Human Cytomegalovirus (HCMV) and Epstein-Barr virus (EBV) are endowed with the ability of establishing lifelong latency in human hosts and reactivating in immunocompromised subjects, including patients suffering from ulcerative colitis (UC). We, therefore, aimed to investigate virus-specific immunity in UC patients. A cohort of 24 UC patients (14 responders and 10 refractory to therapy) and 26 control subjects was prospectively enrolled to undergo virus-specific serology (by ELISA assay) and assessment of both CD4+ and CD8+ virus-specific T-cell response (by interferon-γ enzyme-linked immunospotanalysis). In parallel, mucosal viral load was determined by quantitative real-time PCR and the values were correlated with both clinical and endoscopic indexes of activity. For statistics, the t-test, Mann-Withney test, Fisher's exact test and Spearman rank correlation test were applied; p < 0.05 was considered significant. EBV-specific CD4+ and CD8+ T-cell responses were significantly lower in UC patients compared to controls (p < 0.0001 and p = 0.0006, respectively), whereas no difference was found for HCMV-specific T-cell response. When dividing the UC group according to response to therapy, both responders and refractory UC patients showed a deficient EBV-specific CD4+ T-cell response with respect to controls (p < 0.04 and p = 0.0003, respectively). Moreover, both EBV and HCMV mucosal loads were significantly higher in refractory UC than in responders and controls (p = 0.007 and 0.003; and p = 0.02 and 0.001, respectively), and correlated with activity indexes. Steroid therapy seemed the main risk factor for triggering EBV colitis. Finally, no cases of IgM positivity were found in the study population. An impaired EBV-specific immunity was clearly evident in UC patients, mostly in those refractory to therapy. The ELISPOT assay may serve as new tool for quantifying and monitoring virus-specific T-cell immunity in UC.

1st Department of Internal Medicine Faculty of Medicine Comenius University and University Hospital Bratislava Slovakia

Department of Clinical Surgical Diagnostics and Pediatric Sciences University of Pavia Pavia Italy

Department of Internal Medicine San Matteo Hospital Foundation University of Pavia Pavia Italy

Experimental Research Laboratories Biotechnology Area San Matteo Hospital Foundation Pavia Italy

Gastroenterology Unit Department of Medicine A O U 1 Policlinico G B Rossi University of Verona Piazzale L A Scuro 10 37134 Verona Italy

Molecular Virology Unit Microbiology and Virology Department San Matteo Hospital Foundation Pavia Italy

Nd Department of Surgery Faculty of Medicine Masaryk University and St Anne's University Hospital Pekarska 53 65691 Brno Czech Republic

Zobrazit více v PubMed

Ungaro R, Mehandru S, Allen PB, Peyrin-Biroulet L, Colombel JF. Ulcerative colitis. Lancet. 2017;389:1756–70. PubMed

Rahier JF, Ben-Horin S, Chowers Y, et al. European Crohn’s and Colitis Organisation (ECCO). European evidence-based Consensus on the prevention, diagnosis and management of opportunistic infections in inflammatory bowel disease. J Crohns Colitis. 2009;3:47–91. PubMed

McGeoch DJ, Dolan A, Ralph AC. Toward a comprehensive phylogeny for mammalian and avian herpesviruses. J Virol. 2000;74:10401–6. PubMed PMC

Chiu YF, Sugden B. Epstein-Barr virus: the path from latent to productive infection. Annu Rev Virol. 2016;3:359–72. PubMed

Jarvis MA, Nelson JA. Human cytomegalovirus persistence and latency in endothelial cells and macrophages. Curr Opin Microbiol. 2002;5:403–7. PubMed

Murata T, Tsurumi T. Switching of EBV cycles between latent and lytic states. Rev Med Virol. 2014;24:142–53. PubMed

Vogl BA, Fagin U, Nerbas L, Schlenke P, Lamprecht P, Jabs WJ. Longitudinal analysis of frequency and reactivity of Epstein-Barr virus-specific T lymphocytes and their association with intermittent viral reactivation. J Med Virol. 2012;84:119–31. PubMed

Ljungman P, Griffiths P, Paya C. Definitions of HCMV infection and disease in transplant recipients. Clin Infect Dis. 2002;34:1094–7. PubMed

Yanai H, Shimizu N, Nagasaki S, Mitani N, Okita K. Epstein-Barr virus infection of the colon with inflammatory bowel disease. Am J Gastroenterol. 1999;94:1582–6. PubMed

