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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, IL-12 or IL-15 plus IL-12 induced NK cells from PFCs and PBMCs to expresse and produce IL-22 and IFN-γ.PFCs and PBMCs were stimulated with or without IL-15, IL-12 or IL-15 plus IL-12 for 24 h. The expression of IL-22 (A) and IFN-γ (B) from CD3-CD56+ NK cells was detected by FCM. Representative dot plots of five independent experiments were shown with similar results. Purified NK cells from PFCs and PBMCs were cultured with or without IL-15, IL-12 or IL-15 puls IL-12 for 24 h. The concentration of IL-22 (C) and IFN-γ (D) was detected by ELISA. Statistical results were shown as mean±SD in histogram and error bars represent triplicates within the similar experiment. *P<0.05.
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pone.0151721.g003: IL-15, IL-12 or IL-15 plus IL-12 induced NK cells from PFCs and PBMCs to expresse and produce IL-22 and IFN-γ.PFCs and PBMCs were stimulated with or without IL-15, IL-12 or IL-15 plus IL-12 for 24 h. The expression of IL-22 (A) and IFN-γ (B) from CD3-CD56+ NK cells was detected by FCM. Representative dot plots of five independent experiments were shown with similar results. Purified NK cells from PFCs and PBMCs were cultured with or without IL-15, IL-12 or IL-15 puls IL-12 for 24 h. The concentration of IL-22 (C) and IFN-γ (D) was detected by ELISA. Statistical results were shown as mean±SD in histogram and error bars represent triplicates within the similar experiment. *P<0.05.

Mentions: Our previous study illustrated that NK cells from pleural fluid cells (PFCs) displayed an optimal role on the production of IFN-γ. To evaluate whether NK cells from PFCs induced the expression and production of IL-22 in response to IL-15 or IL-12. PFCs and PBMCs were stimulated with or without IL-15, IL-12 or IL-15 plus IL-12 for 24 h and the cells were harvested for the cytokine evaluation by intracellular cytokine stainings. As illustrated in Fig 3A and 3B, NK cells were gated on CD3-CD56+ cells and the expression of IL-22 and IFN-γ was analyzed. The results showed that NK cells from PFC expressed significantly higher levels of IL-22 compared with NK cells from PBMCs (0.15%±0.06% vs 0.06%±0.009%) in response to IL-15 but not IL-12. More importantly, IL-12 enhancd the expression of IL-22 under the stimulation with IL-15 (0.35%±0.007% vs 0.19%±0.003%). Similarly, NK cells from PFCs produced larger amount of IFN-γ than did NK cells from PBMCs in response to IL-12 (18.7%±2.3% vs 14.6%±2.5%). To confirm our results, NK cells were sorted by flow cytometry from PFCs and PBMCs, and sorted NK cells were cultured with or without IL-15, IL-12 or IL-15 plus IL-12. As illustrated in Fig 3C and 3D, sorted NK cells from PFCs produced higher levels of IL-22 and IFN-γ in response to IL-15 or IL-15 plus IL-12 compared to NK cells form PBMCs.


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, IL-12 or IL-15 plus IL-12 induced NK cells from PFCs and PBMCs to expresse and produce IL-22 and IFN-γ.PFCs and PBMCs were stimulated with or without IL-15, IL-12 or IL-15 plus IL-12 for 24 h. The expression of IL-22 (A) and IFN-γ (B) from CD3-CD56+ NK cells was detected by FCM. Representative dot plots of five independent experiments were shown with similar results. Purified NK cells from PFCs and PBMCs were cultured with or without IL-15, IL-12 or IL-15 puls IL-12 for 24 h. The concentration of IL-22 (C) and IFN-γ (D) was detected by ELISA. Statistical results were shown as mean±SD in histogram and error bars represent triplicates within the similar experiment. *P<0.05.
© Copyright Policy
Related In: Results  -  Collection

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

pone.0151721.g003: IL-15, IL-12 or IL-15 plus IL-12 induced NK cells from PFCs and PBMCs to expresse and produce IL-22 and IFN-γ.PFCs and PBMCs were stimulated with or without IL-15, IL-12 or IL-15 plus IL-12 for 24 h. The expression of IL-22 (A) and IFN-γ (B) from CD3-CD56+ NK cells was detected by FCM. Representative dot plots of five independent experiments were shown with similar results. Purified NK cells from PFCs and PBMCs were cultured with or without IL-15, IL-12 or IL-15 puls IL-12 for 24 h. The concentration of IL-22 (C) and IFN-γ (D) was detected by ELISA. Statistical results were shown as mean±SD in histogram and error bars represent triplicates within the similar experiment. *P<0.05.
Mentions: Our previous study illustrated that NK cells from pleural fluid cells (PFCs) displayed an optimal role on the production of IFN-γ. To evaluate whether NK cells from PFCs induced the expression and production of IL-22 in response to IL-15 or IL-12. PFCs and PBMCs were stimulated with or without IL-15, IL-12 or IL-15 plus IL-12 for 24 h and the cells were harvested for the cytokine evaluation by intracellular cytokine stainings. As illustrated in Fig 3A and 3B, NK cells were gated on CD3-CD56+ cells and the expression of IL-22 and IFN-γ was analyzed. The results showed that NK cells from PFC expressed significantly higher levels of IL-22 compared with NK cells from PBMCs (0.15%±0.06% vs 0.06%±0.009%) in response to IL-15 but not IL-12. More importantly, IL-12 enhancd the expression of IL-22 under the stimulation with IL-15 (0.35%±0.007% vs 0.19%±0.003%). Similarly, NK cells from PFCs produced larger amount of IFN-γ than did NK cells from PBMCs in response to IL-12 (18.7%±2.3% vs 14.6%±2.5%). To confirm our results, NK cells were sorted by flow cytometry from PFCs and PBMCs, and sorted NK cells were cultured with or without IL-15, IL-12 or IL-15 plus IL-12. As illustrated in Fig 3C and 3D, sorted NK cells from PFCs produced higher levels of IL-22 and IFN-γ in response to IL-15 or IL-15 plus IL-12 compared to NK cells form PBMCs.

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