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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: To define Ly-49D-ligand interactions, we transfected the mouse Ly-49D receptor into the rat NK line, RNK-16 (RNK.mLy-49D).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.

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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The Ly-49D receptor activates lysis of H-2d–expressing Con  A blasts. RNK.mLy-49D transfectants were tested as effector cells for lysis  of Con A blast targets prepared from mice that were congenic resistant at  the H-2 locus. Con A blasts were prepared from splenocytes isolated from  mice expressing either H-2d (BALB/c, B10.D2; panels A and B), H-2b  (BALB.B, C57BL/6; panels C and D), or H-2k (BALB.K, B10.BR; panels E and F). Assays were performed after preincubation of effector cells  with media alone (□), anti–Ly-49D mAb (12A8) (♦), or control mAb  (2C7) (○) before addition of targets.
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Figure 3: The Ly-49D receptor activates lysis of H-2d–expressing Con A blasts. RNK.mLy-49D transfectants were tested as effector cells for lysis of Con A blast targets prepared from mice that were congenic resistant at the H-2 locus. Con A blasts were prepared from splenocytes isolated from mice expressing either H-2d (BALB/c, B10.D2; panels A and B), H-2b (BALB.B, C57BL/6; panels C and D), or H-2k (BALB.K, B10.BR; panels E and F). Assays were performed after preincubation of effector cells with media alone (□), anti–Ly-49D mAb (12A8) (♦), or control mAb (2C7) (○) before addition of targets.

Mentions: As shown in Fig. 3, RNK.mLy-49D cells effectively lysed Con A blasts prepared from BALB/c (H-2d; panel A) and B10.D2 (H-2d; panel B) mice, but not Con A blasts prepared from BALB.B10 (H-2b; panel C), C57BL/6 (H-2b; panel D), BALB.K (H-2k; panel E), or B10.BR (H-2k; panel F) mice. In experiments not shown, RNK.mLy-49D effectors also lysed Con A blasts from DBA/2 (H-2d) mice but not blasts from C3H (H-2k) or B10.S (H-2s) mice. Lysis of all H-2d blasts by RNK.mLy-49D effectors was inhibited by anti–Ly-49D mAb 12A8, but not by isotype-matched control mAb 2C7 (panels A and B).


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)

The Ly-49D receptor activates lysis of H-2d–expressing Con  A blasts. RNK.mLy-49D transfectants were tested as effector cells for lysis  of Con A blast targets prepared from mice that were congenic resistant at  the H-2 locus. Con A blasts were prepared from splenocytes isolated from  mice expressing either H-2d (BALB/c, B10.D2; panels A and B), H-2b  (BALB.B, C57BL/6; panels C and D), or H-2k (BALB.K, B10.BR; panels E and F). Assays were performed after preincubation of effector cells  with media alone (□), anti–Ly-49D mAb (12A8) (♦), or control mAb  (2C7) (○) before addition of targets.
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Figure 3: The Ly-49D receptor activates lysis of H-2d–expressing Con A blasts. RNK.mLy-49D transfectants were tested as effector cells for lysis of Con A blast targets prepared from mice that were congenic resistant at the H-2 locus. Con A blasts were prepared from splenocytes isolated from mice expressing either H-2d (BALB/c, B10.D2; panels A and B), H-2b (BALB.B, C57BL/6; panels C and D), or H-2k (BALB.K, B10.BR; panels E and F). Assays were performed after preincubation of effector cells with media alone (□), anti–Ly-49D mAb (12A8) (♦), or control mAb (2C7) (○) before addition of targets.
Mentions: As shown in Fig. 3, RNK.mLy-49D cells effectively lysed Con A blasts prepared from BALB/c (H-2d; panel A) and B10.D2 (H-2d; panel B) mice, but not Con A blasts prepared from BALB.B10 (H-2b; panel C), C57BL/6 (H-2b; panel D), BALB.K (H-2k; panel E), or B10.BR (H-2k; panel F) mice. In experiments not shown, RNK.mLy-49D effectors also lysed Con A blasts from DBA/2 (H-2d) mice but not blasts from C3H (H-2k) or B10.S (H-2s) mice. Lysis of all H-2d blasts by RNK.mLy-49D effectors was inhibited by anti–Ly-49D mAb 12A8, but not by isotype-matched control mAb 2C7 (panels A and B).

Bottom Line: To define Ly-49D-ligand interactions, we transfected the mouse Ly-49D receptor into the rat NK line, RNK-16 (RNK.mLy-49D).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.

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