This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. To develop and implement MHC typing technologies for Cynomolgus macaques (Macaca fascicularis) and Indian Rhesus macaques (Macaca mulatta). We will adapt technologies from HLA typing in humans for MHC class I and class II typing in the macaque. Approximately 66 MHC class I alleles in 25 Cynomolgus macaques of Vietnamese, Chinese and Mauritian origins have been defined. 79 MHC class I sequences have been deposited into Genbank more than doubling the available MHC class I sequence data for Cynomolgus macaques in that database. PCR-SSP typing conditions have been developed for the Cynomolgus alleles Mafa-B*420101, -B*440101, -B*460101, and -B*470101. Reference strand conformation analysis (RSCA) mobility values were determined for 5 Cynomolgus MHC class I alleles. PCR-SSP typing conditions were defined for 20 Indian Rhesus MHC class I alleles and a total of 16 group specific reactions for DRB1, DRB4, DRB5, and DQB1 loci. MHC class I typing was performed for 34 Indian Rhesus macaques by PCR amplification and sequencing of clones cDNA libraries were constructed from a total of 6 Indian Rhesus macaques and their MHC class I and class II genotypes were completely defined by sequence analysis. Stable transfectant cell lines were constructed by gene transfer of Mamu-B*0103 and Mafa-B*440101.
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