Typing for Transplantation Antigens and Their Genes


Safe and effective clinical transplantation requires human leucocyte antigen (HLA) typing of donors and recipients. Various immune diseases are associated with particular HLA types and HLA typing can be of diagnostic value. Finally, HLA typing is used increasingly in peptide vaccine design for cancer treatment. HLA typing is now carried out at the DNA level, which allows testing ranging from high resolution (i.e. DNA sequencing) to low resolution (which defines broadly related allele groups). The clinical application dictates the required resolution and therefore the method. Successful haematopoietic stem cell transplantation usually depends on exact HLA matching and therefore the higher resolution of testing, while for solid organ transplantation, lower resolution testing is generally sufficient. Current DNA testing methods rely on a polymerase chain reaction stage to either generate target DNA for analysis or generate sufficient specific DNA to detect directly. A systematic nomenclature for HLA has been developed and refined to account for the different methods of testing and the extreme genetic variability of the HLA system. The latest approaches to HLA typing use next generation sequencing (NGS) and, although not yet in widespread use in H&I laboratories, these may well provide definitive HLA typing for all applications in the near future.

Key Concepts

  • There are two classes of HLA with multiple members of each class (A, B and C for Class I and DR, DQ and DP for Class II).
  • HLA proteins are determined by the most polymorphic genes in humans – over 12 000 known alleles that determine nearly 9000 genetically different proteins (allotypes).
  • An HLA allele is defined by its entire DNA sequence. There is much sharing of certain polymorphic sequences between different alleles.
  • HLA nomenclature is complex but systematic.
  • Serological typing, the first approach used for HLA, discriminates alleles poorly because of the lack of a full range of antibodies.
  • Rapid HLA typing is currently best achieved using allele‐specific PCR, whereby DNA primers are used to discriminate between selected sequences in different alleles.
  • Typing using DNA probes to discriminate between sequences in different alleles is cost‐effective for large‐scale clinical testing.
  • The highest resolution of HLA typing requires genomic DNA sequencing.
  • Low‐resolution typing can give ambiguous results in heterozygotes (i.e. various allele combinations).

Keywords: HLA; transplant compatibility; polymorphism; polymerase chain reaction; DNA sequence

Figure 1. Comparison of PCR–SSOP and PCR–SSP for HLA typing. (a) PCR–SSP results are obtained quickly by PCR amplicon separation in agarose gels without further downstream processing. Unlike PCR–SSOP, positive results in this technique require simultaneous hybridisation of two primers on the same DNA strand. (b) PCR–SSOP can identify specific sequence variation, but allele resolution can be ambiguous.
Figure 2. Outline protocols of the three commonly used HLA typing methods.


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Further Reading

Arguello JR, Little AM, Bohan E, et al. (1998) High resolution HLA class I typing by reference strand mediated conformation analysis (RSCA). Tissue Antigens 52 (1): 57–66.

Bunce M, Young NT and Welsh KI (1997) Molecular HLA typing – the Brave New World. Transplantation 64: 1505–1513.

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.

Dupont B (ed) (1987) Immunobiology of HLA, vol. 1: Histocompatibility Testing. New York: Springer.

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.

Miller SA, Dykes DD and Polesky HF (1988) A simple salting out procedure for extracting DNA from human nucleated cells. Nucleic Acids Research 16: 1215.

Mitsunaga S, Tokunaga K, Kashiwase K, et al. (1998) A nested PCR‐RFLP method for high resolution typing of HLA‐A alleles. European Journal of Immunogenetics 25: 15–27.

Mizuki N, Ohno S, Sugimura K, et al. (1992) PCR‐RFLP is as sensitive and reliable as PCR‐SSO in HLA class II genotyping. Tissue Antigens 40: 100–103.

Simpson E, Rooperian D and Goulmy E (1998) Much ado about minor histocompatibility antigens. Immunology Today 19 (3): 108–112.

Zhou M, Gao D, Chai X, et al. (2015) Application of high throughput, high resolution and cost effective next‐generation sequencing‐based large‐scale HLA typing in donor registry. Tissue Antigens 85: 20–28.

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Briggs, David, and Jobson, Shirley(Sep 2015) Typing for Transplantation Antigens and Their Genes. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0001245.pub2]