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Reversal of tumor-induced dendritic cell paralysis by CpG immunostimulatory oligonucleotide and anti-interleukin 10 receptor antibody.

Vicari AP, Chiodoni C, Vaure C, Aït-Yahia S, Dercamp C, Matsos F, Reynard O, Taverne C, Merle P, Colombo MP, O'Garra A, Trinchieri G, Caux C - J. Exp. Med. (2002)

Bottom Line: Deficient antitumor immunity could be related to a lack of tumor-associated antigen (TAA) presentation by tumor-infiltrating DCs (TIDCs) or to a functional defect of TIDCs.In particular, CpG plus anti-IL-10R enhanced the TAA-specific immune response and triggered de novo IL-12 production.Subsequently, CpG plus anti-IL-10R treatment showed robust antitumor therapeutic activity exceeding by far that of CpG alone, and elicited antitumor immune memory.

View Article: PubMed Central - PubMed

Affiliation: Schering-Plough Laboratory for Immunological Research, BP11, 27 chemin des Peupliers, 69571 Dardilly, France. alain.vicari@spcorp.com

ABSTRACT
Progressing tumors in man and mouse are often infiltrated by dendritic cells (DCs). Deficient antitumor immunity could be related to a lack of tumor-associated antigen (TAA) presentation by tumor-infiltrating DCs (TIDCs) or to a functional defect of TIDCs. Here we investigated the phenotype and function of TIDCs in transplantable and transgenic mouse tumor models. Although TIDCs could encompass various known DC subsets, most had an immature phenotype. We observed that TIDCs were able to present TAA in the context of major histocompatibility complex class I but that they were refractory to stimulation with the combination of lipopolysaccharide, interferon gamma, and anti-CD40 antibody. We could revert TIDC paralysis, however, by in vitro or in vivo stimulation with the combination of a CpG immunostimulatory sequence and an anti-interleukin 10 receptor (IL-10R) antibody. CpG or anti-IL-10R alone were inactive in TIDCs, whereas CpG triggered activation in normal DCs. In particular, CpG plus anti-IL-10R enhanced the TAA-specific immune response and triggered de novo IL-12 production. Subsequently, CpG plus anti-IL-10R treatment showed robust antitumor therapeutic activity exceeding by far that of CpG alone, and elicited antitumor immune memory.

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Induction of tumor antigen–specific immune responses by TIDCs and modulation by CpG plus anti–IL-10R. (A) TIDC cross-present tumor antigen–derived peptides to T cells. The ability of TIDCs from C26–6CK tumors grown in H-2d × H-2b F1 mice to present tumor-derived antigenic peptides was assessed by measuring the secretion of IFN-γ by the CTL clone E/88 (H-2d CTL, white bars) and CTL cell line TG905 (H-2b CTL, black bars). Positive controls consisted of DCs enriched from H-2d × H-2b spleens (APC) and pulsed with the relevant peptide(s) as well as MCA38 H-2b and C26 H-2d cell lines which both express the antigen. (B and C) TIDCs stimulated with CpG 1668 plus anti–IL-10R induce tumor-associated antigen MHC class I–restricted responses in vivo. TIDCs enriched from C26–6CK tumors were cultured overnight with medium alone (TIDC) or CpG 1668 plus anti–IL-10R antibody (TIDC + CpG + a-IL10R), then injected intracutaneously into naive mice. Controls consisted of uninjected mice (negative control) or mice injected with irradiated C26 cells (irrad. C26). Mice were killed 5 d after injection. Experiments were performed with organs pooled from three naive mice and similar results obtained in two separate experiments. (B) Spleens from injected animals were cultured with irradiated C26 cells and IL-2 and cytotoxicity measured against P815 H-2d target cells, alone (white bars) or loaded with the AH-1 antigenic peptide of C26 (black bars). Results are expressed as the mean cytotoxicity ± SEM per triplicate wells at an effector/target ratio of 1:100. (C) Total cell suspensions from pooled draining popliteal lymph nodes were analyzed for IFN-γ–producing cells after overnight culture without (white bar) or with (black bar) AH-1 peptide. Results are expressed as the mean number of spots ± SD for six different wells.
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fig5: Induction of tumor antigen–specific immune responses by TIDCs and modulation by CpG plus anti–IL-10R. (A) TIDC cross-present tumor antigen–derived peptides to T cells. The ability of TIDCs from C26–6CK tumors grown in H-2d × H-2b F1 mice to present tumor-derived antigenic peptides was assessed by measuring the secretion of IFN-γ by the CTL clone E/88 (H-2d CTL, white bars) and CTL cell line TG905 (H-2b CTL, black bars). Positive controls consisted of DCs enriched from H-2d × H-2b spleens (APC) and pulsed with the relevant peptide(s) as well as MCA38 H-2b and C26 H-2d cell lines which both express the antigen. (B and C) TIDCs stimulated with CpG 1668 plus anti–IL-10R induce tumor-associated antigen MHC class I–restricted responses in vivo. TIDCs enriched from C26–6CK tumors were cultured overnight with medium alone (TIDC) or CpG 1668 plus anti–IL-10R antibody (TIDC + CpG + a-IL10R), then injected intracutaneously into naive mice. Controls consisted of uninjected mice (negative control) or mice injected with irradiated C26 cells (irrad. C26). Mice were killed 5 d after injection. Experiments were performed with organs pooled from three naive mice and similar results obtained in two separate experiments. (B) Spleens from injected animals were cultured with irradiated C26 cells and IL-2 and cytotoxicity measured against P815 H-2d target cells, alone (white bars) or loaded with the AH-1 antigenic peptide of C26 (black bars). Results are expressed as the mean cytotoxicity ± SEM per triplicate wells at an effector/target ratio of 1:100. (C) Total cell suspensions from pooled draining popliteal lymph nodes were analyzed for IFN-γ–producing cells after overnight culture without (white bar) or with (black bar) AH-1 peptide. Results are expressed as the mean number of spots ± SD for six different wells.

