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Memory-Like Antigen-Specific Human NK Cells from TB Pleural Fluids Produced IL-22 in Response to IL-15 or Mycobacterium tuberculosis Antigens.

Fu X, Yu S, Yang B, Lao S, Li B, Wu C - PLoS ONE (2016)

Bottom Line: In the present study, we found that cytokines IL-15 induced and IL-12 enhanced the levels of IL-22 by NK cells from TB PFCs.The result demonstrated that CD45RO+ but not CD45RO- NK cells produced significantly higher level of IL-22.Consistently, BCG specific IL-22-producing NK cells from PFCs expressed CD45ROhighNKG2Dhighgranzyme Bhigh.

View Article: PubMed Central - PubMed

Affiliation: Institute of Immunology, Zhongshan School of Medicine, Key Laboratory of Tropical Disease Control Research of Ministry of Education, Sun Yat-sen University, Guangzhou, China.

ABSTRACT
Our previous result indicated that memory-like human natural killer (NK) cells from TB pleural fluid cells (PFCs) produced large amounts of IFN-γ in response to Bacille Calmette Guerin (BCG). Furthermore, recent studies have shown that human lymphoid tissues harbored a unique NK cell subset that specialized in production of interleukin (IL)-22, a proinflammatory cytokine that mediates host defense against pathogens. Yet little information was available with regard to the properties of IL-22 production by memory-like human NK cells. In the present study, we found that cytokines IL-15 induced and IL-12 enhanced the levels of IL-22 by NK cells from TB PFCs. In addition, IL-22 but not IL-17 was produced by NK cells from PFCs in response to BCG and M.tb-related Ags. More importantly, the subset of specific IL-22-producing NK cells were distinct from IFN-γ-producing NK cells in PFCs. CD45RO+ or CD45RO- NK cells were sorted, co-cultured with autologous monocytes and stimulated with BCG for the production of IL-22. The result demonstrated that CD45RO+ but not CD45RO- NK cells produced significantly higher level of IL-22. Anti-IL-12Rβ1 mAbs (2B10) partially inhibit the expression of IL-22 by NK cells under the culture with BCG. Consistently, BCG specific IL-22-producing NK cells from PFCs expressed CD45ROhighNKG2Dhighgranzyme Bhigh. In conclusion, our data demonstrated that memory-like antigen-specific CD45RO+ NK cells might participate in the recall immune response for M. tb infection via producing IL-22, which display a critical role to fight against M. tb.

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IL-15 and IL-12 induced the expression of IL-22 and IFN-γ by different subsets of NK cells from PBMCs.PBMCs were cultured with or without IL-12 or IL-15 or IL-12 plus IL-15 for 24 h in the presence of BFA. The cells were harvested, gated on lymphocytes and then gated on CD3- and CD3+ cells. NK cells from CD3- cells were divided into three subpopulations of CD56highCD16-, CD56lowCD16- and CD56+CD16+ cells (A). The expression of IFN-γ and IL-22 on three subsets of NK cells and CD3+ T cells were analyzed by FCM (B). Dot plots showed representative results with three separate experiments. (C). Statistical results were shown as mean±SD in histogram and error bars represent triplicates within the similar experiment. *P<0.05, **P<0.01 and ***P<0.001.
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pone.0151721.g002: IL-15 and IL-12 induced the expression of IL-22 and IFN-γ by different subsets of NK cells from PBMCs.PBMCs were cultured with or without IL-12 or IL-15 or IL-12 plus IL-15 for 24 h in the presence of BFA. The cells were harvested, gated on lymphocytes and then gated on CD3- and CD3+ cells. NK cells from CD3- cells were divided into three subpopulations of CD56highCD16-, CD56lowCD16- and CD56+CD16+ cells (A). The expression of IFN-γ and IL-22 on three subsets of NK cells and CD3+ T cells were analyzed by FCM (B). Dot plots showed representative results with three separate experiments. (C). Statistical results were shown as mean±SD in histogram and error bars represent triplicates within the similar experiment. *P<0.05, **P<0.01 and ***P<0.001.

Mentions: We further evaluated the subpopulations of IL-22-producing cells. PBMCs were stimulated in vitro with or without IL-15, IL-12 or IL-15 plus IL-12 in the presence of 10μg/mL brefeldin A. After stimulation for 8 h, the cell-surface markers and intracellular cytokines were stained with anti-CD3, anti-CD56, anti-CD16, anti-IL-22, anti-IFN-γ and isotype-matched controls. For flow cytometric analysis, the cells were gated on CD3- and CD3+ cells. NK cells in CD3- cells were divided into three subpopulations of CD56highCD16-, CD56lowCD16- and CD56+CD16+ NK cells, and the expression of IL-22 and IFN-γ were analyzed (Fig 2). Similarly with the previous study, the expression of IFN-γ in response to IL-12 was largely induced by NK cells but not CD3+ T cells. More importantly, the results demonstrated that the expression of IL-22 in response to IL-15 or IL-15 plus IL-12 was primarily by NK cells, especially by the subpopulation of CD56highCD16- NK cells and CD56lowCD16- but not by CD3+ T cells.


