TCR Repertoire Analysis of Mouse T Follicular Helper Cells and T Follicular Regulatory Cells Following Immunization

Abstract

Abstract Generation of high-affinity and class-switched antibody requires the germinal center (GC) reaction after infection or immunization. Within the B-cell follicles of secondary lymphoid organs, the GC represents a sophisticated collaboration between antigen-specific B cells, follicular dendritic cells, T follicular helper (TFH) cells and T follicular regulatory (TFREG) cells. Despite intensive interest in the development and effector function of TFH and TFREG cells, little is known regarding the selection of T-cell receptor (TCR) repertoire during polyclonal GC reactions. In order to evaluate native, polyclonal TCR responses elicited by a complex antigen, we developed a sorting strategy to isolate TFH/TFREG cell populations that were activated and expanded after s.c. immunization with NP15-OVA. TCRβ VDJ rearrangements were recovered from highly purified TCRβ+CD4+CXCR5hiPD-1+Bcl-6+FoxP3− TFH cells and TCRβ+CD4+ CXCR5hiPD-1+Bcl-6+FoxP3+ TFREG cells, amplified by PCR, and sequenced. Analysis of the antigen-specific TCR repertoire of TFH/TFREG cells provides important insights into the factors influencing T-cell recruitment and clonal expansion following infection or vaccination, especially when linked to contemporary analysis of the GC B-cell repertoire. These findings may inform rational and selective control strategy of the GC reaction. Vaccine development can accordingly focus on modulating TFH/TFREG responses to facilitate optimal adaptive immune responses.