Toll-Like Receptor 3 Ligand and Retinoic Acid Enhance Germinal Center Formation and Increase the Tetanus Toxoid Vaccine Response

Immunizations with T-cell-dependent antigens induce the formation of germinal centers (GC), unique lymphoid microenvironments in which antigen-activated B cells undergo class switching, affinity maturation, and differentiation into memory B cells. Poly(I: C) (PIC), a double-stranded RNA, and retinoic acid (RA), a metabolite of vitamin A which induces cell differentiation, have been shown to augment both primary and memory anti-tetanus toxoid (anti-TT) IgG responses. However, their influence on the GC reaction is unknown. In the present study, 6-week-old C57BL/6 mice were immunized with TT and cotreated with PIC, RA, or both. The splenic GC reaction was evaluated using immunofluorescence staining 10 days after TT priming. Each treatment enhanced the TT-induced GC formation (number of GC/follicle and GC area) about two-to threefold, which correlated with the titers of plasma anti-TT immunoglobulin G (IgG). Isotype switching to IgG1 was dramatically stimulated, with the greatest increase in IgG1-positive GC B cells induced by RA-PIC (P < 0.001). Moreover, PIC alone and RA-PIC robustly promoted the formation of the follicular dendritic cell (FDC) network in the GC light zone. PIC and RA-PIC also increased IgG1-positive B cells in the periarterial lymphatic sheath regions, where most IgG1-positive cells were plasma cells (CD138/ syndecan-1 positive), suggesting that plasma cell generation was also enhanced in non-GC regions. The stimulation of several processes, including antigen-induced GC formation, isotype switching, FDC network formation within GC, and plasma cell differentiation by RA and/or PIC, suggests that this nutritional-immunological combination could be an effective means of promoting a robust vaccine response.