Characterization and predictive value of lymphocyte subsets in peripheral blood of patients with colorectal cancer

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Until now, the role of systemic immune profiles in tumor progression remains unclear. Methods: By using multiparameter flow cytometry, the subset distribution and immunophenotype of lymphocyte subsets were investigated in peripheral blood mononuclear cell samples from 118 colorectal cancer (CRC) patients and 80 healthy donors. Results: Compared with the normal group, the numbers of T lymphocytes and Th lymphocytes were significantly decreased in the CRC group, whereas the number of NK cells was significantly increased. There was no significance between the two groups in the numbers of Ts, γδT, CD3+CD4+CD8+, and CD4+/CD8+ lymphocytes. The number of T lymphocytes was significantly increased after surgical resection, however, the number of B lymphocytes was significantly decreased. There was no significance in the number of other lymphocyte subsets neither before nor after rection. The number of NK cells increased in stage III-IV, as compared to stage I-II. The numbers of Ts and CD4+/CD8+ lymphocytes showed a correlation with the tumor differentiation degree. The number of T lymphocytes was higher in conditions that were without lymph node invasion and distant metastasis. The number of CD4+/CD8+ lymphocytes was significantly higher in the condition without perineural invasion. Conclusion: The altered phenotype of circulating immune cell subsets in CRC and its association with clinical outcome highlights the potential use of lymphocyte subsets as prognostic biomarkers in CRC, thereby contributing to better insight into the role of systemic immune profiles in tumor progression. Lymphocyte subsets Colorectal cancer Cellular immune status Tumor progression Figures Figure 1 Figure 2 Figure 3 INTRODUCTION Colorectal cancer (CRC) is the third most commonly diagnosed cancer worldwide and one of the leading causes of cancer-specific death 1 , with more than 1.1 million cancer deaths expected by 2030 2 . The 5-year survival rate of patients with metastatic CRC is low at approximately 14% 3 , and approximately 50% of patients who receive treatment develop metastases 4 , 5 . Moreover, there is a trend toward a younger age in the incidence of CRC. Despite progress in prevention and therapy, there is still space for new therapies and translational research in colorectal cancer. In recent years, the research on the comprehensive treatment of CRC has continued to increase, and combination therapy with a targeted agent is the main treatment method for metastatic colorectal cancer (mCRC) patients 6 , 7 . In addition, accumulating biomarkers have been proposed to stratify CRC patients according to the risk classification, including miRNAs, circulating tumor cells, circulating tumor DNA, and genetic mutations 8 , 9 . A few biomarkers have been identified for predicting CRC prognosis. Therefore, more biomarkers that can predict the survival and prognosis of patients with CRC are needed. Immunity has become a hot topic of research, which is closely related to tumor development. The immune strength of the host can directly influence tumor development. Many studies also indicated that the immune system has a fundamental role in affecting the development and progression of CRC 10 , 11 . Most studies focus on tumor-infiltrating lymphocytes (TILs), however, immune subsets in the peripheral blood are the primary resources for intratumoral immune events. Therefore, the composition and phenotype of circulating immune cell subsets may potentially play a significant role in predicting tumor progression in CRC. In addition, the impact of CRC on systemic immunity remains to be elucidated. As the development and progression of CRC are known to be affected by the immune system, cell subsets such as T cells and natural killer (NK) cells are considered interesting targets for immunotherapy and clinical biomarker research. Importantly, the phenotype of circulating lymphocyte subsets may reflect the local immune response in the tumor microenvironment (TME), thereby providing potentially important information regarding disease progression in CRC. However, studying the TME alone is insufficient to understand tumor immunity, since the peripheral circulating immune system is an indispensable component of immune regulation in the body 12 . Additional peripheral immunophenotyping is also required, as this provides the chance to identify adaptive immune cell subsets that are altered in patients with solid tumors 12 . As the primary effector cell of humoral immunity, B lymphocytes are poorly investigated because of their controversial role in regulating tumor progression. T lymphocytes are involved in the mechanisms of immunosuppression and immunotolerance, which are often regarded as targets of immunotherapy 13 . Therefore, the investigation of T lymphocyte subsets in colorectal cancer patients is justified. The function of different T-cell subsets has been described over the years, each with distinct functions that promote, or inhibit, antitumor immune responses 14 . Cytotoxic (CD8 + ) T cells recognize tumor-associated antigens presented by classical HLA class I molecules. The cytotoxicity of CD8 + T cells plays an important role in the mechanism of anti-tumor immunity 15 . Natural killer (NK) cells are highly cytotoxic and are considered a component of innate immunity against infections and tumors 16 . Recent studies have shown that adaptive immune responses are involved in regulating the balance between CRC cell invasion and body defense 17 , 18 . However, only a few studies have demonstrated the characterization of peripheral circulating immune cells in CRC patients. Furthermore, comprehensive studies on the connection between oncologic pathology and circulating adaptive immune cells are scarce. In this study, we analyzed the number, phenotype, and function of different subsets of circulating immune cells to gain insight into the changes that occur to these immune cells during the progression of CRC. METHODS 1. Population characteristics The clinical characteristics of patients with CRC are presented in Table 1. The age, gender distribution, total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and body mass index (BMI) between the CRC patients and the healthy people have no significance ( p >0.05). The fasting blood glucose (FBG), uric acid, and triglycerides (TG) have a significant difference between CRC patients and healthy people ( p 0.05 Sex P >0.05 Female 29（36.2%） 41（34.7%） Male 51（63.8%） 77（65.3%） BMI (kg/m 2 ) 22.87±7.62 20.95±9.20 P >0.05 FBG (mmol/L) 6.38±2.09 5.79±1.64* P < 0.05 Serum uric acid (umol/L) 335.29±121.37 288.83±89.85* P 0.05 TG (mmol/L) 1.81±1.08 1.36±0.57* P 0.05 LDL-C (mmol/L) 2.69±0.73 2.50±0.63 P >0.05 Note: * indicates that the difference between the colorectal cancer group and the normal group is statistically significant ( P <0.05); BMI, body mass index; FBG, fasting blood glucose; TC, total cholesterol; TG, triglycerides; HDL-C, high-density lipoprotein cholesterol; LDL-C, low-density lipoprotein cholesterol. 2. Comparison of lymphocyte subsets in the peripheral blood of the CRC and healthy control groups 80 normal people and 118 patients with colorectal cancer were enrolled in this study. Compared with the normal group, the numbers of T lymphocytes and Th lymphocytes were significantly decreased in the CRC group ( p <0.05). In contrast, the number of NK cells was significantly increased in the CRC group as compared with the normal group ( p 0.05) (Fig1). 3. Clinicopathological characteristics of the CRC patients The tumor location, stage, and differentiation grade are presented in Table 3. In addition, the percentages of lymph node invasion, vascular infiltration, distant metastasis, and perineural invasion were also calculated (Table 2). Table 2 Clinicopathological characteristics of the CRC patients Characteristics Number of patients (%) Tumor location Colon 74 （62.7%） Rectum 44 （37.3%） Tumor stage I-II 19 （16.2%） III-IV 98 （83.8%） Tumor differentiation grade Well 6（6.0%） Moderate 60（60.0%） Poor 34 （34.0%） Lymph node invasion Yes 47 （43.1%） No 62（56.9%） Vascular infiltration Yes 22 （29.7%） No 52 （70.3%） Distant metastasis Yes 11（12.6%） No 76（87.4%） Perineural invasion Yes 24（25.0%） No 72（75.0%） 4. Comparison of lymphocyte subsets in the peripheral blood of the CRC patients before and after surgery The peripheral blood was collected before and after surgery within a month. The number of T lymphocytes was significantly increased after surgical resection (Fig2A), however, the number of B lymphocytes was significantly decreased after surgical resection (Fig2H). There was no significance in the number of other lymphocyte subsets (Fig2 B-G). 