Induction of CD4 Th1 cell responses by R/D immunization.

<p>At 12 weeks post-immunization, T lymphocytes were purified from PPD and α-crystallin stimulated splenocytes (pooled from four mice per group) and stained for cell surface marker (CD4) along with intracellular staining for IFNγ, TNFα and IL2 followed by FACS analysis. Frequency of IFNγ, TNFα and IL2 producing cells was determined on CD4 T cell gated population. (A) Total frequency (%) of PPD and α-crystallin specific cytokine producing CD4 T cells in spleen. (B) Frequency (%) of PPD and α-crystallin specific CD4 T cells expressing each of the seven combinations of IFNγ, TNFα and IL2. (C) Proportion of PPD and α-crystallin specific CD4 T cells producing one, two or three cytokines. (D) Frequency (%) of PPD and α-crystallin specific 3⁺ CD4 T cells in spleen along with MFI and iMFI for IFNγ, TNFα and IL2. R/D, rBCG prime – DNAacr boost.