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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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CpG ODN enhance HEL-specific IFN-γ production by BALB/c splenocytes.  Mice were immunized as in Fig.  1 with 100 μg ODN/mouse in  A and 30 μg ODN/mouse in  panel B. After 3 wk, splenocytes  were isolated and incubated with  HEL (closed circles) or medium  alone (open circles). ELISA spot  assay was performed and spots  were quantitated by a computerized image analysis program.  Each point represents the mean  number of spots per well for one  mouse (assayed in duplicate);  horizontal bars indicate the mean  of points for each group of mice.  Similar results were observed in  five independent experiments  with CpG- and non–CpG ODN  in BALB/c mice.
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Figure 2: CpG ODN enhance HEL-specific IFN-γ production by BALB/c splenocytes. Mice were immunized as in Fig. 1 with 100 μg ODN/mouse in A and 30 μg ODN/mouse in panel B. After 3 wk, splenocytes were isolated and incubated with HEL (closed circles) or medium alone (open circles). ELISA spot assay was performed and spots were quantitated by a computerized image analysis program. Each point represents the mean number of spots per well for one mouse (assayed in duplicate); horizontal bars indicate the mean of points for each group of mice. Similar results were observed in five independent experiments with CpG- and non–CpG ODN in BALB/c mice.

Mentions: We used a modified ELISA spot assay (see Materials and Methods) to assess recall antigen-specific IFN-γ secretion as a measure of Th1 memory cells induced after immunization with HEL. Three weeks after immunization of BALB/c mice with HEL in adjuvant, splenocytes were isolated, incubated in vitro with or without HEL and assayed for IFN-γ production. Upon restimulation with HEL, splenocytes from mice immunized with IFA-HEL showed little or no antigen-specific production of IFN-γ (Fig. 2 A), as expected (6). In contrast, splenocytes from mice immunized with CFA-HEL showed antigen-specific production of IFN-γ, demonstrating that CFA-HEL induced a Th1 response to HEL in these mice, as previously observed (6). Prior studies using isolated spleen-derived CD4+ T cells (with irradiated BALB/c-scid splenocytes as antigen-presenting cells) have shown that antigen-specific cytokine secretion measured by this assay is mediated by CD4+ T cells (Yip, H., A. Karulin, M. Tary-Lehmann, P. Heeger, R. Trezza, T. Forsthuber, and P.V. Lehmann, manuscript submitted for publication).


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)

CpG ODN enhance HEL-specific IFN-γ production by BALB/c splenocytes.  Mice were immunized as in Fig.  1 with 100 μg ODN/mouse in  A and 30 μg ODN/mouse in  panel B. After 3 wk, splenocytes  were isolated and incubated with  HEL (closed circles) or medium  alone (open circles). ELISA spot  assay was performed and spots  were quantitated by a computerized image analysis program.  Each point represents the mean  number of spots per well for one  mouse (assayed in duplicate);  horizontal bars indicate the mean  of points for each group of mice.  Similar results were observed in  five independent experiments  with CpG- and non–CpG ODN  in BALB/c mice.
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Related In: Results  -  Collection

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Figure 2: CpG ODN enhance HEL-specific IFN-γ production by BALB/c splenocytes. Mice were immunized as in Fig. 1 with 100 μg ODN/mouse in A and 30 μg ODN/mouse in panel B. After 3 wk, splenocytes were isolated and incubated with HEL (closed circles) or medium alone (open circles). ELISA spot assay was performed and spots were quantitated by a computerized image analysis program. Each point represents the mean number of spots per well for one mouse (assayed in duplicate); horizontal bars indicate the mean of points for each group of mice. Similar results were observed in five independent experiments with CpG- and non–CpG ODN in BALB/c mice.
Mentions: We used a modified ELISA spot assay (see Materials and Methods) to assess recall antigen-specific IFN-γ secretion as a measure of Th1 memory cells induced after immunization with HEL. Three weeks after immunization of BALB/c mice with HEL in adjuvant, splenocytes were isolated, incubated in vitro with or without HEL and assayed for IFN-γ production. Upon restimulation with HEL, splenocytes from mice immunized with IFA-HEL showed little or no antigen-specific production of IFN-γ (Fig. 2 A), as expected (6). In contrast, splenocytes from mice immunized with CFA-HEL showed antigen-specific production of IFN-γ, demonstrating that CFA-HEL induced a Th1 response to HEL in these mice, as previously observed (6). Prior studies using isolated spleen-derived CD4+ T cells (with irradiated BALB/c-scid splenocytes as antigen-presenting cells) have shown that antigen-specific cytokine secretion measured by this assay is mediated by CD4+ T cells (Yip, H., A. Karulin, M. Tary-Lehmann, P. Heeger, R. Trezza, T. Forsthuber, and P.V. Lehmann, manuscript submitted for publication).

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.

Show MeSH
Related in: MedlinePlus