5. The absolute number of lymphocyte subsets in different subgroups, according to tumor characteristics The absolute number of lymphocyte subsets in different subgroups, according to tumor characteristics, was analyzed. There was a statistical difference between NK cell amounts in stage I-II and III-IV. The number of NK cells increased in stage III-IV, as compared to stage I-II (Fig3A). The number of Ts lymphocytes was lower in moderate and well-differentiated than in poor differentiation (Fig3B). The number of CD4+/CD8+ lymphocytes was significantly higher in moderate differentiation than in poor differentiation (Fig3B). The number of CD4+/CD8+ lymphocytes was higher in high differentiation than in poor differentiation but without statistical significance (Fig3B). The number of T lymphocytes was high in conditions without lymph node invasion (Fig3C). There was no significance in the lymphocyte subsets between the group with and without vascular infiltration (Fig3D). The number of T and Th lymphocytes was higher in the condition that was without distant metastasis, however, the number of NK cells was lower in the condition that without distant metastasis (Fig3E). The number of CD4+/CD8+ lymphocytes was significantly higher in the condition without perineural invasion (Fig3F). RESULTS 1. Population characteristics The clinical characteristics of patients with CRC are presented in Table 1 . The age, gender distribution, total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and body mass index (BMI) between the CRC patients and the healthy people have no significance ( p > 0.05). The fasting blood glucose (FBG), uric acid, and triglycerides (TG) have a significant difference between CRC patients and healthy people ( p 0.05 Sex P > 0.05 Female 29(36.2%) 41(34.7%) Male 51(63.8%) 77(65.3%) BMI (kg/m 2 ) 22.87 ± 7.62 20.95 ± 9.20 P > 0.05 FBG (mmol/L) 6.38 ± 2.09 5.79 ± 1.64* P < 0.05 Serum uric acid (umol/L) 335.29 ± 121.37 288.83 ± 89.85* P 0.05 TG (mmol/L) 1.81 ± 1.08 1.36 ± 0.57* P 0.05 LDL-C (mmol/L) 2.69 ± 0.73 2.50 ± 0.63 P > 0.05 Note: * indicates that the difference between the colorectal cancer group and the normal group is statistically significant ( P < 0.05); BMI, body mass index; FBG, fasting blood glucose; TC, total cholesterol; TG, triglycerides; HDL-C, high-density lipoprotein cholesterol; LDL-C, low-density lipoprotein cholesterol. 2. Comparison of lymphocyte subsets in the peripheral blood of the CRC and healthy control groups 80 normal people and 118 patients with colorectal cancer were enrolled in this study. Compared with the normal group, the numbers of T lymphocytes and Th lymphocytes were significantly decreased in the CRC group ( p < 0.05). In contrast, the number of NK cells was significantly increased in the CRC group as compared with the normal group ( p 0.05) (Fig. 1 ). 3. Clinicopathological characteristics of the CRC patients The tumor location, stage, and differentiation grade are presented in Table 3. In addition, the percentages of lymph node invasion, vascular infiltration, distant metastasis, and perineural invasion were also calculated (Table 2 ). Table 2 Clinicopathological characteristics of the CRC patients Characteristics Number of patients (%) Tumor location Colon 74 (62.7%) Rectum 44 (37.3%) Tumor stage I-II 19 (16.2%) III-IV 98 (83.8%) Tumor differentiation grade Well 6(6.0%) Moderate 60(60.0%) Poor 34 (34.0%) Lymph node invasion Yes 47 (43.1%) No 62(56.9%) Vascular infiltration Yes 22 (29.7%) No 52 (70.3%) Distant metastasis Yes 11(12.6%) No 76(87.4%) Perineural invasion Yes 24(25.0%) No 72(75.0%) 4. Comparison of lymphocyte subsets in the peripheral blood of the CRC patients before and after surgery The peripheral blood was collected before and after surgery within a month. The number of T lymphocytes was significantly increased after surgical resection (Fig. 2 A), however, the number of B lymphocytes was significantly decreased after surgical resection (Fig. 2 H). There was no significance in the number of other lymphocyte subsets (Fig. 2 B-G). 5. The absolute number of lymphocyte subsets in different subgroups, according to tumor characteristics The absolute number of lymphocyte subsets in different subgroups, according to tumor characteristics, was analyzed. There was a statistical difference between NK cell amounts in stage I-II and III-IV. The number of NK cells increased in stage III-IV, as compared to stage I-II (Fig. 3 A). The number of Ts lymphocytes was lower in moderate and well-differentiated than in poor differentiation (Fig. 3 B). The number of CD4+/CD8 + lymphocytes was significantly higher in moderate differentiation than in poor differentiation (Fig. 3 B). The number of CD4+/CD8 + lymphocytes was higher in high differentiation than in poor differentiation but without statistical significance (Fig. 3 B). The number of T lymphocytes was high in conditions without lymph node invasion (Fig. 3 C). There was no significance in the lymphocyte subsets between the group with and without vascular infiltration (Fig. 3 D). The number of T and Th lymphocytes was higher in the condition that was without distant metastasis, however, the number of NK cells was lower in the condition that without distant metastasis (Fig. 3 E). The number of CD4+/CD8 + lymphocytes was significantly higher in the condition without perineural invasion (Fig. 3 F). DISCUSSION Immune cell infiltration is a characteristic of almost all malignant tumors, which plays an important role in immune monitoring and contributes to the elimination of tumor cells 21 . Given the minimally invasive nature of blood sample collection compared to tumor-infiltrating tissues, the regularity of various indexes in peripheral blood is more convenient for clinical application, such as T lymphocyte cells, B lymphocyte cells, and NK cells 22 . In this study, we observed significant changes in the circulating adaptive lymphocyte profile of CRC patients compared with those of healthy patients. Furthermore, we assessed differences in the distribution of adaptive immune cells in patients with CRC. The T cell subsets defined as having different proliferative abilities, lymphatic homing, and effector functions have been well established in previous studies 23,24 . Th cells help maintain Tc cell immune responses and regulate the development of Tc cells into functional cells 25 . In our study, colorectal carcinoma patients presented with significantly lower values of absolute circulating T cell (CD45+/CD3+) and Th cell (CD3+/CD4+/CD8-) counts compared to healthy volunteers, which is consistent with prior research 10,26,27 . After the rection of the tumor, the count of T cell counts showed an increased trend, with statistical significance. However, other subtypes of T lymphocyte cells also showed an increase, but without statistical significance. It may be because the suppression of T cells was removed together with the tumor. As our data showed, the number of T lymphocytes was lower in the condition that either with lymph node invasion or with distant metastasis. In peripheral circulation and tissues, both Th and Tc are capable of dynamic metabolic control 28 . The decrease in the Th cell number in CRC patients could be due to a reduction in peripheral circulation produced by adaptive immune cells migrating to tumor tissues. Th cell depletion, which manifests as a decrease in proliferative and functionally active T cells and an increase in senescent T cells, results from persistent metabolic disturbances directed by tumor cells 29 . This is consistent with our findings in patients with distant metastasis. In addition, the density of sT cells (CD3+CD8+) was inversely associated with the degree of tumor differentiation. The previous research showed, that the density of CD3+CD8+ T cells was inversely associated with the extent of tumor budding at the invasive front 30 , and also was significantly associated with high PD-L1 expression scores 31 . However, the specific role of sT cell still need further exploring. B cells contribute to cancer regulation primarily through indirect inhibition of T cell reactivity, resulting in antitumor immunosuppression. In our present study, the values of circulating B cells (CD3-/CD19+) were also lower in CRC patients, as compared to the normal group, but without statistical