Th1 effector cells and macrophages facilitate respiratory IgA after mucosal vaccination
Mucosal immunization and mucosal IgA offer significant promise in protecting against airborne pathogens, including SARS-CoV-2. However, the conditions and mechanisms that lead to the robust induction of mucosal IgA responses following mucosal vaccination remain poorly understood. It is also currently debatable whether mucosal vaccination is still warranted given that most individuals in developed countries have established a hybrid immunity from vaccination and infection. Here we characterized respiratory mucosal immune responses after SARS-CoV-2 infection, vaccination or both in humans. We found that hybrid immunity resulted in moderately increased mucosal IgA and neutralizing antibody responses compared to infection or vaccination alone. However, a direct comparison of hybrid immunity and a mucosal adenovirus-based booster vaccination in animal models revealed that respiratory booster immunization elicited markedly stronger and more durable mucosal IgA, T cell response, and protective immunity against SARS-CoV-2, supporting the promise of respiratory mucosal vaccination. Mechanistically, we found that mucosal booster immunization induced local IgA-secreting cells in the respiratory mucosa, aided by pulmonary CD4+ T cells in situ. Strikingly, local IL-21-producing Blimp-1+ Th1 effector cells were critical in mediating the CD4+ T cell help for IgA production. Furthermore, lung macrophages were important for this mucosal IgA response via the production of TGF-ß. Consequently, Spike mRNA coated with pulmonary surfactant (PS) containing liposomes, rationally designed to target lung macrophages, elicits a stronger mucosal IgA response. Collectively, our results uncover a local cellular network supporting enhanced mucosal IgA responses, with implications for the development of optimal mucosal immunization strategies against SARS-CoV-2 and other respiratory pathogens. Overall design: For library preparation, BAL cells or sorted BAL CD4+ T cells from immunized mice in each group were pooled together and each group was hashtagged using BioLegend TotalSeq™-A antibodies according to the provided protocol. The library of hashtagged pooled groups was then constructed using the 10x Genomics Chromium Next GEM Single Cell 3' Reagent Kit with the v3.1 (Dual Index) protocol. Sequencing was performed using the NovaSeq X Plus platform, a service provided by Novogene. The sequenced data were aligned using Cell Ranger. The curated data then underwent demultiplexing, quality assurance, filtering, differential gene expression analysis, and cluster identification, all conducted using the standard Seurat protocol with Seurat (v 5.1.0) in R (R-4.3.3).