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Blood-derived inflammatory dendritic cells in lymph nodes stimulate acute T helper type 1 immune responses.

Nakano H, Lin KL, Yanagita M, Charbonneau C, Cook DN, Kakiuchi T, Gunn MD - Nat. Immunol. (2009)

Bottom Line: Here we found after viral infection or immunization, inflammatory monocytes were recruited into lymph nodes directly from the blood to become CD11c(+)CD11b(hi)Gr-1(+) inflammatory DCs, which produced abundant interleukin 12p70 and potently stimulated T(H)1 responses.This monocyte extravasation required the chemokine receptor CCR2 but not the chemokine CCL2 or receptor CCR7.We conclude that blood-derived inflammatory DCs are important in the development of T(H)1 immune responses.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Duke University Medical Center, Durham, North Carolina, USA.

ABSTRACT
T helper type 1 (T(H)1)-polarized immune responses, which confer protection against intracellular pathogens, are thought to be initiated by dendritic cells (DCs) that enter lymph nodes from peripheral tissues. Here we found after viral infection or immunization, inflammatory monocytes were recruited into lymph nodes directly from the blood to become CD11c(+)CD11b(hi)Gr-1(+) inflammatory DCs, which produced abundant interleukin 12p70 and potently stimulated T(H)1 responses. This monocyte extravasation required the chemokine receptor CCR2 but not the chemokine CCL2 or receptor CCR7. Thus, the accumulation of inflammatory DCs and T(H)1 responses were much lower in Ccr2(-/-) mice, were preserved in Ccl2(-/-) mice and were relatively higher in CCL19-CCL21-Ser-deficient plt mutant mice, in which all other lymph node DC types were fewer in number. We conclude that blood-derived inflammatory DCs are important in the development of T(H)1 immune responses.

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Increased immune responses in plt mice. (a–c) DTH responses in plt mice. WT and plt mice were sensitized with OVA/CFA (a,b) or OVA/RIBI x1 or x2 (c). Responses were elicited after 7 days and ear swelling measured at 24 h. (d) Time course of contact hypersensitivity (CHS) responses. BALB/c and plt mice were sensitized by oxazolone skin painting and ear swelling measured 24 h after elicitation. Data points represent mean ± SD for 5 mice (a–d). (e) IFN-γ intracellular staining of BALB/c and plt lymph node CD4+ T cells 9 days after immunization with OVA/CFA. Numbers indicate percentage of CD44+IFN-γ+ cells among CD4+ T cells. (f) Percentage of CD44+IFN-γ+ cells among CD4+ T cells after restimulation with increasing concentrations of OVA. (g) DTH responses, performed as above, after reciprocal bone marrow transplantation between BALB/c and BALB/c-plt mice. Bars represent mean ear swelling as a percentage of the responses in BALB/c mice. n = 5 mice/group, P<0.02 for BALB/c vs. plt recipients. (h) Allogeneic T cell proliferation induced by lymph node CD11c+ cells from untreated mice. (i,j) OVA-specific proliferation of a DO11.10 CD4 T cell line induced by total lymph node DCs 9 days (i) or 28 days (j) after immunization with OVA/CFA. Points represent mean ± SD for 3 mice (h j). * P < 0.05; ** P < 0.005; *** P < 0.0005 by Student’s t-test.
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Figure 1: Increased immune responses in plt mice. (a–c) DTH responses in plt mice. WT and plt mice were sensitized with OVA/CFA (a,b) or OVA/RIBI x1 or x2 (c). Responses were elicited after 7 days and ear swelling measured at 24 h. (d) Time course of contact hypersensitivity (CHS) responses. BALB/c and plt mice were sensitized by oxazolone skin painting and ear swelling measured 24 h after elicitation. Data points represent mean ± SD for 5 mice (a–d). (e) IFN-γ intracellular staining of BALB/c and plt lymph node CD4+ T cells 9 days after immunization with OVA/CFA. Numbers indicate percentage of CD44+IFN-γ+ cells among CD4+ T cells. (f) Percentage of CD44+IFN-γ+ cells among CD4+ T cells after restimulation with increasing concentrations of OVA. (g) DTH responses, performed as above, after reciprocal bone marrow transplantation between BALB/c and BALB/c-plt mice. Bars represent mean ear swelling as a percentage of the responses in BALB/c mice. n = 5 mice/group, P<0.02 for BALB/c vs. plt recipients. (h) Allogeneic T cell proliferation induced by lymph node CD11c+ cells from untreated mice. (i,j) OVA-specific proliferation of a DO11.10 CD4 T cell line induced by total lymph node DCs 9 days (i) or 28 days (j) after immunization with OVA/CFA. Points represent mean ± SD for 3 mice (h j). * P < 0.05; ** P < 0.005; *** P < 0.0005 by Student’s t-test.

