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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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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.
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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.

Mentions: Recombinant murine IL-12 (rmIL-12) was generously provided by Dr. Maurice K. Gately (Hoffmann-LaRoche, Nutley, NJ) and was given in various doses to mice by the i.p. or oral route (see Fig. 1). For i.p. delivery, rmIL-12 (10, 100, or 1000 ng/dose) was diluted in sterile PBS containing 1% normal mouse serum and given daily (5 times/wk) as indicated in Fig. 1. For oral delivery, rmIL-12 (1,000 ng/dose) was complexed with preformed cationic liposomes (DOTAP; Boehringer Mannheim Corp., Indianapolis, IN). Preliminary experiments established that administration of liposomes alone did not affect immune responses to the vaccine and that rmIL-12 complexed to DOTAP liposomes retained its biologic activity in vitro and in vivo. Moreover, dose-response experiments established that optimal liposome uptake of rmIL-12 was obtained at a 50:1 ratio (wt/wt) of DOTAP liposome mixed with IL-12. Mice receiving oral DOTAP-IL-12 (liposome-IL-12) were deprived of food for 2 h followed by neutralization of stomach acidity as described above. On days where mice were given oral liposome-IL-12 and TT with CT, two separate gavages were made first with oral TT plus CT followed by oral liposome-IL-12 at an ∼30 min interval (Fig. 1).


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)

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.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196042&req=5

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.
Mentions: Recombinant murine IL-12 (rmIL-12) was generously provided by Dr. Maurice K. Gately (Hoffmann-LaRoche, Nutley, NJ) and was given in various doses to mice by the i.p. or oral route (see Fig. 1). For i.p. delivery, rmIL-12 (10, 100, or 1000 ng/dose) was diluted in sterile PBS containing 1% normal mouse serum and given daily (5 times/wk) as indicated in Fig. 1. For oral delivery, rmIL-12 (1,000 ng/dose) was complexed with preformed cationic liposomes (DOTAP; Boehringer Mannheim Corp., Indianapolis, IN). Preliminary experiments established that administration of liposomes alone did not affect immune responses to the vaccine and that rmIL-12 complexed to DOTAP liposomes retained its biologic activity in vitro and in vivo. Moreover, dose-response experiments established that optimal liposome uptake of rmIL-12 was obtained at a 50:1 ratio (wt/wt) of DOTAP liposome mixed with IL-12. Mice receiving oral DOTAP-IL-12 (liposome-IL-12) were deprived of food for 2 h followed by neutralization of stomach acidity as described above. On days where mice were given oral liposome-IL-12 and TT with CT, two separate gavages were made first with oral TT plus CT followed by oral liposome-IL-12 at an ∼30 min interval (Fig. 1).

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