¿p#1 Cytomegalovirus IL-10 in plasma as a marker of active infection in allogeneic hematopoietic transplant recipients: An exploratory study

¿p#1 Cytomegalovirus IL-10 in plasma as a marker of active infection in allogeneic hematopoietic transplant recipients: An exploratory study | Authorea try { document.documentElement.classList.add('js'); } catch (e) { } var _gaq = _gaq || []; _gaq.push(['_setAccount', 'G-8VDV14Y67G']); _gaq.push(['_trackPageview']); (function() { var ga = document.createElement('script'); ga.type = 'text/javascript'; ga.async = true; ga.src = ('https:' == document.location.protocol ? 'https://ssl' : 'http://www') + '.google-analytics.com/ga.js'; var s = document.getElementsByTagName('script')[0]; s.parentNode.insertBefore(ga, s); })(); Skip to main content Preprints Collections Wiley Open Research IET Open Research Ecological Society of Japan All Collections About About Authorea FAQs Contact Us Quick Search anywhere Search for preprint articles, keywords, etc. Search Search ADVANCED SEARCH SCROLL Journal of Medical Virology This is a preprint and has not been peer reviewed. Data may be preliminary. 17 November 2025 V1 Latest version Share on ¿p#1 Cytomegalovirus IL-10 in plasma as a marker of active infection in allogeneic hematopoietic transplant recipients: An exploratory study Authors : Ángela Sánchez , Eliseo Albert , Estela Giménez 0000-0003-1774-9790 , Ester Colomer , Ariadna Pérez , José PIñana 0000-0001-8533-2562 , Carlos Solano , and David Navarro 0000-0003-3010-4110 [email protected] Authors Info & Affiliations https://doi.org/10.22541/au.176338490.03500282/v1 229 views 128 downloads Contents Abstract Supplementary Material Information & Authors Metrics & Citations View Options References Figures Tables Media Share Abstract ¿p#1 We investigated whether plasma Cytomegalovirus (CMV) IL-10 (cmvIL-10) levels could serve as a biomarker of active CMV replication in allogeneic hematopoietic transplant recipients (allo-HCT) in the presence or absence of letermovir (LMV) prophylaxis. A total of 189 leftover plasma samples that tested positive for CMV DNA (Alinity m CMV assay), representing 33 episodes of CMV DNAemia were run on a laboratory developed enzyme linked immunosorbent assay for cmvIL-10 quantification. Eighteen episodes developed during LMV prophylaxis. Overall, 16 episodes of CMV DNAemia were classified as clinically significant (CsCMVi). There was an overall very weak correlation between the two biomarkers (Rho = 0.10; P = 0.16). Overall, the median cmvIL-10 area under the curve (AUC) until CMV DNA levels reached their peak was significantly higher ( P < 0.001) in CsCMVi episodes than in non-CsCMVi episodes. cmvIL-10 AUC between days 14 and 23 after allo-HCT (AUC 14-23 ) values were significantly higher in CsCMVi episodes compared with non-CsCMVi episodes among patients receiving LMV therapy ( P =0.008). An AUC 14-23 cutoff value of log 10 3.06 discriminated anticipately between CsCMVi and non-CsCMVi with a sensitivity and specificity of 100%. Plasma cmvIL-10 levels may reflect true CMV replication and thus provide a unique perspective on viral dynamics, serving as an ancillary marker to CMV DNA monitoring. ¿p#1 INTRODUCTION Monitoring CMV DNA load in blood is a cornerstone of cytomegalovirus (CMV) infection management in allogeneic hematopoietic stem cell transplant (allo-HCT) recipients, as it enables timely initiation of antiviral therapy to control viral replication and thereby minimize the risk of CMV end-organ disease¹. However, the presence of CMV DNA in blood does not always indicate an ongoing episode of viral replication, since free viral DNA may transiently leak into the bloodstream and be rapidly cleared (i.e., “blips” or self-resolving episodes). This phenomenon is particularly frequent among patients receiving letermovir (LMV) prophylaxis² , ³, as LMV inhibits the production of infectious viral particles but does not block CMV DNA replication⁴. While the kinetic pattern of CMV DNA in plasma or whole blood often provides valuable information in this context⁵ , ⁶, novel biomarkers of active CMV infection, such as CMV UL25.1 RNAemia, have been proposed⁷ - ⁹. The CMV gene UL111a encodes a homolog of human interleukin-10 (hIL-10), referred to as cmvIL-10, which is predominantly expressed during lytic infection¹⁰. cmvIL-10 exerts a broad range of immunosuppressive functions through its interaction with the human IL-10 receptor (IL-10R1)¹¹ , ¹². Deep sequencing of the UL111a gene directly from clinical samples has revealed the presence of viral variants that may differentially modulate host immune responses¹³. Furthermore, it was recently shown that expression of cmvIL-10 RNA in peripheral blood from kidney transplant recipients was positively associated with an increase in viral DNA detection in subsequent specimens, suggesting that monitoring cmvIL-10 may be ancillary to viral DNA to allow early detection of active CMV infection in transplant recipients 14 . In this study, we investigated whether plasma cmvIL-10 levels could serve as a biomarker of active CMV replication in patients with CMV DNAemia, occurring either during or outside LMV prophylaxis. ¿p#1 MATERIAL AND METHODS ¿p#1 Patients and specimens A convenience panel of 189 leftover plasma samples that tested positive for CMV DNA using the Alinity m CMV assay (Abbott Molecular Inc., Des Plaines, IL, USA)⁵ was assembled. These samples represented 33 episodes of CMV DNAemia occurring in unique CMV-seropositive allo-HCT recipients that underwent primary LMV prophylaxis. Eighteen episodes developed during LMV prophylaxis (n=110 specimens), and 15 occurred outside LMV prophylaxis (n= 79 specimens). Sixteen episodes of CMV DNAemia were classified as clinically significant (CsCMVi), defined as those in which peak CMV DNA levels exceeded the institutional threshold for preemptive antiviral therapy (1,500 IU/mL, regardless of LMV prophylaxis status). Seven of these 16 CsCMVi episodes occurred in patients receiving LMV therapy. All plasma samples had been cryopreserved at -80°C within 24 h of collection between January, 2023 and February 2025 and had not been thawed before cmvIL-10 testing. The current study was approved by the Institutional Review Board (IRB) Research Ethics Committee of Hospital Clínico Universitario INCLIVA (2024/153). The IRB issued an informed consent waiver. ¿p#1 Quantification of cmvIL-10 in plasma Quantification of cmvIL-10 in plasma was performed using an enzyme-linked immunosorbent assay (ELISA) as previously described 15 . Briefly, 96-well Nunc MaxiSorp™ plates were coated with 100 μL of capture antibody (cmvIL-10 Affinity Purified Polyclonal Antibody, Goat IgG; R&D Systems, Minneapolis, MN) at a final concentration of 2 μg/mL and incubated overnight at 4°C. Following removal of the capture antibody, plates were washed three times with wash buffer (1× PBS + 0.05% Tween 20) and blocked with blocking buffer (PBS + 3% bovine serum albumin) for 1 hour at room temperature. After three additional washes, 100 μL of plasma samples (diluted 1:5) and standards were added in duplicate and incubated for 2 hours at room temperature. Protein standards were prepared by serial two-fold dilutions of recombinant cmvIL-10 (R&D Systems) in protein standard dilution buffer (PBS + 0.1% BSA), ranging from 1000 pg/mL to 15.625 pg/mL. Following incubation and washing, detection antibody (cmvIL-10 Biotinylated Affinity Purified Polyclonal Antibody, Goat IgG; R&D Systems) was added at a final concentration of 0.2 μg/mL and incubated for 2 hours at room temperature. After three washes, streptavidin-HRP (R&D Systems) was added at working concentration