Mentions: We identified lymph node moDCs in the course of examining immune response abnormalities in plt mice. Plt mice display a >95% reduction in the migration of naive T cells into lymph nodes and a 50–60% decrease in total lymph node DCs21,22. However, despite these defects, plt mice display increased antigen-specific T cell numbers and increased antigen-specific T cell proliferation for at least 16 months after immunization23,24. On both BALB/c (Fig. 1a) and C57BL/6 (Fig. 1b) genetic backgrounds, plt mice displayed delayed-type hypersensitivity (DTH) responses that are about 2-fold greater than seen in wild-type mice. This increased response was independent of whether Complete Freund’s adjuvant (CFA), monophosphoryl lipid A-trehalose dicorynomycolate (RIBI), or lipopolysaccharide (LPS) was used as the adjuvant for sensitization and of the number of sensitizing immunizations (Fig. 1c and data not shown). Plt mice also displayed contact hypersensitivity (CHS) responses that were markedly increased by day 7 and were maintained without reduction for at least eight weeks (Fig. 1d). After immunization with ovalbumin (OVA) in CFA, plt mice displayed a 6-fold increase in the proportion of CD4+ T cells that produced IFN-γ (Fig. 1e, f), indicating the development of markedly increased TH1 T cell responses. These increased T cell responses were not due to an intrinsic defect in T cells or DCs. After reciprocal bone marrow transplantations between wild-type and plt mice, the plt phenotype of increased DTH responses was seen in plt recipients, irrespective of the bone marrow donor genotype (Fig. 1g). In our hands, bone marrow transplantation typically leads to some reduction in DTH response relative to non-transplanted mice (data not shown). Like plt mice, CCR7-deficient mice display severe defects in the accumulation of T cells and DCs in draining lymph nodes25, but display increased T cell responses in contact hypersensitivity assays26. The cause of the paradoxically increased T cell responses seen in plt and CCR7-deficient mice has not been previously determined.


Blood-derived inflammatory dendritic cells in lymph nodes stimulate acute T helper type 1 immune responses.

Nakano H, Lin KL, Yanagita M, Charbonneau C, Cook DN, Kakiuchi T, Gunn MD - Nat. Immunol. (2009)

