessment\u003c/p\u003e \u003cp\u003eSlides were deparaffinized and rehydrated following standard protocols, then stained using hematoxylin and eosin. The parenchyma-to-stroma ratio in chordoma samples was interpreted by a dedicated pathologist.\u003c/p\u003e \u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eStatistical analysis\u003c/h2\u003e \u003cp\u003eStatistical analyses were conducted using SPSS Statistics 26.0 software (IBM Corp., Armonk, NY, USA) and GraphPad Prism 10. Continuous variables were expressed as mean\u0026thinsp;\u0026plusmn;\u0026thinsp;standard deviation (SD), and categorical as frequencies and percentages. Association between CD3L1 expression and clinicopathological parameters were evaluated using chi-square or Fisher\u0026rsquo;s exact tests. Progression-free survival (PFS) was assessed using Kaplan-Meier survival analysis, and differences between groups were compared using the log-rank test. Statistical significance was set at \u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05.\u003c/p\u003e \u003c/div\u003e"},{"header":"Results","content":"\u003cp\u003eCD3L1 protein expression in tumor tissues\u003c/p\u003e \u003cp\u003eA total of 42 patients with osteosarcoma and 22 with chordoma were enrolled for CD3L1 IHC analysis. Two chordoma samples were excluded due to tissue loss, resulting in 62 valid cases. Among the osteosarcoma patients, the median age was 16 years old (range: 12\u0026ndash;59). Of the 42 osteosarcoma samples, 47.6% (20/42) originated from bone, 47.6% (20/42) from pulmonary metastases, and 4.8% (2/42) from lymph node or skin lesions. In the chordoma group, the median age was 56 years old (range: 10\u0026ndash;74), and all of the samples were derived from the sacrococcygeal region. Among them, 5% (1/20) were classified as dedifferentiated chordoma and 95% (19/20) as conventional chordoma. Additionally, 65% (13/20) were primary and 35% (7/20) were recurrent cases (Tables\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e and \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eCD3L1 protein was detected via brown cytoplasmic staining, with occasional nuclear localization, in IHC (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Of the 62 evaluable samples, 39 (62.9%) showed positive expression. CD3L1 was expressed in 70.0% (14/20) of chordoma samples and 59.6% (25/42) of osteosarcoma samples (Table\u0026nbsp;\u003cspan refid=\"Tab3\" class=\"InternalRef\"\u003e3\u003c/span\u003e). Specimens were categorized as negative, low-positive, or moderately positive, with no cases exhibiting high-intensity staining (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab3\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eThe positive expression of CD3L1 in osteosarcoma and chordoma\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eCases\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003ePositivity\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"3\" nameend=\"c6\" namest=\"c4\"\u003e \u003cp\u003eStaining intensity\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eLow\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eModerate\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eHigh\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eAll sarcoma\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e62\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e39 (62.9%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e33 (53.2%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e6 (9.7%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eOsteosarcoma\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e42\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e25 (59.6%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e22 (52.4%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3 (7.2%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eChordomas\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c2\"\u003e \u003cp\u003e20\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e \u003cp\u003e14 (70.0%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c4\"\u003e \u003cp\u003e11 (55.5%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c5\"\u003e \u003cp\u003e3 (15.0%)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c6\"\u003e \u003cp\u003e0\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003eThese results demonstrate that CD3L1 is broadly expressed in both osteosarcoma and chordoma, suggesting its potential as a disease marker.