Memory-Like Antigen-Specific Human NK Cells from TB Pleural Fluids Produced IL-22 in Response to IL-15 or Mycobacterium tuberculosis Antigens.

Fu X, Yu S, Yang B, Lao S, Li B, Wu C - PLoS ONE (2016)

IL-15 and IL-12 induced the expression of IL-22 and IFN-γ by different subsets of NK cells from PBMCs.PBMCs were cultured with or without IL-12 or IL-15 or IL-12 plus IL-15 for 24 h in the presence of BFA. The cells were harvested, gated on lymphocytes and then gated on CD3- and CD3+ cells. NK cells from CD3- cells were divided into three subpopulations of CD56highCD16-, CD56lowCD16- and CD56+CD16+ cells (A). The expression of IFN-γ and IL-22 on three subsets of NK cells and CD3+ T cells were analyzed by FCM (B). Dot plots showed representative results with three separate experiments. (C). Statistical results were shown as mean±SD in histogram and error bars represent triplicates within the similar experiment. *P<0.05, **P<0.01 and ***P<0.001.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4816314&req=5

pone.0151721.g002: IL-15 and IL-12 induced the expression of IL-22 and IFN-γ by different subsets of NK cells from PBMCs.PBMCs were cultured with or without IL-12 or IL-15 or IL-12 plus IL-15 for 24 h in the presence of BFA. The cells were harvested, gated on lymphocytes and then gated on CD3- and CD3+ cells. NK cells from CD3- cells were divided into three subpopulations of CD56highCD16-, CD56lowCD16- and CD56+CD16+ cells (A). The expression of IFN-γ and IL-22 on three subsets of NK cells and CD3+ T cells were analyzed by FCM (B). Dot plots showed representative results with three separate experiments. (C). Statistical results were shown as mean±SD in histogram and error bars represent triplicates within the similar experiment. *P<0.05, **P<0.01 and ***P<0.001.
Mentions: We further evaluated the subpopulations of IL-22-producing cells. PBMCs were stimulated in vitro with or without IL-15, IL-12 or IL-15 plus IL-12 in the presence of 10μg/mL brefeldin A. After stimulation for 8 h, the cell-surface markers and intracellular cytokines were stained with anti-CD3, anti-CD56, anti-CD16, anti-IL-22, anti-IFN-γ and isotype-matched controls. For flow cytometric analysis, the cells were gated on CD3- and CD3+ cells. NK cells in CD3- cells were divided into three subpopulations of CD56highCD16-, CD56lowCD16- and CD56+CD16+ NK cells, and the expression of IL-22 and IFN-γ were analyzed (Fig 2). Similarly with the previous study, the expression of IFN-γ in response to IL-12 was largely induced by NK cells but not CD3+ T cells. More importantly, the results demonstrated that the expression of IL-22 in response to IL-15 or IL-15 plus IL-12 was primarily by NK cells, especially by the subpopulation of CD56highCD16- NK cells and CD56lowCD16- but not by CD3+ T cells.

Bottom Line: In the present study, we found that cytokines IL-15 induced and IL-12 enhanced the levels of IL-22 by NK cells from TB PFCs.The result demonstrated that CD45RO+ but not CD45RO- NK cells produced significantly higher level of IL-22.Consistently, BCG specific IL-22-producing NK cells from PFCs expressed CD45ROhighNKG2Dhighgranzyme Bhigh.

View Article: PubMed Central - PubMed

Affiliation: Institute of Immunology, Zhongshan School of Medicine, Key Laboratory of Tropical Disease Control Research of Ministry of Education, Sun Yat-sen University, Guangzhou, China.

ABSTRACT
Our previous result indicated that memory-like human natural killer (NK) cells from TB pleural fluid cells (PFCs) produced large amounts of IFN-γ in response to Bacille Calmette Guerin (BCG). Furthermore, recent studies have shown that human lymphoid tissues harbored a unique NK cell subset that specialized in production of interleukin (IL)-22, a proinflammatory cytokine that mediates host defense against pathogens. Yet little information was available with regard to the properties of IL-22 production by memory-like human NK cells. In the present study, we found that cytokines IL-15 induced and IL-12 enhanced the levels of IL-22 by NK cells from TB PFCs. In addition, IL-22 but not IL-17 was produced by NK cells from PFCs in response to BCG and M.tb-related Ags. More importantly, the subset of specific IL-22-producing NK cells were distinct from IFN-γ-producing NK cells in PFCs. CD45RO+ or CD45RO- NK cells were sorted, co-cultured with autologous monocytes and stimulated with BCG for the production of IL-22. The result demonstrated that CD45RO+ but not CD45RO- NK cells produced significantly higher level of IL-22. Anti-IL-12Rβ1 mAbs (2B10) partially inhibit the expression of IL-22 by NK cells under the culture with BCG. Consistently, BCG specific IL-22-producing NK cells from PFCs expressed CD45ROhighNKG2Dhighgranzyme Bhigh. In conclusion, our data demonstrated that memory-like antigen-specific CD45RO+ NK cells might participate in the recall immune response for M. tb infection via producing IL-22, which display a critical role to fight against M. tb.

Show MeSH
Related in: MedlinePlus