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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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Related in: MedlinePlus

BCG and M.tb-related Ags induced more production of IL-22 and IFN-γ but not IL-17 by NK cells from PFCs but not PBMCs.(A) PFCs and PBMCs were stimulated with or without BCG. The expression of IL-22 and IL-17 by CD3-CD56+ NK cells was evaluated by FCM. Representative dot plots of five independent experiments were shown with similar results. (B) Statistical results of IL-22 and IL-17 expression by by CD3-CD56+ NK cells from PFCs and PBMCs. The median and individual frequencies for each patient and healthy volunteer were performed. (C) Analysis for the relationship of IL-22, IFN-γ and IL-17 expression by FCM. The dot plots were representative of four separate experiments with similar results. (D) PFCs were incubated with BCG and M.tb-related Ags for 8 h in the presence of BFA. The expression of IL-22 and IFN-γ by CD3-CD56+ cells were evaluated by FCM. The dot plots were representative of four separate experiments with similar results. (E) The statistical histograms were shown as mean±SD and error bars represent triplicates within the same experiment. (F) PFCs were incubated with BCG and M.tb-related Ags for 8 hours. The cells were harvested and stained for the expression of IL-22 and IFN-γ by FCM. The representative dot plots were shown from three separate experiments with similar results. (G) Statistical results of IL-22 and IFN-γ expression on NK cells from PFCs. Data were expressed in box plots as medians, minimum and maximum values and statistical significance was determined with the Student’s t-test, *P<0.05 or **P<0.01.
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pone.0151721.g004: BCG and M.tb-related Ags induced more production of IL-22 and IFN-γ but not IL-17 by NK cells from PFCs but not PBMCs.(A) PFCs and PBMCs were stimulated with or without BCG. The expression of IL-22 and IL-17 by CD3-CD56+ NK cells was evaluated by FCM. Representative dot plots of five independent experiments were shown with similar results. (B) Statistical results of IL-22 and IL-17 expression by by CD3-CD56+ NK cells from PFCs and PBMCs. The median and individual frequencies for each patient and healthy volunteer were performed. (C) Analysis for the relationship of IL-22, IFN-γ and IL-17 expression by FCM. The dot plots were representative of four separate experiments with similar results. (D) PFCs were incubated with BCG and M.tb-related Ags for 8 h in the presence of BFA. The expression of IL-22 and IFN-γ by CD3-CD56+ cells were evaluated by FCM. The dot plots were representative of four separate experiments with similar results. (E) The statistical histograms were shown as mean±SD and error bars represent triplicates within the same experiment. (F) PFCs were incubated with BCG and M.tb-related Ags for 8 hours. The cells were harvested and stained for the expression of IL-22 and IFN-γ by FCM. The representative dot plots were shown from three separate experiments with similar results. (G) Statistical results of IL-22 and IFN-γ expression on NK cells from PFCs. Data were expressed in box plots as medians, minimum and maximum values and statistical significance was determined with the Student’s t-test, *P<0.05 or **P<0.01.

Mentions: Our previous study indicated that NK cells from PFCs produced significantly higher levels of IFN-γ under the stimulation with BCG compared to PBMCs. Therefore, we further characterized the production of other cytokines both by PFCs and PBMCs in response to BCG. The results showed that NK cells from PFCs but not from PBMCs expressed IL-22 in response to BCG. However, IL-17 was not produced by NK cells from PFCs or PBMCs under the stimulation with BCG (Fig 4A). Statistical results demonstrated that significantly higher level of IL-22 was expressed by NK cells from PFCs compared to NK cells from PBMCs (0.17%±0.06% vs 0.05%±0.009%, P<0.01) (Fig 4B). In addition, NK cells from PFCs could be divided into three distinct populations (Fig 4C). Further evaluation of cytokine expression showed that IL-22-producing NK cells did not co-expresse IFN-γ and IL-17 by NK cells from PFCs in response to BCG. These results demonstrated that a subset of the specific IL-22-producing NK cells was distinct from IFN-γ-producing NK cells in PFCs. Furthermore, we analyzed IL-22 and IFN-γ producing NK cells from PFCs in response to various M. tb-related antigens, including BCG, M. tb-HAg, M. tb -SoAg and M. tb -ScAg. As showed in the Fig 4D and 4E, addition of BCG, M. tb -HAg, M. tb -SoAg or M. tb-ScAg to cell cultures markedly induced the expression of IL-22 and IFN-γ by NK cells from PFCs. The statistical results indicated that significantly higher levels of IL-22 expression by NK cells from PFCs were induced with different TB antigens compared with medium (Fig 4E). Moreover, IL-12 and IL-15 significantly enhanced the expression of IL-22 and IFN-γ by NK cells from PFCs induced by TB antigens (Fig 4F). Most cells expressed either IL-22 or IFN-γ alone, and very few cells expressed both IL-22 and IFN-γ. The statistical results of three subpopulations of cytokine expression from five independent experiments were showed (Fig 4G).