Vega R, Bertrán X, Menacho M, et al. Cytomegalovirus infection in patients with inflammatory bowel disease. Am J Gastroenterol. 1999;94:1053–6. PubMed

Magro F, Gionchetti P, Eliakim R, et al. European Crohn’s and Colitis Organisation [ECCO]. Third European Evidence-based Consensus on Diagnosis and Management of Ulcerative Colitis. Part 1: definitions, diagnosis, extra-intestinal manifestations, pregnancy, cancer surveillance, surgery, and ileo-anal pouch disorders. J Crohns Colitis. 2017;11:649–70. PubMed

Harbord M, Eliakim R, Bettenworth D, et al. European Crohn’s and Colitis Organisation [ECCO]. Third European evidence-based consensus on diagnosis and management of ulcerative colitis. Part 2: current management. J Crohns Colitis. 2017;11:769–84. PubMed

Yanai H, Hanauer SB. Assessing response and loss of response to biological therapies in IBD. Am J Gastroenterol. 2011;106:685–98. PubMed

Bradford K, Shih DQ. Optimizing 6-mercaptopurine and azathioprine therapy in the management of inflammatory bowel disease. World J Gastroenterol. 2011;17:4166–73. PubMed PMC

Satsangi J, Silverberg MS, Vermeir S, Colombel JF. The Montreal classification of inflammatory bowel disease: controversies, consensus, and implications. Gut. 2006;55:749–53. PubMed PMC

Walmsley RS, Ayres RC, Pounder RE, Allan R. A simple clinical colitis activity index. Gut. 1998;43:29–32. PubMed PMC

Travis SP, Schnell D, Krzeski P, et al. Developing an instrument to assess the endoscopic severity of ulcerative colitis: the Ulcerative Colitis Endoscopic Index of Severity (UCEIS). Gut. 2012;61:535–42. PubMed

Calarota SA, Chiesa A, Zelini P, Comolli G, Minoli L, Baldanti F. Detection of Epstein-Barr virus-specific memory CD4 PubMed PMC

Calarota SA, Chiesa A, Scaramuzzi L, et al. Normalizing ELISPOT responses to T-cell counts: a novel approach for quantification of HCMV-specific CD4(+) and CD8(+) T-cell responses in kidney transplant recipients. J Clin Virol. 2014;61:65–73. PubMed

Furione M, Rognoni V, Cabano E, Baldanti F. Kinetics of human cytomegalovirus (HCMV) DNAemia in transplanted patients expressed in international units as determined with the Abbott RealTime CMV assay and an in-house assay. J Clin Virol. 2012;55:317–22. PubMed

Baldanti F, Gatti M, Furione M, et al. Kinetics of Epstein-Barr virus DNA load in different blood compartments of pediatric recipients of T-cell-depleted HLA-haploidentical stem cell transplantation. J Clin Microbiol. 2008;46:3672–7. PubMed PMC

Watzinger F, Suda M, Preuner S, et al. Real-time quantitative PCR assays for detection and monitoring of pathogenic human viruses in immunosuppressed pediatric patients. J Clin Microbiol. 2004;42:5189–98. PubMed PMC

Cottone M, Pietrosi G, Martorana G, et al. Pagliaro, Prevalence of cytomegalovirus infection in severe refractory ulcerative and Crohn’s colitis. Am J Gastroenterol. 2001;96:773–5. PubMed

Kambham N, Vij R, Cartwright CA, Longacre T. Cytomegalovitus infection in steroid-refractory ulcerative colitis: a case-control study. Am J Surg Pathol. 2004;28:365–73. PubMed

Roblin X, Pillet S, Oussalah A, et al. Cytomegalovirus load in inflamed intestinal tissue is predictive of resistance to immunosuppressive therapy in ulcerative colitis. Am J Gastroenterol. 2011;106:2001–8. PubMed

Dimitroulia E, Pitiriga VC, Piperaki ET, Spanakis NE, Tsakris A. Inflammatory bowel disease exacerbation associated with Epstein-Barr virus infection. Dis Colon Rectum. 2013;56:322–7. PubMed

Sylwester AW, Mitchell BL, Edgar JB, et al. Broadly targeted human cytomegalovirus-specific CD4+ and CD8+ T cells dominate the memory compartments of exposed subjects. J Exp Med. 2005;202:673–85. PubMed PMC

