Activation and Induction of Antigen-Specific T Follicular Helper Cells Play a Critical Role in Live-Attenuated Influenza Vaccine-Induced Human Mucosal Anti-influenza Antibody Response

Abstract

There is increasing interest recently in developing intranasal vaccines against respiratory tract infections. The antibody response is critical for vaccine-induced protection, and T follicular helper cells (T_FH) are considered important for mediating the antibody response. Most data supporting the role for T_FH in the antibody response are from animal studies, and direct evidence from humans is limited, apart from the presence of T_FH-like cells in blood. We studied the activation and induction of T_FH and their role in the anti-influenza antibody response induced by a live-attenuated influenza vaccine (LAIV) in human nasopharynx-associated lymphoid tissue (NALT). T_FH activation in adenotonsillar tissues was analyzed by flow cytometry, and anti-hemagglutinin (anti-HA) antibodies were examined following LAIV stimulation of tonsillar mononuclear cells (MNC). Induction of antigen-specific T_FH by LAIV was studied by flow cytometry analysis of induced T_FH and CD154 expression. LAIV induced T_FH proliferation, which correlated with anti-HA antibody production, and T_FH were shown to be critical for the antibody response. Induction of T_FH from naive T cells by LAIV was shown in newly induced T_FH expressing BCL6 and CD21, followed by the detection of anti-HA antibodies. Antigen specificity of LAIV-induced T_FH was demonstrated by expression of the antigen-specific T cell activation marker CD154 upon challenge by H1N1 virus antigen or HA. LAIV-induced T_FH differentiation was inhibited by BCL6, interleukin-21 (IL-21), ICOS, and CD40 signaling blocking, and that diminished anti-HA antibody production. In conclusion, we demonstrated the induction by LAIV of antigen-specific T_FH in human NALT that provide critical support for the anti-influenza antibody response. Promoting antigen-specific T_FH in NALT by use of intranasal vaccines may provide an effective vaccination strategy against respiratory infections in humans.<b>IMPORTANCE</b> Airway infections, such as influenza, are common in humans. Intranasal vaccination has been considered a biologically relevant and effective way of immunization against airway infection. The vaccine-induced antibody response is crucial for protection against infection. Recent data from animal studies suggest that one type of T cells, T_FH, are important for the antibody response. However, data on whether T_FH-mediated help for antibody production operates in humans are limited due to the lack of access to human immune tissue containing T_FH In this study, we demonstrate the induction of T_FH in human immune tissue, providing critical support for the anti-influenza antibody response, by use of an intranasal influenza vaccine. Our findings provide direct evidence that T_FH play a critical role in vaccine-induced immunity in humans and suggest a novel strategy for promoting such cells by use of intranasal vaccines against respiratory infections.