Limits...
Association with FcRgamma is essential for activation signal through NKR-P1 (CD161) in natural killer (NK) cells and NK1.1+ T cells.

Arase N, Arase H, Park SY, Ohno H, Ra C, Saito T - J. Exp. Med. (1997)

Bottom Line: In this study, we analyzed the cell surface complex associated with NKR-P1 on NK cells and found that NKR-P1 associates with the FcRgamma chain which is an essential component of Fc receptors for IgG and IgE.Similarly, NK1.1+ T cells from FcRgamma-deficient mice did not produce IFN-gamma upon NKR-P1 crosslinking.These findings demonstrate that the FcRgamma chain plays an important role in activation of NK cells via the NKR-P1 molecule.

View Article: PubMed Central - PubMed

Affiliation: Division of Molecular Genetics, Center for Biological Science, Chiba University School of Medicine, Chiba 260, Japan.

ABSTRACT
Natural killer (NK) cells exhibit cytotoxicity against variety of tumor cells and virus-infected cells without prior sensitization and represent unique lymphocytes involved in primary host defense. NKR-P1 is thought to be one of NK receptors mediating activation signals because cross-linking of NKR-P1 activates NK cells to exhibit cytotoxicity and IFN-gamma production. However, molecular mechanism of NK cell activation via NKR-P1 is not well elucidated. In this study, we analyzed the cell surface complex associated with NKR-P1 on NK cells and found that NKR-P1 associates with the FcRgamma chain which is an essential component of Fc receptors for IgG and IgE. The association between FcRgamma and NKR-P1 is independent of Fc receptor complexes. Furthermore, NK cells from FcRgamma-deficient mice did not show cytotoxicity or IFN-gamma production upon NKR-P1 cross-linking. Similarly, NK1.1+ T cells from FcRgamma-deficient mice did not produce IFN-gamma upon NKR-P1 crosslinking. These findings demonstrate that the FcRgamma chain plays an important role in activation of NK cells via the NKR-P1 molecule.

Show MeSH

Related in: MedlinePlus

Association of the FcRγ chain with the NKR-P1 molecule  on the cell surface of CTLL-2 cell line. (A) CTLL-2 cells were surface-biotinylated and the cell lysate was immunoprecipitated with anti-NK1.1  mAb or anti-CD3ε mAb and analyzed on two-dimensional nonreducing  (NR) and reducing (R) SDS-PAGE. Biotinylated proteins were detected  by an ECL. (B) FcRγ chain was detected by blotting the membrane with  anti-FcRγ Ab after immunoprecipitation with anti-NK1.1 or anti-CD3ε  mAbs. FcRγ homodimer (γ-γ) and CD3ζ-FcRγ heterodimers (ζ-γ)  were indicated.
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC2199168&req=5

Figure 1: Association of the FcRγ chain with the NKR-P1 molecule on the cell surface of CTLL-2 cell line. (A) CTLL-2 cells were surface-biotinylated and the cell lysate was immunoprecipitated with anti-NK1.1 mAb or anti-CD3ε mAb and analyzed on two-dimensional nonreducing (NR) and reducing (R) SDS-PAGE. Biotinylated proteins were detected by an ECL. (B) FcRγ chain was detected by blotting the membrane with anti-FcRγ Ab after immunoprecipitation with anti-NK1.1 or anti-CD3ε mAbs. FcRγ homodimer (γ-γ) and CD3ζ-FcRγ heterodimers (ζ-γ) were indicated.

Mentions: As shown in Fig. 1 A, immunoprecipitation with anti-NK1.1 mAb revealed that a 9-kD homodimer was coprecipitated with NK1.1 molecule. In contrast, when the TCR complex was precipitated with anti-CD3ε mAb, CD3ζ homodimers, CD3ζ-FcRγ heterodimers and a small amount of FcRγ homodimers were observed as previously reported (21). Interestingly, the 9-kD homodimers coprecipitated with NK1.1 appeared to be identical to FcRγ within the TCR complex. Indeed, the homodimers coprecipitated with NK1.1 were blotted with anti-FcRγ Ab similarly to FcRγ observed in the CD3ε-immunoprecipitates (Fig. 1 B). The association of the FcRγ chain with the NKR-P1 molecule was also observed in a rat NK cell line, RNK-16 (data not shown). We next analyzed normal NK cells to generalize the physical association between FcRγ and NKR-P1 in normal NK cell population. Similar to CTLL-2, when NK1.1 was immunoprecipitated from IL-2 activated NK cells, the FcRγ homodimer was coprecipitated with NK1.1 (Fig. 2). Collectively, these data indicate that the association of FcRγ with NKR-P1 is generally observed for NK cells in vivo.


