Tissue Typing for Transplantation Antigens

Robert W Vaughan, Guy's Hospital, London, UK

Published online: April 2001

DOI: 10.1038/npg.els.0001245

Tissue typing usually refers to methods for detecting human leucocyte antigens (HLAs), although it is applicable to other species. The major practical need for tissue typing is to facilitate the transplantation of organs.

Keywords: tissue typing; HLA; transplantation

Figure 1. Use of linkage in the analysis of human leucocyte antigen (HLA) polymorphism: an advantage of PCR-SSP over PCR-SSO. Polymerase chain reaction–sequence-specific priming (PCR-SSP) over PCR–sequence-specific oligonucleotide probe (PCR-SSOP).

(a) PCR-SSOP. In this example a gene to be analysed has two polymorphic sites separated by 100 nucleotides. Each polymorphic site has two variants, in this individual labelled A/C and B/D. Although DNA is double stranded, only one strand from each chromosome is represented for clarity. The first stage is a generic PCR priming reaction across the site shown. The product is then blotted on to a membrane; probes complementary to the polymorphic sites at A, B, C and D are used and all combine with the heterozygote DNA sample represented to produce four possible combinations (or alleles). It is not known whether A is linked to (on the same chromosome as) B or D, and similarly C may be linked to B or D. The alleles are defined only as AB and CD or AD and CB.

(b) PCR-SSP. PCR primers can be made to amplify exclusively the polymorphism at A (___A) and C (___C), with a reverse primer for B (B___ ) and D (D___ ). If PCR-SSP reactions are set up using all combinations of these primers, products will be seen only when the polymorphic sites are linked. Only reactions (1) and (4) will give a product, and the alleles are correctly identified as AB and CD. In this way a series of mutations can be placed on one chromosome. A limitation to this type of analysis is the range of nucleotides over which it is possible to obtain a PCR reaction. There are practical difficulties in amplifying more than about 1000 nucleotides.
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 References
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    proceedings Bignon JD, Fernandez-Vina MA, Cheneau ML et al. (1997) HLA DNA class II typing by PCR-SSOP: 12th International Histocompatibility Workshop experience. In: Charron D (ed.) Proceedings of the Twelfth International Histocompatibility Workshop and Conference, vol. 1, pp. 21–25. Sevres, France: EDK.
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    book Inoko H and Ota M (1993) "PCR-RFLP". In: Hui KM and Bidwell JL (eds) Handbook of HLA Typing Techniques, pp. 9–70. Raton, FL: CRC Press.
    proceedings Kennedy LJ, Poulton KV, Thomson W et al. (1997) HLA class I DNA typing using sequence specific oligonucleotide probes (SSOP). In: Charron D (ed.) Proceedings of the Twelfth International Histocompatibility Workshop and Conference, vol. 1, pp. 216–225. Sevres, France: EDK.
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 Further Reading
    book Bignon JD, Fernandez-Vina MA, Cheneau ML et al. (1997) "HLA DNA class II typing by PCR-SSOP: 12th International Histocompatibility Workshop experience". In: Charron D (ed.) Proceedings of the Twelfth International Histocompatibility Workshop and Conference, vol. 1, pp. 21–25. Sevres, France: EDK.
    Bunce M, Young NT and Welsh KI (1997) Molecular HLA typing – the Brave New World. Transplantation 64: 1505–1513.
    book Darke C and Dyer P (1993) "Clinical HLA typing by cytotoxicity". In: Dyer P and Middleton D (eds) Histocompatibility Testing. A Practical Approach. Oxford: IRL Press at Oxford University Press.
    book Dupont B (ed.) (1987) Immunobiology of HLA, vol. 1. "Histocompatibility Testing". New York: Springer.
    proceedings Kennedy LJ, Poulton, KV, Thomson W et al. (1997) HLA class I DNA typing using sequence specific oligonucleotide probes (SSOP). In: Charron D (ed.) Proceedings of the Twelfth International Histocompatibility Workshop and Conference, vol. 1, pp. 216–225. Sevres, France: EDK.
    Simpson E, Rooperian D and Goulmy E (1998) Much ado about minor histocompatibility antigens. Immunology Today 19(3): 108–112.

Related Sub-Topics:

Diagnostics, Therapeutics and Methods | Antigens | Transplantation
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Article Title: Tissue Typing for Transplantation Antigens
 
How to Cite close
Vaughan, Robert W(Apr 2001) Tissue Typing for Transplantation Antigens. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1038/npg.els.0001245]