significance. Waidhauser et al. 32 found that B cell counts were lower in CRC patients, whereas Spacek et al. 33 demonstrated that it was higher in colon cancer patients than those in control groups. It is interesting that after the rection of the tumor, the count of B cells was decreased significantly. In addition, the circulating B lymphocytes showed no significant correction with tumor pathological characteristics. According to the previous study, there were no significant differences in the value changes of B cell subsets in advanced-stage CRC patients compared with those of patients in the early stage 10 . However, an increasing number of studies have shown that cytokines secreted by B cells have dual roles in tumors during tumorigenesis and progression 34,35 . Whether the role of B cell subsets in CRC can be further explored from the perspective of cytokine secretion remains to be confirmed by further studies. NK cells participate in tumor immunosurveillance by monitoring and killing tumor cells in an antigen-independent and antigen-dependent manner 36 . Several studies have revealed that low NK cell counts predict a higher risk of tumors 37 . Although NK cells possess cytotoxic capabilities, the specific activity of each NK cell subset is complex. It is interesting in our study that, the number of NK cells increased in the CRC patients, as compared with normal group, which is opposite to the previous studies 32,33 . We speculated that the NK immunity of CRC patients might be strengthened to fight against the tumor. Due to the small sample size, this study may not fully represent the broader population, which introduces certain limitations to the findings. The mechanisms underlying the observed changes in lymphocyte subsets profiles require further investigation. In summary, our study revealed significant alterations in circulating lymphocyte subsets that can be attributed to the presence of colorectal cancer. Additionally, certain tumor characteristics like tumor differentiation seem to make a difference in the individual immune response. Whether these circulating lymphocyte alterations play a role in predicting the prognosis of colorectal cancer and whether the alterations are the cause or consequence of the tumor disease will need further investigations. Declarations ACKNOWLEDGMENTS This work was supported by the Tianjin Natural Science Foundation (no. 21JCYBJC01070), the Foundation of Tianjin Union Medical Center (2022PWXK04)， and the Tianjin Key Medical Discipline Construction Project (TJYXZDXK-3-009B). CONFLICTS OF INTEREST The authors claim that none of the material in the paper has been published or is under consideration for publication elsewhere, and there is no conflict of interests. All authors have approved the manuscript for submission. Ethics approval and consent to participate Not applicable. Authors' contributions Guanghai Wu and Kunming Zheng performed the statistical analyses and wrote the paper. Wang Han collected the data, and Shichao Xu contributed to the experiment. Jing Xu revised this paper and designed the study. All authors contributed to the development of this manuscript and read and approved the final version. The Data Availability The data used and/or analyzed during the current study are available from the corresponding authors on reasonable request. References Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A. Global cancer statistics, 2012. CA: a cancer journal for clinicians. Mar 2015;65(2):87-108. Arnold M, Sierra MS, Laversanne M, Soerjomataram I, Jemal A, Bray F. Global patterns and trends in colorectal cancer incidence and mortality. Gut. Apr 2017;66(4):683-691. Siegel RL, Miller KD, Fedewa SA, et al. Colorectal cancer statistics, 2017. CA Cancer J Clin. May 6 2017;67(3):177-193. Siegel RL, Miller KD, Jemal A. Cancer statistics, 2020. CA Cancer J Clin. Jan 2020;70(1):7-30. Tolba MF. Revolutionizing the landscape of colorectal cancer treatment: The potential role of immune checkpoint inhibitors. Int J Cancer. 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Cite Share Download PDF Status: Published Journal Publication published 26 Feb, 2026 Read the published version in Journal of Gastrointestinal Cancer → Version 1 posted Editorial decision: Revision requested 25 Oct, 2025 Reviews received at journal 18 Oct, 2025 Reviews received at journal 15 Oct, 2025 Reviewers agreed at journal 30 Sep, 2025 Reviewers agreed at journal 30 Sep, 2025 Reviewers agreed at journal 29 Sep, 2025 Reviewers invited by journal 25 Sep, 2025 Editor assigned by journal 24 Sep, 2025 Submission checks completed at journal 24 Sep, 2025 First submitted to journal 22 Sep, 2025 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. 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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-7672745","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":524888460,"identity":"b31faa09-6c7b-4468-8048-00467e39d8fb","order_by":0,"name":"Guanghai Wu","email":"","orcid":"","institution":"Tianjin Union Medical Center, The First Affiliated Hospital of Nankai University","correspondingAuthor":false,"prefix":"","firstName":"Guanghai","middleName":"","lastName":"Wu","suffix":""},{"id":524888461,"identity":"003cccc9-f381-4e53-9d33-3c4b848afc35","order_by":1,"name":"Kunming Zheng","email":"","orcid":"","institution":"Tianjin Union 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Xu","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAAy0lEQVRIiWNgGAWjYHCChAMMDMwMDOyNjQ8+kKaF53Cz4QwSbAJqkUhvk+YgRq3BjYSHhwt+Wcvxz3zYIM3AYCen20BAi+SMhITDM/vSjSVuJzYYFzAkG5sdIKCFXwKohbfncOIG6cSG5BkMBxK3EdLCBtciebDhMA8xWsC28PwAapFgbGwmSotkzwOgLQ1Av5xJbGacYUCEXwyO5yR/5vkDDLH2489/fKiwkyOoBRiFCQyMbXATCCoHAXagqX+IUjkKRsEoGAUjFQAAKfNG8W3r2ioAAAAASUVORK5CYII=","orcid":"","institution":"Tianjin Union Medical Center, The First Affiliated Hospital of Nankai University","correspondingAuthor":true,"prefix":"","firstName":"Jing","middleName":"","lastName":"Xu","suffix":""}],"badges":[],"createdAt":"2025-09-22 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16:38:00","extension":"xml","order_by":9,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":85144,"visible":true,"origin":"","legend":"","description":"","filename":"f1865b79341e4cf1852d0c5a7bf052cf1structuring.xml","url":"https://assets-eu.researchsquare.com/files/rs-7672745/v1/aa25e3682f2d1a4333c88672.xml"},{"id":93062897,"identity":"dcd2995c-e0fe-44fa-9f4a-878c221f4cf6","added_by":"auto","created_at":"2025-10-08 16:38:05","extension":"html","order_by":10,"title":"","display":"","copyAsset":false,"role":"acdc-reference","size":91616,"visible":true,"origin":"","legend":"","description":"","filename":"earlyproof.html","url":"https://assets-eu.researchsquare.com/files/rs-7672745/v1/8dc9197f22b449dc14bd0496.html"},{"id":93063254,"identity":"e39c2546-f004-4335-a5a9-cfcfef305531","added_by":"auto","created_at":"2025-10-08 16:38:11","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":562473,"visible":true,"origin":"","legend":"\u003cp\u003eComparison of lymphocyte subsets in the peripheral blood. The number of T (A), Th (B), Ts (C), γδT (D), CD3+CD4+CD8+ (E), and CD4+/CD8+ lymphocytes (F). G: The number of NK cells. H: The number of B lymphocytes. *\u003cem\u003eP\u003c/em\u003e\u0026lt;0.05;\u003c/p\u003e","description":"","filename":"floatimage1.png","url":"https://assets-eu.researchsquare.com/files/rs-7672745/v1/a59088ff0fe64c9fe38b55b1.png"},{"id":93062728,"identity":"c7281539-25c8-4106-a9b4-be9b07b7ee14","added_by":"auto","created_at":"2025-10-08 16:38:02","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":657980,"visible":true,"origin":"","legend":"\u003cp\u003eComparison of lymphocyte subsets in CRC patients before and after surgery. The number of T (A), Th (B), Ts (C), γδT (D), CD3+CD4+CD8+ (E), and CD4+/CD8+ lymphocytes (F). G: The number of NK cells. H: The number of B lymphocytes. *\u003cem\u003eP\u003c/em\u003e\u0026lt;0.05;\u003c/p\u003e","description":"","filename":"floatimage2.png","url":"https://assets-eu.researchsquare.com/files/rs-7672745/v1/33f84c76089300d2b0451b6d.png"},{"id":93062938,"identity":"15d8a4c3-8089-4eaa-8b41-d45453aacc31","added_by":"auto","created_at":"2025-10-08 16:38:05","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":373784,"visible":true,"origin":"","legend":"\u003cp\u003eThe absolute number of lymphocyte subsets in different subgroups, according to tumor characteristics. A: The number of lymphocyte subsets in different tumor stage. B: The number of lymphocyte subsets in different tumor differentiation grade. C: The number of lymphocyte subsets was compared according to lymph node invasion. D: The number of lymphocyte subsets was compared according to vascular infiltration. E: The number of lymphocyte subsets was compared according to distant metastasis. F: The number of lymphocyte subsets was compared according to perineural invasion. *\u003cem\u003eP\u003c/em\u003e\u0026lt;0.05\u003c/p\u003e","description":"","filename":"floatimage3.png","url":"https://assets-eu.researchsquare.com/files/rs-7672745/v1/e347f4fbce90a0a82d99393d.png"},{"id":103765743,"identity":"cf56ccc8-13c3-4984-9a7f-bfd06867bd57","added_by":"auto","created_at":"2026-03-02 16:08:33","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":2377795,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-7672745/v1/424c66db-22a0-4db7-a9a6-60a7309b536f.