Figure 2.

Sex bias in B and T cell responses to influenza antigens in human tonsil organoids. (A) Tonsil organoids were prepared from 35 cryopreserved tonsil samples (n=18 female, n=17 male) and stimulated with live-attenuated influenza vaccine (LAIV), inactivated influenza vaccine (IIV), or wild-type H1N1 and H3N2 viruses. Cells were harvested on day 7 for analysis by flow cytometry. Created with Biorender. (B) Representative flow cytometry plots showing the distribution of major B cell subsets in unstimulated and LAIV-stimulated tonsil organoids. (C) B cell phenotype quantification in male- and female-derived organoids. (D) Frequencies of atypical memory B cells (IgD- CD27-) in male and female-derived organoids. (E) Frequencies of activated (CD83+) total B cells and activated pre-GC B cells (as measured by CD83 and CD39). Sex differences in the frequency of (F) light zone and (G) dark zone GC B cells in unstimulated and influenza antigen-stimulated organoids. (H) Th1 CD4 T cell frequencies in male and female-derived organoids. (I) Linear correlation analysis of Th1 cell frequency and donor age in LAIV-stimulated organoids from male and female participants. Each point represents one donor. Mann Whitney U tests followed by multiple hypothesis correction (using the Benjamini & Hochberg method) were used to calculate p values. Spearman’s rank correlation test and multiple linear regression was performed to calculate the linear regression. Boxplots indicate the median value, with hinges denoting the first and third quartiles and whiskers denoting the highest and lowest value within 1.5 times the interquartile range of the hinges.