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The p110 delta of PI3K plays a critical role in NK cell terminal maturation and cytokine/chemokine generation.

Guo H, Samarakoon A, Vanhaesebroeck B, Malarkannan S - J. Exp. Med. (2008)

Bottom Line: Further, p110 delta(D910A/D910A) NK cell-mediated antiviral responses through natural cytotoxicity receptor 1 were reduced.Analysis of signaling events demonstrates that p110 delta(D910A/D910A) NK cells had a reduced c-Jun N-terminal kinase 1/2 phosphorylation in response to NKG2D-mediated activation.These results reveal a previously unrecognized role of PI3K-p110 delta in NK cell development and effector functions.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Immunology, Blood Research Institute, Medical College of Wisconsin, Milwaukee, WI 53226, USA.

ABSTRACT
Phosphatidylinositol 3-kinases (PI3Ks) play a critical role in regulating B cell receptor- and T cell receptor-mediated signaling. However, their role in natural killer (NK) cell development and functions is not well understood. Using mice expressing p110 delta(D910A), a catalytically inactive p110 delta, we show that these mice had reduced NK cellularity, defective Ly49C and Ly49I NK subset maturation, and decreased CD27(High) NK numbers. p110 delta inactivation marginally impaired NK-mediated cytotoxicity against tumor cells in vitro and in vivo. However, NKG2D, Ly49D, and NK1.1 receptor-mediated cytokine and chemokine generation by NK cells was severely affected in these mice. Further, p110 delta(D910A/D910A) NK cell-mediated antiviral responses through natural cytotoxicity receptor 1 were reduced. Analysis of signaling events demonstrates that p110 delta(D910A/D910A) NK cells had a reduced c-Jun N-terminal kinase 1/2 phosphorylation in response to NKG2D-mediated activation. These results reveal a previously unrecognized role of PI3K-p110 delta in NK cell development and effector functions.

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Catalytic inactivation of p110δ reduces CD27High NK subsets. BM- (a) or spleen- (b) derived CD3−NK1.1+ NK cells were analyzed for CD11b and CD27 expression. The percentages of CD27HighCD11bLow, CD27HighCD11bHigh, and CD27LowCD11bHigh cells were calculated. Data are means ± SD. (c) Alteration in CD27High NK subsets correlates with the reduction in Ly49C/I NK subsets. Spleen-derived single-cell suspensions were stained with anti-NK1.1, anti-CD3ε, and each of the indicated anti-Ly49 or CD11b and CD27 mAbs. NK cells as specified by CD3−NK1.1+ positivity were gated and analyzed. Data shown were obtained using cells from five to seven mice of each genotype. One representative out of three independent experiments is shown.
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fig4: Catalytic inactivation of p110δ reduces CD27High NK subsets. BM- (a) or spleen- (b) derived CD3−NK1.1+ NK cells were analyzed for CD11b and CD27 expression. The percentages of CD27HighCD11bLow, CD27HighCD11bHigh, and CD27LowCD11bHigh cells were calculated. Data are means ± SD. (c) Alteration in CD27High NK subsets correlates with the reduction in Ly49C/I NK subsets. Spleen-derived single-cell suspensions were stained with anti-NK1.1, anti-CD3ε, and each of the indicated anti-Ly49 or CD11b and CD27 mAbs. NK cells as specified by CD3−NK1.1+ positivity were gated and analyzed. Data shown were obtained using cells from five to seven mice of each genotype. One representative out of three independent experiments is shown.

Mentions: Recent studies have shown that the mature CD11b+ NK cells can be further divided into CD27High and CD27Low with distinct functional abilities (39–41). These studies demonstrate that CD27HighCD11bHigh NK cells were superior in their ability to lyse target cells compared with that of CD27LowCD11bHigh cells. Further, CD27HighCD11bHigh NK cells produced considerably higher amounts of IFN-γ (39). Because NK cells from p110δD910A/D910A mice showed a specific defect in Ly49C/I expression, we extended our analyses to CD27-based NK maturation. Fresh BM and splenocytes were gated for CD3−NK1.1+ cells and analyzed for CD27 and CD11b markers. Fig. 4 shows that a lack of functional p110δ altered the ratio of CD27- and CD11b-expressing NK subsets. There was a significant reduction in the CD27HighCD11bLow NK cells in both fresh BM (Fig. 4 a) and spleen (Fig. 4 b) of p110δD910A/D910A mice. The CD27HighCD11bHigh NK cell numbers were also affected in the spleen of p110δD910A/D910A mice. More importantly, the decrease in the CD27HighCD11bHigh population was correlated with a concomitant increase in the CD27LowCD11bHigh NK subsets in both fresh BM and spleen of the p110δD910A/D910A NK population. Earlier studies have also indicated a skewed expression of Ly49C/I receptors in CD27LowCD11bhigh NK cells (39). Because we observed a specific reduction in the Ly49C/I subsets in the absence of functional p110δ, we analyzed the fresh NK cells from p110δD910A/D910A mice for CD27 and Ly49 expressions. Results presented in Fig. 4 c indicate that Ly49A, Ly49I, Ly49D, and Ly49G2 NK subsets with CD27High positivity were reduced more in p110δD910A/D910A NK cells than those of WT/WT. However, the reduction in the Ly49I+CD27High NK subset was more severe compared with other subsets in p110δD910A/D910A NK cells. Based on these results, we conclude that p110δ is needed for both Ly49C/I subset specification and proliferation, and CD27HighCD11bHigh NK maturation.


