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CpG oligodeoxynucleotides act as adjuvants that switch on T helper 1 (Th1) immunity.

Chu RS, Targoni OS, Krieg AM, Lehmann PV, Harding CV - J. Exp. Med. (1997)

Bottom Line: Since these cytokines can induce T helper 1 (Th1) differentiation, we examined the effects of coadministered CpG ODN on the differentiation of Th responses to hen egg lysozyme (HEL).IFA-HEL plus CpG ODN also induced anti-HEL IgG2a (a Th1-associated isotype), which was not induced by IFA-HEL alone.Control non-CpG ODN did not induce IFN-gamma or IgG2a, excepting lesser increases in B10.D2 (Th1-biased) mice.

View Article: PubMed Central - PubMed

Affiliation: Institute of Pathology, Case Western Reserve University, Cleveland, Ohio 44106, USA.

ABSTRACT
Synthetic oligodeoxynucleotides (ODN) that contain unmethylated CpG motifs (CpG ODN) induce macrophages to secrete IL-12, which induces interferon (IFN)-gamma secretion by natural killer (NK) cells. Since these cytokines can induce T helper 1 (Th1) differentiation, we examined the effects of coadministered CpG ODN on the differentiation of Th responses to hen egg lysozyme (HEL). In both BALB/c (Th2-biased) and B10.D2 (Th1-biased) mice, immunization with HEL in incomplete Freund's adjuvant (IFA) resulted in Th2-dominated immune responses characterized by HEL-specific secretion of IL-5 but not IFN-gamma. In contrast, immunization with IFA-HEL plus CpG ODN switched the immune response to a Th1-dominated cytokine pattern, with high levels of HEL-specific IFN-gamma secretion and decreased HEL-specific IL-5 production. IFA-HEL plus CpG ODN also induced anti-HEL IgG2a (a Th1-associated isotype), which was not induced by IFA-HEL alone. Control non-CpG ODN did not induce IFN-gamma or IgG2a, excepting lesser increases in B10.D2 (Th1-biased) mice. Thus, CpG ODN provide a signal to switch on Th1-dominated responses to coadministered antigen and are potential adjuvants for human vaccines to elicit protective Th1 immunity.

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Th1-associated antigen-specific IgG2a responses are induced  by immunization of BALB/c mice with IFA-HEL-CpG ODN but not  IFA-HEL-non–CpG ODN. (A–C). Mice were injected i.p. with CFA-HEL (a control for a Th1-dominated response), IFA-HEL (a control for a  Th2-dominated response), or IFA-HEL with 100 μg of CpG ODN 1826  or non–CpG ODN 1745. Sera were collected from mice 15–18 d after  injection and assayed by ELISA for: (A) anti-HEL IgG2a, an isotype associated with Th1-dominated responses; (B) anti-HEL IgG1; and (C) anti-HEL total Ig response. A–C represent data from a single experiment representative of three similar experiments. (D) BALB/c mice were immunized as above, except that 30 μg of CpG ODN 1585, non–CpG ODN  1972, CpG ODN 1760, or non–CpG ODN 1908 was used for each  mouse. Anti-HEL IgG2a antibodies were detected by serum ELISA. Data  shown in D are representative of three similar experiments.
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Figure 1: Th1-associated antigen-specific IgG2a responses are induced by immunization of BALB/c mice with IFA-HEL-CpG ODN but not IFA-HEL-non–CpG ODN. (A–C). Mice were injected i.p. with CFA-HEL (a control for a Th1-dominated response), IFA-HEL (a control for a Th2-dominated response), or IFA-HEL with 100 μg of CpG ODN 1826 or non–CpG ODN 1745. Sera were collected from mice 15–18 d after injection and assayed by ELISA for: (A) anti-HEL IgG2a, an isotype associated with Th1-dominated responses; (B) anti-HEL IgG1; and (C) anti-HEL total Ig response. A–C represent data from a single experiment representative of three similar experiments. (D) BALB/c mice were immunized as above, except that 30 μg of CpG ODN 1585, non–CpG ODN 1972, CpG ODN 1760, or non–CpG ODN 1908 was used for each mouse. Anti-HEL IgG2a antibodies were detected by serum ELISA. Data shown in D are representative of three similar experiments.

Mentions: BALB/c mice were injected i.p. with 200 μg of HEL in the following adjuvants: CFA, IFA, IFA with CpG ODN 1826, or IFA with a similar ODN lacking the CpG motif, ODN 1745. ODN 1826 and ODN 1745 are phosphorothioate-modified for the entire length of the backbone (S ODN; see Table 1), which greatly increases resistance to nuclease degradation (19). Based on preliminary dose titration studies, ODN were initially used at 100 μg/mouse. Sera were collected 15–18 d after immunization and assayed for anti-HEL Ig (total or specific isotype) by ELISA. Consistent with previous results demonstrating that IFA induces a Th2 response while CFA induces a Th1 response to antigen (6), mice injected with IFA-HEL did not produce detectable IgG2a responses (Fig. 1 A). In contrast, mice injected with CFA-HEL produced high levels of IgG2a. The addition of non–CpG ODN 1745 to the IFA-HEL protocol did not induce IgG2a production. However, immunization with IFA-HEL-CpG ODN 1826 altered the isotype profile of the antibody response, causing a marked increase in anti-HEL IgG2a. Furthermore, in three independent experiments, the production of HEL-specific IgG2a was consistently higher in mice treated with IFA-HEL-CpG ODN 1826 than in mice treated with CFA-HEL.


