Contributions
Abstract: P449
Type: Poster Sessions
Abstract Category: Pathology and pathogenesis of MS - Environmental factors
EBV infection is strongly associated with MS and increasing evidence indicates that EBV deregulation might be implicated in the dysimmune process that targets the brain and spinal cord. Some studies reported that EBV DNA load is higher in peripheral blood mononuclear cells (PBMC) of patients with MS compared to control subjects. To investigate this issue further, droplet digital PCR and pre-amplification real time RT-PCR were used to quantify EBV DNA and RNA, respectively, in PBMC from healthy donors (HD; n = 28), therapy-free patients with relapsing-remitting MS (RRMS; n = 61), and CIS patients (n = 25). Patients included in the study were enrolled at the MS centers of San Luigi Gonzaga Hospital/Orbassano, Florence University and Cagliari University; PBMC samples from HD were obtained from CRESM/San Luigi Gonzaga Hospital.
EBV DNA was detected in PBMC from 46% of therapy-free RRMS patients, 36% of CIS patients and 39% of HD, without significant differences in EBV DNA frequency or amount between patients and controls. EBV transcripts were detected in PBMC from 22/61 RRMS patients (36%), 6/25 CIS patients (24%), and 7/28 (25%) HD. However, the EBV transcriptional profile markedly differed between patients and HD. Six out of 7 EBV+ samples from the 28 HD contained only EBER1 (latency 0) and only one sample displayed very low amounts of both EBER1 and LMP1 (latency II program). Interestingly, EBV transcripts associated with EBV latency activation (EBNA3A, EBNA1, LMP1, LMP2A: latency III and latency II programs) and lytic cycle (BZLF1, gp350/220) were detected in 18/22 (82%) and in 3/6 (50%) EBV+ RNA PBMC samples from RRMS and CIS patients, respectively. EBV transcripts were also quantified in CSF cell samples (n=76) matched to PBMC from CIS and RRMS patients. Despite the percentage of patient CSF samples with detectable EBV RNA was very low (7%, 5/76 samples), transcripts associated with EBV latency disruption and/or entry into the lytic cycle were present in all 5 EBV RNA+ CSF cell samples.
This is the first study to show EBV RNA alterations in peripheral blood of CIS/MS patients compared to HD, strengthening the idea that altered control of EBV infection is associated with the disease. These data also suggest that RNA markers of EBV deregulation can be monitored in the peripheral blood during therapy and could provide useful information on the relationship between drug efficacy and changes in EBV-immune system interaction.
Disclosure:
- This study is funded by Italian Multiple Sclerosis Foundation (grant FISM GRANT 2014/R/1 to F. Aloisi)
- C. Veroni has nothing to disclose
- F. Marnetto received financial support from Euroimmun, Biogen Idec.
- A. Bertolotto declares the following: Advisory boards and/or speaker honoraria (Biogen, Novartis, Sanofi, and Teva); grant support (Almirall, Associazione San Luigi Gonzaga ONLUS, Bayer, Biogen, Fondazione per la Ricerca Biomedica ONLUS, Merck, Novartis, Sanofi, Teva, and Italian Multiple Sclerosis Foundation).
- J. Frau serves on scientific advisory boards for Biogen and Genzyme, has received honoraria for speaking from Merck Serono, Genzyme, Biogen and Teva.
- E. Cocco has served on advisory boards and/or has received travel grants and/or speaker honoraria from Bayer, Merck, Roche, Teva, Biogen, Almirall, Novartis, Genzyme.
- A. Repice has nothing to disclose
- C. Ballerini declares the following grant support: FISM, Fondazione Careggi, Serono, Fondazione San Michele all'Adige, Angelini, Lundbeck.
- F. Aloisi declares the following grant support: Italian Ministry of Health, Italian Multiple Sclerosis Foundation/FISM, Biogen.
Abstract: P449
Type: Poster Sessions
Abstract Category: Pathology and pathogenesis of MS - Environmental factors
EBV infection is strongly associated with MS and increasing evidence indicates that EBV deregulation might be implicated in the dysimmune process that targets the brain and spinal cord. Some studies reported that EBV DNA load is higher in peripheral blood mononuclear cells (PBMC) of patients with MS compared to control subjects. To investigate this issue further, droplet digital PCR and pre-amplification real time RT-PCR were used to quantify EBV DNA and RNA, respectively, in PBMC from healthy donors (HD; n = 28), therapy-free patients with relapsing-remitting MS (RRMS; n = 61), and CIS patients (n = 25). Patients included in the study were enrolled at the MS centers of San Luigi Gonzaga Hospital/Orbassano, Florence University and Cagliari University; PBMC samples from HD were obtained from CRESM/San Luigi Gonzaga Hospital.
EBV DNA was detected in PBMC from 46% of therapy-free RRMS patients, 36% of CIS patients and 39% of HD, without significant differences in EBV DNA frequency or amount between patients and controls. EBV transcripts were detected in PBMC from 22/61 RRMS patients (36%), 6/25 CIS patients (24%), and 7/28 (25%) HD. However, the EBV transcriptional profile markedly differed between patients and HD. Six out of 7 EBV+ samples from the 28 HD contained only EBER1 (latency 0) and only one sample displayed very low amounts of both EBER1 and LMP1 (latency II program). Interestingly, EBV transcripts associated with EBV latency activation (EBNA3A, EBNA1, LMP1, LMP2A: latency III and latency II programs) and lytic cycle (BZLF1, gp350/220) were detected in 18/22 (82%) and in 3/6 (50%) EBV+ RNA PBMC samples from RRMS and CIS patients, respectively. EBV transcripts were also quantified in CSF cell samples (n=76) matched to PBMC from CIS and RRMS patients. Despite the percentage of patient CSF samples with detectable EBV RNA was very low (7%, 5/76 samples), transcripts associated with EBV latency disruption and/or entry into the lytic cycle were present in all 5 EBV RNA+ CSF cell samples.
This is the first study to show EBV RNA alterations in peripheral blood of CIS/MS patients compared to HD, strengthening the idea that altered control of EBV infection is associated with the disease. These data also suggest that RNA markers of EBV deregulation can be monitored in the peripheral blood during therapy and could provide useful information on the relationship between drug efficacy and changes in EBV-immune system interaction.
Disclosure:
- This study is funded by Italian Multiple Sclerosis Foundation (grant FISM GRANT 2014/R/1 to F. Aloisi)
- C. Veroni has nothing to disclose
- F. Marnetto received financial support from Euroimmun, Biogen Idec.
- A. Bertolotto declares the following: Advisory boards and/or speaker honoraria (Biogen, Novartis, Sanofi, and Teva); grant support (Almirall, Associazione San Luigi Gonzaga ONLUS, Bayer, Biogen, Fondazione per la Ricerca Biomedica ONLUS, Merck, Novartis, Sanofi, Teva, and Italian Multiple Sclerosis Foundation).
- J. Frau serves on scientific advisory boards for Biogen and Genzyme, has received honoraria for speaking from Merck Serono, Genzyme, Biogen and Teva.
- E. Cocco has served on advisory boards and/or has received travel grants and/or speaker honoraria from Bayer, Merck, Roche, Teva, Biogen, Almirall, Novartis, Genzyme.
- A. Repice has nothing to disclose
- C. Ballerini declares the following grant support: FISM, Fondazione Careggi, Serono, Fondazione San Michele all'Adige, Angelini, Lundbeck.
- F. Aloisi declares the following grant support: Italian Ministry of Health, Italian Multiple Sclerosis Foundation/FISM, Biogen.