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Characterization and predictive value of lymphocyte subsets in peripheral blood of patients with colorectal cancer","fulltext":[{"header":"INTRODUCTION","content":"\u003cp\u003eColorectal cancer (CRC) is the third most commonly diagnosed cancer worldwide and one of the leading causes of cancer-specific death \u003csup\u003e\u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1\u003c/span\u003e\u003c/sup\u003e, with more than 1.1\u0026nbsp;million cancer deaths expected by 2030 \u003csup\u003e2\u003c/sup\u003e. The 5-year survival rate of patients with metastatic CRC is low at approximately 14% \u003csup\u003e3\u003c/sup\u003e, and approximately 50% of patients who receive treatment develop metastases \u003csup\u003e\u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e4\u003c/span\u003e,\u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e5\u003c/span\u003e\u003c/sup\u003e. Moreover, there is a trend toward a younger age in the incidence of CRC. Despite progress in prevention and therapy, there is still space for new therapies and translational research in colorectal cancer.\u003c/p\u003e\u003cp\u003eIn recent years, the research on the comprehensive treatment of CRC has continued to increase, and combination therapy with a targeted agent is the main treatment method for metastatic colorectal cancer (mCRC) patients \u003csup\u003e\u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e6\u003c/span\u003e,\u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e7\u003c/span\u003e\u003c/sup\u003e. In addition, accumulating biomarkers have been proposed to stratify CRC patients according to the risk classification, including miRNAs, circulating tumor cells, circulating tumor DNA, and genetic mutations \u003csup\u003e\u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e8\u003c/span\u003e,\u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e9\u003c/span\u003e\u003c/sup\u003e. A few biomarkers have been identified for predicting CRC prognosis. Therefore, more biomarkers that can predict the survival and prognosis of patients with CRC are needed. Immunity has become a hot topic of research, which is closely related to tumor development. The immune strength of the host can directly influence tumor development. Many studies also indicated that the immune system has a fundamental role in affecting the development and progression of CRC \u003csup\u003e\u003cspan citationid=\"CR10\" class=\"CitationRef\"\u003e10\u003c/span\u003e,\u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e11\u003c/span\u003e\u003c/sup\u003e. Most studies focus on tumor-infiltrating lymphocytes (TILs), however, immune subsets in the peripheral blood are the primary resources for intratumoral immune events. Therefore, the composition and phenotype of circulating immune cell subsets may potentially play a significant role in predicting tumor progression in CRC. In addition, the impact of CRC on systemic immunity remains to be elucidated.\u003c/p\u003e\u003cp\u003eAs the development and progression of CRC are known to be affected by the immune system, cell subsets such as T cells and natural killer (NK) cells are considered interesting targets for immunotherapy and clinical biomarker research. Importantly, the phenotype of circulating lymphocyte subsets may reflect the local immune response in the tumor microenvironment (TME), thereby providing potentially important information regarding disease progression in CRC. However, studying the TME alone is insufficient to understand tumor immunity, since the peripheral circulating immune system is an indispensable component of immune regulation in the body \u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eAdditional peripheral immunophenotyping is also required, as this provides the chance to identify adaptive immune cell subsets that are altered in patients with solid tumors \u003csup\u003e\u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e12\u003c/span\u003e\u003c/sup\u003e. As the primary effector cell of humoral immunity, B lymphocytes are poorly investigated because of their controversial role in regulating tumor progression. T lymphocytes are involved in the mechanisms of immunosuppression and immunotolerance, which are often regarded as targets of immunotherapy \u003csup\u003e\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e13\u003c/span\u003e\u003c/sup\u003e. Therefore, the investigation of T lymphocyte subsets in colorectal cancer patients is justified. The function of different T-cell subsets has been described over the years, each with distinct functions that promote, or inhibit, antitumor immune responses \u003csup\u003e\u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e14\u003c/span\u003e\u003c/sup\u003e. Cytotoxic (CD8\u003csup\u003e+\u003c/sup\u003e) T cells recognize tumor-associated antigens presented by classical HLA class I molecules. The cytotoxicity of CD8\u003csup\u003e+\u003c/sup\u003e T cells plays an important role in the mechanism of anti-tumor immunity \u003csup\u003e\u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e15\u003c/span\u003e\u003c/sup\u003e. Natural killer (NK) cells are highly cytotoxic and are considered a component of innate immunity against infections and tumors \u003csup\u003e\u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e16\u003c/span\u003e\u003c/sup\u003e.\u003c/p\u003e\u003cp\u003eRecent studies have shown that adaptive immune responses are involved in regulating the balance between CRC cell invasion and body defense \u003csup\u003e\u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e17\u003c/span\u003e,\u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e18\u003c/span\u003e\u003c/sup\u003e. However, only a few studies have demonstrated the characterization of peripheral circulating immune cells in CRC patients. Furthermore, comprehensive studies on the connection between oncologic pathology and circulating adaptive immune cells are scarce. In this study, we analyzed the number, phenotype, and function of different subsets of circulating immune cells to gain insight into the changes that occur to these immune cells during the progression of CRC.\u003c/p\u003e"},{"header":"METHODS","content":"\u003cp\u003e\u003cstrong\u003e1. Population characteristics\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe clinical characteristics of patients with CRC are presented in Table 1. The age, gender distribution, total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and body mass index (BMI) between the CRC patients and the healthy people have no significance (\u003cem\u003ep\u003c/em\u003e\u0026gt;0.05). The fasting blood glucose (FBG), uric acid, and triglycerides (TG) have a significant difference between CRC patients and healthy people (\u003cem\u003ep\u003c/em\u003e\u0026lt;0.05).