Human natural killer (NK) cells are capable of spontaneously killing certain tumor and virus-infected cells. In addition, NK cells can kill IgG antibody-coated target cells via antibody- dependent cellular cytotoxicity (ADCC) and it is the Fc portion of the IgG binding to an Fc receptor (FcgammaR) on the surface of the NK cell that provides the signal for the activation of the lytic machinery. Almost all NK cells express FcgammaRIII, and this receptor has been shown to be involved in NK cell-mediated ADCC. Until recently this was thought to be the only FcgammaR expressed on NK cells. However, we have recently shown that NK cells, from certain individuals, also express high levels of FcgammaRII, and found these to be products of the FcgammaRIIC gene, an isoform that has not been well characterized in any cell. We have further shown that an allelic polymorphism of aa 13 in the first extracellular (EC1) domain results in individuals whose NK cells are CD32neg or CD32pos. This molecule, when expressed on NK cells, can trigger ADCC and increases in intracellular CA+2 flux. In addition, using Jurkat cells transfected with two FcgammaRIIc isoforms we have defined the ligand binding specificity and signal transduction pathway of these FcgammaRs. These studies are of potential significance because NK cells have not been previously shown to express CD32 and thus this potentially represents a new means of activating and/or regulating NK cell lysis. Based on these data we hypothesize that the presence of high levels of FcgammaRII expression on NK cells of some individuals influences the function of their NK cells by activating and/or regulating the lytic and immunoregulatory function of NK cells. The present proposal is aimed at determining the functional significance of the novel CD32 isoforms expressed by human NK cells and to test our hypothesis, by assessing their role in NK cell functions.
The specific aims of the proposal are: 1) To determine the functional significance of novel CD32 isoforms in normal NK cells; 2. To determine the role of MHC class I-specific inhibitory receptors in the modulation of functions mediated by CD32; 3) To assess the possible differences in ADCC activity of NK cells against tumor targets from CD32pos or CD32neg individuals.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI042204-05
Application #
6649270
Study Section
Immunobiology Study Section (IMB)
Program Officer
Miller, Lara R
Project Start
1999-09-30
Project End
2005-06-30
Budget Start
2003-07-01
Budget End
2005-06-30
Support Year
5
Fiscal Year
2003
Total Cost
$180,096
Indirect Cost
Name
University of Pittsburgh
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
004514360
City
Pittsburgh
State
PA
Country
United States
Zip Code
15213
Stewart-Akers, A M; Cunningham, A; Wasko, M C et al. (2004) Fc gamma R expression on NK cells influences disease severity in rheumatoid arthritis. Genes Immun 5:521-9
Kepley, Christopher L; Taghavi, Sharven; Mackay, Graham et al. (2004) Co-aggregation of FcgammaRII with FcepsilonRI on human mast cells inhibits antigen-induced secretion and involves SHIP-Grb2-Dok complexes. J Biol Chem 279:35139-49
Ernst, Linda K; Metes, Diana; Herberman, Ronald B et al. (2002) Allelic polymorphisms in the FcgammaRIIC gene can influence its function on normal human natural killer cells. J Mol Med 80:248-57
Metes, D; Gambotto, A A; Nellis, J et al. (2001) Identification of the CD32/FcgammaRIIc-Q13/STP13 polymorphism using an allele-specific restriction enzyme digestion assay. J Immunol Methods 258:85-95