CpG oligodeoxynucleotides act as adjuvants that switch on T helper 1 (Th1) immunity.

Chu RS, Targoni OS, Krieg AM, Lehmann PV, Harding CV - J. Exp. Med. (1997)

Th1-associated antigen-specific IgG2a responses are induced  by immunization of BALB/c mice with IFA-HEL-CpG ODN but not  IFA-HEL-non–CpG ODN. (A–C). Mice were injected i.p. with CFA-HEL (a control for a Th1-dominated response), IFA-HEL (a control for a  Th2-dominated response), or IFA-HEL with 100 μg of CpG ODN 1826  or non–CpG ODN 1745. Sera were collected from mice 15–18 d after  injection and assayed by ELISA for: (A) anti-HEL IgG2a, an isotype associated with Th1-dominated responses; (B) anti-HEL IgG1; and (C) anti-HEL total Ig response. A–C represent data from a single experiment representative of three similar experiments. (D) BALB/c mice were immunized as above, except that 30 μg of CpG ODN 1585, non–CpG ODN  1972, CpG ODN 1760, or non–CpG ODN 1908 was used for each  mouse. Anti-HEL IgG2a antibodies were detected by serum ELISA. Data  shown in D are representative of three similar experiments.
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Figure 1: Th1-associated antigen-specific IgG2a responses are induced by immunization of BALB/c mice with IFA-HEL-CpG ODN but not IFA-HEL-non–CpG ODN. (A–C). Mice were injected i.p. with CFA-HEL (a control for a Th1-dominated response), IFA-HEL (a control for a Th2-dominated response), or IFA-HEL with 100 μg of CpG ODN 1826 or non–CpG ODN 1745. Sera were collected from mice 15–18 d after injection and assayed by ELISA for: (A) anti-HEL IgG2a, an isotype associated with Th1-dominated responses; (B) anti-HEL IgG1; and (C) anti-HEL total Ig response. A–C represent data from a single experiment representative of three similar experiments. (D) BALB/c mice were immunized as above, except that 30 μg of CpG ODN 1585, non–CpG ODN 1972, CpG ODN 1760, or non–CpG ODN 1908 was used for each mouse. Anti-HEL IgG2a antibodies were detected by serum ELISA. Data shown in D are representative of three similar experiments.
Mentions: BALB/c mice were injected i.p. with 200 μg of HEL in the following adjuvants: CFA, IFA, IFA with CpG ODN 1826, or IFA with a similar ODN lacking the CpG motif, ODN 1745. ODN 1826 and ODN 1745 are phosphorothioate-modified for the entire length of the backbone (S ODN; see Table 1), which greatly increases resistance to nuclease degradation (19). Based on preliminary dose titration studies, ODN were initially used at 100 μg/mouse. Sera were collected 15–18 d after immunization and assayed for anti-HEL Ig (total or specific isotype) by ELISA. Consistent with previous results demonstrating that IFA induces a Th2 response while CFA induces a Th1 response to antigen (6), mice injected with IFA-HEL did not produce detectable IgG2a responses (Fig. 1 A). In contrast, mice injected with CFA-HEL produced high levels of IgG2a. The addition of non–CpG ODN 1745 to the IFA-HEL protocol did not induce IgG2a production. However, immunization with IFA-HEL-CpG ODN 1826 altered the isotype profile of the antibody response, causing a marked increase in anti-HEL IgG2a. Furthermore, in three independent experiments, the production of HEL-specific IgG2a was consistently higher in mice treated with IFA-HEL-CpG ODN 1826 than in mice treated with CFA-HEL.

Bottom Line: Since these cytokines can induce T helper 1 (Th1) differentiation, we examined the effects of coadministered CpG ODN on the differentiation of Th responses to hen egg lysozyme (HEL).IFA-HEL plus CpG ODN also induced anti-HEL IgG2a (a Th1-associated isotype), which was not induced by IFA-HEL alone.Control non-CpG ODN did not induce IFN-gamma or IgG2a, excepting lesser increases in B10.D2 (Th1-biased) mice.

View Article: PubMed Central - PubMed

Affiliation: Institute of Pathology, Case Western Reserve University, Cleveland, Ohio 44106, USA.

ABSTRACT
Synthetic oligodeoxynucleotides (ODN) that contain unmethylated CpG motifs (CpG ODN) induce macrophages to secrete IL-12, which induces interferon (IFN)-gamma secretion by natural killer (NK) cells. Since these cytokines can induce T helper 1 (Th1) differentiation, we examined the effects of coadministered CpG ODN on the differentiation of Th responses to hen egg lysozyme (HEL). In both BALB/c (Th2-biased) and B10.D2 (Th1-biased) mice, immunization with HEL in incomplete Freund's adjuvant (IFA) resulted in Th2-dominated immune responses characterized by HEL-specific secretion of IL-5 but not IFN-gamma. In contrast, immunization with IFA-HEL plus CpG ODN switched the immune response to a Th1-dominated cytokine pattern, with high levels of HEL-specific IFN-gamma secretion and decreased HEL-specific IL-5 production. IFA-HEL plus CpG ODN also induced anti-HEL IgG2a (a Th1-associated isotype), which was not induced by IFA-HEL alone. Control non-CpG ODN did not induce IFN-gamma or IgG2a, excepting lesser increases in B10.D2 (Th1-biased) mice. Thus, CpG ODN provide a signal to switch on Th1-dominated responses to coadministered antigen and are potential adjuvants for human vaccines to elicit protective Th1 immunity.

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