Oral but not parenteral interleukin (IL)-12 redirects T helper 2 (Th2)-type responses to an oral vaccine without altering mucosal IgA responses - PubMed (original) (raw)

Oral but not parenteral interleukin (IL)-12 redirects T helper 2 (Th2)-type responses to an oral vaccine without altering mucosal IgA responses

M Marinaro et al. J Exp Med. 1997.

Abstract

Our past studies have shown that the mucosal adjuvant cholera toxin (CT) induces T helper type 2 (Th2) responses with systemic IgG1, IgE and mucosal secretory IgA (S-IgA) antibodies (Abs). In this study, recombinant murine IL-12 (rmIL-12) was given either parenterally or orally to mice orally immunized with tetanus toxoid (TT) and CT to determine whether this cytokine could redirect the CT-induced Th2-type responses and what effect this shift would have on S-IgA Ab responses. Intraperitoneal administration of rmIL-12 shifted TT-specific responses toward Th1-type and resulted in CD4+ T cells producing IFN-gamma and IL-2 with markedly reduced levels of Th2-type cytokines. This cytokine profile was accompanied by increased delayed-type hypersensitivity (DTH) and shifts in serum IgG1 to IgG2a and IgG3 anti-TT Ab responses. Further, serum IgE and S-IgA Ab responses were markedly reduced by parenteral IL-12. When IL-12 complexed to liposomes was given orally both shifts to IgG2a and IgG3 and low IgE Abs again occurred concomitant with enhanced serum IFN-gamma and DTH responses. Interestingly, oral rmIL-12 did not result in significant levels of serum IL-12 nor altered S-IgA Ab responses and resulted in higher levels of some Th2-type cytokines both in vitro and in vivo when compared with parenteral IL-12. Our results show that the shifts in systemic immune responses with intact S-IgA Abs which occur after oral delivery of IL-12-liposomes are due to cytokine effects in the Peyer's patches and suggest new strategies for the targeted manipulation of Th1- and Th2-type responses to mucosal vaccines.

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Figures

Figure 1

rmIL-12 treatment schedules for mice which received a combined oral vaccine. Four groups of C57BL/6 mice were given TT plus CT as adjuvant on days 0, 7, and 14 by the oral route. Group A were positive controls receiving oral vaccine but not rmIL-12. Group B received 15 doses of rmIL-12 (day 0 to 5, 7 to 11, and 14 to 18) by the intraperitoneal route. This group was subdivided into mice receiving 10, 100, or 1,000 ng of rmIL-12/dose. Groups C and D received three (days 0, 7, 14) or six (days 0, 3, 7, 10, 14, 17) oral doses of rIL-12 complexed to liposomes.

Figure 2

The effect of parenteral administration of rmIL-12 on polyclonal and TT-specific serum Ab responses in mice orally immunized with TT and CT as mucosal adjuvant. Mice received the oral vaccine alone (unshaded) or together with rmIL-12 by the i.p. route (shaded, 10 ng/dose or striped, 100 ng/dose). (A) Polyclonal and TT-specific serum IgE Abs were measured on day 14 by ELISA and PCA assays, respectively. (B) The distribution of TT-specific serum IgG subclasses were analyzed on day 21 by ELISA. (C) Serum TT-specific IgM, IgA, and IgG Abs were measured by ELISA on day 21. Results are expressed as the mean ± one SD from three different experiments (five mice per group).

Figure 3

The effect of oral administration of rmIL-12 on polyclonal and TT-specific serum Ab responses. Mice received the oral combined vaccine alone (unshaded) or vaccine together with rmIL-12 complexed to liposomes by the oral route (striped, 1,000 ng/dose three times and shaded, 1,000 ng/dose six times). (A) Polyclonal and TT-specific serum IgE Abs were measured on day 14 by ELISA and PCA assays, respectively. (B) The distribution of TT-specific serum IgG subclasses were determined on day 21 by ELISA. (C) Serum TT-specific IgM, IgA, and IgG Abs were measured by ELISA on day 21. Results are expressed as the mean ± one SD from four different experiments (five mice per group).

Figure 4

The effect of oral and parenteral administration of rmIL-12 on mucosal IgA responses to an oral vaccine consisting of TT and CT as mucosal adjuvant. TT-specific S-IgA Ab titers in fecal extracts were measured on day 21 by ELISA. The frequency of TT-specific IgA AFCs in LPLs was determined on day 21 by ELISPOT. Mice received the oral vaccine alone (unshaded) or together with rmIL-12 by i.p. injection (striped, 100 ng/dose) or six oral administrations (shaded, 1,000 ng/dose). The results are expressed as the mean ± one SD from four different experiments (five mice per group).

Figure 5

The induction of Th1-type (IL-2 and IFN-γ) and Th2-type (IL-4, IL-5, IL-6, and IL-10) cytokine secretion by CD4+ T cells isolated from mucosally immunized mice which had been given oral (shaded) or parenteral (striped) rmIL-12. SP and PP CD4+ T cells, were purified from mice receiving the combined oral vaccine only (unshaded) or combined vaccine together with rmIL-12 orally (six times, 1,000 ng/dose) or parenterally (15 times, 100 ng/dose) and were restimulated in vitro for 6 d in the presence of feeder cells and TT-coated beads. Th1type (A) and Th2-type (B) cytokine production in culture supernatants were analyzed by ELISA. Cytokine levels are representative of three separate experiments.

Figure 5

Figure 6

The induction of Th1-type (IFN-γ) and Th2-type (IL-5 and IL-10) cytokines in sera of mice receiving rmIL-12 and orally immunized with TT and CT as adjuvant. Mice received the combined oral vaccine (unshaded) together with rmIL-12 by the i.p. route (striped, 15 times, 100 ng/dose) or by the oral route (shaded six times, 1,000 ng/dose). Levels of IFN-γ, IL-5, and IL-10 in the serum were determined on day 21 by ELISA. The results are expressed as the mean ± one SD and are representative of three different experiments.

Figure 7

Induction of delayed-type hypersensitivity responses in rmIL-12–treated mice given the combined oral vaccine. Three groups of mice were assessed and included combined oral vaccine only (unshaded), those receiving combined oral vaccine and rmIL-12 by the i.p. (striped; 15 times 100 ng/dose) route, or mice given rmIL-12 by the oral route (shaded; six times 1,000 ng/dose). The results are expressed as the mean ± one SD of the difference in the ear swelling between the TT-injected and OVA-injected ear pinna and are representative of three separate experiments.

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Oral but not parenteral interleukin (IL)-12 redirects T helper 2 (Th2)-type responses to an oral vaccine without altering mucosal IgA responses - PubMed (original) (raw)