STUDYING HCV-SPECIFIC B CELL REPERTOIRES FOR IMMUNOTHERAPEUTICS

The potent adaptive immune system is fundamentally reliant upon the generation of a diverse repertoire of B-lymphocyte antigen specific. The earliest response of the adaptive immune against infection threats will likely determine the outcome of infection. Hepatitis C virus (HCV) has emerged as a major etiological agent of liver disease. Of the persons acutely infected with HCV, 30% spontaneously recover, while 70% develop chronic infection. In this context, using HCV as a model for comparing B cell responses between spontaneous clearance and chronic patients will enable identifying unique humoral immune responses controlling infection, and will point to the mechanisms that determine outcomes of human diseases.

We aimed to establish a platform that will enable to study HCV-specific antibody response by identifying and isolating human HCV-specific B cells from infected patients. We have isolated peripheral blood mononuclear cells (PBMCs) from chronic infected donors and sorted for B cells (CD19 positive staining). These cells were activated with HCV envelop protein (rE2), IL2,IL21 and CD40L feeder cells for 14 days. To isolate HCV-specific B cells, we sorted by identifying B cells that binds to rE2. The antibodies that were produced from these B cells demonstrated high HCV-neutralization capacity which supports the successful separation of HCV specific B cells. Identifying antigen-specific sequences from total repertoire data can aid our understanding B cells driven immunity, and be used for disease diagnostics and vaccine evaluation.