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Identification of human NK17/NK1 cells.

Pandya AD, Al-Jaderi Z, Høglund RA, Holmøy T, Harbo HF, Norgauer J, Maghazachi AA - PLoS ONE (2011)

Bottom Line: They also express CD161, NKp30, NKp44, NKp46, NKG2D, CD158, CCL22, IL-2Rβ and the common γ chain but not CD127 or IL-23R.Antibodies to IL-1β, IL-6, IL-21, or TGF-β1 do not inhibit IL-2-induced generation of NK17/NK1 cells, suggesting that IL-2 has the capacity to polarize these cells.NK17/NK1 cells identified here have not been previously described in healthy or MS patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, Faculty of Medicine, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway.

ABSTRACT

Background: Natural killer (NK) cells have both cytolytic and immunoregulatory functions. We recently described that these cells release the inflammatory cytokines IL-17 and IFN-γ. However, the precise identity of the NK cell subset(s) that secrete these cytokines is not known.

Methodology/principal findings: To isolate the cells secreting IL-17 and IFN-γ, we took advantage of the findings that Th17/Th1 cells express chemokine receptors. Therefore, CD56(+)NK cells were stained with antibodies against various chemokine receptors and intracellularly with antibodies toward IL-17 and IFN-γ. Consequently, we identified previously unrecognized subset of NK cells generated from normal human peripheral blood after activation with IL-2 but not PMA plus ionomycin. The cells are characterized by the expression of CD56(+) and CCR4(+), produce IL-17 and IFN-γ and are consequently named NK17/NK1 cells. They also express CD161, NKp30, NKp44, NKp46, NKG2D, CD158, CCL22, IL-2Rβ and the common γ chain but not CD127 or IL-23R. Further, they possess T-bet and RORγt transcription factors. Antibodies to IL-1β, IL-6, IL-21, or TGF-β1 do not inhibit IL-2-induced generation of NK17/NK1 cells, suggesting that IL-2 has the capacity to polarize these cells. Notably, NK17/NK1 cells are abundant in the cerebrospinal fluid (CSF) of patients with multiple sclerosis (MS) without activation, and are generated from the peripheral blood of these patients after activation with IL-2.

Conclusions/significance: NK17/NK1 cells identified here have not been previously described in healthy or MS patients.

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Related in: MedlinePlus

Expression of various markers by NK17/NK1 cells.A) IL-2-activated CD56+ NK cells were sorted by antibody-coated beads into CCR4+ or CCR6+ and then labeled with antibodies to various cell markers. The numbers are generated from 3 donors and are shown as mean±SEM. P value compares the frequency of CCR4+ and CCR6+ positive cells expressing IL-23R. B) CD56+ NK cells were sorted into CCR4+ or CCR4−and then labeled intracellularly with antibodies to RORγt and T-bet. Background using isotype control antibodies are shown in black. Numbers indicate the percentage of positive cells.
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pone-0026780-g004: Expression of various markers by NK17/NK1 cells.A) IL-2-activated CD56+ NK cells were sorted by antibody-coated beads into CCR4+ or CCR6+ and then labeled with antibodies to various cell markers. The numbers are generated from 3 donors and are shown as mean±SEM. P value compares the frequency of CCR4+ and CCR6+ positive cells expressing IL-23R. B) CD56+ NK cells were sorted into CCR4+ or CCR4−and then labeled intracellularly with antibodies to RORγt and T-bet. Background using isotype control antibodies are shown in black. Numbers indicate the percentage of positive cells.

Mentions: To gain insights into the expression of NK cell markers among different NK cell subsets based on their chemokine receptor possession, we double sorted IL-2-activated NK cells, first into CD56+ and then into CCR4+ or CCR6+ cells by antibody-conjugated beads. These subsets were labeled with antibodies to various surface receptors. The results show that both NK cell subsets expressed the mature NK cell molecules NKp30, NKp44, NKp46, NKG2D, CD158 and CD161, but lacked the expression of immature cell marker CD127. The most obvious difference between the two subsets is the expression of IL-23 on the surface of CCR6+ NK cells and not on NK17/NK1 (CD56+CCR4+) cells (P<0.03, Figure 4A). Notably, NK17/NK1 cells expressed the ligand for CCR4, i.e. CCL22/MDC, suggesting that this chemokine may play a role in the maintenance and/or survival of these cells.


Identification of human NK17/NK1 cells.