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTable 1\u0026nbsp;Clinical Characteristics of Study Population\u003c/p\u003e\n\u003cdiv align=\"\"\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eVariables\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eNC（n=80）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eCRC（n=118）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e-value\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eAge (yr)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e59.8\u0026plusmn;12.6\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e62.9\u0026plusmn;11.0\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e \u0026gt;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eSex\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u003cem\u003eP\u0026nbsp;\u003c/em\u003e\u0026gt;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eFemale\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e29（36.2%）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e41（34.7%）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eMale\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e51（63.8%）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e77（65.3%）\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eBMI (kg/m\u003csup\u003e2\u003c/sup\u003e)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e22.87\u0026plusmn;7.62\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e20.95\u0026plusmn;9.20\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e \u0026gt;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eFBG (mmol/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e6.38\u0026plusmn;2.09\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e5.79\u0026plusmn;1.64*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eSerum uric acid (umol/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e335.29\u0026plusmn;121.37\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e288.83\u0026plusmn;89.85*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eTC (mmol/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e4.83\u0026plusmn;1.14\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e4.66\u0026plusmn;1.00\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e \u0026gt;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eTG (mmol/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e1.81\u0026plusmn;1.08\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e1.36\u0026plusmn;0.57*\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e \u0026lt; 0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eHDL-C (mmol/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e1.35\u0026plusmn;0.36\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e1.41\u0026plusmn;0.45\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e \u0026gt;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eLDL-C (mmol/L)\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e2.69\u0026plusmn;0.73\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e2.50\u0026plusmn;0.63\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u003cem\u003eP\u003c/em\u003e \u0026gt;0.05\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003eNote: * indicates that the difference between the colorectal cancer group and the normal group is statistically significant (\u003cem\u003eP\u003c/em\u003e\u0026lt;0.05); BMI, body mass index; FBG, fasting blood glucose; TC, total cholesterol; TG, triglycerides; HDL-C, high-density lipoprotein cholesterol; LDL-C, low-density lipoprotein cholesterol.\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e2.\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eComparison of lymphocyte subsets in the peripheral blood of the CRC and healthy control groups\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003e80 normal people and 118 patients with colorectal cancer were enrolled in this study. Compared with the normal group, the numbers of T lymphocytes and Th lymphocytes were significantly decreased in the CRC group\u0026nbsp;(\u003cem\u003ep\u003c/em\u003e\u0026lt;0.05). In contrast, the number of NK cells was significantly increased in the CRC group as compared with the normal group\u0026nbsp;(\u003cem\u003ep\u003c/em\u003e\u0026lt;0.05). However, there was no significance between the two groups in the numbers of\u0026nbsp;Ts, \u0026gamma;\u0026delta;T, CD3+CD4+CD8+, and CD4+/CD8+ lymphocytes (\u003cem\u003ep\u003c/em\u003e\u0026gt;0.05) (Fig1).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e3.\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;Clinicopathological characteristics of the CRC patients\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe tumor location, stage, and differentiation grade are presented in Table 3. In addition, the percentages of lymph node invasion, vascular infiltration, distant metastasis, and perineural invasion were also calculated (Table 2).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003eTable 2\u0026nbsp;Clinicopathological characteristics of the CRC patients\u003c/p\u003e\n\u003cdiv align=\"\"\u003e\n \u003ctable border=\"0\" cellspacing=\"0\" cellpadding=\"0\"\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eCharacteristics\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eNumber of\u0026nbsp;patients (%)\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eTumor location\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eColon\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e74\u0026nbsp;（62.7%）\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eRectum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e44\u0026nbsp;（37.3%）\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eTumor stage\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eI-II\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e19\u0026nbsp;（16.2%）\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eIII-IV\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e98\u0026nbsp;（83.8%）\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eTumor differentiation grade\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eWell\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e6（6.0%）\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eModerate\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e60（60.0%）\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003ePoor\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e34\u0026nbsp;（34.0%）\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eLymph node invasion\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e47\u0026nbsp;（43.1%）\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e62（56.9%）\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eVascular infiltration\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e22\u0026nbsp;（29.7%）\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e52\u0026nbsp;（70.3%）\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eDistant metastasis\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e11（12.6%）\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e76（87.4%）\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003ePerineural invasion\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eYes\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e24（25.0%）\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e\u0026nbsp;\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003eNo\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd valign=\"bottom\"\u003e\n \u003cp\u003e72（75.0%）\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n\u003c/div\u003e\n\u003cp\u003e\u003cstrong\u003e4.\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eComparison of lymphocyte subsets in the peripheral blood of the CRC patients before and after surgery\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe peripheral blood was collected before and after surgery within a month. The number of T lymphocytes was significantly increased after surgical resection\u0026nbsp;(Fig2A), however, the number of B lymphocytes was significantly decreased after surgical resection\u0026nbsp;(Fig2H). There was no significance in the number of other\u0026nbsp;lymphocyte subsets\u0026nbsp;(Fig2 B-G).\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003e5.\u003c/strong\u003e\u003cstrong\u003e\u0026nbsp;\u003c/strong\u003e\u003cstrong\u003eThe absolute number of lymphocyte subsets in different subgroups, according to tumor characteristics\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe absolute number of lymphocyte subsets in different subgroups, according to tumor characteristics, was analyzed. There was a statistical difference between NK cell amounts in stage I-II and III-IV. The number of NK cells increased in stage III-IV, as compared to stage I-II (Fig3A). The number of Ts lymphocytes was lower in moderate and well-differentiated than in poor differentiation (Fig3B). The number of CD4+/CD8+ lymphocytes was significantly higher in moderate differentiation than in poor differentiation (Fig3B). The number of CD4+/CD8+ lymphocytes was higher in high differentiation than in poor differentiation but without statistical significance (Fig3B). The number of T lymphocytes was high in conditions without lymph node invasion (Fig3C). There was no significance in the lymphocyte subsets between the group with and without vascular infiltration (Fig3D). The number of T and Th lymphocytes was higher in the condition that was without distant metastasis, however, the number of NK cells was lower in the condition that without distant metastasis (Fig3E). The number of CD4+/CD8+ lymphocytes was significantly higher in the condition without perineural invasion (Fig3F).