Background: Epstein-Barr virus (EBV) has a key role in the development of multiple sclerosis (MS), however, the mechanisms linking EBV infection to MS pathology are still unclear and it remains to be determined whether EBV and its interactions with the host-immune system have a role in ongoing disease. To test possible associations between EBV reactivation and host immune activation, we analyzed EBV serology, EBV DNA load and viral and cellular transcripts in the peripheral blood of people with MS (PwMS) and healthy donors (HD). Methods: PBMCs and sera from people with newly diagnosed relapsing-remitting MS (n=79) and HD (n=45) were collected in three MS Centers. Clinical and MRI data were available for PwMS. Droplet-digital PCR system was used for EBV DNA quantification in PBMC. Targeted Pre-Amp real time RT-PCR was used to analyze the expression of 7 genes associated with EBV latency and lytic cycle and 57 cellular immune-related genes. The antibody response to EBV (anti-EBNA1 IgG) was assessed using ELISA. Results were analysed in combination with clinical, radiological and demographic data using univariate and multivariate statistical approaches. Results: Consistent with previous studies, levels of EBNA1 IgG and EBV DNA were significantly higher (p=0.04) and tended to be higher (p=0.056), respectively, in PwMS compared to HD. The prevalence of viral transcripts associated with disrupted EBV latency and/or lytic cycle was significantly higher in PwMS than in HD (25.3% vs 4.4%; p=0.002), supporting EBV dysregulation in MS. Factor analysis identified groups of co-ordinately regulated genes. Anti-EBNA1 IgG and EBV DNA levels did not associate to any of the artificial factors. Scores of Factors 2, 3 and 4, containing cellular genes involved in T-cell activation and migration, innate immunity activation, antigen presentation, and type-I IFN signalling together with viral genes associated with EBV dysregulation, were significantly higher in PBMCs of PwMS compared to HD. Neither viral nor cellular variables analyzed associated with sex, clinical or MRI activity in PwMS. Conclusions: Our study shows for the first time an increase in viral transcripts associated with EBV dysregulation in the peripheral blood of PwMS compared to HD, and a positive correlation between expression of EBV latent/lytic genes and cellular genes involved in immune activation in PwMS. Although these two processes do not appear to be related to clinical and radiological signs of MS disease activity, their association could be relevant for MS progression and should be examined further in longitudinal studies
Study of the interactons between Epstein-Barr virus and the host immune system in multiple sclerosis: insights from the analysis of EBV serology, EBV nucleic acids and their association with immune-related gene expression / Chiara, Meloni1; Corrado, Fagnani2; Fabiana, Marnetto3; Paola, Valentino3; 4, ; Antonio, Bertolotto3; Anna, Repice5; Clara, Ballerini6; Jessica, Frau7; Eleonora, Cocco7; 1Department of Neuroscience, and Caterina Veroni1.; Istituto Superiore di, Sanità; Rome, PROPERTIES LTD; Italy, 2 Reference Centre for Behavioural Sciences and Mental Health; Istituto Superiore di, Sanità; Rome, PROPERTIES LTD; Italy, 3 CRESM (Multiple Sclerosis Regional Reference Center); Neuroscience Institute Cavalieri Ottolenghi (NICO) and AOU San Luigi, Gonzaga; Orbassano, (TO); Italy., 4 Department of Clinical and Biological Sciences; University of, Turin; Italy., 5 Department of Neuroscience; Drug and Child Health, (NEUROFARBA); University of, Florence; Italy., 6 Department of Experimental and Clinical Medicine (DMSC); University of, Florence; Italy., 7 Department of Medical Science and Public Health; University of, Cagliari; Ms, Centre; Binaghi, Hospital; Cagliari, Italy. - (2024). (Intervento presentato al convegno 18th European school neuroimmunology course ESNI 2024 tenutosi a Hasselt, Belgium).
Study of the interactons between Epstein-Barr virus and the host immune system in multiple sclerosis: insights from the analysis of EBV serology, EBV nucleic acids and their association with immune-related gene expression
Chiara Meloni1;Rome;Rome;
2024
Abstract
Background: Epstein-Barr virus (EBV) has a key role in the development of multiple sclerosis (MS), however, the mechanisms linking EBV infection to MS pathology are still unclear and it remains to be determined whether EBV and its interactions with the host-immune system have a role in ongoing disease. To test possible associations between EBV reactivation and host immune activation, we analyzed EBV serology, EBV DNA load and viral and cellular transcripts in the peripheral blood of people with MS (PwMS) and healthy donors (HD). Methods: PBMCs and sera from people with newly diagnosed relapsing-remitting MS (n=79) and HD (n=45) were collected in three MS Centers. Clinical and MRI data were available for PwMS. Droplet-digital PCR system was used for EBV DNA quantification in PBMC. Targeted Pre-Amp real time RT-PCR was used to analyze the expression of 7 genes associated with EBV latency and lytic cycle and 57 cellular immune-related genes. The antibody response to EBV (anti-EBNA1 IgG) was assessed using ELISA. Results were analysed in combination with clinical, radiological and demographic data using univariate and multivariate statistical approaches. Results: Consistent with previous studies, levels of EBNA1 IgG and EBV DNA were significantly higher (p=0.04) and tended to be higher (p=0.056), respectively, in PwMS compared to HD. The prevalence of viral transcripts associated with disrupted EBV latency and/or lytic cycle was significantly higher in PwMS than in HD (25.3% vs 4.4%; p=0.002), supporting EBV dysregulation in MS. Factor analysis identified groups of co-ordinately regulated genes. Anti-EBNA1 IgG and EBV DNA levels did not associate to any of the artificial factors. Scores of Factors 2, 3 and 4, containing cellular genes involved in T-cell activation and migration, innate immunity activation, antigen presentation, and type-I IFN signalling together with viral genes associated with EBV dysregulation, were significantly higher in PBMCs of PwMS compared to HD. Neither viral nor cellular variables analyzed associated with sex, clinical or MRI activity in PwMS. Conclusions: Our study shows for the first time an increase in viral transcripts associated with EBV dysregulation in the peripheral blood of PwMS compared to HD, and a positive correlation between expression of EBV latent/lytic genes and cellular genes involved in immune activation in PwMS. Although these two processes do not appear to be related to clinical and radiological signs of MS disease activity, their association could be relevant for MS progression and should be examined further in longitudinal studiesI documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
