Abstract

CD28 functions as a cytotoxicity activation receptor in the natural killer (NK) cell line YT-Indy. To analyze the requirement of p56lck kinase in the function of killer inhibitory receptors, we transfected the p56lck negative YT-Indy cell line with the c143 gene encoding for KIR2DL2. Prevanadate treatment revealed KIR2DL2 phosphorylation in the YT-Indy-c143 cell line,as well as SHP1/SHP2 recruitment. YT-Indy-c143 cells were inhibited in their ability to lyse target cells expressing HLA-Cw3, a ligand for KIR2DL2. This inhibition was blocked by anti-KIR2DL2 or anti-HLA class I mAb. CD28 crosslinking on YT-Indy-c143 cells enhanced tyrosine phosphorylation of PLC-gamma 1. The simultaneous ligation of KIR2DL2 with mAb resulted in a decrease in CD28-induced tyrosine phosphorylation of PLC-gamma 1 confirming that dephosphorylation of this protein is involved in the KIR2DL2 induced inhibition of CD28-mediated cytotoxicity. As YT-Indy-c143 cells do not express detectable levels of p56lck, these results indicate that this kinase is not required for transmitting the negative signals generated by KIR2DL2 ligation.

  Funding Agency

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Intramural Research (Z01)
Project #
1Z01AI000543-14
Application #
6521376
Study Section
(LAD)
Project Start
Project End
Budget Start
Budget End
Support Year
14
Fiscal Year
2001
Total Cost
Indirect Cost

  Institution

Name
Niaid Extramural Activities
Department
Type
DUNS #
City
State
Country
United States
Zip Code

  Publications

Burgess, Steven J; Marusina, Alina I; Pathmanathan, Ishani et al. (2006) IL-21 down-regulates NKG2D/DAP10 expression on human NK and CD8+ T cells. J Immunol 176:1490-7
Lieto, L D; Maasho, K; West, D et al. (2006) The human CD94 gene encodes multiple, expressible transcripts including a new partner of NKG2A/B. Genes Immun 7:36-43
Marusina, Alina I; Kim, Dae-Ki; Lieto, Louis D et al. (2005) GATA-3 is an important transcription factor for regulating human NKG2A gene expression. J Immunol 174:2152-9
Borrego, Francisco; Masilamani, Madhan; Kabat, Juraj et al. (2005) The cell biology of the human natural killer cell CD94/NKG2A inhibitory receptor. Mol Immunol 42:485-8
Maasho, Kerima; Marusina, Alina; Reynolds, Nicole M et al. (2004) Efficient gene transfer into the human natural killer cell line, NKL, using the Amaxa nucleofection system. J Immunol Methods 284:133-40
Kim, Dae-Ki; Kabat, Juraj; Borrego, Francisco et al. (2004) Human NKG2F is expressed and can associate with DAP12. Mol Immunol 41:53-62
Sanni, Tolib B; Masilamani, Madhan; Kabat, Juraj et al. (2004) Exclusion of lipid rafts and decreased mobility of CD94/NKG2A receptors at the inhibitory NK cell synapse. Mol Biol Cell 15:3210-23
Lieto, Louis D; Borrego, Francisco; You, Chi-Hyun et al. (2003) Human CD94 gene expression: dual promoters differing in responsiveness to IL-2 or IL-15. J Immunol 171:5277-86
Kabat, Juraj; Borrego, Francisco; Brooks, Andrew et al. (2002) Role that each NKG2A immunoreceptor tyrosine-based inhibitory motif plays in mediating the human CD94/NKG2A inhibitory signal. J Immunol 169:1948-58
Tarazona, Raquel; Borrego, Francisco; Galiani, Maria D et al. (2002) Inhibition of CD28-mediated natural cytotoxicity by KIR2DL2 does not require p56(lck) in the NK cell line YT-Indy. Mol Immunol 38:495-503

Showing the most recent 10 out of 11 publications