Selective induction of Th2 cells in murine Peyer's patches by oral immunization

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Abstract
We have used three different methods to determine the T helper (Th) cell response, Including Th1 and Th2 types, in murine Peyer's patches (PP) following oral immunization with sheep red blood cells (SRBC). These Include: (I) use of cytokine-spectfic (IFN-γ and IL-5), single cell assays to estimate the frequencies of Th1 and Th2 cells respectively, (II) cytokine-speciflc mRNA-cDNA dot blot and Northern gel hybridizations to detect levels of specific mRNA, and (ill) T cell cloning techniques to determine the frequency of Th1 and Th2 clones. Mice were immunized with SRBC by either the oral or i.p. route. The PP and splenic (SP) CD3+ and CD3+ CD4+ T cell subsets were Isolated and cultured with antigen, feeder cells, and IL-2, and were assessed at various intervals (days 0, 1, 3, and 6) for numbers of T cells producing either IFN-γ or IL-5 by use of an enzyme-linked immunospot (ELISPOT) procedure. Cultures of T cells from PP or SP of mice given SRBC by the oral route had a high frequency of IL-5 spot forming cells (SFC), with lower numbers of IFN-γ SFC. However, cultures of CD3+ T cells and CD3+ CD4+ Th cells from spleens of i.p. immunized mice exhibited predominantly IFN-γ SFC, with smaller but significant numbers of IL-5 SFC. This distinct pattern of cytokine production was supported by mRNA analysis where high IL-5 specific mRNA levels were noted in PP T cell cultures of orally primed mice, while IFN-γ mRNA was predominant In the SP CD3+ T cell and CD3+ CD4+ Th cell cultures from i.p. immunized mice. When the frequencies of IFN-γ or IL-5 SFC were assessed among cloned Th cells from orally- or systemically-lmmunlzed mice, 74% of Th cell clones from PP of mice orally immunized with SRBC were IL-5 producers (Th2 type), while 67% of Th cell clones from SP of mice Immunized by the i.p. route were IFN-γ producers (Th1 type). Our studies show that higher frequencies of IFN-γ producing Th1-type cells occur in SP of mice given antigen by the systemic route, while oral immunization results in predominantly IL-5 producing, Th2-type cells in PP.

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