Mentions: To test whether TIDCs from C26–6CK tumors had been able to capture and process TAAs in the MHC class I pathway, we examined their capacity to present TAA-derived peptides to CTL, as described previously for a C26 tumor engineered to express GM-CSF and CD40L (9; Fig. 5 A). The C26 (H-2d) colon carcinoma expresses an immunodominant TAA which contains the Ld-restricted peptide AH-1 (18), recognized by the E/88 CTL clone. The MC38 (H-2b) colon carcinoma expresses the same TAA and contains a Kb-restricted peptide recognized by the TG905 CTL line (17). Stimulation of E/88 or TG905 cells by C26 and MC38 cells, respectively, induced IFN-γ production in an MHC-restricted fashion (Fig. 5A). TIDCs purified from BALB/c × C57BL/6 (H-2dxb) F1 mice bearing C26–6CK tumors were able to stimulate both CTL lines in a cell dose-dependent fashion, whereas spleen DCs purified from naive H-2dxb F1 mice stimulated IFN-γ production only when pulsed with the relevant peptide (Fig. 5 A). In particular, the stimulation of the H-2b TG905 CTL clearly indicates that TIDCs have been able to capture and present exogenous TAA in the MHC class I pathway.


Reversal of tumor-induced dendritic cell paralysis by CpG immunostimulatory oligonucleotide and anti-interleukin 10 receptor antibody.

Vicari AP, Chiodoni C, Vaure C, Aït-Yahia S, Dercamp C, Matsos F, Reynard O, Taverne C, Merle P, Colombo MP, O'Garra A, Trinchieri G, Caux C - J. Exp. Med. (2002)

Induction of tumor antigen–specific immune responses by TIDCs and modulation by CpG plus anti–IL-10R. (A) TIDC cross-present tumor antigen–derived peptides to T cells. The ability of TIDCs from C26–6CK tumors grown in H-2d × H-2b F1 mice to present tumor-derived antigenic peptides was assessed by measuring the secretion of IFN-γ by the CTL clone E/88 (H-2d CTL, white bars) and CTL cell line TG905 (H-2b CTL, black bars). Positive controls consisted of DCs enriched from H-2d × H-2b spleens (APC) and pulsed with the relevant peptide(s) as well as MCA38 H-2b and C26 H-2d cell lines which both express the antigen. (B and C) TIDCs stimulated with CpG 1668 plus anti–IL-10R induce tumor-associated antigen MHC class I–restricted responses in vivo. TIDCs enriched from C26–6CK tumors were cultured overnight with medium alone (TIDC) or CpG 1668 plus anti–IL-10R antibody (TIDC + CpG + a-IL10R), then injected intracutaneously into naive mice. Controls consisted of uninjected mice (negative control) or mice injected with irradiated C26 cells (irrad. C26). Mice were killed 5 d after injection. Experiments were performed with organs pooled from three naive mice and similar results obtained in two separate experiments. (B) Spleens from injected animals were cultured with irradiated C26 cells and IL-2 and cytotoxicity measured against P815 H-2d target cells, alone (white bars) or loaded with the AH-1 antigenic peptide of C26 (black bars). Results are expressed as the mean cytotoxicity ± SEM per triplicate wells at an effector/target ratio of 1:100. (C) Total cell suspensions from pooled draining popliteal lymph nodes were analyzed for IFN-γ–producing cells after overnight culture without (white bar) or with (black bar) AH-1 peptide. Results are expressed as the mean number of spots ± SD for six different wells.
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Related In: Results  -  Collection

