Abstract
Normal human lymphoblastoid cell lines (LCLs) express six classical class I HLA genes from three gene loci, HLA-A, HLA-B, and HLA-C, which are located within the major histocompatibility complex (MHC). Because of the extensive allelic polymorphism of HLA class I genes, cell lines from heterozygous individuals can express up to six different class I alleles (1). Class I phenotypes, as well as other properties of different lymphoblastoid target cells, can therefore be very heterogeneous. Hence, in investigating the effects on and specificity of class I recognition by natural killer (NK) cells, it has proved useful to use transfected target cells that express only one class I allele, so that the effects of individual class I allotypes can be compared directly using the same target cell background.
Keywords
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.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Parham P., Adams E. J., and Arnett K. L. (1995) The origins of HLA-A, B, C polymorphism. Immunol. Rev. 143, 141–180.
Kavathas P., Bach F., and DeMars R. (1980) Gamma ray-induced loss of expression of HLA and glyoxalase I alleles in lymphoblastoid cells. PNAS 77, 4251–4255.
Shimizu Y., Geraghty D. E., Koller B. H., Orr H. T., and DeMars R. (1988) Transfer and expression of three cloned human non-HLA-A, B, C class I major histocompatibility complex genes in mutant lymphoblastoid cells. PNAS 85, 227–231.
Litwin V., Gumperz J., Parham P., Phillips J. H., and Lanier L. L. (1994) NKB1: a natural killer cell receptor involved in the recognition of polymorphic HLA-B molecules. J. Exp. Med. 180, 537–543.
Moretta A., Vitale M., Bottino C., et al. (1993) p58 molecules as putative receptors for major histocompatibility complex (MHC) class I molecules in human natural killer (NK) cells. Anti-p58 antibodies reconstitute lysis of MHC class I-protected cells in NK clones displaying different specificities. J. Exp. Med. 178, 597–604.
Biassoni R., Falco M., Cambiaggi A., et al. (1995) Single amino acid substitutions can influence the NK-mediated recognition of HLA-C molecules. Role of serine-77 and lysine-80 in the target cell protection from lysis mediated by “group 2” or “group 1” NK clones. J. Exp. Med. 182, 605–609.
Gumperz J. E., Barber L. D., Valiante N. M., et al. (1997) Conserved and variable residues within the Bw4 motif of HLA-B make separable contributions to recognition by the NKB1 killer cell inhibitory recptor. J. Immunol. 158, 5237–5241.
Takebem Y., Seiki M., Fujisawa J. Hoy P., Yokota K., Arai K., et al. (1988) SR alpha promoter: an efficient and versatile mammalian cDNA expression system composed of the simian virus 40 early promoter and the R-U5 segment of human T-cell leukemia virus type 1 long terminal repeat. Mol. Cell. Biol. 8, 466–472.
Lin A., Devaux B., Green A., Sagerström C., Elliot J. F., and Davis M. M. (1990) Expression of T cell antigen receptor heterodimers in a lipid-linked form. Science 249, 677–679.
Sugden B, Marsh K, Yates J. (1985) A vector that replicates as a plasmid and can be efficiently selected in B-lymphoblasts transformed by Epstein-Barr virus. Mol. Cell. Biol. 5, 410–413.
Chowdhury K. (1991) One step ‘miniprep’ method for the isolation of plasmid DNA. Nucleic Acids Res. 19(10), 2792.
Little A-M. and Parham P. (1993) HLA class I gene and protein sequence polymorphisms, in Histocompatibility Testing: A Practical Approach (Rickwood D. and Hames B. D., eds.), IRL Press, Oxford, New York, Tokyo, pp. 159–190.
DeMars R., Rudersdorf R., Chang C., et al. (1985) Mutations that impair a post-translational step in expression of HLA-A and-B antigens. PNAS. 82, 8183–8187.
Edwards P. A., Smith C. M., Neville A. M., and O’Hare M. J. (1982) A human hybridoma system based on a fast growing mutant of the ARH-77 plasma cell leukemia derived cell line. Eur. J. Immunol. 12, 641–648.
Zemmour J., Little A-M., Schendel D. J., and Parham P. (1992) The HLA-A,B “negative” mutant cell line C1R expresses a novel HLA-B35 allele, which also has a point mutation in the translation initiation codon. J. Immunol. 148(6), 1941–1948.
Riberdy J. M. and Cresswell P. (1992) The antigen-processing mutant T2 suggests a role for MHC-linked genes class II antigen presentation. J. Immunol. 148(8), 2586–2590.
Malnati M. S., Peruzzi M., Parker K. C., et al. Peptide specificity in the recognition of MHC class I by natural killer cell clones. (1995) Science. 267(5200), 1016–1018.
Lozzio B. B. and Lozzio C. B. (1979) Properties and usefulness of the original K-562 human myelogenous leukemia cell line. Leukemia Res. 3(6), 363–370.
Rosa F., Berissi H., Weissenbach J., Maroteaux L., Fellous M., and Revel M. (1983) The β2-microglobulin mRNA in human Daudi cells has a mutated initiation codon but is still inducible by interferon. EMBO J. 2, 239–243.
Lee L., McHugh L., Ribaudo R. K., Kozlowski S., Margulies D. H., and Mage M. G. (1994) Functional cell surface expression by a recombinant single-chain class I major histocompatibility complex molecule with a cis-active beta2-microglobulin domain. Eur. J. Immunol. 24(11), 2633–2639.
Alber G., Kent U. M., and Metzger H. (1992) Functional comparison of Fc epsilon RI, Fc gamma RII, and Fc gamma RIII in mast cells. J. Immunol. 149(7), 2428–2436.
Perez-Villar J. J., Melero I., Rodriguez A, et al. (1995) Functional ambivalence of the Kp43 (CD94) NK cell-associated surface antigen. J. Immunol. 154, 5779–5788.
McCutcheon J. A., Gumperz J., Smith K. D., Lutz C. T., and Parham P. (1995) Low HLA-C expression at cell surfaces correlates with increased turnover of heavy chain mRNA. J. Exp. Med. 181(6), 2085–2095.
Marsh S. G. (1997) Nomenclature for factors of the HLA system, update July 1996. The WHO Nomenclature Committee for factors of the HLA system. Hum. Immunol. 52(1), 72.
van den Hoff M. J. B., Moorman A. F. M., and Lamers W. H. (1992) Electroporation in ‘intracellular’ buffer increases cell survival. Nucleic Acids Res. 20(11), 2902.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1999 Humana Press Inc.
About this protocol
Cite this protocol
Gumperz, J.E. (1999). Generation of HLA Class I Transfected Target Cell Lines. In: Campbell, K.S., Colonna, M. (eds) Natural Killer Cell Protocols. Methods in Molecular Biology, vol 121. Humana Press. https://doi.org/10.1385/1-59259-044-6:49
Download citation
DOI: https://doi.org/10.1385/1-59259-044-6:49
Publisher Name: Humana Press
Print ISBN: 978-0-89603-683-3
Online ISBN: 978-1-59259-044-5
eBook Packages: Springer Protocols