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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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TIDCs are not activated by LPS plus IFN-γ plus anti-CD40 stimulation. (A) Control BM-DCs or enriched C26–6CK TIDCs were cultured overnight with medium alone or with the combination of LPS, IFN-γ plus anti-CD40 antagonist antibody. Flow cytometry histograms (log scale) show the expression of MHC class II, CD40, and CD86 among gated CD11c+ cells (solid lines) in unstimulated (top panels) or stimulated (bottom panels) DC populations. The percentage of positive cells was determined by comparison with isotype control (gray histogram) and is indicated in the top right corner of each histogram. Results are representative of more than five experiments. (B) The ability to respond to LPS plus IFN-γ and anti-CD40 activation was analyzed by measuring IL-12p70 levels in 18 h culture supernatants in various DC populations: DCs isolated from C26–6CK tumors or axillary draining lymph nodes from the same tumor-bearing animals; DCs enriched from hepatocarcinoma developing in x/myc transgenic mice or from normal liver; BM-DCs cultured in the presence or absence of supernatant from C26 tumors. ELISA results are expressed as the mean concentration ± SEM of triplicate cultures. Similar results were obtained in two to five experiments, depending upon the conditions tested.
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fig2: TIDCs are not activated by LPS plus IFN-γ plus anti-CD40 stimulation. (A) Control BM-DCs or enriched C26–6CK TIDCs were cultured overnight with medium alone or with the combination of LPS, IFN-γ plus anti-CD40 antagonist antibody. Flow cytometry histograms (log scale) show the expression of MHC class II, CD40, and CD86 among gated CD11c+ cells (solid lines) in unstimulated (top panels) or stimulated (bottom panels) DC populations. The percentage of positive cells was determined by comparison with isotype control (gray histogram) and is indicated in the top right corner of each histogram. Results are representative of more than five experiments. (B) The ability to respond to LPS plus IFN-γ and anti-CD40 activation was analyzed by measuring IL-12p70 levels in 18 h culture supernatants in various DC populations: DCs isolated from C26–6CK tumors or axillary draining lymph nodes from the same tumor-bearing animals; DCs enriched from hepatocarcinoma developing in x/myc transgenic mice or from normal liver; BM-DCs cultured in the presence or absence of supernatant from C26 tumors. ELISA results are expressed as the mean concentration ± SEM of triplicate cultures. Similar results were obtained in two to five experiments, depending upon the conditions tested.

Mentions: A feature of immature DCs is a response to stimulation with LPS plus IFN-γ plus anti-CD40 antibody by increasing the expression of CD40 and CD86 (Fig. 2 A) as well as by producing IL-12 p70 (Fig. 2 B). In contrast, TIDCs from C26–6CK tumors maintained a similar phenotype under activation and did not produce detectable IL-12 p70 (Fig. 2). Similarly, DCs from normal liver produced IL-12 p70 in response to LPS plus IFN-γ plus anti-CD40, whereas TIDCs from hepatocarcinoma did not (Fig. 2 B). Last, a supernatant from C26 tumors added at the time of activation abolished the secretion of IL-12 p70 in BM-DCs (Fig. 2 B). These results indicate that tumors induced DCs to be refractory to LPS plus IFN-γ plus anti-CD40 stimulation.


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)

TIDCs are not activated by LPS plus IFN-γ plus anti-CD40 stimulation. (A) Control BM-DCs or enriched C26–6CK TIDCs were cultured overnight with medium alone or with the combination of LPS, IFN-γ plus anti-CD40 antagonist antibody. Flow cytometry histograms (log scale) show the expression of MHC class II, CD40, and CD86 among gated CD11c+ cells (solid lines) in unstimulated (top panels) or stimulated (bottom panels) DC populations. The percentage of positive cells was determined by comparison with isotype control (gray histogram) and is indicated in the top right corner of each histogram. Results are representative of more than five experiments. (B) The ability to respond to LPS plus IFN-γ and anti-CD40 activation was analyzed by measuring IL-12p70 levels in 18 h culture supernatants in various DC populations: DCs isolated from C26–6CK tumors or axillary draining lymph nodes from the same tumor-bearing animals; DCs enriched from hepatocarcinoma developing in x/myc transgenic mice or from normal liver; BM-DCs cultured in the presence or absence of supernatant from C26 tumors. ELISA results are expressed as the mean concentration ± SEM of triplicate cultures. Similar results were obtained in two to five experiments, depending upon the conditions tested.
© Copyright Policy
Related In: Results  -  Collection

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

fig2: TIDCs are not activated by LPS plus IFN-γ plus anti-CD40 stimulation. (A) Control BM-DCs or enriched C26–6CK TIDCs were cultured overnight with medium alone or with the combination of LPS, IFN-γ plus anti-CD40 antagonist antibody. Flow cytometry histograms (log scale) show the expression of MHC class II, CD40, and CD86 among gated CD11c+ cells (solid lines) in unstimulated (top panels) or stimulated (bottom panels) DC populations. The percentage of positive cells was determined by comparison with isotype control (gray histogram) and is indicated in the top right corner of each histogram. Results are representative of more than five experiments. (B) The ability to respond to LPS plus IFN-γ and anti-CD40 activation was analyzed by measuring IL-12p70 levels in 18 h culture supernatants in various DC populations: DCs isolated from C26–6CK tumors or axillary draining lymph nodes from the same tumor-bearing animals; DCs enriched from hepatocarcinoma developing in x/myc transgenic mice or from normal liver; BM-DCs cultured in the presence or absence of supernatant from C26 tumors. ELISA results are expressed as the mean concentration ± SEM of triplicate cultures. Similar results were obtained in two to five experiments, depending upon the conditions tested.
Mentions: A feature of immature DCs is a response to stimulation with LPS plus IFN-γ plus anti-CD40 antibody by increasing the expression of CD40 and CD86 (Fig. 2 A) as well as by producing IL-12 p70 (Fig. 2 B). In contrast, TIDCs from C26–6CK tumors maintained a similar phenotype under activation and did not produce detectable IL-12 p70 (Fig. 2). Similarly, DCs from normal liver produced IL-12 p70 in response to LPS plus IFN-γ plus anti-CD40, whereas TIDCs from hepatocarcinoma did not (Fig. 2 B). Last, a supernatant from C26 tumors added at the time of activation abolished the secretion of IL-12 p70 in BM-DCs (Fig. 2 B). These results indicate that tumors induced DCs to be refractory to LPS plus IFN-γ plus anti-CD40 stimulation.

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