\u003c/p\u003e"},{"header":"RESULTS","content":"\u003cp\u003e\u003cb\u003e1. Population characteristics\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe clinical characteristics of patients with CRC are presented in Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e. The age, gender distribution, total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), and body mass index (BMI) between the CRC patients and the healthy people have no significance (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05). The fasting blood glucose (FBG), uric acid, and triglycerides (TG) have a significant difference between CRC patients and healthy people (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05).\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\u003eClinical Characteristics of Study Population\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"4\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" 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=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eVariables\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u003cp\u003eNC(n\u0026thinsp;=\u0026thinsp;80)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eCRC(n\u0026thinsp;=\u0026thinsp;118)\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cem\u003eP\u003c/em\u003e-value\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eAge (yr)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e59.8\u0026thinsp;\u0026plusmn;\u0026thinsp;12.6\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e62.9\u0026thinsp;\u0026plusmn;\u0026thinsp;11.0\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSex\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\u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eFemale\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e29(36.2%)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e41(34.7%)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\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=\"left\" colname=\"c2\"\u003e\u003cp\u003e51(63.8%)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e77(65.3%)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u0026nbsp;\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eBMI (kg/m\u003csup\u003e2\u003c/sup\u003e)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e22.87\u0026thinsp;\u0026plusmn;\u0026thinsp;7.62\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e20.95\u0026thinsp;\u0026plusmn;\u0026thinsp;9.20\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eFBG (mmol/L)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e6.38\u0026thinsp;\u0026plusmn;\u0026thinsp;2.09\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e5.79\u0026thinsp;\u0026plusmn;\u0026thinsp;1.64*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eSerum uric acid (umol/L)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e335.29\u0026thinsp;\u0026plusmn;\u0026thinsp;121.37\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e288.83\u0026thinsp;\u0026plusmn;\u0026thinsp;89.85*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTC (mmol/L)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e4.83\u0026thinsp;\u0026plusmn;\u0026thinsp;1.14\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e4.66\u0026thinsp;\u0026plusmn;\u0026thinsp;1.00\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTG (mmol/L)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1.81\u0026thinsp;\u0026plusmn;\u0026thinsp;1.08\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1.36\u0026thinsp;\u0026plusmn;\u0026thinsp;0.57*\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eHDL-C (mmol/L)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e1.35\u0026thinsp;\u0026plusmn;\u0026thinsp;0.36\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e1.41\u0026thinsp;\u0026plusmn;\u0026thinsp;0.45\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLDL-C (mmol/L)\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003e2.69\u0026thinsp;\u0026plusmn;\u0026thinsp;0.73\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c3\"\u003e\u003cp\u003e2.50\u0026thinsp;\u0026plusmn;\u0026thinsp;0.63\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c4\"\u003e\u003cp\u003e\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003c/tbody\u003e\u003c/colgroup\u003e\u003ctfoot\u003e\u003ctr\u003e\u003ctd colspan=\"4\"\u003eNote: * indicates that the difference between the colorectal cancer group and the normal group is statistically significant (\u003cem\u003eP\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05); BMI, body mass index; FBG, fasting blood glucose; TC, total cholesterol; TG, triglycerides; HDL-C, high-density lipoprotein cholesterol; LDL-C, low-density lipoprotein cholesterol.\u003c/td\u003e\u003c/tr\u003e\u003c/tfoot\u003e\u003c/table\u003e\u003c/div\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003e2. Comparison of lymphocyte subsets in the peripheral blood of the CRC and healthy control groups\u003c/b\u003e\u003c/p\u003e\u003cp\u003e80 normal people and 118 patients with colorectal cancer were enrolled in this study. Compared with the normal group, the numbers of T lymphocytes and Th lymphocytes were significantly decreased in the CRC group (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). In contrast, the number of NK cells was significantly increased in the CRC group as compared with the normal group (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026lt;\u0026thinsp;0.05). However, there was no significance between the two groups in the numbers of Ts, γδT, CD3\u0026thinsp;+\u0026thinsp;CD4\u0026thinsp;+\u0026thinsp;CD8+, and CD4+/CD8\u0026thinsp;+\u0026thinsp;lymphocytes (\u003cem\u003ep\u003c/em\u003e\u0026thinsp;\u0026gt;\u0026thinsp;0.05) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003e3. Clinicopathological characteristics of the CRC patients\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe tumor location, stage, and differentiation grade are presented in Table\u0026nbsp;3. In addition, the percentages of lymph node invasion, vascular infiltration, distant metastasis, and perineural invasion were also calculated (Table\u0026nbsp;\u003cspan refid=\"Tab2\" class=\"InternalRef\"\u003e2\u003c/span\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\u003eClinicopathological characteristics of the CRC patients\u003c/p\u003e\u003c/div\u003e\u003c/caption\u003e\u003ccolgroup cols=\"3\"\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e\u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e\u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e\u003cthead\u003e\u003ctr\u003e\u003cth align=\"left\" colname=\"c1\"\u003e\u003cp\u003eCharacteristics\u003c/p\u003e\u003c/th\u003e\u003cth align=\"left\" colname=\"c2\"\u003e\u0026nbsp;\u003c/th\u003e\u003cth align=\"left\" colname=\"c3\"\u003e\u003cp\u003eNumber of patients (%)\u003c/p\u003e\u003c/th\u003e\u003c/tr\u003e\u003c/thead\u003e\u003ctbody\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTumor location\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eColon\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e74 (62.7%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eRectum\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e44 (37.3%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTumor stage\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eI-II\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e19 (16.2%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eIII-IV\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e98 (83.8%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eTumor differentiation grade\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eWell\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e6(6.0%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eModerate\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e60(60.0%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003ePoor\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e34 (34.0%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eLymph node invasion\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eYes\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e47 (43.1%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eNo\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e62(56.9%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eVascular infiltration\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eYes\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e22 (29.7%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eNo\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e52 (70.3%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003eDistant metastasis\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eYes\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e11(12.6%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eNo\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e76(87.4%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u003cp\u003ePerineural invasion\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eYes\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e24(25.0%)\u003c/p\u003e\u003c/td\u003e\u003c/tr\u003e\u003ctr\u003e\u003ctd align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/td\u003e\u003ctd align=\"left\" colname=\"c2\"\u003e\u003cp\u003eNo\u003c/p\u003e\u003c/td\u003e\u003ctd align=\"char\" char=\".