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Mouse Ly-49D recognizes H-2Dd and activates natural killer cell cytotoxicity.

Nakamura MC, Linnemeyer PA, Niemi EC, Mason LH, Ortaldo JR, Ryan JC, Seaman WE - J. Exp. Med. (1999)

Bottom Line: As expected, anti- Ly-49D monoclonal antibody 12A8 specifically stimulated redirected lysis of the Fc receptor- bearing rat target YB2/0 by RNK.mLy-49D transfectants.RNK.mLy-49D cells lysed YB2/0.Dd targets more efficiently than untransfected YB2/0 or YB2/0 transfected with Db, Kk, or Kb.These experiments show that the activating receptor Ly-49D specifically interacts with the MHC I antigen, H-2Dd, demonstrating the existence of alloactivating receptors on murine NK cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, University of California San Francisco, San Francisco, California 94143, USA. marynak@itsa.ucsf.edu

ABSTRACT
Although activation of natural killer (NK) cytotoxicity is generally inhibited by target major histocompatibility complex (MHC) class I expression, subtle features of NK allorecognition suggest that NK cells possess receptors that are activated by target MHC I. The mouse Ly-49D receptor has been shown to activate NK cytotoxicity, although recognition of MHC class I has not been demonstrated previously. To define Ly-49D-ligand interactions, we transfected the mouse Ly-49D receptor into the rat NK line, RNK-16 (RNK.mLy-49D). As expected, anti- Ly-49D monoclonal antibody 12A8 specifically stimulated redirected lysis of the Fc receptor- bearing rat target YB2/0 by RNK.mLy-49D transfectants. RNK.mLy-49D effectors were tested against YB2/0 targets transfected with the mouse MHC I alleles H-2Dd, Db, Kk, or Kb. RNK.mLy-49D cells lysed YB2/0.Dd targets more efficiently than untransfected YB2/0 or YB2/0 transfected with Db, Kk, or Kb. This augmented lysis of H-2Dd targets was specifically inhibited by F(ab')2 anti-Ly-49D (12A8) and F(ab')2 anti-H-2Dd (34-5-8S). RNK.mLy-49D effectors were also able to specifically lyse Concanavalin A blasts isolated from H-2(d) mice (BALB/c, B10.D2, and DBA/2) but not from H-2(b) or H-2(k) mice. These experiments show that the activating receptor Ly-49D specifically interacts with the MHC I antigen, H-2Dd, demonstrating the existence of alloactivating receptors on murine NK cells.

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Activation of Ly-49D by H-2Dd  is inhibited by an antibody specific for the  α1/α2 domains of H-2Dd. Effector cells  RNK-16 (A), RNK.mLy-49A transfectants  (B), and RNK.mLy-49D transfectants (C)  were tested in a standard 4-h cytotoxicity  assay against YB2/0 targets expressing Dd  (YB2/0.Dd) in the presence of media alone  (□) or anti–H-2Dd F(ab′)2 fragments (34-5-8S) (♦).
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Figure 5: Activation of Ly-49D by H-2Dd is inhibited by an antibody specific for the α1/α2 domains of H-2Dd. Effector cells RNK-16 (A), RNK.mLy-49A transfectants (B), and RNK.mLy-49D transfectants (C) were tested in a standard 4-h cytotoxicity assay against YB2/0 targets expressing Dd (YB2/0.Dd) in the presence of media alone (□) or anti–H-2Dd F(ab′)2 fragments (34-5-8S) (♦).

Mentions: Previous studies have shown that the mLy-49A/H-2Dd interaction can be blocked by one anti–H-2Dd antibody (34-5-8S), which is specific for a conformationally dependent epitope in the H-2Dd α1/α2 domains. Other mAbs to H-2Dd do not interrupt its interaction with Ly-49A (32). We, therefore, determined whether F(ab′)2 34-5-8S could inhibit the Ly-49D/H-2Dd interaction as well. As shown in Fig. 5, activation of RNK.mLy-49D effectors by YB2/ 0.Dd was inhibited by F(ab′)2 34-5-8S (panel C), whereas lysis of YB2/0.Dd by RNK.mLy-49A effectors was restored in the presence of F(ab′)2 34-5-8S. RNK-16 lysis of YB2/0.Dd was unaffected by F(ab′)2 34-5-8S.