\u003c/p\u003e \u003cp\u003eAssessment stability and repeatability of the CD3L1-targeting antibody in chordoma tissue microarrays\u003c/p\u003e \u003cp\u003eThe CD3L1 humanized antibody, developed and provided by the Shanghai Key Laboratory of Medical Epigenetics [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e], was tested for stability and reproducibility. Two sequential tissue microarray (TMA) slides from 20 chordoma specimens were stained in parallel using the anti-CD3L1 antibody. The staining patterns between the two slides exhibited high consistency (Supplementary Fig. S1), confirming the excellent stability of the antibody.\u003c/p\u003e \u003cp\u003eAssociation between CD3L1 expression and clinicopathologic characteristics of osteosarcoma and chordoma\u003c/p\u003e \u003cp\u003eTo assess the clinical relevance of CD3L1, the association between its expression and patient characteristics were analyzed (Tables\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e and \u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e). In osteosarcoma, CD3L1 expression was significantly correlated with the presence of chondroid stroma (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003e), a histological hallmark of chondroblastic osteosarcoma. Additionally, a positive association was observed between CD3L1 expression and prior PD-1 immunotherapy (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05), suggesting a potential role in modulating immune responses. No significant associations were found with other clinical variables, such as age, sex, or tumor site.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eIn the chordoma cohort, CD3L1 expression did not significantly correlate with gender, age, preoperative treatment, local invasion, disease state, or presence of mucus matrix (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eRelationship between CD3L1 expression and immune cells infiltration in chordoma\u003c/p\u003e \u003cp\u003eGiven the role of CD3L1 in regulating the tumor immune microenvironment [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e], we assessed immune cell infiltration in chordoma using mIF staining (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA). Increased CD3L1 expression was associated with higher infiltration of CD20\u003csup\u003e+\u003c/sup\u003e B cells, CD8A\u003csup\u003e+\u003c/sup\u003e T cells, and CD68\u003csup\u003e+\u003c/sup\u003e macrophages. Specifically, CD3L1 expression positively correlated with the presence of CD206\u003csup\u003e+\u003c/sup\u003e and CD163\u003csup\u003e+\u003c/sup\u003e M2-type macrophages infiltration, suggesting a potential role in macrophage polarization and tumor-associated immune modulation (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eB and \u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eC).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003eTo further assess the clinical relevance of immune infiltration, we assessed its relationship with prognosis. Notably, recurrent chordoma cases exhibited higher levels of CD206\u003csup\u003e+\u003c/sup\u003e macrophages, supporting our previous findings that M2 macrophages promote tumor progression in chordoma [\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e] (Fig.\u0026nbsp;\u003cspan refid=\"Fig4\" class=\"InternalRef\"\u003e4\u003c/span\u003e).\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003ePrognostic significance of CD3L1 expression in osteosarcoma and chordoma\u003c/p\u003e \u003cp\u003eTo further validate whether CD3L1 expression impacted clinical outcomes, we performed Kaplan\u0026ndash;Meier survival analysis using follow-up data from all 62 patients. CD3L1 expression was not significantly associated with PFS in osteosarcoma or chordoma. No differences in PFS were observed between patients with moderate or low CD3L1 expression, or between those with positive or negative expression (Supplementary Fig. S2A and S2B).