and incubated for 20 minutes in the dark. Following three final washes, substrate solution (R&D Systems) was added and incubated in the dark for 20 minutes. The reaction was stopped with 50 μL of stop solution (1 M H₂SO₄), and optical density was measured at 450 nm using a Virclia instrument (Vircell, Granada, Spain). Standard curves were generated using GraphPad Prism software by plotting optical density values against recombinant cmvIL-10 concentrations using four-parameter logistic curve fitting. Plasma cmvIL-10 concentrations (pg/mL) were interpolated from the standard curve. ¿p#1 Statistical analyses Differences between medians were compared using the Mann–Whitney U test. The degree of correlation between continuous variables was analyzed using Spearman’s Rank test. The cmvIL-10 area under a curve (AUC) was calculated when appropriate, and required 2 or more specimens/patient. The Youden index was used to determine the optimal AUC threshold to maximize the difference between the true positive rate (sensitivity) and the false positive rate (1-specificity). Two-sided exact P -values are reported. A P -value < 0.05 was considered statistically significant. The analyses were performed using the GraphPad Prism 9.0.2 statistical package. ¿p#1 Correlation between cmvIL-10 and CMV DNA levels in plasma We first investigated whether plasma cmvIL-10 levels correlated with CMV DNA loads as measured by a real-time PCR assay. A total of 189 plasma specimens with quantifiable CMV DNA, representing 33 episodes of active CMV infection that developed a median of 26 days after allo-HCT (IQR, 8.5–65), were available for cmvIL-10 testing (median of 5 samples per episode; range, 3–12). The timing of specimen collection relative to allo-HCT is presented in Supplementary Table 1. Among the 189 specimens, 135 tested positive for cmvIL-10 (median value, 190 pg/ml; IQR, 82.3-373.2), while 54 were undetectable. Median cmvIL-10 peak value was 396.5 pg/mL (range, 83-5,657). As shown in Figure 1, there was an overall very weak correlation between the two biomarkers (Rho = 0.10; P = 0.16). The degree of correlation was poor regardless of whether patients were under LMV prophylaxis (Figure 1B) or not (Figure 1C). ¿p#1 cmvIL-10 as a marker of active infection To assess whether cmvIL-10 measurement could identify CsCMVi episodes, we calculated the AUCs using cmvIL-10 values from specimens collected until the CMV DNA peak level. This parameter was available for 29 of the 33 episodes (CsCMVi, n= 15; non-CsCMVi, n=14). As shown in Figure 2A, the median cmvIL-10 AUC was significantly higher ( P < 0.001) in CsCMVi episodes than in non-CsCMVi episodes (log 10 3.1 vs. log 10 0.56). Notably, the number of specimens used for AUC calculations was comparable across groups (median, 3 specimens; P ≥ 0.5), as was the timing of the first specimen collection after allo-HCT (median, 18 days for CsCMVi vs. 19 days for non-CsCMVi). A similar pattern was observed when only episodes occurring during LMV prophylaxis were analyzed separately (Figure 2B). We next investigated whether cmvIL-10 AUCs could predict the occurrence of CsCMVi. To this end, we calculated AUCs between days 14 and 23 post–allo-HCT (cmvIL-10 AUC₁₄₋₂₃), encompassing measurements from plasma specimens collected prior to the CMV DNAemia peak (median, 7 days; range, 6–10 days earlier). As shown in Figure 3A, there was a trend toward higher cmvIL-10 AUC₁₄₋₂₃ values in CsCMVi episodes (median, log 10 3.03) compared with non-CsCMVi episodes (median, log 10 2.4; P= 0.08). The median number of specimens used for AUC calculations was similar across groups (n = 3). In this setting, an AUC cutoff value of log 10 2.45 provided the most efficient discrimination between CsCMVi and non-.CsCMVi, with a sensitivity of 50% and a specificity of 89% (AUC=0.73; P =0.06). Interestingly, cmvIL-10 AUC₁₄₋₂₃ values were significantly higher in CsCMVi episodes compared with non-CsCMVi episodes among patients receiving LMV therapy ( P =0.008). In this setting, an AUC₁₄₋₂₃ cutoff value of log 10 3.06 discriminated perfectly across comparison groups, with a sensitivity and specificity of 100% (AUC=1; P =0.009). ¿p#1 DISCUSSION The kinetics of CMV DNA in blood has proven to be a useful parameter for anticipating the occurrence of clinically significant CMV infection (CsCMVi) in allo-HCT recipients irrespective of the real-time PCR assay employed. 16,17 It is well documented that the presence of CMV DNA in blood may not necessarily reflect active CMV infection in tissues or in the blood compartment due to the mechanism of action of LMV 2,3 . In this context, identifying episodes of CMV DNAemia that represent true viral replication, as opposed to abortive infection, is particularly relevant in patients receiving LMV, in order to avoid unnecessary interruption of therapy. Our group has demonstrated the potential value of CMV DNA doubling time (CMV dt) for the early identification of true episodes of active CMV infection, both in the presence and absence of LMV treatment 5,6,16,17 ; nevertheless, the clinical performance of this parameter, in terms of its predictive value, is not maximally accurate. Monitoring of CMV UL25.1 RNAemia using a commercially available assay has been proposed as a reliable marker of active CMV infection, since this late CMV transcript appears to be virion-associated in plasma⁷˒⁸. Nevertheless, because LMV does not inhibit either CMV DNA replication or the synthesis of late CMV mRNAs, we believe that this marker does not substantially enhance the information already provided by quantitative CMV DNA testing in patients receiving LMV¹ 8 . Here, we reasoned that monitoring plasma cmvIL-10 levels might aid in the early identification of CsCMVi, as this viral human-homologue cytokine is mainly synthesized during viral replication and exerts immunosuppressive effects that may vary across strains 10-13 . We found a negligible correlation between CMV DNA and cmvIL-10 levels, in contrast to what has been reported for CMV UL25.1 RNAemia. Furthermore, monitoring cmvIL-10 levels could reliably identify and, more importantly, anticipate clinically significant CMV infection (CsCMVi) both in the presence and absence of LMV therapy. Notably, an AUC cutoff value of log₁₀ 3.06 perfectly discriminated between CsCMVi and non-CsCMVi groups, with this distinction occurring a median of one week before CMV DNAemia reached levels sufficient for classification as such. Taken together, our findings support the notion that plasma cmvIL-10 levels may reflect true CMV replication and thus provide a unique perspective on viral dynamics, serving as an ancillary marker to CMV DNA monitoring. Nevertheless, well-designed prospective studies are warranted to further assess the clinical utility of cnvIL-10 testing, particularly in LMV-treated patients. ¿p#1 ACKNOWLEDGEMENTS This research was supported by grant from the Fondo de Investigaciones Sanitarias, Ministerio de Sanidad y Consumo, Spain (FIS PI24/01844). ¿p#1 ETHICAL APPROVAL The current study was approved by the Institutional Review Board (IRB) Research Ethics Committee of Hospital Clínico Universitario INCLIVA (2024/153). The IRB issued an informed consent waiver. ¿p#1 CONFLICTS OF INTEREST JLP, CS and DN received honoraria for conferences sponsored by MSD. ¿p#1 AUTHOR CONTRIBUTION AS-S, EA, EG, EC, AP, JLP, and CS, data curation and analysis, methodology. DN, Conceptualization, Supervision, Data analysis and writing. ¿p#1 AVAILABILITY OF DATA AND MATERIALS The datasets generated and/or analyzed during the current study are available from the corresponding author upon reasonable request. ¿p#1 REFERENCES 1. Ljungman P, Alain S, Chemaly RF, Einsele H, Galaverna F, Hirsch HH, Sadowska-Klasa A, Navarro D, Styczynski J, de la Camara R. Recommendations from the 10th European Conference on Infections in Leukaemia for the management of cytomegalovirus in patients after allogeneic haematopoietic cell transplantation and other T-cell-engaging therapies. Lancet Infect Dis . 