Pandya AD, Al-Jaderi Z, Høglund RA, Holmøy T, Harbo HF, Norgauer J, Maghazachi AA - PLoS ONE (2011)

Expression of various markers by NK17/NK1 cells.A) IL-2-activated CD56+ NK cells were sorted by antibody-coated beads into CCR4+ or CCR6+ and then labeled with antibodies to various cell markers. The numbers are generated from 3 donors and are shown as mean±SEM. P value compares the frequency of CCR4+ and CCR6+ positive cells expressing IL-23R. B) CD56+ NK cells were sorted into CCR4+ or CCR4−and then labeled intracellularly with antibodies to RORγt and T-bet. Background using isotype control antibodies are shown in black. Numbers indicate the percentage of positive cells.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3198820&req=5

pone-0026780-g004: Expression of various markers by NK17/NK1 cells.A) IL-2-activated CD56+ NK cells were sorted by antibody-coated beads into CCR4+ or CCR6+ and then labeled with antibodies to various cell markers. The numbers are generated from 3 donors and are shown as mean±SEM. P value compares the frequency of CCR4+ and CCR6+ positive cells expressing IL-23R. B) CD56+ NK cells were sorted into CCR4+ or CCR4−and then labeled intracellularly with antibodies to RORγt and T-bet. Background using isotype control antibodies are shown in black. Numbers indicate the percentage of positive cells.
Mentions: To gain insights into the expression of NK cell markers among different NK cell subsets based on their chemokine receptor possession, we double sorted IL-2-activated NK cells, first into CD56+ and then into CCR4+ or CCR6+ cells by antibody-conjugated beads. These subsets were labeled with antibodies to various surface receptors. The results show that both NK cell subsets expressed the mature NK cell molecules NKp30, NKp44, NKp46, NKG2D, CD158 and CD161, but lacked the expression of immature cell marker CD127. The most obvious difference between the two subsets is the expression of IL-23 on the surface of CCR6+ NK cells and not on NK17/NK1 (CD56+CCR4+) cells (P<0.03, Figure 4A). Notably, NK17/NK1 cells expressed the ligand for CCR4, i.e. CCL22/MDC, suggesting that this chemokine may play a role in the maintenance and/or survival of these cells.

Bottom Line: They also express CD161, NKp30, NKp44, NKp46, NKG2D, CD158, CCL22, IL-2Rβ and the common γ chain but not CD127 or IL-23R.Antibodies to IL-1β, IL-6, IL-21, or TGF-β1 do not inhibit IL-2-induced generation of NK17/NK1 cells, suggesting that IL-2 has the capacity to polarize these cells.NK17/NK1 cells identified here have not been previously described in healthy or MS patients.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, Faculty of Medicine, Institute of Basic Medical Sciences, University of Oslo, Oslo, Norway.

ABSTRACT

Background: Natural killer (NK) cells have both cytolytic and immunoregulatory functions. We recently described that these cells release the inflammatory cytokines IL-17 and IFN-γ. However, the precise identity of the NK cell subset(s) that secrete these cytokines is not known.

Methodology/principal findings: To isolate the cells secreting IL-17 and IFN-γ, we took advantage of the findings that Th17/Th1 cells express chemokine receptors. Therefore, CD56(+)NK cells were stained with antibodies against various chemokine receptors and intracellularly with antibodies toward IL-17 and IFN-γ. Consequently, we identified previously unrecognized subset of NK cells generated from normal human peripheral blood after activation with IL-2 but not PMA plus ionomycin. The cells are characterized by the expression of CD56(+) and CCR4(+), produce IL-17 and IFN-γ and are consequently named NK17/NK1 cells. They also express CD161, NKp30, NKp44, NKp46, NKG2D, CD158, CCL22, IL-2Rβ and the common γ chain but not CD127 or IL-23R. Further, they possess T-bet and RORγt transcription factors. Antibodies to IL-1β, IL-6, IL-21, or TGF-β1 do not inhibit IL-2-induced generation of NK17/NK1 cells, suggesting that IL-2 has the capacity to polarize these cells. Notably, NK17/NK1 cells are abundant in the cerebrospinal fluid (CSF) of patients with multiple sclerosis (MS) without activation, and are generated from the peripheral blood of these patients after activation with IL-2.

Conclusions/significance: NK17/NK1 cells identified here have not been previously described in healthy or MS patients.

Show MeSH
Related in: MedlinePlus