\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eCD3L1 is a single-pass type I membrane protein that functions as a ligand for CD3ε and plays a regulatory role in tumor-immune interactions. Previous studies have reported CD3L1 overexpression in several solid tumors, including NSCLC, breast cancer, rectal cancer, and thyroid cancer, compared to corresponding normal tissues [\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e]. This overexpression may contribute impaired T cell activation and immune evasion. Analysis of public databases has also shown that CD3L1 mRNA levels are associated with reduced overall survival in patients with sarcoma [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e]. However, given the substantial histological diversity across more than 100 sarcoma subtypes [\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e], the role of CD3L1 in specific bone tumors, such as osteosarcoma and chordoma, remains largely unexplored.\u003c/p\u003e \u003cp\u003eTo address this knowledge gap, our study provides the first comprehensive assessment of CD3L1 protein expression in osteosarcoma and chordoma using IHC. We also examined its clinicopathological relevance and potential prognostic significance. Our findings revealed that CD3L1 was positivity expressed in 59.6% osteosarcoma and 70.0% of chordoma cases, levels comparable to or exceeding those of established immune checkpoints such as B7-H3 and CD47, and markedly higher than PD-L1 [\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e]. These data suggest that CD3L1 may serve as a novel biomarker or therapeutic target in these malignancies.\u003c/p\u003e \u003cp\u003eAlthough the function of CD3L1 in sarcoma remains poorly understood, investigating its association with pathological risk factors could provide mechanistic insights. We found no significant correlation between CD3L1 expression and demographic variables such as age, sex, or tumor site. However, CD3L1 expression was significantly associated with the presence of chondroid stroma in osteosarcoma (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;=\u0026thinsp;0.003). Chondroid stroma is a defining feature of chondroblastic osteosarcoma, a subtype linked to increased chemoresistance, metastasis, and poorer prognosis, as demonstrated in long-term follow-up studies such as EURAMOS-1 [\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e]. Some reports also suggest that this subtype has a higher risk of venous tumor thrombus (VTTs), contributing to pulmonary metastases and worse outcomes [\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e]. In our study, we observed tumor thrombus formation in samples with chondroid stroma (Fig.\u0026nbsp;\u003cspan refid=\"Fig5\" class=\"InternalRef\"\u003e5\u003c/span\u003e), supporting these findings. Therefore, CD3L1 may be involved in mesenchymal remodeling and the pathogenesis of aggressive osteosarcoma variants, warranting further investigation.\u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003ePrevious studies have demonstrated an inverse relationship between CD3L1 and PD-L1 expression in tumor cells [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e]. Our data further show that CD3L1 expression was positively associated with prior PD-1 immunotherapy in osteosarcoma, suggesting a compensatory immune escape mechanism. It is possible that anti-PD-1 therapy selectively eradicates PD-L1-expressing tumor cells, leaving behind cells with upregulated CD3L1 expression. Deng et al. [\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e] reported the anti-tumor efficacy of CD3L1 inhibition in osteosarcoma mouse models, highlighting the therapeutic potential. Given the limited efficacy of anti-PD-1 monotherapy in osteosarcoma, it may be worthwhile to explore combination strategies involving CD3L1 inhibition and PD-1/PD-L1 blockade.