Abstract
Human histocompatibility antigens HLA-A, HLA-B, and HLA-C are a complex of two noncovalently associated subunits: a heavy chain glycoprotein (alpha) carrying the genetic polymorphism and an invariant light chain, beta 2-microglobulin (beta 2m). Upon incubation of papain-solubilized HLA with radiolabeled urinary beta 2m, the latter is incorporated into HLA, where it substitutes for the preexisting beta 2m that has dissociated from the complex. The association-dissociation equilibrium that governs this beta 2m exchange reaction was investigated and found to be characterized by a long lifetime of the complex (half-life of 80 min at 37 degrees C) and a relatively low Kd (4 nM). The beta 2m exchange was used as the basis of a radioimmunoassay for HLA antigens with radiolabeled beta 2m as a unique label for all HLA specificities. In a similar fashion, radiolabeled beta 2m can be incorporated into HLA at the cell surface. Although the process is slower and less extensive than in solution, it can be used as a means to tag cells with specific probes for HLA antigens.
Full text
PDF![5834](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a71/411746/a551316b21c2/pnas00011-0428.png)
![5835](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a71/411746/d2b620c145e2/pnas00011-0429.png)
![5836](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a71/411746/2fae248c36a5/pnas00011-0430.png)
![5837](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a71/411746/56549f688759/pnas00011-0431.png)
![5838](https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2a71/411746/702b94f7f184/pnas00011-0432.png)
Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Anderson C. L., Kubo R. T., Grey H. M. Studies on the cytophilic properties of human beta2 microglobulin. J Immunol. 1975 Mar;114(3):997–1000. [PubMed] [Google Scholar]
- Arce-Gomez B., Jones E. A., Barnstable C. J., Solomon E., Bodmer W. F. The genetic control of HLA-A and B antigens in somatic cell hybrids: requirement for beta2 microglobulin. Tissue Antigens. 1978 Feb;11(2):96–112. doi: 10.1111/j.1399-0039.1978.tb01233.x. [DOI] [PubMed] [Google Scholar]
- Bach F. H., Bach M. L., Sondel P. M. Differential function of major histocompatibility complex antigens in T-lymphocyte activation. Nature. 1976 Jan 29;259(5541):273–281. doi: 10.1038/259273a0. [DOI] [PubMed] [Google Scholar]
- Bach F. H., van Rood J. J. The major histocompatibility complex - genetics and biology (third of three parts). N Engl J Med. 1976 Oct 21;295(17):927–936. doi: 10.1056/NEJM197610212951705. [DOI] [PubMed] [Google Scholar]
- Barnstable C. J., Jones E. A., Crumpton M. J. Isolation, structure and genetics of HLA-A, -B, -C and -DRw (Ia) antigens. Br Med Bull. 1978 Sep;34(3):241–246. doi: 10.1093/oxfordjournals.bmb.a071504. [DOI] [PubMed] [Google Scholar]
- Bolton A. E., Hunter W. M. The labelling of proteins to high specific radioactivities by conjugation to a 125I-containing acylating agent. Biochem J. 1973 Jul;133(3):529–539. doi: 10.1042/bj1330529. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Brown J. L., Nathenson S. G. Structural differences between parent and mutant H-2K glycoproteins from two H-2K gene mutants: b6.c-h-2ba (Hzl) and B6-H-2bd (M505). J Immunol. 1977 Jan;118(1):98–102. [PubMed] [Google Scholar]
- Cejka J., Kithier K. Isolation and characterization of 2 -microglobulin from urine of kidney transplant and multiple myeloma patients. Arch Biochem Biophys. 1972 Dec;153(2):854–857. doi: 10.1016/0003-9861(72)90409-2. [DOI] [PubMed] [Google Scholar]
