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. Author manuscript; available in PMC: 2023 Jan 15.
Published in final edited form as: J Immunol. 2021 Dec 29;208(2):407–419. doi: 10.4049/jimmunol.2100029

Figure 1. Intranasal immunization with H1/CDN enhances CD4 T cell responses in the lungs of vaccinated mice both pre and post-challenge.

Figure 1.

Mice were vaccinated with intranasal M. tuberculosis H1 antigen and ML-RR-cGAMP cyclic-di-nucleotide adjuvant (H1/CDN) 12 weeks before challenge, then boosted twice at 4-week intervals after priming, before M. tuberculosis aerosol challenge with 50–100 CFU. Mice were mock primed and boosted with PBS as a control. (a) Cellular percentages were enumerated in the lungs using flow cytometry in vaccinated and mock vaccinated mice on day 0 prior to challenge with M. tuberculosis. (b) Intracellular cytokine staining (ICS) for percentage of lung CD4 T cells that produce IFN-γ (Th1), (c) IL-17 (Th17), (d) both IFN-γ and IL-17 (Th1-Th17) after restimulation ex vivo with recombinant Ag85b peptide pool on day 0 prior to challenge with M. tuberculosis. Percentage of CD4 T cells that express (e) CXCR3 (parenchymal T cells) or (f) KLRG1 (vascular T cells) on day 0 prior to challenge with M. tuberculosis. Mice were vaccinated with s.c. BCG or i.n. H1/CDN as above and (g) lung bacterial burden was enumerated at 1 – 4 weeks post M. tuberculosis challenge. (h) Intracellular cytokine staining (ICS) for percentage of lung CD4 T cells that produce IFN-γ (Th1) (i) IL-17 (Th17), or (j) both IFN-γ and IL-17 (Th1-Th17) after restimulation ex vivo with recombinant Ag85b peptide pool at indicated timepoints post challenge. (k) Enumeration of TCRɣδ and (l) IL-17-producing TCRɣδ T cells at 4 weeks post challenge. Data are expressed as mean (± SD) of eight to ten mice per group from two independent experiments; in (b-f; k, l) each animal is represented by a point. Mann-Whitney t test p values calculated in comparison to PBS vaccinated controls except where indicated; *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001, ****p ≤ 0.0001. Significance is relative to PBS control unless otherwise indicated. All experiments were performed a minimum of two times; for a-d two pooled experiments are shown; otherwise a representative experiment is shown.