Exosomes From MS Patient Serum Carry EBV Early Antigens not Detected in Exosomes From Healthy Donor Patient Serum
Monique Anderson1, Fiona Salas2, Natalia Drosu3, Philippe-Antoine Bilodeau4, Joao Mahler Ferreira Oliveira3, Mulan Jiang5, Taka Mikami3, Rebecca Salky3, James Nguyen3, Gabriela Romanow4, Michael Levy6
1Mass General Hospital, 2Skidmore College, 3Mass General Brigham, 4Massachusetts General Hospital, 5Harvard University, 6Massachusetts General Hospital/Harvard Medical School
Objective:
To characterize EBV latent and lytic protein contents in MS serum exosomes compared to healthy controls and assess for any relationships to disease activity.
Background:
Epstein-Barr Virus (EBV) has be shown to be key risk factor in the development of MS. However, the exact relationship to MS pathogenesis is still under investigation. Prior work has focused on potential relationships between EBV latent proteins and MS disease activity. exosomes can play a role in the immune response.
Design/Methods:
We collected and stored MS and healthy control (HC) serum samples at -80C. We included 24 MS patients and 5 healthy controls within this study. Exosomes were isolated following initial filtration with 0.22um filter, using ExoQuick reagent (SBI). Samples were run on Western blot and probed for EBNA1, EBNA2A, Early antigen D (EaD), LMP1, LMP2A, BZLF, BHRF1. The presence or absence of antigen was assessed using ImageJ software.
Results:
Here, results for EBNA1, EaD and BZLF are presented. EBNA1 was detected in 23 of 24 MS serum samples and in all 5 healthy controls tested (Fisher exact test, p-value=1). EaD was detected in 7 of 24 MS serum samples tested but not detected in healthy controls (Fisher exact test, p-value=.1608). BZLF was detected in 7 of 21 MS serum exosome samples while 1 of 5 healthy controls exosomes carried it (Fisher exact test, p-value=.1414) Based on results, clinical evaluation was undertaken in patients positive for EaD, and for those which the information was recorded, EaD exosome detection was found to be detectable in samples drawn within 3 mos of flare, while those that were negative were at least 3 mos removed from a flare.
Conclusions:
EaD, an early lytic protein of EBV, was only detected in exosomes from MS patients and appeared within a subset of patients whose samples were collected within 3 mos of flare.
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