The p110 delta of PI3K plays a critical role in NK cell terminal maturation and cytokine/chemokine generation.

Guo H, Samarakoon A, Vanhaesebroeck B, Malarkannan S - J. Exp. Med. (2008)

Catalytic inactivation of p110δ reduces CD27High NK subsets. BM- (a) or spleen- (b) derived CD3−NK1.1+ NK cells were analyzed for CD11b and CD27 expression. The percentages of CD27HighCD11bLow, CD27HighCD11bHigh, and CD27LowCD11bHigh cells were calculated. Data are means ± SD. (c) Alteration in CD27High NK subsets correlates with the reduction in Ly49C/I NK subsets. Spleen-derived single-cell suspensions were stained with anti-NK1.1, anti-CD3ε, and each of the indicated anti-Ly49 or CD11b and CD27 mAbs. NK cells as specified by CD3−NK1.1+ positivity were gated and analyzed. Data shown were obtained using cells from five to seven mice of each genotype. One representative out of three independent experiments is shown.
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fig4: Catalytic inactivation of p110δ reduces CD27High NK subsets. BM- (a) or spleen- (b) derived CD3−NK1.1+ NK cells were analyzed for CD11b and CD27 expression. The percentages of CD27HighCD11bLow, CD27HighCD11bHigh, and CD27LowCD11bHigh cells were calculated. Data are means ± SD. (c) Alteration in CD27High NK subsets correlates with the reduction in Ly49C/I NK subsets. Spleen-derived single-cell suspensions were stained with anti-NK1.1, anti-CD3ε, and each of the indicated anti-Ly49 or CD11b and CD27 mAbs. NK cells as specified by CD3−NK1.1+ positivity were gated and analyzed. Data shown were obtained using cells from five to seven mice of each genotype. One representative out of three independent experiments is shown.
Mentions: Recent studies have shown that the mature CD11b+ NK cells can be further divided into CD27High and CD27Low with distinct functional abilities (39–41). These studies demonstrate that CD27HighCD11bHigh NK cells were superior in their ability to lyse target cells compared with that of CD27LowCD11bHigh cells. Further, CD27HighCD11bHigh NK cells produced considerably higher amounts of IFN-γ (39). Because NK cells from p110δD910A/D910A mice showed a specific defect in Ly49C/I expression, we extended our analyses to CD27-based NK maturation. Fresh BM and splenocytes were gated for CD3−NK1.1+ cells and analyzed for CD27 and CD11b markers. Fig. 4 shows that a lack of functional p110δ altered the ratio of CD27- and CD11b-expressing NK subsets. There was a significant reduction in the CD27HighCD11bLow NK cells in both fresh BM (Fig. 4 a) and spleen (Fig. 4 b) of p110δD910A/D910A mice. The CD27HighCD11bHigh NK cell numbers were also affected in the spleen of p110δD910A/D910A mice. More importantly, the decrease in the CD27HighCD11bHigh population was correlated with a concomitant increase in the CD27LowCD11bHigh NK subsets in both fresh BM and spleen of the p110δD910A/D910A NK population. Earlier studies have also indicated a skewed expression of Ly49C/I receptors in CD27LowCD11bhigh NK cells (39). Because we observed a specific reduction in the Ly49C/I subsets in the absence of functional p110δ, we analyzed the fresh NK cells from p110δD910A/D910A mice for CD27 and Ly49 expressions. Results presented in Fig. 4 c indicate that Ly49A, Ly49I, Ly49D, and Ly49G2 NK subsets with CD27High positivity were reduced more in p110δD910A/D910A NK cells than those of WT/WT. However, the reduction in the Ly49I+CD27High NK subset was more severe compared with other subsets in p110δD910A/D910A NK cells. Based on these results, we conclude that p110δ is needed for both Ly49C/I subset specification and proliferation, and CD27HighCD11bHigh NK maturation.

Bottom Line: Further, p110 delta(D910A/D910A) NK cell-mediated antiviral responses through natural cytotoxicity receptor 1 were reduced.Analysis of signaling events demonstrates that p110 delta(D910A/D910A) NK cells had a reduced c-Jun N-terminal kinase 1/2 phosphorylation in response to NKG2D-mediated activation.These results reveal a previously unrecognized role of PI3K-p110 delta in NK cell development and effector functions.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Molecular Immunology, Blood Research Institute, Medical College of Wisconsin, Milwaukee, WI 53226, USA.

ABSTRACT
Phosphatidylinositol 3-kinases (PI3Ks) play a critical role in regulating B cell receptor- and T cell receptor-mediated signaling. However, their role in natural killer (NK) cell development and functions is not well understood. Using mice expressing p110 delta(D910A), a catalytically inactive p110 delta, we show that these mice had reduced NK cellularity, defective Ly49C and Ly49I NK subset maturation, and decreased CD27(High) NK numbers. p110 delta inactivation marginally impaired NK-mediated cytotoxicity against tumor cells in vitro and in vivo. However, NKG2D, Ly49D, and NK1.1 receptor-mediated cytokine and chemokine generation by NK cells was severely affected in these mice. Further, p110 delta(D910A/D910A) NK cell-mediated antiviral responses through natural cytotoxicity receptor 1 were reduced. Analysis of signaling events demonstrates that p110 delta(D910A/D910A) NK cells had a reduced c-Jun N-terminal kinase 1/2 phosphorylation in response to NKG2D-mediated activation. These results reveal a previously unrecognized role of PI3K-p110 delta in NK cell development and effector functions.

Show MeSH
Related in: MedlinePlus