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Dysregulated cytokine expression by CD4+ T cells from post-septic mice modulates both Th1 and Th2-mediated granulomatous lung inflammation.

Carson WF, Ito T, Schaller M, Cavassani KA, Chensue SW, Kunkel SL - PLoS ONE (2011)

Bottom Line: Previous epidemiological studies in humans and experimental studies in animals indicate that survivors of severe sepsis exhibit deficiencies in the activation and effector function of immune cells.These include increased production of T(H)2 cytokines in TH1 inflammation, and increased production of T(H)1 cytokines in TH2 inflammation.These results suggest that cell-intrinsic defects in CD4+ T cell effector function can have deleterious effects on inflammatory processes post-sepsis, due to a defect in the proper regulation of TH-specific cytokine expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, University of Michigan Medical School, University of Michigan, Ann Arbor, Michigan, United States of America. wfcarson@umich.edu

ABSTRACT
Previous epidemiological studies in humans and experimental studies in animals indicate that survivors of severe sepsis exhibit deficiencies in the activation and effector function of immune cells. In particular, CD4+ T lymphocytes can exhibit reduced proliferative capacity and improper cytokine responses following sepsis. To further investigate the cell-intrinsic defects of CD4+ T cells following sepsis, splenic CD4+ T cells from sham surgery and post-septic mice were transferred into lymphopenic mice. These recipient mice were then subjected to both TH1-(purified protein derivative) and TH2-(Schistosoma mansoni egg antigen) driven models of granulomatous lung inflammation. Post-septic CD4+ T cells mediated smaller TH1 and larger TH2 lung granulomas as compared to mice receiving CD4+ T cells from sham surgery donors. However, cytokine production by lymph node cells in antigen restimulation assays indicated increased pan-specific cytokine expression by post-septic CD4+ T cell recipient mice in both TH1 and TH2 granuloma models. These include increased production of T(H)2 cytokines in TH1 inflammation, and increased production of T(H)1 cytokines in TH2 inflammation. These results suggest that cell-intrinsic defects in CD4+ T cell effector function can have deleterious effects on inflammatory processes post-sepsis, due to a defect in the proper regulation of TH-specific cytokine expression.

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Cytokine expression in SEA-restimulated lymph node cultures from sham and CLP RAG mice.Single-cell suspensions of lymph nodes from S. mansoni-sensitized sham and CLP RAG mice four days following SEA-bead challenge were restimulated with soluble SEA in cell culture for 48 hours, and cytokine expression in cell culture supernatants was analyzed via multiplex bead assay (Luminex). Data presented is representative of two separate experiments, with triplicate wells of samples from sham and CLP RAG mice, n = 5 mice per group. The limit of detection for each cytokine was routinely <5 pg/ml. (*) = p<0.05 vs. sham RAG, SEA-stimulated.
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pone-0020385-g006: Cytokine expression in SEA-restimulated lymph node cultures from sham and CLP RAG mice.Single-cell suspensions of lymph nodes from S. mansoni-sensitized sham and CLP RAG mice four days following SEA-bead challenge were restimulated with soluble SEA in cell culture for 48 hours, and cytokine expression in cell culture supernatants was analyzed via multiplex bead assay (Luminex). Data presented is representative of two separate experiments, with triplicate wells of samples from sham and CLP RAG mice, n = 5 mice per group. The limit of detection for each cytokine was routinely <5 pg/ml. (*) = p<0.05 vs. sham RAG, SEA-stimulated.

Mentions: For analysis of antigen-dependent cytokine production by lymph node-resident lymphocytes, single cell suspensions of mediastynal lymph nodes from sham and CLP RAG mice were restimulated with SEA and supernatants were analyzed for the presence of various proinflammatory cytokines. In response to SEA stimulation, CLP RAG lymph node cultures produced significantly decreased levels of the CD4+ T cell proliferative factor IL-2 as compared to sham RAG cultures (Fig. 6A). In accordance with the increased size of SEA-bead granulomas in CLP RAG mice, TH2 cytokine production by SEA-restimulated lymph node cultures from CLP RAG mice was increased as compared to sham RAG lymph node cultures (Fig. 3). CLP RAG lymph node restimulation assays exhibited SEA-specific increases in the TH2 cytokines IL-4 (Fig. 6B), IL-5 (Fig. 6C) and IL-13 (Fig. 6E), along with the TH2/immunosuppressive cytokine IL-10 (Fig. 6D). Surprisingly, CLP RAG lymph nodes also exhibited significantly elevated levels of the TH17 cytokine IL-17 (Fig. 6F), as well as levels of the TH1 cytokine IFN-γ (Fig. 6G). While sham RAG lymph node cultures also exhibited SEA-specific production of both IL-17 and IFN-γ, the levels of these cytokines were below the levels observed for TH2 cytokines from the same cultures. In a similar fashion as the RAG PPD lymph nodes, flow cytometric analysis indicated no significant difference in the percentage (Figures S2C and S2D) or total number (2.20×104±1.1 for sham LN, 1.55×104±0.9 for CLP LN, p>0:05) of CD3+ CD4+ T cells present in either sham or CLP RAG SEA lymph nodes, suggesting that the modulations in cytokine production by these lymph node cultures was not due to modulations in CD4+ T cells.