\" colname=\"c3\"\u003e\u003cp\u003e72(75.0%)\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\u003cb\u003e4. Comparison of lymphocyte subsets in the peripheral blood of the CRC patients before and after surgery\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe peripheral blood was collected before and after surgery within a month. The number of T lymphocytes was significantly increased after surgical resection (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eA), however, the number of B lymphocytes was significantly decreased after surgical resection (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eH). There was no significance in the number of other lymphocyte subsets (Fig.\u0026nbsp;\u003cspan refid=\"Fig2\" class=\"InternalRef\"\u003e2\u003c/span\u003eB-G).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e\u003cp\u003e\u003cb\u003e5. The absolute number of lymphocyte subsets in different subgroups, according to tumor characteristics\u003c/b\u003e\u003c/p\u003e\u003cp\u003eThe absolute number of lymphocyte subsets in different subgroups, according to tumor characteristics, was analyzed. There was a statistical difference between NK cell amounts in stage I-II and III-IV. The number of NK cells increased in stage III-IV, as compared to stage I-II (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eA). The number of Ts lymphocytes was lower in moderate and well-differentiated than in poor differentiation (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eB). The number of CD4+/CD8\u0026thinsp;+\u0026thinsp;lymphocytes was significantly higher in moderate differentiation than in poor differentiation (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eB). The number of CD4+/CD8\u0026thinsp;+\u0026thinsp;lymphocytes was higher in high differentiation than in poor differentiation but without statistical significance (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eB). The number of T lymphocytes was high in conditions without lymph node invasion (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eC). There was no significance in the lymphocyte subsets between the group with and without vascular infiltration (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eD). The number of T and Th lymphocytes was higher in the condition that was without distant metastasis, however, the number of NK cells was lower in the condition that without distant metastasis (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eE). The number of CD4+/CD8\u0026thinsp;+\u0026thinsp;lymphocytes was significantly higher in the condition without perineural invasion (Fig.\u0026nbsp;\u003cspan refid=\"Fig3\" class=\"InternalRef\"\u003e3\u003c/span\u003eF).\u003c/p\u003e\u003cp\u003e\u003c/p\u003e"},{"header":"DISCUSSION","content":"\u003cp\u003eImmune cell infiltration is a characteristic of almost all malignant tumors, which plays an important role in immune monitoring and contributes to the elimination of tumor cells \u003csup\u003e21\u003c/sup\u003e. Given the minimally invasive nature of blood sample collection compared to tumor-infiltrating tissues, the regularity of various indexes in peripheral blood is more convenient for clinical application, such as T lymphocyte cells, B lymphocyte cells, and NK cells \u003csup\u003e22\u003c/sup\u003e. In this study, we observed significant changes in the circulating adaptive lymphocyte profile of CRC patients compared with those of healthy patients. Furthermore, we assessed differences in the distribution of adaptive immune cells in patients with CRC. \u003c/p\u003e\n\u003cp\u003eThe T cell subsets defined as having different proliferative abilities, lymphatic homing, and effector functions have been well established in previous studies \u003csup\u003e23,24\u003c/sup\u003e. Th cells help maintain Tc cell immune responses and regulate the development of Tc cells into functional cells \u003csup\u003e25\u003c/sup\u003e. In our study, colorectal carcinoma patients presented with significantly lower values of absolute circulating T cell (CD45+/CD3+) and Th cell (CD3+/CD4+/CD8-) counts compared to healthy volunteers, which is consistent with prior research \u003csup\u003e10,26,27\u003c/sup\u003e. After the rection of the tumor, the count of T cell counts showed an increased trend, with statistical significance. However, other subtypes of T lymphocyte cells also showed an increase, but without statistical significance. It may be because the suppression of T cells was removed together with the tumor. \u003c/p\u003e\n\u003cp\u003eAs our data showed, the number of T lymphocytes was lower in the condition that either with lymph node invasion or with distant metastasis. In peripheral circulation and tissues, both Th and Tc are capable of dynamic metabolic control \u003csup\u003e28\u003c/sup\u003e. The decrease in the Th cell number in CRC patients could be due to a reduction in peripheral circulation produced by adaptive immune cells migrating to tumor tissues. Th cell depletion, which manifests as a decrease in proliferative and functionally active T cells and an increase in senescent T cells, results from persistent metabolic disturbances directed by tumor cells \u003csup\u003e29\u003c/sup\u003e. This is consistent with our findings in patients with distant metastasis. In addition, the density of sT cells (CD3+CD8+) was inversely associated with the degree of tumor differentiation. The previous research showed, that the density of CD3+CD8+ T cells was inversely associated with the extent of tumor budding at the invasive front \u003csup\u003e30\u003c/sup\u003e, and also was significantly associated with high PD-L1 expression scores \u003csup\u003e31\u003c/sup\u003e. However, the specific role of sT cell still need further exploring.\u003c/p\u003e\n\u003cp\u003eB cells contribute to cancer regulation primarily through indirect inhibition of T cell reactivity, resulting in antitumor immunosuppression. In our present study, the values of circulating B cells (CD3-/CD19+) were also lower in CRC patients, as compared to the normal group, but without statistical significance. Waidhauser et al.\u003csup\u003e32\u003c/sup\u003e found that B cell counts were lower in CRC patients, whereas Spacek et al.\u003csup\u003e33\u003c/sup\u003e demonstrated that it was higher in colon cancer patients than those in control groups. It is interesting that after the rection of the tumor, the count of B cells was decreased significantly. In addition, the circulating B lymphocytes showed no significant correction with tumor pathological characteristics. According to the previous study, there were no significant differences in the value changes of B cell subsets in advanced-stage CRC patients compared with those of patients in the early stage \u003csup\u003e10\u003c/sup\u003e. However, an increasing number of studies have shown that cytokines secreted by B cells have dual roles in tumors during tumorigenesis and progression \u003csup\u003e34,35\u003c/sup\u003e. Whether the role of B cell subsets in CRC can be further explored from the perspective of cytokine secretion remains to be confirmed by further studies.\u003c/p\u003e\n\u003cp\u003eNK cells participate in tumor immunosurveillance by monitoring and killing tumor cells in an antigen-independent and antigen-dependent manner \u003csup\u003e36\u003c/sup\u003e. Several studies have revealed that low NK cell counts predict a higher risk of tumors \u003csup\u003e37\u003c/sup\u003e. Although NK cells possess cytotoxic capabilities, the specific activity of each NK cell subset is complex. It is interesting in our study that, the number of NK cells increased in the CRC patients, as compared with normal group, which is opposite to the previous studies \u003csup\u003e32,33\u003c/sup\u003e. We speculated that the NK immunity of CRC patients might be strengthened to fight against the tumor.