[Molecular cloning and three-dimensional structure modelling of HLA-DRalpha * 0101, DRbeta1 * 04051 and DRbeta1 *...

To study the correlation between the differences of three-dimensional structures among different HLA loci and their functions, and to investigate the potential usage of HLA conformational differences in predicting the occurrence of graft-versus-host disease (GVHD).

Total RNA were extracted from human peripheral blood, and reverse-transcription PCR (RT-PCR) were used to amplify the whole gene sequences of HLA-DR loci, and the PCR products were linked to vector and transformed to E.coli, and then sequencing. HLA-DR second exon amino acid sequences were inputted to computer, through internet to SWISS-MODELLING molecular modelling server to model their three-dimensional structure.

The whole HLA-DR loci sequences were successfully reversibly transcript from 2 normal unrelated donors. The sequencing results were completely identical to the published sequences of HLA-DRalpha * 0101, DRbeta1 * 04051 and DRbeta1 * 07011. The modelling results of the later two indicated that there were significant differences between their structures.

The sequencing analysis of the whole HLA loci could clearly designate HLA allelic types, and three-dimensional structure modelling could directly identify the differences among them. Therefore, this study provide experimental foundation to the study of HLA structure and function, and uo the study of predicting GVHD after HLA incompletely matched hematopoietic stem cell transplantation (HSCT).