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Generation of a novel regulatory NK cell subset from peripheral blood CD34+ progenitors promoted by membrane-bound IL-15.

Giuliani M, Giron-Michel J, Negrini S, Vacca P, Durali D, Caignard A, Le Bousse-Kerdiles C, Chouaib S, Devocelle A, Bahri R, Durrbach A, Taoufik Y, Ferrini S, Croce M, Mingari MC, Moretta L, Azzarone B - PLoS ONE (2008)

Bottom Line: Through these factors, NK-ireg cells act as effectors of the down-regulation of the immune response: reconverting mature myeloid DC (mDC) into immature/tolerogenic DC, blocking cytolytic functions on conventional NK cells and inducing HLA-G membrane expression on PB-derived monocytes.In conclusion, NK-ireg cells represent a novel truly differentiated non-cytolytic NK subset with a self-sustainable phenotype (CD56(+) CD16(+) NKp30(+) NKp44(+) NKp46(+) CD94(+) CD69(+) CCR7(+)) generated from specific pSTAT6(+) GATA3(+) precursors.In addition, NK-ireg cells can be easily derived from peripheral blood of the patients and could constitute an autologous biotherapic tool to be used combined or in alternative to other immuno-regulatory cells.

View Article: PubMed Central - PubMed

Affiliation: INSERM, UMR 542, Université de Paris XI, Hôpital Paul Brousse, Villejuif, France.

ABSTRACT

Background: NK cells have been long time considered as cytotoxic lymphocytes competent in killing virus-infected cells and tumors. However, NK cells may also play essential immuno-regulatory functions. In this context, the real existence of a defined NK subset with negative regulatory properties has been hypothesized but never clearly demonstrated.

Methodology/principal findings: Herein, we show the in vitro generation from human peripheral blood haematopoietic progenitors (PB-HP), of a novel subset of non-cytolytic NK cells displaying a mature phenotype and remarkable immuno-regulatory functions (NK-ireg). The main functional hallmark of these NK-ireg cells is represented by the surface expression/release of HLA-G, a major immunosuppressive molecule. In addition, NK-ireg cells secrete two powerful immuno-regulatory factors: IL-10 and IL-21. Through these factors, NK-ireg cells act as effectors of the down-regulation of the immune response: reconverting mature myeloid DC (mDC) into immature/tolerogenic DC, blocking cytolytic functions on conventional NK cells and inducing HLA-G membrane expression on PB-derived monocytes. The generation of "NK-ireg" cells is obtained, by default, in culture conditions favouring cell-to-cell contacts, and it is strictly dependent on reciprocal trans-presentation of membrane-bound IL-15 forms constitutively and selectively expressed by human CD34(+) PB-HP. Finally, a small subset of NKp46(+) HLA-G(+) IL-10(+) is detected within freshly isolated decidual NK cells, suggesting that these cells could represent an in vivo counterpart of the NK-ireg cells.

Conclusions/significance: In conclusion, NK-ireg cells represent a novel truly differentiated non-cytolytic NK subset with a self-sustainable phenotype (CD56(+) CD16(+) NKp30(+) NKp44(+) NKp46(+) CD94(+) CD69(+) CCR7(+)) generated from specific pSTAT6(+) GATA3(+) precursors. NK-ireg cells could be employed to develop new immuno-suppressive strategies in autoimmune diseases, transplant rejection or graft versus host diseases. In addition, NK-ireg cells can be easily derived from peripheral blood of the patients and could constitute an autologous biotherapic tool to be used combined or in alternative to other immuno-regulatory cells.

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Related in: MedlinePlus

NKPSG cells interfere on the lytic functions of NK cells.Dot plot analysis shows CD107a and IFNγ surface expression in conventional IL-2 activated NK cells triggered (+ anti-NCR) or not (untreated) by NCR cross-linking and after 24 hours pre-incubation with NKPSG cells. In order to distinguish co-cultured NK cells from NKPSG cells, these latter were stained with CFSE and excluded from cytometric analysis. The data are representative of three separate experiments.
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pone-0002241-g007: NKPSG cells interfere on the lytic functions of NK cells.Dot plot analysis shows CD107a and IFNγ surface expression in conventional IL-2 activated NK cells triggered (+ anti-NCR) or not (untreated) by NCR cross-linking and after 24 hours pre-incubation with NKPSG cells. In order to distinguish co-cultured NK cells from NKPSG cells, these latter were stained with CFSE and excluded from cytometric analysis. The data are representative of three separate experiments.

Mentions: Flow cytometric analysis (Fig. 7) showed that about 20% of human NK cells stimulated by the cross-linking of NCR expressed on their surface CD107a within 3 hours, and IFN-γ within 24 hours. When conventional PB-derived NK cells were incubated overnight with NKPSG cells, a complete inhibition of CD107a expression and IFN-γ production could be detected. Remarkably, the cell supernatants obtained from NKPSG cells had similar effect (data not shown).