2025; S1473-3099(25)00069-6. 2. Cassaniti I, Colombo AA, Bernasconi P, Malagola M, Russo D, Iori AP, et al . Positive HCMV DNAemia in stem cell recipients undergoing letermovir prophylaxis is expression of abortive infection. Am J Transplant . 2021; 21(4):1622-1628. 3. Giménez E, Guerreiro M, Torres I, Aguilar C, Albert E, Hernández-Boluda JC, et al. Features of cytomegalovirus DNAemia and virus-specific T-cell responses in allogeneic hematopoietic stem-cell transplant recipients during prophylaxis with letermovir. Transpl Infect Dis . 2023; 25:e1402. 4. Piret J, Boivin G. Antiviral Drugs Against Herpesviruses. Adv Exp Med Biol . 2025; 1322:1-30. 5. Esteve M, Albert E, Giménez E, Colomer E, Hernández-Boluda JC, Hernani R, et al. Assessment of Cytomegalovirus DNA doubling time and virus-specific T-cell responses in the management of CMV infection in allogeneic hematopoietic stem cell transplant recipients undergoing Letermovir prophylaxis. Bone Marrow Transplant . 2024; 59(10):1480-1482. 6. Gimenez E, García Cadenas I, Piñana JL, Albert E, Vázquez L, Avendaño L, et al. Cytomegalovirus DNA doubling time for early identification of clinically significant infection episodes in allogeneic hematopoietic stem cell transplant recipients undergoing primary Letermovir prophylaxis: A multicenter study. Transplant Infect Dis . 2025. e70080 7. Piccirilli G, Lanna F, Gabrielli L, Motta V, Franceschiello M, Cantiani A, et al. CMV-RNAemia as new marker of active viral replication in transplant recipients. J Clin Microbiol. 2024; 62(5):e0163023. 8. Giardina F, Paolucci S, Mele D, Mura O, Ramus M, Sammartino JC, et al . Human Cytomegalovirus Virion-Associated mRNA as a Marker of Productive Infection in Immunocompromised Patients. J Med Virol . 2025; 97(5):e70378. 9. de la Asunción CS, Ocete MD, Giménez E, Albert E, Gimeno C, Piñana JL, et al . Cytomegalovirus UL21-5 mRNAemia as a Marker of Active Virus Replication in Allogeneic Hematopoietic Transplant Recipients. J Med Virol. 2025;97(10):e70617. 10. Kotenko SV, Saccani S, Izotova LS, Mirochnitchenko OV, Pestka S. Human cytomegalovirus harbors its own unique IL-10 homolog (cmvIL-10). Proc Natl Acad Sci USA . 2000; 97 (4) 1695–1700. 11. Poole E, Neves TC, Oliveira MT, Sinclair J, da Silva MCC. Human Cytomegalovirus Interleukin 10 Homologs: Facing the Immune System. Front Cell Infect Microbiol. 2020;10:245. 12. Jones BC, Logsdon NJ, Josephson K, Cook J, Barry PA, Walter MR. Crystal structure of human cytomegalovirus IL-10 bound to soluble human IL-10R1. Proc Natl Acad Sci U S A. 2002;99(14):9404-9409. 13, Waters S, Lee S, Ariyanto I, Kresoje N, Leary S, Munyard K, et al. Sequencing of the Viral UL111a Gene Directly from Clinical Specimens Reveals Variants of HCMV-Encoded IL-10 That Are Associated with Altered Immune Responses to HCMV. Int J Mol Sci. 2022;23(9):4644. 14. Almeida GWC, Oliveira MT, Martines IGL, Fiori GC, Nevels MM, Groves IJ, et al. Expression Profile of Human Cytomegalovirus UL111A cmvIL-10 and LAcmvIL-10 Transcripts in Primary Cells and Cells from Renal Transplant Recipients. Viruses. 2025;17(4):501. 15. Young VP, Mariano MC, Faure L, Spencer JV. Detection of Cytomegalovirus Interleukin 10 (cmvIL-10) by Enzyme-Linked Immunosorbent Assay (ELISA). Methods Mol Biol. 2021;2244:291-299. 16. Giménez E, Muñoz-Cobo B, Solano C, Amat P, Navarro D. Early kinetics of plasma cytomegalovirus DNA load in allogeneic stem cell transplant recipients in the era of highly sensitive real-time PCR assays: does it have any clinical value? J Clin Microbiol. 