Increased immune responses in plt mice. (a–c) DTH responses in plt mice. WT and plt mice were sensitized with OVA/CFA (a,b) or OVA/RIBI x1 or x2 (c). Responses were elicited after 7 days and ear swelling measured at 24 h. (d) Time course of contact hypersensitivity (CHS) responses. BALB/c and plt mice were sensitized by oxazolone skin painting and ear swelling measured 24 h after elicitation. Data points represent mean ± SD for 5 mice (a–d). (e) IFN-γ intracellular staining of BALB/c and plt lymph node CD4+ T cells 9 days after immunization with OVA/CFA. Numbers indicate percentage of CD44+IFN-γ+ cells among CD4+ T cells. (f) Percentage of CD44+IFN-γ+ cells among CD4+ T cells after restimulation with increasing concentrations of OVA. (g) DTH responses, performed as above, after reciprocal bone marrow transplantation between BALB/c and BALB/c-plt mice. Bars represent mean ear swelling as a percentage of the responses in BALB/c mice. n = 5 mice/group, P<0.02 for BALB/c vs. plt recipients. (h) Allogeneic T cell proliferation induced by lymph node CD11c+ cells from untreated mice. (i,j) OVA-specific proliferation of a DO11.10 CD4 T cell line induced by total lymph node DCs 9 days (i) or 28 days (j) after immunization with OVA/CFA. Points represent mean ± SD for 3 mice (h j). * P < 0.05; ** P < 0.005; *** P < 0.0005 by Student’s t-test.
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Figure 1: Increased immune responses in plt mice. (a–c) DTH responses in plt mice. WT and plt mice were sensitized with OVA/CFA (a,b) or OVA/RIBI x1 or x2 (c). Responses were elicited after 7 days and ear swelling measured at 24 h. (d) Time course of contact hypersensitivity (CHS) responses. BALB/c and plt mice were sensitized by oxazolone skin painting and ear swelling measured 24 h after elicitation. Data points represent mean ± SD for 5 mice (a–d). (e) IFN-γ intracellular staining of BALB/c and plt lymph node CD4+ T cells 9 days after immunization with OVA/CFA. Numbers indicate percentage of CD44+IFN-γ+ cells among CD4+ T cells. (f) Percentage of CD44+IFN-γ+ cells among CD4+ T cells after restimulation with increasing concentrations of OVA. (g) DTH responses, performed as above, after reciprocal bone marrow transplantation between BALB/c and BALB/c-plt mice. Bars represent mean ear swelling as a percentage of the responses in BALB/c mice. n = 5 mice/group, P<0.02 for BALB/c vs. plt recipients. (h) Allogeneic T cell proliferation induced by lymph node CD11c+ cells from untreated mice. (i,j) OVA-specific proliferation of a DO11.10 CD4 T cell line induced by total lymph node DCs 9 days (i) or 28 days (j) after immunization with OVA/CFA. Points represent mean ± SD for 3 mice (h j). * P < 0.05; ** P < 0.005; *** P < 0.0005 by Student’s t-test.
Mentions: We identified lymph node moDCs in the course of examining immune response abnormalities in plt mice. Plt mice display a >95% reduction in the migration of naive T cells into lymph nodes and a 50–60% decrease in total lymph node DCs21,22. However, despite these defects, plt mice display increased antigen-specific T cell numbers and increased antigen-specific T cell proliferation for at least 16 months after immunization23,24. On both BALB/c (Fig. 1a) and C57BL/6 (Fig. 1b) genetic backgrounds, plt mice displayed delayed-type hypersensitivity (DTH) responses that are about 2-fold greater than seen in wild-type mice. This increased response was independent of whether Complete Freund’s adjuvant (CFA), monophosphoryl lipid A-trehalose dicorynomycolate (RIBI), or lipopolysaccharide (LPS) was used as the adjuvant for sensitization and of the number of sensitizing immunizations (Fig. 1c and data not shown). Plt mice also displayed contact hypersensitivity (CHS) responses that were markedly increased by day 7 and were maintained without reduction for at least eight weeks (Fig. 1d). After immunization with ovalbumin (OVA) in CFA, plt mice displayed a 6-fold increase in the proportion of CD4+ T cells that produced IFN-γ (Fig. 1e, f), indicating the development of markedly increased TH1 T cell responses. These increased T cell responses were not due to an intrinsic defect in T cells or DCs. After reciprocal bone marrow transplantations between wild-type and plt mice, the plt phenotype of increased DTH responses was seen in plt recipients, irrespective of the bone marrow donor genotype (Fig. 1g). In our hands, bone marrow transplantation typically leads to some reduction in DTH response relative to non-transplanted mice (data not shown). Like plt mice, CCR7-deficient mice display severe defects in the accumulation of T cells and DCs in draining lymph nodes25, but display increased T cell responses in contact hypersensitivity assays26. The cause of the paradoxically increased T cell responses seen in plt and CCR7-deficient mice has not been previously determined.

Bottom Line: Here we found after viral infection or immunization, inflammatory monocytes were recruited into lymph nodes directly from the blood to become CD11c(+)CD11b(hi)Gr-1(+) inflammatory DCs, which produced abundant interleukin 12p70 and potently stimulated T(H)1 responses.This monocyte extravasation required the chemokine receptor CCR2 but not the chemokine CCL2 or receptor CCR7.We conclude that blood-derived inflammatory DCs are important in the development of T(H)1 immune responses.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, Duke University Medical Center, Durham, North Carolina, USA.

ABSTRACT
T helper type 1 (T(H)1)-polarized immune responses, which confer protection against intracellular pathogens, are thought to be initiated by dendritic cells (DCs) that enter lymph nodes from peripheral tissues. Here we found after viral infection or immunization, inflammatory monocytes were recruited into lymph nodes directly from the blood to become CD11c(+)CD11b(hi)Gr-1(+) inflammatory DCs, which produced abundant interleukin 12p70 and potently stimulated T(H)1 responses. This monocyte extravasation required the chemokine receptor CCR2 but not the chemokine CCL2 or receptor CCR7. Thus, the accumulation of inflammatory DCs and T(H)1 responses were much lower in Ccr2(-/-) mice, were preserved in Ccl2(-/-) mice and were relatively higher in CCL19-CCL21-Ser-deficient plt mutant mice, in which all other lymph node DC types were fewer in number. We conclude that blood-derived inflammatory DCs are important in the development of T(H)1 immune responses.

Show MeSH
Related in: MedlinePlus