Thorley-Lawson DA, Gross A. Persistence of the Epstein-Barr virus and the origins of associated lymphoma. N Engl J Med. 2004;350:1328–37. PubMed

Ciccocioppo R, Racca F, Paolucci S, et al. Human cytomegalovirus and Epstein-Barr virus infection in inflammatory bowel disease: need for mucosal viral load measurement. World J Gastroenterol. 2015;21:1915–26. PubMed PMC

Nebbia G, Mattes FM, Sabin CA, et al. Differential effects of prednisolone and azathioprine on the development of human cytomegalovirus replication post liver transplantation. Transplantation. 2007;84:605–10. PubMed

Goodrum F, Caviness K, Zagallo P. Human cytomegalovirus persistence. Cell Microbiol. 2012;14:644–55. PubMed PMC

Kirman I, Whelan RL, Nielsen OH. Infliximab: mechanism of action beyond anti-TNF-alpha neutralization in inflammatory bowel disease. Eur J Gastroenterol Hepatol. 2004;16:639–41. PubMed

Chang CM, Yu KJ, Mbulaiteye SM, Hildesheim A, Bhatia K. The extent of genetic diversity of Epstein-Barr virus and its geographic and disease patterns: a need for reappraisal. Virus Res. 2009;143:209–21. PubMed PMC

Forbes BA, Bonville CA, Dock NL. The effects of a promoter of cell differentiation and selected hormones on human cytomegalovirus infection using an in vitro cell system. J Infect Dis. 1990;162:39–45. PubMed

Sester M, Sester U, Gärtner BC, Girndt M, Meyerhans A, Köhler H. Dominance of virus-specific CD8 T cells in human primary cytomegalovirus infection. J Am Soc Nephrol. 2002;13:2577–84. PubMed

Sester U, Gärtner BC, Wilkens H, et al. Differences in CMV-specific T-cell levels and long-term susceptibility to CMV infection after kidney, heart and lung transplantation. Am J Transplant. 2005;5:1483–9. PubMed

Reusser P, Riddell SR, Meyers JD, Greenberg PD. Cytotoxic T-lymphocyte response to cytomegalovirus after human allogeneic bone marrow transplantation: pattern of recovery and correlation with cytomegalovirus infection and disease. Blood. 1991;78:1373–80. PubMed

Kotton CN, Kumar D, Caliendo AM, et al. The Transplantation Society International CMV Consensus Group, The Third International Consensus Guidelines on the Management of Cytomegalovirus in Solid-organ Transplantation. Transplantation. 2018;102:900–31. PubMed

Cannon MJ, Schmid DS, Hyde TB. Review of cytomegalovirus seroprevalence and demographic characteristics associated with infection. Rev Med Virol. 2010;20:202–13. PubMed

Rasmussen L, Matkin C, Spaete R, Pachl C, Merigan TC. Antibody response to human cytomegalovirus glycoproteins gB and gH after natural infection in humans. J Infect Dis. 1991;164:835–42. PubMed

Jonjić S, Pavić I, Polić B, Crnković I, Lucin P, Koszinowski UH. Antibodies are not essential for the resolution of primary cytomegalovirus infection but limit dissemination of recurrent virus. J Exp Med. 1994;179:1713–7. PubMed

Boppana SB, Britt WJ. Antiviral antibody responses and intrauterine transmission after primary maternal cytomegalovirus infection. J Infect Dis. 1995;171:1115–21. PubMed

Hakki M. Moving Past Ganciclovir and Foscarnet: Advances in CMV Therapy. Curr Hematol Malig Rep. 2020;15:90–102. PubMed

Keller MD, Bollard CM. Virus-specific T-cell therapies for oatients with primary immune deficiency. Blood. 2020;135:620–8. PubMed PMC

Yoshino T, Nakase H, Ueno S, et al. Usefulness of quantitative real-time PCR assay for early detection of cytomegalovirus infection in patients with ulcerative colitis refractory to immunosuppressive therapies. Inflamm Bowel Dis. 2007;13:1516–21. PubMed

Ganzenmueller T, Henke-Gendo C, Schlué J, Wedemeyer J, Huebner S, Heim A. Quantification of cytomegalovirus DNA levels in intestinal biopsies as a diagnostic tool for CMV intestinal disease. J Clin Virol. 2009;46:254–8. PubMed