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fig5: Induction of tumor antigen–specific immune responses by TIDCs and modulation by CpG plus anti–IL-10R. (A) TIDC cross-present tumor antigen–derived peptides to T cells. The ability of TIDCs from C26–6CK tumors grown in H-2d × H-2b F1 mice to present tumor-derived antigenic peptides was assessed by measuring the secretion of IFN-γ by the CTL clone E/88 (H-2d CTL, white bars) and CTL cell line TG905 (H-2b CTL, black bars). Positive controls consisted of DCs enriched from H-2d × H-2b spleens (APC) and pulsed with the relevant peptide(s) as well as MCA38 H-2b and C26 H-2d cell lines which both express the antigen. (B and C) TIDCs stimulated with CpG 1668 plus anti–IL-10R induce tumor-associated antigen MHC class I–restricted responses in vivo. TIDCs enriched from C26–6CK tumors were cultured overnight with medium alone (TIDC) or CpG 1668 plus anti–IL-10R antibody (TIDC + CpG + a-IL10R), then injected intracutaneously into naive mice. Controls consisted of uninjected mice (negative control) or mice injected with irradiated C26 cells (irrad. C26). Mice were killed 5 d after injection. Experiments were performed with organs pooled from three naive mice and similar results obtained in two separate experiments. (B) Spleens from injected animals were cultured with irradiated C26 cells and IL-2 and cytotoxicity measured against P815 H-2d target cells, alone (white bars) or loaded with the AH-1 antigenic peptide of C26 (black bars). Results are expressed as the mean cytotoxicity ± SEM per triplicate wells at an effector/target ratio of 1:100. (C) Total cell suspensions from pooled draining popliteal lymph nodes were analyzed for IFN-γ–producing cells after overnight culture without (white bar) or with (black bar) AH-1 peptide. Results are expressed as the mean number of spots ± SD for six different wells.
Mentions: To test whether TIDCs from C26–6CK tumors had been able to capture and process TAAs in the MHC class I pathway, we examined their capacity to present TAA-derived peptides to CTL, as described previously for a C26 tumor engineered to express GM-CSF and CD40L (9; Fig. 5 A). The C26 (H-2d) colon carcinoma expresses an immunodominant TAA which contains the Ld-restricted peptide AH-1 (18), recognized by the E/88 CTL clone. The MC38 (H-2b) colon carcinoma expresses the same TAA and contains a Kb-restricted peptide recognized by the TG905 CTL line (17). Stimulation of E/88 or TG905 cells by C26 and MC38 cells, respectively, induced IFN-γ production in an MHC-restricted fashion (Fig. 5A). TIDCs purified from BALB/c × C57BL/6 (H-2dxb) F1 mice bearing C26–6CK tumors were able to stimulate both CTL lines in a cell dose-dependent fashion, whereas spleen DCs purified from naive H-2dxb F1 mice stimulated IFN-γ production only when pulsed with the relevant peptide (Fig. 5 A). In particular, the stimulation of the H-2b TG905 CTL clearly indicates that TIDCs have been able to capture and present exogenous TAA in the MHC class I pathway.

Bottom Line: Deficient antitumor immunity could be related to a lack of tumor-associated antigen (TAA) presentation by tumor-infiltrating DCs (TIDCs) or to a functional defect of TIDCs.In particular, CpG plus anti-IL-10R enhanced the TAA-specific immune response and triggered de novo IL-12 production.Subsequently, CpG plus anti-IL-10R treatment showed robust antitumor therapeutic activity exceeding by far that of CpG alone, and elicited antitumor immune memory.

View Article: PubMed Central - PubMed

Affiliation: Schering-Plough Laboratory for Immunological Research, BP11, 27 chemin des Peupliers, 69571 Dardilly, France. alain.vicari@spcorp.com

ABSTRACT
Progressing tumors in man and mouse are often infiltrated by dendritic cells (DCs). Deficient antitumor immunity could be related to a lack of tumor-associated antigen (TAA) presentation by tumor-infiltrating DCs (TIDCs) or to a functional defect of TIDCs. Here we investigated the phenotype and function of TIDCs in transplantable and transgenic mouse tumor models. Although TIDCs could encompass various known DC subsets, most had an immature phenotype. We observed that TIDCs were able to present TAA in the context of major histocompatibility complex class I but that they were refractory to stimulation with the combination of lipopolysaccharide, interferon gamma, and anti-CD40 antibody. We could revert TIDC paralysis, however, by in vitro or in vivo stimulation with the combination of a CpG immunostimulatory sequence and an anti-interleukin 10 receptor (IL-10R) antibody. CpG or anti-IL-10R alone were inactive in TIDCs, whereas CpG triggered activation in normal DCs. In particular, CpG plus anti-IL-10R enhanced the TAA-specific immune response and triggered de novo IL-12 production. Subsequently, CpG plus anti-IL-10R treatment showed robust antitumor therapeutic activity exceeding by far that of CpG alone, and elicited antitumor immune memory.

Show MeSH
Related in: MedlinePlus