Generation of Antibodies to Cell Surface Markers on Mature Natural Killer Cells
Part of the
Methods in Molecular Biology
book series (MIMB, volume 121)
A limitation in the ability to analyze the function of the natural killer cell Ly49 and other receptors is the lack of antibodies to most of them. Even when monoclonal antibodies (MAbs) have been generated, they do not always react with a given receptor from all strains of mice. For example, 5E6 binds to Ly49C and/or I from B6, C3H, and NZB strains, but fails to react with any NK cells of 129, C57BR, C58, or FVB mice. Besides Ly49 receptors, NKR-P1 receptor gene products (NKR-P1B) also contain an (immunoreceptor tyrosine-based inhibiting motif (ITIM) in the cytoplasmic domain (1). In the human system, the NKG2 family of receptor molecules with or without ITIMs associate as heterodimers with CD94 molecules (2), and are therefore candidates for murine non-Ly49 receptors for class I antigens. Molecules similar to human killer cell immunoglobulin-like receptors (KIRs) have not been cloned in mice. Other receptors receive activating or positive signals and therefore may have important NK cell functions (3,4). We describe here our approaches to develop anti-NK cell antibodies to study their receptors.
KeywordsAffinity Purification Intrasplenic Injection Ly49 Receptor Mature Natural Killer Cell NKG2 Family
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
Ryan J. C. and Seaman W. E. (1997) Divergent functions of lectin-like receptors on NK cells. Immunol. Rev.
, 79–89.PubMedCrossRefGoogle Scholar
López-Botet M. Pérez-Villar J. J., Carretero M. Rodriguez A., Meleero I., Bellón T., Llano M., and Navarro F. (1997) Structure and function of the CD94 C-type lectin receptor complex involved in recognition of HLA class I molecules. Immunol. Rev.
, 165–174.PubMedCrossRefGoogle Scholar
Garni-Wagner B. A., Purohit A, Mathew P. A., Bennett M., and Kumar V. (1993) A novel function-associated molecule related to non-MHC-restricted cytotoxicity mediated by activated natural killer cells and T cells. J. Immunol.
, 60–70.PubMedGoogle Scholar
Karlhofer F. M. and Yokoyama W. M. (1991) Stimulation of murine natural killer (NK) cells by a monclonal antibody specific for the NK1.1 antigen. IL-2 activatied NK cells possess additional specific stimulation pathways. J. Immunol.
, 3662–3673.PubMedGoogle Scholar
Sentman C. L., Hackett J. Jr., Moore T. A., Kumar V., and Bennett M. (1989) Pan natural killer cell monoclonal antibodies and their relationship to the NK1.1 antigen. Hybridoma
, 605–614.PubMedCrossRefGoogle Scholar
Posner M. R. (1982) An enzyme-linked immunosorbent assay (ELISA) for the detection of monclonal antibodies to cell surface antigens on viable cells. J. Immunol. Methods
, 23–31.PubMedCrossRefGoogle Scholar
Lai W. C., Bennett M., Pakes S. P., and Murphree S. S. (1991) Potential subunit vaccine against Mycoplasma pulmonis
purified by a protective monclonal antibody. Vaccine
, 177–184.PubMedCrossRefGoogle Scholar