\u003c/p\u003e \u003cp\u003eChordoma, being a rare malignancy, has an understudied immune microenvironment. Understanding immune regulatory mechanisms in chordoma is crucial for therapeutic development. Currently, chordomas typically exhibit an immune-excluded or immunosuppressive phenotype, characterized by infiltration of T cells and macrophages. In our study, high CD3L1 expression was significantly associated with increased infiltration of CD8\u0026thinsp;+\u0026thinsp;T cells, B cells, and macrophages, particularly M2-polarized macrophages (CD163\u003csup\u003e+\u003c/sup\u003e and CD206\u003csup\u003e+\u003c/sup\u003e) [\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e, \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e19\u003c/span\u003e]. While B cells and CD8\u0026thinsp;+\u0026thinsp;T cells are generally associated with anti-tumor responses in solid tumors, M2 macrophages have been implicated in tumor promotion, including in chordoma [\u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e20\u003c/span\u003e]. These findings suggest that CD3L1 plays a dual role in immune modulation, promoting both immune activation and immune suppression, depending on the cellular context.\u003c/p\u003e \u003cp\u003eDespite the novel insights provided by this study, several limitations must be acknowledged. Kaplan\u0026ndash;Meier survival analysis did not reveal a significant association between CD3L1 expression and PFS. This may be attributed to limited sample size, patient heterogeneity, or confounding clinical factors. Furthermore, the complex biology of osteosarcoma and chordoma suggest that CD3L1 alone may not be a sufficient prognostic prognosis. Nonetheless, our findings underscore the potential relevance of CD3L1 in these tumors and support its further evaluation in larger, more homogenous cohorts, ideally with multi-omics integration and immune profiling.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eProject Administration: XDT, TTR, JX, LX. Funding Acquisition: XDT, JX, LX. Methodology: YHW, JHX, SYD. Data Curation: YHW, JHX, KKS. Software: YQW. Writing-Original Draft Preparation: YHW, JHX. Writing\u0026ndash;Review and Editing: SYD, LX, JX. Accountable for all aspects of the work: YHW, JHX, SYD, KKS, YQW, TTR, XDT, JX, LX. All authors have read and approved the manuscript.\u003c/p\u003e\u003ch2\u003eData Availability\u003c/h2\u003e\u003cp\u003eWe promised to maintain patient data confidentially. However, patient data will be made disguisedly available (including data dictionaries) for approved data sharing requests. Individual data that underlie the results reported in this article will be shared after de-identification and normalization of information (text, tables, figures, and appendices). The statistical analysis plan will also be available upon request. Anonymized data will be available beginning 3 months after and ending 2 years after publication of this article to researchers after a methodological review of the proposed analysis plan by the directors of the Musculoskeletal Tumor Center of Peking University People\u0026rsquo;s Hospital. Proposals should be directed to [email protected]. To gain access, data requestors will need to sign a data access agreement.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eDorfman HD, Czerniak B (1995) Bone cancers. Cancer 75(S1):203\u0026ndash;210. https://doi.org/10.1002/1097-0142(19950101)75:1+\u0026lt;203::aid-cncr2820751308\u0026gt;3.0.co;2-v\u003c/li\u003e\n\u003cli\u003eSchott C, Shah AT, Sweet-Cordero EA (2020) Genomic complexity of osteosarcoma and its implication for preclinical and clinical targeted therapies. In: Kleinerman E, Gorlick R (eds) Current advances in the science of osteosarcoma. Springer, Cham. pp 1\u0026ndash;19. https://doi.org/10.1007/978-3-030-43085-6_1\u003c/li\u003e\n\u003cli\u003eGill J, Gorlick R (2021) Advancing therapy for osteosarcoma. Nat Rev Clin Oncol 18(10):609\u0026ndash;624. https://doi.org/10.1038/s41571-021-00519-8\u003c/li\u003e\n\u003cli\u003eDuchman KR, Gao Y, Miller BJ (2015) Prognostic factors for survival in patients with high-grade osteosarcoma using the Surveillance, Epidemiology, and End Results (SEER) Program database. Cancer Epidemiol 39(4):593\u0026ndash;599. https://doi.org/10.1016/j.canep.2015.05.001\u003c/li\u003e\n\u003cli\u003eMcMaster