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. 2021 Nov 18;12:785457. doi: 10.3389/fimmu.2021.785457

Figure 5.

Figure 5

Pulmonary innate and adaptive immune responses induced by influenza infection in WT and CXCL5-/- mice. (A) Single lung cells were collected from WT and CXCL5-/- mice at 3 d.p.i. and analyzed by flow cytometry to detect macrophages (CD64+CD45+), neutrophils (Ly6G+CD45+), NK cells (NK1.1+CD45+), CD4 T cells (CD4+CD45+), CD8 T cells (CD8+CD45+), and CD19 B cells (CD19+CD45+). (B) Numbers of different leukocyte subsets among pulmonary CD45+ leukocytes at 3 d.p.i. (n=4). (C) Microscopic view of BALF leukocytes from WT and CXCL5-/- mice collected at 3 d.p.i. and cytospun. Black arrows, neutrophils (×400 magnification). (D) Single lung cells were collected from WT and CXCL5-/- mice at 8 d.p.i. and analyzed by flow cytometry to detect CD4 T cells (CD4+CD45+), CD8 T cells (CD8+CD45+), CD19 B cells (CD19+CD45+), and influenza-specific CD8 T cells (APC-conjugated class I tetramers loaded with the influenza virus NP peptide ASNENMETM). (E) Numbers of different leukocyte subsets among pulmonary CD45+ leukocytes at 8 d.p.i. (n=4). (F) Lung sections from WT and CXCL5-/- mice were stained at 3 d.p.i. with an anti-CD19 antibody and hematoxylin (×200 magnification). (G) Numbers of NP tetramer-positive CD8+ T cells among pulmonary CD45+ leukocytes at 8 d.p.i. (n=4). (H) The levels of IFN-γ in BALF from WT and CXCL5-/- mice at 8 d.p.i. were determined by ELISA (n=5). (I) The total numbers of CD4+ T cells and B220+ B cells in the MLNs at 8 d.p.i. were determined by cell counting together with flow cytometry (n=4). (J) The HI titers of the BALF samples from WT and CXCL5-/- mice at 8 d.p.i. against H1N1 virus were evaluated (n=4). (K) Measurements of influenza-specific total IgG in the BALF of WT and CXCL5-/- mice infected with the H1N1 virus at 8 d.p.i. (n = 5). (L) The numbers of plasmablasts among pulmonary B cells were detected using anti-CD19 and anti-CD138 antibodies by flow cytometry of samples from WT and CXCL5-/- mice at 8 d.p.i. (n=4). (M) The total numbers of CD138+CD19+ B cells in the MLNs at 8 d.p.i. were determined by cell counting together with flow cytometry (n=4). The error bars represent the SDs. *P < 0.05 based on Student’s t-test.