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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.

Marinaro M, Boyaka PN, Finkelman FD, Kiyono H, Jackson RJ, Jirillo E, McGhee JR - J. Exp. Med. (1997)

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, University of Alabama at Birmingham 35294-2170, USA.

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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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).
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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).

Mentions: Since IL-12 plays an important role in the induction of IFN-γ production and subsequent Th1-type responses, we queried whether IL-12 treatment would alter Th2-type responses and the resultant serum IgE and IgG subclass responses in mice receiving oral TT plus CT as adjuvant (31, 32, 40, 41). Frequent i.p. injections of rmIL-12 downregulated both polyclonal and TT-specific IgE Abs (see Fig. 2 A). The effect of rmIL-12 was dose dependent because IgE responses were not significantly affected by administration of 10 ng of rmIL-12, while a significant decrease in both polyclonal and TT-specific IgE Abs was observed in mice receiving 100 ng/dose (see Fig. 2 A). The distribution of anti-TT IgG subclass Ab responses was also affected by parenteral rmIL-12 treatment and small increases in IgG2a anti-TT Abs were observed in mice given a 10 ng/dose of rmIL-12 (see Fig. 2 B). When the dose of rmIL-12 was increased to 100 ng, a significant change in the profile of IgG subclasses was observed with concomitant down regulation of IgG1 and increases in IgG2a and IgG3 Ab titers. Despite this shift in IgG subclass distribution, serum TT-specific IgG, IgA, and IgM titers were similar in mice orally immunized with TT plus CT in the presence or in the absence of IL-12, suggesting that rmIL-12 did not affect the potency of CT as adjuvant for serum Ab responses (see Fig. 2 C). Antibody responses of all isotypes were abrogated in mice receiving 1,000 ng/dose of rmIL-12, indicating that high doses of rmIL-12 impaired the ability of CT to act as a mucosal adjuvant (results not shown). However, ∼40% of mice which received 10–14 doses of 1,000 ng of rmIL-12 did not survive, perhaps due to combined toxic effects of CT and IL-12.


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

Marinaro M, Boyaka PN, Finkelman FD, Kiyono H, Jackson RJ, Jirillo E, McGhee JR - J. Exp. Med. (1997)

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).
© Copyright Policy
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC2196042&req=5

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).
Mentions: Since IL-12 plays an important role in the induction of IFN-γ production and subsequent Th1-type responses, we queried whether IL-12 treatment would alter Th2-type responses and the resultant serum IgE and IgG subclass responses in mice receiving oral TT plus CT as adjuvant (31, 32, 40, 41). Frequent i.p. injections of rmIL-12 downregulated both polyclonal and TT-specific IgE Abs (see Fig. 2 A). The effect of rmIL-12 was dose dependent because IgE responses were not significantly affected by administration of 10 ng of rmIL-12, while a significant decrease in both polyclonal and TT-specific IgE Abs was observed in mice receiving 100 ng/dose (see Fig. 2 A). The distribution of anti-TT IgG subclass Ab responses was also affected by parenteral rmIL-12 treatment and small increases in IgG2a anti-TT Abs were observed in mice given a 10 ng/dose of rmIL-12 (see Fig. 2 B). When the dose of rmIL-12 was increased to 100 ng, a significant change in the profile of IgG subclasses was observed with concomitant down regulation of IgG1 and increases in IgG2a and IgG3 Ab titers. Despite this shift in IgG subclass distribution, serum TT-specific IgG, IgA, and IgM titers were similar in mice orally immunized with TT plus CT in the presence or in the absence of IL-12, suggesting that rmIL-12 did not affect the potency of CT as adjuvant for serum Ab responses (see Fig. 2 C). Antibody responses of all isotypes were abrogated in mice receiving 1,000 ng/dose of rmIL-12, indicating that high doses of rmIL-12 impaired the ability of CT to act as a mucosal adjuvant (results not shown). However, ∼40% of mice which received 10–14 doses of 1,000 ng of rmIL-12 did not survive, perhaps due to combined toxic effects of CT and IL-12.

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Department of Microbiology, University of Alabama at Birmingham 35294-2170, USA.

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.

Show MeSH
Related in: MedlinePlus