ML, Goldstein AM, Bromley CM, Ishibe N, Parry DM (2001) Chordoma: incidence and survival patterns in the United States, 1973-1995. Cancer Causes Control 12(1):1\u0026ndash;11. https://doi.org/10.1023/a:1008947301735\u003c/li\u003e\n\u003cli\u003eWalcott BP, Nahed BV, Mohyeldin A, Coumans J-V, Kahle KT, Ferreira MJ (2012) Chordoma: current concepts, management, and future directions. Lancet Oncol 13(2):e69\u0026ndash;e76. https://doi.org/10.1016/s1470-2045(11)70337-0\u003c/li\u003e\n\u003cli\u003eDeng S, Zhang Y, Wang H, Liang W, Xie L, Li N, Fang Y, Wang Y, Liu J, Chi H, Sun Y, Ye R, Shan L, Shi J, Shen Z, Wang Y, Wang S, Brosseau J-P, Wang F, Liu G, Quan Y, Xu J (2024) ITPRIPL1 binds CD3\u0026epsilon; to impede T cell activation and enable tumor immune evasion. Cell 187(9):2305\u0026ndash;2323.e33. https://doi.org/10.1016/j.cell.2024.03.019\u003c/li\u003e\n\u003cli\u003eDeng S, Shi J, Sun Y, Quan Y, Shen Z, Wang Y, Li H, Xu J (2023) Development of a monoclonal antibody to ITPRIPL1 for immunohistochemical diagnosis of non-small cell lung cancers: accuracy and correlation with CD8(+) T cell infiltration. Front Cell Dev Biol 11:1297211. https://doi.org/10.3389/fcell.2023.1297211\u003c/li\u003e\n\u003cli\u003eEisenhauer EA, Therasse P, Bogaerts J, Schwartz LH, Sargent D, Ford R, Dancey J, Arbuck S, Gwyther S, Mooney M, Rubinstein L, Shankar L, Dodd L, Kaplan R, Lacombe D, Verweij J (2009) New response evaluation criteria in solid tumours: revised RECIST guideline (version 1.1). Eur J Cancer 45(2):228\u0026ndash;247. https://doi.org/10.1016/j.ejca.2008.10.026\u003c/li\u003e\n\u003cli\u003eXie L, Chen C, Liang X, Xu J, Sun X, Sun K, Yang R, Tang X, Guo W (2023) Expression and clinical significance of various checkpoint molecules in advanced osteosarcoma: possibilities for novel immunotherapy. Orthop Surg 15(3):829\u0026ndash;838. https://doi.org/10.1111/os.13620\u003c/li\u003e\n\u003cli\u003eXu J, Shi Q, Lou J, Wang B, Wang W, Niu J, Guo L, Chen C, Yu Y, Huang Y, Guo W, Lan J, Zhu Y, Ren T, Tang X (2023) Chordoma recruits and polarizes tumor-associated macrophages via secreting CCL5 to promote malignant progression. J Immunother Cancer 11(4):e006808. https://doi.org/10.1136/jitc-2023-006808\u003c/li\u003e\n\u003cli\u003eDuan W, Tian W, Li Z, Liu Y, Xu L (2024) A comprehensive pan-cancer analysis revealing the role of ITPRIPL1 as a prognostic and immunological biomarker. Front Mol Biosci 11:1452290. https://doi.org/10.3389/fmolb.2024.1452290\u003c/li\u003e\n\u003cli\u003eSmeland S, Bielack SS, Whelan J, Bernstein M, Hogendoorn P, Krailo MD, Gorlick R, Janeway KA, Ingleby FC, Anninga J, Antal I, Arndt C, Brown KLB, Butterfass-Bahloul T, Calaminus G, Capra M, Dhooge C, Eriksson M, Flanagan AM, Friedel G, Gebhardt MC, Gelderblom H, Goldsby R, Grier HE, Grimer R, Hawkins DS, Hecker-Nolting S, Sundby Hall K, Isakoff MS, Jovic G, K\u0026uuml;hne T, Kager L, von Kalle T, Kabickova E, Lang S, Lau CC, Leavey PJ, Lessnick SL, Mascarenhas L, Mayer-Steinacker R, Meyers PA, Nagarajan R, Randall RL, Reichardt P, Renard M, Rechnitzer C, Schwartz CL, Strauss S, Teot L, Timmermann B, Sydes MR, Marina N (2019) Survival and prognosis with osteosarcoma: outcomes in more than 2000 patients in the EURAMOS-1 (European and American Osteosarcoma Study) cohort. Eur J Cancer 109:36\u0026ndash;50. https://doi.org/10.1016/j.ejca.2018.11.027\u003c/li\u003e\n\u003cli\u003eSbaraglia M, Bellan E, Dei Tos AP (2021) The 2020 WHO Classification of Soft Tissue Tumours: news and perspectives. Pathologica 113(2):70\u0026ndash;84. https://doi.org/10.32074/1591-951X-213\u003c/li\u003e\n\u003cli\u003eTsagozis P, Laitinen MK, Stevenson JD, Jeys LM, Abudu A, Parry MC (2019) Treatment outcome of patients with chondroblastic osteosarcoma of the limbs and pelvis. Bone Joint J 101-B(6):739\u0026ndash;744. https://doi.org/10.1302/0301-620x.101b6.bjj-2018-1090.r1\u003c/li\u003e\n\u003cli\u003eMamdani H, Grethlein SJ (2018) Pulmonary metastases from chondroblastic osteosarcoma. N Engl J Med 378(15):1429. https://doi.org/10.1056/nejmicm1711874\u003c/li\u003e\n\u003cli\u003eLiang H, Guo W, Yang R, Tang X, Yan T, Ji T, Yang Y, Li D, Xie L, Xu J (2018) Radiological