New HLA-A, -B, and -C locus-specific primers for PCR amplification from cDNA: application in clinical immunology.
The individual cellular immune response to intracellular antigens is modeled by the highly polymorphic major histocompatibility complex (HLA) class I molecules. The epitopes presented and the T cell repertoire that recognizes them depend on the HLA constitution of the individual. Therefore, to monitor and to modify an individual's HLA class I-driven cellular immune response, it is necessary to know the HLA class I alleles of the person and the possible epitopes of the target antigen presented by those alleles. In particular, this is necessary in order to design peptide-based vaccines and immune therapies for the treatment of diseases caused by viruses, intracellular parasites or cancer, and to monitor the immune response during those treatments. We describe a new set of HLA-A, -B, and -C locus-specific primers for the polymerase chain reaction (PCR) amplification of the whole coding sequence of these genes from complementary DNA (cDNA). We describe their use for typing and for the production of a library of recombinant HLA class I genes. We discuss two downstream applications of this gene collection: production of soluble HLA molecules and discovery of new epitopes.
Bettinotti
Maria P
MP
Clinical Center, National Institutes of Health, Bethesda, MD 20892-1184, USA. mbettinotti@dtm.cc.nih.gov
Hadzikadic
Lejla
L
Ruppe
Erin
E
Dhillon
Gitika
G
Stroncek
David S
DS
Marincola
Francesco M
FM
D017931:DNA Primers; D018076:DNA, Complementary / Q000378:metabolism*; D015723:Gene Library; D005805:Genes, MHC Class I; D005838:Genotype; D006680:HLA Antigens / Q000235:genetics*; D006801:Humans; D007158:Immunologic Techniques; D016133:Polymerase Chain Reaction / Q000379:methods*
D017931:DNA Primers; D018076:DNA, Complementary; D006680:HLA Antigens