\u003c/p\u003e\n\u003cp\u003eDue to the small sample size, this study may not fully represent the broader population, which introduces certain limitations to the findings. The mechanisms underlying the observed changes in lymphocyte subsets profiles require further investigation.\u003c/p\u003e\n\u003cp\u003eIn summary, our study revealed significant alterations in circulating lymphocyte subsets that can be attributed to the presence of colorectal cancer. Additionally, certain tumor characteristics like tumor differentiation seem to make a difference in the individual immune response. Whether these circulating lymphocyte alterations play a role in predicting the prognosis of colorectal cancer and whether the alterations are the cause or consequence of the tumor disease will need further investigations.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003e\u003cstrong\u003eACKNOWLEDGMENTS\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThis work was supported by the Tianjin Natural Science Foundation (no. 21JCYBJC01070), the\u0026nbsp;Foundation\u0026nbsp;of Tianjin Union Medical Center (2022PWXK04)， and the Tianjin Key Medical Discipline Construction Project (TJYXZDXK-3-009B).\u0026nbsp;\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eCONFLICTS OF INTEREST\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe authors claim that none of the material in the paper has been published or is under consideration for publication elsewhere, and there is no conflict of interests. All authors have approved the manuscript for submission.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthics approval and consent to participate\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eNot applicable.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eAuthors\u0026apos; contributions\u0026nbsp;\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eGuanghai Wu and Kunming Zheng performed the statistical analyses and wrote the paper. Wang Han collected the data, and Shichao Xu contributed to the experiment. Jing Xu revised this paper and designed the study. All authors contributed to the development of this manuscript and read and approved the final version.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eThe Data Availability\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eThe data used and/or analyzed during the current study are available from the corresponding authors on reasonable request.\u0026nbsp;\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eTorre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A. Global cancer statistics, 2012. \u003cem\u003eCA: a cancer journal for clinicians. \u003c/em\u003eMar 2015;65(2):87-108.\u003c/li\u003e\n\u003cli\u003eArnold M, Sierra MS, Laversanne M, Soerjomataram I, Jemal A, Bray F. Global patterns and trends in colorectal cancer incidence and mortality. \u003cem\u003eGut. \u003c/em\u003eApr 2017;66(4):683-691.\u003c/li\u003e\n\u003cli\u003eSiegel RL, Miller KD, Fedewa SA, et al. 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Abnormal distribution of peripheral lymphocyte subsets induced by PDAC modulates overall survival. \u003cem\u003ePancreatology. \u003c/em\u003eJul-Aug 2014;14(4):295-301.\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"journal-of-gastrointestinal-cancer","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"ijgc","sideBox":"Learn more about [Journal of Gastrointestinal Cancer](https://www.springer.com/journal/12029)","snPcode":"12029","submissionUrl":"https://submission.nature.com/new-submission/12029/3","title":"Journal of Gastrointestinal Cancer","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Lymphocyte subsets, Colorectal cancer, Cellular immune status, Tumor progression","lastPublishedDoi":"10.21203/rs.3.rs-7672745/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-7672745/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003e\u003cstrong\u003ePurpose:\u003c/strong\u003e Cellular immune response to cancer is known to be of great importance for tumor control. Until now, the role of systemic immune profiles in tumor progression remains unclear.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eMethods: \u003c/strong\u003eBy using multiparameter flow cytometry, the subset distribution and immunophenotype of lymphocyte subsets were investigated in peripheral blood mononuclear cell samples from 118 colorectal cancer (CRC) patients and 80 healthy donors.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eResults:\u003c/strong\u003e Compared with the normal group, the numbers of T lymphocytes and Th lymphocytes were significantly decreased in the CRC group, whereas the number of NK cells was significantly increased. There was no significance between the two groups in the numbers of Ts, γδT, CD3+CD4+CD8+, and CD4+/CD8+ lymphocytes. The number of T lymphocytes was significantly increased after surgical resection, however, the number of B lymphocytes was significantly decreased. There was no significance in the number of other lymphocyte subsets neither before nor after rection. The number of NK cells increased in stage III-IV, as compared to stage I-II. The numbers of Ts and CD4+/CD8+ lymphocytes showed a correlation with the tumor differentiation degree. The number of T lymphocytes was higher in conditions that were without lymph node invasion and distant metastasis. The number of CD4+/CD8+ lymphocytes was significantly higher in the condition without perineural invasion.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eConclusion:\u003c/strong\u003e The altered phenotype of circulating immune cell subsets in CRC and its association with clinical outcome highlights the potential use of lymphocyte subsets as prognostic biomarkers in CRC, thereby contributing to better insight into the role of systemic immune profiles in tumor progression.\u003c/p\u003e","manuscriptTitle":"Characterization and predictive value of lymphocyte subsets in peripheral blood of patients with colorectal cancer","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-10-08 15:58:32","doi":"10.21203/rs.3.rs-7672745/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Revision requested","date":"2025-10-25T14:33:30+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-18T18:28:54+00:00","index":"hide","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2025-10-15T11:12:28+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"103250383974641740587442147188830526862","date":"2025-09-30T10:30:19+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"90863371734763501606434519294928424490","date":"2025-09-30T10:20:16+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"221417754808285487510735337969314423633","date":"2025-09-29T14:02:06+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2025-09-25T11:52:09+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2025-09-24T22:06:44+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2025-09-24T07:32:27+00:00","index":"","fulltext":""},{"type":"submitted","content":"Journal of Gastrointestinal Cancer","date":"2025-09-22T09:33:24+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"journal-of-gastrointestinal-cancer","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"ijgc","sideBox":"Learn more about [Journal of Gastrointestinal Cancer](https://www.springer.com/journal/12029)","snPcode":"12029","submissionUrl":"https://submission.nature.com/new-submission/12029/3","title":"Journal of Gastrointestinal Cancer","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"5c4125cb-ab35-4dac-a105-74d55dcc2b1f","owner":[],"postedDate":"October 8th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2026-03-02T16:05:44+00:00","versionOfRecord":{"articleIdentity":"rs-7672745","link":"https://doi.org/10.1007/s12029-026-01402-4","journal":{"identity":"journal-of-gastrointestinal-cancer","isVorOnly":false,"title":"Journal of Gastrointestinal Cancer"},"publishedOn":"2026-02-26 15:59:10","publishedOnDateReadable":"February 26th, 2026"},"versionCreatedAt":"2025-10-08 15:58:32","video":"","vorDoi":"10.1007/s12029-026-01402-4","vorDoiUrl":"https://doi.org/10.1007/s12029-026-01402-4","workflowStages":[]},"version":"v1","identity":"rs-7672745","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-7672745","identity":"rs-7672745","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}