characteristics and predisposing factors of venous tumor thrombus in pelvic osteosarcoma: a mono-institutional retrospective study of 115 cases. Cancer Med 7(10):4903\u0026ndash;4913. https://doi.org/10.1002/cam4.1739\u003c/li\u003e\n\u003cli\u003eLopez DC, Robbins YL, Kowalczyk JT, Lassoued W, Gulley JL, Miettinen MM, Gallia GL, Allen CT, Hodge JW, London NR, Jr. (2022) Multi-spectral immunofluorescence evaluation of the myeloid, T cell, and natural killer cell tumor immune microenvironment in chordoma may guide immunotherapeutic strategies. Front Oncol 12:1012058. https://doi.org/10.3389/fonc.2022.1012058\u003c/li\u003e\n\u003cli\u003eDuan W, Zhang B, Li X, Chen W, Jia S, Xin Z, Jian Q, Jian F, Chou D, Chen Z (2022) Single-cell transcriptome profiling reveals intra-tumoral heterogeneity in human chordomas. Cancer Immunol Immunother 71(9):2185\u0026ndash;2195. https://doi.org/10.1007/s00262-022-03152-1\u003c/li\u003e\n\u003cli\u003eHuo X, Ma S, Wang C, Song L, Yao B, Zhu S, Li P, Wang L, Wu Z, Wang K (2023) Unravelling the role of immune cells and FN1 in the recurrence and therapeutic process of skull base chordoma. Clin Transl Med 13(10):e1429. https://doi.org/10.1002/ctm2.1429\u003c/li\u003e\n\u003c/ol\u003e"},{"header":"Supplementary Table and Figure","content":"\u003cp\u003eSupplementary Table and Figure are not available with this version. \u003cp\u003e\n"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":false,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"CD3L1 (ITPRIPL1), Osteosarcoma, Chordoma, Immunohistochemistry, Prognosis","lastPublishedDoi":"10.21203/rs.3.rs-6689428/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6689428/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003ch2\u003ePurpose\u003c/h2\u003e \u003cp\u003eCD3L1 (ITPRIPL1), a natural ligand of CD3ε, is implicated in tumor-immune interactions and is overexpressed in several solid tumors. However, its expression and functional significance in osteosarcoma and chordoma remain uncharacterized. This study aimed to evaluate CD3L1 expression in osteosarcoma and chordoma and assess its association with clinical features and outcomes.\u003c/p\u003e\u003ch2\u003eMethods\u003c/h2\u003e \u003cp\u003eFormalin-fixed, paraffin-embedded tissue samples from 42 osteosarcoma and 20 chordoma patients were analyzed for CD3L1 expression using immunohistochemistry. Multiplex immunofluorescence staining was performed to evaluate immune cell infiltration in chordoma. Associations between CD3L1 expression and clinicopathological parameters were examined using chi-square tests, Fisher\u0026rsquo;s exact test, and Kaplan-Meier survival analysis.\u003c/p\u003e\u003ch2\u003eResults\u003c/h2\u003e \u003cp\u003eCD3L1 was detected in 62.9% (39/62) of cases, with positivity rates of 59.6% in osteosarcoma and 70.0% in chordoma. CD3L1 expression was independent of patient age, sex, or tumor location but was significantly associated with chondroid stroma and prior PD-1 immunotherapy in osteosarcoma. In chordoma, higher CD3L1 expression correlated with increased infiltration of CD8\u003csup\u003e+\u003c/sup\u003e T cells, B cells, and M2 macrophages.\u003c/p\u003e\u003ch2\u003eConclusion\u003c/h2\u003e \u003cp\u003eCD3L1 is expressed in osteosarcoma and chordoma and may play a pivotal role in regulating tumor stroma and the immune microenvironment, representing a potential target for future therapeutic strategies.\u003c/p\u003e","manuscriptTitle":"CD3L1 expression in osteosarcoma and chordoma: a retrospective analysis of 62 cases","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-05-22 13:20:30","doi":"10.21203/rs.3.rs-6689428/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"00d6d75a-2892-4e7b-a6f4-3c287b8951d2","owner":[],"postedDate":"May 22nd, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-05-28T06:39:02+00:00","versionOfRecord":[],"versionCreatedAt":"2025-05-22 13:20:30","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6689428","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6689428","identity":"rs-6689428","version":["v1"]},"buildId":"XKTyCvWXoU3ODBz1xrDgd","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}