IRF4-regulated transcriptional and functional heterogeneity of lung-resident CD11b+ cDC2 subsets during influenza virus infection.

Lung-resident conventional dendritic cells (cDCs) coordinate immune responses to respiratory viruses in the respiratory tract or after migration to mediastinal lymph nodes (mLN). Migratory DCs include cDC1s (CD103+XCR1+CD24hi) expressing IRF8 or cDC2s (CD11b+SIRPα+CD24+) expressing IRF4. IRF4+ cDC2s are divided into a CD24hi subset that requires IRF4 for differentiation and a CD24int subset that is present in the absence of IRF4. During influenza A virus (IAV) infection of mice, we characterized the kinetics of cDC2 subset accumulation in the lung and mLN and their differences in IRF4-dependent gene expression and function. We found that the 2 IRF4-expressing cDC2 subsets upregulated CD86 to high levels, produced IL-12p40 and the chemokines CCL17 and CCL22, and were capable of acquiring antigen in vivo and activating antigen-specific CD8+ T cells. Notably, the CD11b+CD24int cDC2 subset expressed canonical cDC markers and transcription factors and expanded to high numbers in the lung and mLN by d 6 postinfection. Transcriptome analyses on d 5 postinfection revealed that the CD11b+CD24int cDC2 subset expressed both IRF4 and IRF8 and harbored an elevated IFN response signature compared to the CD11b+CD24hi subset. Analyses of mice lacking Irf4 in CD11c+ cells showed that IRF4 promoted the function of CD11b+CD24int cDC2s, including the capacity to migrate to mLN and to produce CCL17 and CCL22, consistent with their altered gene expression profile in the absence of IRF4. In sum, our data show that the 2 lung-resident CD11b+ cDC2 subsets present in naïve mice elaborated distinct and common functional responses regulated by IRF4 during IAV infection.