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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 and BCG enhanced the expression of NKG2D and granzyme B by IL-22+ NK cells from PFCs.(A) PFCs were incubated with or without IL-15, IL-12 or IL-15 plus IL-12 and the cells were collected for the detection of NKG2D expression on CD3-CD56+ NK cells from PFCs by FCM. The representative histogram of NKG2D expression (open histogram) and isotype control (filled histogram) on CD3-CD56+ NK cells. (B) Statistical results were shown for NKG2D percentage and MFI on NK cells (n = 6). (C) PFCs were stimulated with BCG for 8 hours in the presence of BFA and the cells were harvested for the detection and characterizytion of IL-22 on NK cells by FCM. The cells were gated on CD3-CD56+IL-22- (left side) and CD3-CD56+IL-22+ (right side) cells. The representative histogram of the phenotypes of NK cells were shown from three separate experiments with similar results. (D) Mean values of the results were shown as Mean±SD). Student’s t-test was used for statistical analysis. *P<0.05, **P<0.01 and ***P<0.001.
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pone.0151721.g006: IL-15, IL-12 and BCG enhanced the expression of NKG2D and granzyme B by IL-22+ NK cells from PFCs.(A) PFCs were incubated with or without IL-15, IL-12 or IL-15 plus IL-12 and the cells were collected for the detection of NKG2D expression on CD3-CD56+ NK cells from PFCs by FCM. The representative histogram of NKG2D expression (open histogram) and isotype control (filled histogram) on CD3-CD56+ NK cells. (B) Statistical results were shown for NKG2D percentage and MFI on NK cells (n = 6). (C) PFCs were stimulated with BCG for 8 hours in the presence of BFA and the cells were harvested for the detection and characterizytion of IL-22 on NK cells by FCM. The cells were gated on CD3-CD56+IL-22- (left side) and CD3-CD56+IL-22+ (right side) cells. The representative histogram of the phenotypes of NK cells were shown from three separate experiments with similar results. (D) Mean values of the results were shown as Mean±SD). Student’s t-test was used for statistical analysis. *P<0.05, **P<0.01 and ***P<0.001.

Mentions: To investigate the possible mechanism of the response by CD45RO+ NK cells from PFCs under the stimulation with BCG or cytokines, we evaluated the expression of NK cell activation molecule-NKG2D. The results in Fig 6A and 6B demonstrated that IL-15, IL-12 or IL-15 plus IL-12 stimulation significantly enhanced the expression of NKG2D on NK cells compared with medium. Consistently, the MFI of NKG2D on NK cells was significantly increased in response to IL-15, IL-12 or IL-15 plus IL-12 compared with medium (Fig 6B). To further evaluate the phenotype of BCG-specific NK cells from PFCs on the basis of their ability to produce IL-22, we analyzed the expression of CD45RO as a marker for memory, NKG2D as NK cell activation molecule, granzyme B as a cytotoxic-related marker, CD69 and CD25 as activation molecules on IL-22-producing NK cells from PFCs after stimulation with BCG. As shown in Fig 6C, very low level of CD45RO was observed on IL-22- NK cells but approximately 50% IL-22+ NK cells expressed CD45RO. Consistently, statistical result demonstrated that significantly enhanced expression of NKG2D, CD69, CD25 and granzyme B was detected on IL-22+ NK cells compared with IL-22- NK cells (Fig 6D).


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 and BCG enhanced the expression of NKG2D and granzyme B by IL-22+ NK cells from PFCs.(A) PFCs were incubated with or without IL-15, IL-12 or IL-15 plus IL-12 and the cells were collected for the detection of NKG2D expression on CD3-CD56+ NK cells from PFCs by FCM. The representative histogram of NKG2D expression (open histogram) and isotype control (filled histogram) on CD3-CD56+ NK cells. (B) Statistical results were shown for NKG2D percentage and MFI on NK cells (n = 6). (C) PFCs were stimulated with BCG for 8 hours in the presence of BFA and the cells were harvested for the detection and characterizytion of IL-22 on NK cells by FCM. The cells were gated on CD3-CD56+IL-22- (left side) and CD3-CD56+IL-22+ (right side) cells. The representative histogram of the phenotypes of NK cells were shown from three separate experiments with similar results. (D) Mean values of the results were shown as Mean±SD). Student’s t-test was used for statistical analysis. *P<0.05, **P<0.01 and ***P<0.001.
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pone.0151721.g006: IL-15, IL-12 and BCG enhanced the expression of NKG2D and granzyme B by IL-22+ NK cells from PFCs.(A) PFCs were incubated with or without IL-15, IL-12 or IL-15 plus IL-12 and the cells were collected for the detection of NKG2D expression on CD3-CD56+ NK cells from PFCs by FCM. The representative histogram of NKG2D expression (open histogram) and isotype control (filled histogram) on CD3-CD56+ NK cells. (B) Statistical results were shown for NKG2D percentage and MFI on NK cells (n = 6). (C) PFCs were stimulated with BCG for 8 hours in the presence of BFA and the cells were harvested for the detection and characterizytion of IL-22 on NK cells by FCM. The cells were gated on CD3-CD56+IL-22- (left side) and CD3-CD56+IL-22+ (right side) cells. The representative histogram of the phenotypes of NK cells were shown from three separate experiments with similar results. (D) Mean values of the results were shown as Mean±SD). Student’s t-test was used for statistical analysis. *P<0.05, **P<0.01 and ***P<0.001.
Mentions: To investigate the possible mechanism of the response by CD45RO+ NK cells from PFCs under the stimulation with BCG or cytokines, we evaluated the expression of NK cell activation molecule-NKG2D. The results in Fig 6A and 6B demonstrated that IL-15, IL-12 or IL-15 plus IL-12 stimulation significantly enhanced the expression of NKG2D on NK cells compared with medium. Consistently, the MFI of NKG2D on NK cells was significantly increased in response to IL-15, IL-12 or IL-15 plus IL-12 compared with medium (Fig 6B). To further evaluate the phenotype of BCG-specific NK cells from PFCs on the basis of their ability to produce IL-22, we analyzed the expression of CD45RO as a marker for memory, NKG2D as NK cell activation molecule, granzyme B as a cytotoxic-related marker, CD69 and CD25 as activation molecules on IL-22-producing NK cells from PFCs after stimulation with BCG. As shown in Fig 6C, very low level of CD45RO was observed on IL-22- NK cells but approximately 50% IL-22+ NK cells expressed CD45RO. Consistently, statistical result demonstrated that significantly enhanced expression of NKG2D, CD69, CD25 and granzyme B was detected on IL-22+ NK cells compared with IL-22- NK cells (Fig 6D).

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