Expression cloning and function of the rat NK activating and inhibitory receptors NKR-P1A and -P1B.
We have characterized the rat NK receptors NKR-P1A and -P1B. A cDNA library was constructed from the rat NK cell line, RNK-16. Using the pMX retroviral cloning system, the library was expressed in the human NK cell line, YTSeco, and cells staining with the anti-rat mAb 10/78 identified, FACS sorted and cloned. Two genes, corresponding to rat NK receptors NKR-P1A and -P1B, were identified. YTSeco clones expressing either NKR-P1A or -P1B were functionally tested using (51)Cr-release redirected lysis assays and calcium flux experiments. This demonstrated that NKR-P1A functions as an activation receptor, as previously shown, and that NKR-P1B functions as an inhibitory receptor, as predicted by the presence of an immunoreceptor tyrosine-based inhibition motif. Although annotated as NKR-P1A specific, we found that mAb 10/78 stained YTSeco clones expressing NKR-P1A or -P1B equally well, as did the mAb 3.2.3 used for the original cloning of rat NKR-P1A.
Li
Jennifer
J
Department of Medical Biophysics, Ontario Cancer Institute, University of Toronto, 610 University Avenue, Toronto, Ontario M5G 2M9, Canada.
Rabinovich
Brian A
BA
Hurren
Rose
R
Shannon
John
J
Miller
Richard G
RG
D000818:Animals; D000954:Antigens, Surface / Q000235:genetics* / Q000502:physiology; D003001:Cloning, Molecular; D018076:DNA, Complementary; D007694:Killer Cells, Natural / Q000502:physiology*; D037181:Lectins, C-Type / Q000235:genetics* / Q000502:physiology; D051379:Mice; D055651:NK Cell Lectin-Like Receptor Subfamily B; D051381:Rats; D011971:Receptors, Immunologic / Q000235:genetics
D000954:Antigens, Surface; D018076:DNA, Complementary; C527203:KLRB1 protein, human; C479632:Klrb1b protein, rat; D037181:Lectins, C-Type; D055651:NK Cell Lectin-Like Receptor Subfamily B; D011971:Receptors, Immunologic