Association with FcRgamma is essential for activation signal through NKR-P1 (CD161) in natural killer (NK) cells and NK1.1+ T cells.

Arase N, Arase H, Park SY, Ohno H, Ra C, Saito T - J. Exp. Med. (1997)

Association of the FcRγ chain with the NKR-P1 molecule  on the cell surface of CTLL-2 cell line. (A) CTLL-2 cells were surface-biotinylated and the cell lysate was immunoprecipitated with anti-NK1.1  mAb or anti-CD3ε mAb and analyzed on two-dimensional nonreducing  (NR) and reducing (R) SDS-PAGE. Biotinylated proteins were detected  by an ECL. (B) FcRγ chain was detected by blotting the membrane with  anti-FcRγ Ab after immunoprecipitation with anti-NK1.1 or anti-CD3ε  mAbs. FcRγ homodimer (γ-γ) and CD3ζ-FcRγ heterodimers (ζ-γ)  were indicated.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2199168&req=5

Figure 1: Association of the FcRγ chain with the NKR-P1 molecule on the cell surface of CTLL-2 cell line. (A) CTLL-2 cells were surface-biotinylated and the cell lysate was immunoprecipitated with anti-NK1.1 mAb or anti-CD3ε mAb and analyzed on two-dimensional nonreducing (NR) and reducing (R) SDS-PAGE. Biotinylated proteins were detected by an ECL. (B) FcRγ chain was detected by blotting the membrane with anti-FcRγ Ab after immunoprecipitation with anti-NK1.1 or anti-CD3ε mAbs. FcRγ homodimer (γ-γ) and CD3ζ-FcRγ heterodimers (ζ-γ) were indicated.
Mentions: As shown in Fig. 1 A, immunoprecipitation with anti-NK1.1 mAb revealed that a 9-kD homodimer was coprecipitated with NK1.1 molecule. In contrast, when the TCR complex was precipitated with anti-CD3ε mAb, CD3ζ homodimers, CD3ζ-FcRγ heterodimers and a small amount of FcRγ homodimers were observed as previously reported (21). Interestingly, the 9-kD homodimers coprecipitated with NK1.1 appeared to be identical to FcRγ within the TCR complex. Indeed, the homodimers coprecipitated with NK1.1 were blotted with anti-FcRγ Ab similarly to FcRγ observed in the CD3ε-immunoprecipitates (Fig. 1 B). The association of the FcRγ chain with the NKR-P1 molecule was also observed in a rat NK cell line, RNK-16 (data not shown). We next analyzed normal NK cells to generalize the physical association between FcRγ and NKR-P1 in normal NK cell population. Similar to CTLL-2, when NK1.1 was immunoprecipitated from IL-2 activated NK cells, the FcRγ homodimer was coprecipitated with NK1.1 (Fig. 2). Collectively, these data indicate that the association of FcRγ with NKR-P1 is generally observed for NK cells in vivo.

Bottom Line: In this study, we analyzed the cell surface complex associated with NKR-P1 on NK cells and found that NKR-P1 associates with the FcRgamma chain which is an essential component of Fc receptors for IgG and IgE.Similarly, NK1.1+ T cells from FcRgamma-deficient mice did not produce IFN-gamma upon NKR-P1 crosslinking.These findings demonstrate that the FcRgamma chain plays an important role in activation of NK cells via the NKR-P1 molecule.

View Article: PubMed Central - PubMed

Affiliation: Division of Molecular Genetics, Center for Biological Science, Chiba University School of Medicine, Chiba 260, Japan.

ABSTRACT
Natural killer (NK) cells exhibit cytotoxicity against variety of tumor cells and virus-infected cells without prior sensitization and represent unique lymphocytes involved in primary host defense. NKR-P1 is thought to be one of NK receptors mediating activation signals because cross-linking of NKR-P1 activates NK cells to exhibit cytotoxicity and IFN-gamma production. However, molecular mechanism of NK cell activation via NKR-P1 is not well elucidated. In this study, we analyzed the cell surface complex associated with NKR-P1 on NK cells and found that NKR-P1 associates with the FcRgamma chain which is an essential component of Fc receptors for IgG and IgE. The association between FcRgamma and NKR-P1 is independent of Fc receptor complexes. Furthermore, NK cells from FcRgamma-deficient mice did not show cytotoxicity or IFN-gamma production upon NKR-P1 cross-linking. Similarly, NK1.1+ T cells from FcRgamma-deficient mice did not produce IFN-gamma upon NKR-P1 crosslinking. These findings demonstrate that the FcRgamma chain plays an important role in activation of NK cells via the NKR-P1 molecule.

Show MeSH
Related in: MedlinePlus