2014;52(2):654-656. 17. Vinuesa V, Giménez E, Solano C, Gimeno C, Navarro D. Would Kinetic Analyses of Plasma Cytomegalovirus DNA Load Help to Reach Consensus Criteria for Triggering the Initiation of Preemptive Antiviral Therapy in Transplant Recipients? Clin Infect Dis. 2016;63(11):1533-1535. 18. de la Asunción CS, Ocete MD, Giménez E, Albert E, Gimeno C, Piñana JL, et al . Cytomegalovirus UL21-5 mRNAemia as a Marker of Active Virus Replication in Allogeneic Hematopoietic Transplant Recipients. J Med Virol. 2025;97(10):e70617. ¿p#1 FIGURE LEGENDS Figure 1. Correlation between Cytomegalovirus (CMV) IL-10 (cmvIL-10) and CMV DNA levels in plasma from allogeneic hematopoietic stem cell transplant recipients. (A) all specimens; (B) specimens collected while patients were under letermovir prophylaxis; (C) specimens collected while patients were not under letermovir prophylaxis. Spearman rank (Rho) and P values are shown. Figure 2. Area under the curve (AUC) of Cytomegalovirus (CMV) IL-10 levels (cmvIL-10) measured in plasma up to the CMV DNA peak in allogeneic hematopoietic stem cell transplant recipients who either developed or did not develop clinically significant CMV infection (CsCMVi). (A) All patients in the cohort. (B) Patients receiving letermovir therapy at the time of CMV DNAemia occurrence. Bars represent medians and interquartile ranges. P values for group comparisons are shown. Figure 3 . Area under the curve (AUC) of Cytomegalovirus (CMV) IL-10 levels (cmvIL-10) measured in plasma between days 14 and 23 (AUC₁₄₋₂₃) after allogeneic hematopoietic stem cell transplant in patients who either developed or did not develop clinically significant CMV infection (CsCMVi). (A) All patients in the cohort. (B) Patients receiving letermovir therapy at the time of CMV DNAemia occurrence. Bars represent medians and interquartile ranges. P values for group comparisons are shown. Supplementary Material File (angela sanchez jmv 2025 table 1.docx) Download 16.31 KB Information & Authors Information Version history V1 Version 1 17 November 2025 Copyright This work is licensed under a Non Exclusive No Reuse License. Collection Journal of Medical Virology Keywords cytomegalovirus epidemiology infection transplantation virus classification Authors Affiliations Ángela Sánchez Fundacion para la Investigacion del Hospital Clinico de la Comunitat Valenciana View all articles by this author Eliseo Albert Fundacion para la Investigacion del Hospital Clinico de la Comunitat Valenciana View all articles by this author Estela Giménez 0000-0003-1774-9790 Fundacion para la Investigacion del Hospital Clinico de la Comunitat Valenciana View all articles by this author Ester Colomer Fundacion para la Investigacion del Hospital Clinico de la Comunitat Valenciana View all articles by this author Ariadna Pérez Fundacion para la Investigacion del Hospital Clinico de la Comunitat Valenciana View all articles by this author José PIñana 0000-0001-8533-2562 Fundacion para la Investigacion del Hospital Clinico de la Comunitat Valenciana View all articles by this author Carlos Solano Fundacion para la Investigacion del Hospital Clinico de la Comunitat Valenciana View all articles by this author David Navarro 0000-0003-3010-4110 [email protected] Fundacion para la Investigacion del Hospital Clinico de la Comunitat Valenciana View all articles by this author Metrics & Citations Metrics Article Usage 229 views 128 downloads .FvxKWukQNSOunydq8rnd { width: 100px; } Citations Download citation Ángela Sánchez, Eliseo Albert, Estela Giménez, et al. ¿p#1 Cytomegalovirus IL-10 in plasma as a marker of active infection in allogeneic hematopoietic transplant recipients: An exploratory study. 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