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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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Combination of CpG 1668 and anti–IL-10R antibody overcome TIDC paralysis in vitro. (A and B) BM-DCs and TIDCs enriched from C26–6CK tumors were activated overnight with either LPS, IFN-γ, and anti-CD40 (white bars), or CpG 1668 (black bars) in the presence (anti–IL-10R) or absence (none) of anti–IL-10R antibody. Culture supernatants were assayed for IL-12p70 and TNFα content. Results are expressed as the mean concentration ± SEM of triplicate cultures and are representative of more than three experiments. (C) Mixed leukocyte reaction. Irradiated populations of enriched TIDCs, previously activated overnight with none (□), CpG 1668 (○), CpG plus anti–IL-10R (▪), LPS plus IFN-γ plus anti-CD40 (•), or anti–IL-10R plus LPS plus IFN-γ plus anti-CD40 (♦), were cultured with allogeneic purified T cells. Proliferation is expressed as the mean cpm incorporation ± SEM for triplicates.
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fig3: Combination of CpG 1668 and anti–IL-10R antibody overcome TIDC paralysis in vitro. (A and B) BM-DCs and TIDCs enriched from C26–6CK tumors were activated overnight with either LPS, IFN-γ, and anti-CD40 (white bars), or CpG 1668 (black bars) in the presence (anti–IL-10R) or absence (none) of anti–IL-10R antibody. Culture supernatants were assayed for IL-12p70 and TNFα content. Results are expressed as the mean concentration ± SEM of triplicate cultures and are representative of more than three experiments. (C) Mixed leukocyte reaction. Irradiated populations of enriched TIDCs, previously activated overnight with none (□), CpG 1668 (○), CpG plus anti–IL-10R (▪), LPS plus IFN-γ plus anti-CD40 (•), or anti–IL-10R plus LPS plus IFN-γ plus anti-CD40 (♦), were cultured with allogeneic purified T cells. Proliferation is expressed as the mean cpm incorporation ± SEM for triplicates.

Mentions: We tested different combinations of substances with the aim of relieving tumor-mediated inhibition and simultaneously mediating DC activation, including combinations of the TLR-9 ligand CpG 1668 (21) and an anti–IL-10R blocking antibody (15). In control BM-DCs, CpG alone was able to induce the secretion of IL-12p70 and TNFα in amounts similar to that obtained with LPS plus IFN-γ plus anti-CD40 (Fig. 3 A). The addition of anti–IL-10R antibody to BM-DC culture increased the production of IL-12p70 by ∼30% but did not modify TNFα secretion (Fig. 3 A). Of note, we could not detect endogenous production of IL-10 by BM-DCs (detection limit: 15 pg/ml). An anti–IL-10 antibody also increased IL-12p70 production by BM-DCs (unpublished data), suggesting that indeed IL-10 and not another ligand of IL-10Rα was involved and that low levels of autocrine or paracrine IL-10 were probably sufficient to mediate the effect.


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)

Combination of CpG 1668 and anti–IL-10R antibody overcome TIDC paralysis in vitro. (A and B) BM-DCs and TIDCs enriched from C26–6CK tumors were activated overnight with either LPS, IFN-γ, and anti-CD40 (white bars), or CpG 1668 (black bars) in the presence (anti–IL-10R) or absence (none) of anti–IL-10R antibody. Culture supernatants were assayed for IL-12p70 and TNFα content. Results are expressed as the mean concentration ± SEM of triplicate cultures and are representative of more than three experiments. (C) Mixed leukocyte reaction. Irradiated populations of enriched TIDCs, previously activated overnight with none (□), CpG 1668 (○), CpG plus anti–IL-10R (▪), LPS plus IFN-γ plus anti-CD40 (•), or anti–IL-10R plus LPS plus IFN-γ plus anti-CD40 (♦), were cultured with allogeneic purified T cells. Proliferation is expressed as the mean cpm incorporation ± SEM for triplicates.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196048&req=5

fig3: Combination of CpG 1668 and anti–IL-10R antibody overcome TIDC paralysis in vitro. (A and B) BM-DCs and TIDCs enriched from C26–6CK tumors were activated overnight with either LPS, IFN-γ, and anti-CD40 (white bars), or CpG 1668 (black bars) in the presence (anti–IL-10R) or absence (none) of anti–IL-10R antibody. Culture supernatants were assayed for IL-12p70 and TNFα content. Results are expressed as the mean concentration ± SEM of triplicate cultures and are representative of more than three experiments. (C) Mixed leukocyte reaction. Irradiated populations of enriched TIDCs, previously activated overnight with none (□), CpG 1668 (○), CpG plus anti–IL-10R (▪), LPS plus IFN-γ plus anti-CD40 (•), or anti–IL-10R plus LPS plus IFN-γ plus anti-CD40 (♦), were cultured with allogeneic purified T cells. Proliferation is expressed as the mean cpm incorporation ± SEM for triplicates.
Mentions: We tested different combinations of substances with the aim of relieving tumor-mediated inhibition and simultaneously mediating DC activation, including combinations of the TLR-9 ligand CpG 1668 (21) and an anti–IL-10R blocking antibody (15). In control BM-DCs, CpG alone was able to induce the secretion of IL-12p70 and TNFα in amounts similar to that obtained with LPS plus IFN-γ plus anti-CD40 (Fig. 3 A). The addition of anti–IL-10R antibody to BM-DC culture increased the production of IL-12p70 by ∼30% but did not modify TNFα secretion (Fig. 3 A). Of note, we could not detect endogenous production of IL-10 by BM-DCs (detection limit: 15 pg/ml). An anti–IL-10 antibody also increased IL-12p70 production by BM-DCs (unpublished data), suggesting that indeed IL-10 and not another ligand of IL-10Rα was involved and that low levels of autocrine or paracrine IL-10 were probably sufficient to mediate the effect.

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