- Engelhard V. H., Strominger J. L., Mescher M., Burakoff S. Induction of secondary cytotoxic T lymphocytes by purified HLA-A and HLA-B antigens reconstituted into phospholipid vesicles. Proc Natl Acad Sci U S A. 1978 Nov;75(11):5688–5691. doi: 10.1073/pnas.75.11.5688. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Fellous M., Günther E., Kemler R., Wiels J., Berger R., Guenet J. L., Jakob H., Jacob F. Association of the H-Y male antigen with beta2-microglobulin on human lymphoid and differentiated mouse teratocarcinoma cell lines. J Exp Med. 1978 Jul 1;148(1):58–70. doi: 10.1084/jem.148.1.58. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Helenius A., Morein B., Fries E., Simons K., Robinson P., Schirrmacher V., Terhorst C., Strominger J. L. Human (HLA-A and HLA-B) and murine (H-2K and H-2D) histocompatibility antigens are cell surface receptors for Semliki Forest virus. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3846–3850. doi: 10.1073/pnas.75.8.3846. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Kessler S. W. Rapid isolation of antigens from cells with a staphylococcal protein A-antibody adsorbent: parameters of the interaction of antibody-antigen complexes with protein A. J Immunol. 1975 Dec;115(6):1617–1624. [PubMed] [Google Scholar]
- Klareskog L., Banck G., Forsgren A., Peterson P. A. Binding of HLA antigen-containing liposomes to bacteria. Proc Natl Acad Sci U S A. 1978 Dec;75(12):6197–6201. doi: 10.1073/pnas.75.12.6197. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
- Maron R., Cohen I. R. Mutation at H-2K locus influences susceptibility to autoimmune thyroiditis. Nature. 1979 Jun 21;279(5715):715–716. doi: 10.1038/279715a0. [DOI] [PubMed] [Google Scholar]
- McCune J. M., Humphreys R. E., Yocum R. R., Strominger J. L. Enhanced representation of HL-A antigens on human lymphocytes after mitogenesis induced by phytohemagglutinin or Epstein-Barr virus. Proc Natl Acad Sci U S A. 1975 Aug;72(8):3206–3209. doi: 10.1073/pnas.72.8.3206. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Morello D., Gachelin G., Dubois P., Tanigaki N., Pressman D., Jacob F. Absence of reaction of a xenogenic anti-H-2 serum with mouse embryonal carcinoma cells. Transplantation. 1978 Aug;26(2):119–125. doi: 10.1097/00007890-197808000-00012. [DOI] [PubMed] [Google Scholar]
- Munro A., Bright S. Products of the major histocompatibility complex and their relationship to the immune response. Nature. 1976 Nov 11;264(5582):145–152. doi: 10.1038/264145a0. [DOI] [PubMed] [Google Scholar]
- Munro A., Waldmann H. The major histocompatibility system and the immune response. Br Med Bull. 1978 Sep;34(3):253–258. doi: 10.1093/oxfordjournals.bmb.a071506. [DOI] [PubMed] [Google Scholar]
- Parham P., Alpert B. N., Orr H. T., Strominger J. L. Carbohydrate moiety of HLA antigens. Antigenic properties and amino acid sequences around the site of glycosylation. J Biol Chem. 1977 Nov 10;252(21):7555–7567. [PubMed] [Google Scholar]
- Parham P., Terhorst C., Herrmann H., Humphreys R. E., Waterfield M. D., Strominger J. L. Immunological and chemical purity of papain-solubilized HL-A antigens. Proc Natl Acad Sci U S A. 1975 Apr;72(4):1594–1598. doi: 10.1073/pnas.72.4.1594. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Rice R. H., Means G. E. Radioactive labeling of proteins in vitro. J Biol Chem. 1971 Feb 10;246(3):831–832. [PubMed] [Google Scholar]
- Robb R. J., Strominger J. L. Rapid purification of detergent-solubilized HLA antigen by affinity chromatography employing anti-beta2-microglobulin serum. J Biol Chem. 1976 Sep 10;251(17):5427–5428. [PubMed] [Google Scholar]
- Robb R. J., Terhorst C., Strominger J. L. Sequence of the COOH-terminal hydrophilic region of histocompatibility antigens HLA-A2 and HLA-B7. J Biol Chem. 1978 Aug 10;253(15):5319–5324. [PubMed] [Google Scholar]
- Shearer G. M., Schmitt-Verhulst A. M. Major histocompatibility complex restricted cell-mediated immunity. Adv Immunol. 1977;25:55–91. doi: 10.1016/s0065-2776(08)60931-1. [DOI] [PubMed] [Google Scholar]
- Thomson D. M., Tataryn D. N., O'Connor R., Rauch J., Friedlander P., Gold P., Shuster J. Evidence for the expression of human tumor-specific antigens associated with beta2-microglobulin in human cancer and in some colon adenomas and benign breast lesions. Cancer Res. 1979 Feb;39(2 Pt 2):604–611. [PubMed] [Google Scholar]
- Vitetta E. S., Capra J. D. The protein products of the murine 17th chromosome: genetics and structure. Adv Immunol. 1978;26:147–193. doi: 10.1016/s0065-2776(08)60230-8. [DOI] [PubMed] [Google Scholar]
- Ziegler A., Harrison S. C., Leberman R. The minor proteins in tomato bushy stunt and turnip crinkle viruses. Virology. 1974 Jun;59(2):509–515. doi: 10.1016/0042-6822(74)90460-7. [DOI] [PubMed] [Google Scholar]