Mouse Ly-49D recognizes H-2Dd and activates natural killer cell cytotoxicity.

Nakamura MC, Linnemeyer PA, Niemi EC, Mason LH, Ortaldo JR, Ryan JC, Seaman WE - J. Exp. Med. (1999)

Activation of Ly-49D by H-2Dd  is inhibited by an antibody specific for the  α1/α2 domains of H-2Dd. Effector cells  RNK-16 (A), RNK.mLy-49A transfectants  (B), and RNK.mLy-49D transfectants (C)  were tested in a standard 4-h cytotoxicity  assay against YB2/0 targets expressing Dd  (YB2/0.Dd) in the presence of media alone  (□) or anti–H-2Dd F(ab′)2 fragments (34-5-8S) (♦).
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Related In: Results  -  Collection

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Figure 5: Activation of Ly-49D by H-2Dd is inhibited by an antibody specific for the α1/α2 domains of H-2Dd. Effector cells RNK-16 (A), RNK.mLy-49A transfectants (B), and RNK.mLy-49D transfectants (C) were tested in a standard 4-h cytotoxicity assay against YB2/0 targets expressing Dd (YB2/0.Dd) in the presence of media alone (□) or anti–H-2Dd F(ab′)2 fragments (34-5-8S) (♦).
Mentions: Previous studies have shown that the mLy-49A/H-2Dd interaction can be blocked by one anti–H-2Dd antibody (34-5-8S), which is specific for a conformationally dependent epitope in the H-2Dd α1/α2 domains. Other mAbs to H-2Dd do not interrupt its interaction with Ly-49A (32). We, therefore, determined whether F(ab′)2 34-5-8S could inhibit the Ly-49D/H-2Dd interaction as well. As shown in Fig. 5, activation of RNK.mLy-49D effectors by YB2/ 0.Dd was inhibited by F(ab′)2 34-5-8S (panel C), whereas lysis of YB2/0.Dd by RNK.mLy-49A effectors was restored in the presence of F(ab′)2 34-5-8S. RNK-16 lysis of YB2/0.Dd was unaffected by F(ab′)2 34-5-8S.

Bottom Line: As expected, anti- Ly-49D monoclonal antibody 12A8 specifically stimulated redirected lysis of the Fc receptor- bearing rat target YB2/0 by RNK.mLy-49D transfectants.RNK.mLy-49D cells lysed YB2/0.Dd targets more efficiently than untransfected YB2/0 or YB2/0 transfected with Db, Kk, or Kb.These experiments show that the activating receptor Ly-49D specifically interacts with the MHC I antigen, H-2Dd, demonstrating the existence of alloactivating receptors on murine NK cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Medicine, University of California San Francisco, San Francisco, California 94143, USA. marynak@itsa.ucsf.edu

ABSTRACT
Although activation of natural killer (NK) cytotoxicity is generally inhibited by target major histocompatibility complex (MHC) class I expression, subtle features of NK allorecognition suggest that NK cells possess receptors that are activated by target MHC I. The mouse Ly-49D receptor has been shown to activate NK cytotoxicity, although recognition of MHC class I has not been demonstrated previously. To define Ly-49D-ligand interactions, we transfected the mouse Ly-49D receptor into the rat NK line, RNK-16 (RNK.mLy-49D). As expected, anti- Ly-49D monoclonal antibody 12A8 specifically stimulated redirected lysis of the Fc receptor- bearing rat target YB2/0 by RNK.mLy-49D transfectants. RNK.mLy-49D effectors were tested against YB2/0 targets transfected with the mouse MHC I alleles H-2Dd, Db, Kk, or Kb. RNK.mLy-49D cells lysed YB2/0.Dd targets more efficiently than untransfected YB2/0 or YB2/0 transfected with Db, Kk, or Kb. This augmented lysis of H-2Dd targets was specifically inhibited by F(ab')2 anti-Ly-49D (12A8) and F(ab')2 anti-H-2Dd (34-5-8S). RNK.mLy-49D effectors were also able to specifically lyse Concanavalin A blasts isolated from H-2(d) mice (BALB/c, B10.D2, and DBA/2) but not from H-2(b) or H-2(k) mice. These experiments show that the activating receptor Ly-49D specifically interacts with the MHC I antigen, H-2Dd, demonstrating the existence of alloactivating receptors on murine NK cells.

Show MeSH
Related in: MedlinePlus