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Blood, 1 March 2005, Vol. 105, No. 5, pp. 2028-2035. Prepublished online as a Blood First Edition Paper on November 4, 2004; DOI 10.1182/blood-2004-08-3174. This Article Full Text (PDF) All Versions of this Article: 2004-08-3174v1 105/5/2028    most recent Alert me when this article is cited Alert me if a correction is posted Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Rights and Permissions Citing Articles Citing Articles via HighWire Citing Articles via CrossRef Citing Articles via Google Scholar Google Scholar Articles by Draghi, M. Articles by Parham, P. Search for Related Content PubMed PubMed Citation Articles by Draghi, M. Articles by Parham, P. Social Bookmarking                   What's this?  Previous Article  |  Next Article  Submitted August 17, 2004 Accepted October 6, 2004 Single cell analysis of the human NK cell response to missing self and its inhibition by HLA class I Monia Draghi, Nobuyo Yawata, Michael Gleimer, Makoto Yawata, Nicholas M Valiante, and Peter Parham* Departments of Structural Biology and Microbiology & Immunology, Stanford University School of Medicine, Stanford, CA, USA Vaccines Research, Chiron Corporation, Emeryville, CA, USA * Corresponding author; email: peropa{at}stanford.edu. Natural killer (NK) cells activate quickly in response to pathogens, tumors and allogeneic hematopoietic cell transplants. Modulating the NK cell response are clonally distributed NK cell receptors that survey cells for change in the expression of MHC class I and structurally related ligands. Here the ELISpot assay, intracellular cytokine staining (ICS) and short-term culture were used to quantify the response of bulk NK cell populations from human donors to HLA class I deficient 221 cells and to 221 cells transfected with single HLA class I allotypes. NK cells in cultures containing IL-2 or IL-12 exhibited specificities of HLA class I mediated inhibition that correlated well with those previously defined using NK cell clones in long-term culture and with the frequencies of cells expressing particular inhibitory HLA class I receptors. Culture with IL-12, but not IL-2, gave an increased frequency of cells expressing CD94:NKG2A, but no change in KIR expression. For some heterozygote combinations of KIR3DL1 alleles, ICS can be used to compare the functional properties of the two allotypes. Thus both the low expressing KIR3DL1*005 and the high expressing KIR3DL1*002 gave similar inhibitory response on challenge with an HLA-B*5801 ligand. 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