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)

BCG and M.tb-related Ags induced more production of IL-22 and IFN-γ but not IL-17 by NK cells from PFCs but not PBMCs.(A) PFCs and PBMCs were stimulated with or without BCG. The expression of IL-22 and IL-17 by CD3-CD56+ NK cells was evaluated by FCM. Representative dot plots of five independent experiments were shown with similar results. (B) Statistical results of IL-22 and IL-17 expression by by CD3-CD56+ NK cells from PFCs and PBMCs. The median and individual frequencies for each patient and healthy volunteer were performed. (C) Analysis for the relationship of IL-22, IFN-γ and IL-17 expression by FCM. The dot plots were representative of four separate experiments with similar results. (D) PFCs were incubated with BCG and M.tb-related Ags for 8 h in the presence of BFA. The expression of IL-22 and IFN-γ by CD3-CD56+ cells were evaluated by FCM. The dot plots were representative of four separate experiments with similar results. (E) The statistical histograms were shown as mean±SD and error bars represent triplicates within the same experiment. (F) PFCs were incubated with BCG and M.tb-related Ags for 8 hours. The cells were harvested and stained for the expression of IL-22 and IFN-γ by FCM. The representative dot plots were shown from three separate experiments with similar results. (G) Statistical results of IL-22 and IFN-γ expression on NK cells from PFCs. Data were expressed in box plots as medians, minimum and maximum values and statistical significance was determined with the Student’s t-test, *P<0.05 or **P<0.01.
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Related In: Results  -  Collection

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pone.0151721.g004: BCG and M.tb-related Ags induced more production of IL-22 and IFN-γ but not IL-17 by NK cells from PFCs but not PBMCs.(A) PFCs and PBMCs were stimulated with or without BCG. The expression of IL-22 and IL-17 by CD3-CD56+ NK cells was evaluated by FCM. Representative dot plots of five independent experiments were shown with similar results. (B) Statistical results of IL-22 and IL-17 expression by by CD3-CD56+ NK cells from PFCs and PBMCs. The median and individual frequencies for each patient and healthy volunteer were performed. (C) Analysis for the relationship of IL-22, IFN-γ and IL-17 expression by FCM. The dot plots were representative of four separate experiments with similar results. (D) PFCs were incubated with BCG and M.tb-related Ags for 8 h in the presence of BFA. The expression of IL-22 and IFN-γ by CD3-CD56+ cells were evaluated by FCM. The dot plots were representative of four separate experiments with similar results. (E) The statistical histograms were shown as mean±SD and error bars represent triplicates within the same experiment. (F) PFCs were incubated with BCG and M.tb-related Ags for 8 hours. The cells were harvested and stained for the expression of IL-22 and IFN-γ by FCM. The representative dot plots were shown from three separate experiments with similar results. (G) Statistical results of IL-22 and IFN-γ expression on NK cells from PFCs. Data were expressed in box plots as medians, minimum and maximum values and statistical significance was determined with the Student’s t-test, *P<0.05 or **P<0.01.
Mentions: Our previous study indicated that NK cells from PFCs produced significantly higher levels of IFN-γ under the stimulation with BCG compared to PBMCs. Therefore, we further characterized the production of other cytokines both by PFCs and PBMCs in response to BCG. The results showed that NK cells from PFCs but not from PBMCs expressed IL-22 in response to BCG. However, IL-17 was not produced by NK cells from PFCs or PBMCs under the stimulation with BCG (Fig 4A). Statistical results demonstrated that significantly higher level of IL-22 was expressed by NK cells from PFCs compared to NK cells from PBMCs (0.17%±0.06% vs 0.05%±0.009%, P<0.01) (Fig 4B). In addition, NK cells from PFCs could be divided into three distinct populations (Fig 4C). Further evaluation of cytokine expression showed that IL-22-producing NK cells did not co-expresse IFN-γ and IL-17 by NK cells from PFCs in response to BCG. These results demonstrated that a subset of the specific IL-22-producing NK cells was distinct from IFN-γ-producing NK cells in PFCs. Furthermore, we analyzed IL-22 and IFN-γ producing NK cells from PFCs in response to various M. tb-related antigens, including BCG, M. tb-HAg, M. tb -SoAg and M. tb -ScAg. As showed in the Fig 4D and 4E, addition of BCG, M. tb -HAg, M. tb -SoAg or M. tb-ScAg to cell cultures markedly induced the expression of IL-22 and IFN-γ by NK cells from PFCs. The statistical results indicated that significantly higher levels of IL-22 expression by NK cells from PFCs were induced with different TB antigens compared with medium (Fig 4E). Moreover, IL-12 and IL-15 significantly enhanced the expression of IL-22 and IFN-γ by NK cells from PFCs induced by TB antigens (Fig 4F). Most cells expressed either IL-22 or IFN-γ alone, and very few cells expressed both IL-22 and IFN-γ. The statistical results of three subpopulations of cytokine expression from five independent experiments were showed (Fig 4G).

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