Generation of a novel regulatory NK cell subset from peripheral blood CD34+ progenitors promoted by membrane-bound IL-15.

Giuliani M, Giron-Michel J, Negrini S, Vacca P, Durali D, Caignard A, Le Bousse-Kerdiles C, Chouaib S, Devocelle A, Bahri R, Durrbach A, Taoufik Y, Ferrini S, Croce M, Mingari MC, Moretta L, Azzarone B - PLoS ONE (2008)

NKPSG cells interfere on the lytic functions of NK cells.Dot plot analysis shows CD107a and IFNγ surface expression in conventional IL-2 activated NK cells triggered (+ anti-NCR) or not (untreated) by NCR cross-linking and after 24 hours pre-incubation with NKPSG cells. In order to distinguish co-cultured NK cells from NKPSG cells, these latter were stained with CFSE and excluded from cytometric analysis. The data are representative of three separate experiments.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2376096&req=5

pone-0002241-g007: NKPSG cells interfere on the lytic functions of NK cells.Dot plot analysis shows CD107a and IFNγ surface expression in conventional IL-2 activated NK cells triggered (+ anti-NCR) or not (untreated) by NCR cross-linking and after 24 hours pre-incubation with NKPSG cells. In order to distinguish co-cultured NK cells from NKPSG cells, these latter were stained with CFSE and excluded from cytometric analysis. The data are representative of three separate experiments.
Mentions: Flow cytometric analysis (Fig. 7) showed that about 20% of human NK cells stimulated by the cross-linking of NCR expressed on their surface CD107a within 3 hours, and IFN-γ within 24 hours. When conventional PB-derived NK cells were incubated overnight with NKPSG cells, a complete inhibition of CD107a expression and IFN-γ production could be detected. Remarkably, the cell supernatants obtained from NKPSG cells had similar effect (data not shown).

Bottom Line: Through these factors, NK-ireg cells act as effectors of the down-regulation of the immune response: reconverting mature myeloid DC (mDC) into immature/tolerogenic DC, blocking cytolytic functions on conventional NK cells and inducing HLA-G membrane expression on PB-derived monocytes.In conclusion, NK-ireg cells represent a novel truly differentiated non-cytolytic NK subset with a self-sustainable phenotype (CD56(+) CD16(+) NKp30(+) NKp44(+) NKp46(+) CD94(+) CD69(+) CCR7(+)) generated from specific pSTAT6(+) GATA3(+) precursors.In addition, NK-ireg cells can be easily derived from peripheral blood of the patients and could constitute an autologous biotherapic tool to be used combined or in alternative to other immuno-regulatory cells.

View Article: PubMed Central - PubMed

Affiliation: INSERM, UMR 542, Université de Paris XI, Hôpital Paul Brousse, Villejuif, France.

ABSTRACT

Background: NK cells have been long time considered as cytotoxic lymphocytes competent in killing virus-infected cells and tumors. However, NK cells may also play essential immuno-regulatory functions. In this context, the real existence of a defined NK subset with negative regulatory properties has been hypothesized but never clearly demonstrated.

Methodology/principal findings: Herein, we show the in vitro generation from human peripheral blood haematopoietic progenitors (PB-HP), of a novel subset of non-cytolytic NK cells displaying a mature phenotype and remarkable immuno-regulatory functions (NK-ireg). The main functional hallmark of these NK-ireg cells is represented by the surface expression/release of HLA-G, a major immunosuppressive molecule. In addition, NK-ireg cells secrete two powerful immuno-regulatory factors: IL-10 and IL-21. Through these factors, NK-ireg cells act as effectors of the down-regulation of the immune response: reconverting mature myeloid DC (mDC) into immature/tolerogenic DC, blocking cytolytic functions on conventional NK cells and inducing HLA-G membrane expression on PB-derived monocytes. The generation of "NK-ireg" cells is obtained, by default, in culture conditions favouring cell-to-cell contacts, and it is strictly dependent on reciprocal trans-presentation of membrane-bound IL-15 forms constitutively and selectively expressed by human CD34(+) PB-HP. Finally, a small subset of NKp46(+) HLA-G(+) IL-10(+) is detected within freshly isolated decidual NK cells, suggesting that these cells could represent an in vivo counterpart of the NK-ireg cells.

Conclusions/significance: In conclusion, NK-ireg cells represent a novel truly differentiated non-cytolytic NK subset with a self-sustainable phenotype (CD56(+) CD16(+) NKp30(+) NKp44(+) NKp46(+) CD94(+) CD69(+) CCR7(+)) generated from specific pSTAT6(+) GATA3(+) precursors. NK-ireg cells could be employed to develop new immuno-suppressive strategies in autoimmune diseases, transplant rejection or graft versus host diseases. In addition, NK-ireg cells can be easily derived from peripheral blood of the patients and could constitute an autologous biotherapic tool to be used combined or in alternative to other immuno-regulatory cells.

Show MeSH
Related in: MedlinePlus