Dysregulated cytokine expression by CD4+ T cells from post-septic mice modulates both Th1 and Th2-mediated granulomatous lung inflammation.

Carson WF, Ito T, Schaller M, Cavassani KA, Chensue SW, Kunkel SL - PLoS ONE (2011)

Cytokine expression in SEA-restimulated lymph node cultures from sham and CLP RAG mice.Single-cell suspensions of lymph nodes from S. mansoni-sensitized sham and CLP RAG mice four days following SEA-bead challenge were restimulated with soluble SEA in cell culture for 48 hours, and cytokine expression in cell culture supernatants was analyzed via multiplex bead assay (Luminex). Data presented is representative of two separate experiments, with triplicate wells of samples from sham and CLP RAG mice, n = 5 mice per group. The limit of detection for each cytokine was routinely <5 pg/ml. (*) = p<0.05 vs. sham RAG, SEA-stimulated.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3105020&req=5

pone-0020385-g006: Cytokine expression in SEA-restimulated lymph node cultures from sham and CLP RAG mice.Single-cell suspensions of lymph nodes from S. mansoni-sensitized sham and CLP RAG mice four days following SEA-bead challenge were restimulated with soluble SEA in cell culture for 48 hours, and cytokine expression in cell culture supernatants was analyzed via multiplex bead assay (Luminex). Data presented is representative of two separate experiments, with triplicate wells of samples from sham and CLP RAG mice, n = 5 mice per group. The limit of detection for each cytokine was routinely <5 pg/ml. (*) = p<0.05 vs. sham RAG, SEA-stimulated.
Mentions: For analysis of antigen-dependent cytokine production by lymph node-resident lymphocytes, single cell suspensions of mediastynal lymph nodes from sham and CLP RAG mice were restimulated with SEA and supernatants were analyzed for the presence of various proinflammatory cytokines. In response to SEA stimulation, CLP RAG lymph node cultures produced significantly decreased levels of the CD4+ T cell proliferative factor IL-2 as compared to sham RAG cultures (Fig. 6A). In accordance with the increased size of SEA-bead granulomas in CLP RAG mice, TH2 cytokine production by SEA-restimulated lymph node cultures from CLP RAG mice was increased as compared to sham RAG lymph node cultures (Fig. 3). CLP RAG lymph node restimulation assays exhibited SEA-specific increases in the TH2 cytokines IL-4 (Fig. 6B), IL-5 (Fig. 6C) and IL-13 (Fig. 6E), along with the TH2/immunosuppressive cytokine IL-10 (Fig. 6D). Surprisingly, CLP RAG lymph nodes also exhibited significantly elevated levels of the TH17 cytokine IL-17 (Fig. 6F), as well as levels of the TH1 cytokine IFN-γ (Fig. 6G). While sham RAG lymph node cultures also exhibited SEA-specific production of both IL-17 and IFN-γ, the levels of these cytokines were below the levels observed for TH2 cytokines from the same cultures. In a similar fashion as the RAG PPD lymph nodes, flow cytometric analysis indicated no significant difference in the percentage (Figures S2C and S2D) or total number (2.20×104±1.1 for sham LN, 1.55×104±0.9 for CLP LN, p>0:05) of CD3+ CD4+ T cells present in either sham or CLP RAG SEA lymph nodes, suggesting that the modulations in cytokine production by these lymph node cultures was not due to modulations in CD4+ T cells.

Bottom Line: Previous epidemiological studies in humans and experimental studies in animals indicate that survivors of severe sepsis exhibit deficiencies in the activation and effector function of immune cells.These include increased production of T(H)2 cytokines in TH1 inflammation, and increased production of T(H)1 cytokines in TH2 inflammation.These results suggest that cell-intrinsic defects in CD4+ T cell effector function can have deleterious effects on inflammatory processes post-sepsis, due to a defect in the proper regulation of TH-specific cytokine expression.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology, University of Michigan Medical School, University of Michigan, Ann Arbor, Michigan, United States of America. wfcarson@umich.edu

ABSTRACT
Previous epidemiological studies in humans and experimental studies in animals indicate that survivors of severe sepsis exhibit deficiencies in the activation and effector function of immune cells. In particular, CD4+ T lymphocytes can exhibit reduced proliferative capacity and improper cytokine responses following sepsis. To further investigate the cell-intrinsic defects of CD4+ T cells following sepsis, splenic CD4+ T cells from sham surgery and post-septic mice were transferred into lymphopenic mice. These recipient mice were then subjected to both TH1-(purified protein derivative) and TH2-(Schistosoma mansoni egg antigen) driven models of granulomatous lung inflammation. Post-septic CD4+ T cells mediated smaller TH1 and larger TH2 lung granulomas as compared to mice receiving CD4+ T cells from sham surgery donors. However, cytokine production by lymph node cells in antigen restimulation assays indicated increased pan-specific cytokine expression by post-septic CD4+ T cell recipient mice in both TH1 and TH2 granuloma models. These include increased production of T(H)2 cytokines in TH1 inflammation, and increased production of T(H)1 cytokines in TH2 inflammation. These results suggest that cell-intrinsic defects in CD4+ T cell effector function can have deleterious effects on inflammatory processes post-sepsis, due to a defect in the proper regulation of TH-specific cytokine expression.

Show MeSH
Related in: MedlinePlus