TY - JOUR KW - Amino Acid Sequence KW - Animals KW - Antigen-Presenting Cells KW - Antigens, Bacterial KW - Clone Cells KW - DNA KW - Genes, MHC Class II KW - HLA-D Antigens KW - HLA-DR Antigens KW - HLA-DR2 Antigen KW - Humans KW - L Cells (Cell Line) KW - leprosy KW - Mice KW - Molecular Sequence Data KW - Mycobacterium leprae KW - Recombinant Fusion Proteins KW - T-Lymphocytes AU - Wilkinson D AU - Vries R R AU - Madrigal J A AU - Lock C B AU - Morgenstern J P AU - Trowsdale J AU - Altmann D M AB -

We have used DNA-mediated gene transfer to express HLA class II molecules in mouse L cells for serological, biochemical, and functional analysis. cDNA clones encoding the DR2 beta a and DR2 beta b products of the DR2Dw2 haplotype were subcloned into a mouse Moloney leukemia virus-based expression vector (pJ4) and transfected separately into mouse L cells together with a HLA-DR alpha/pJ4 construct. These transfectants have allowed differential analysis of the two DR2 beta products in a manner normally prohibited by the concomitant expression seen in B cells. Two-dimensional SDS-PAGE analysis of the transfectants defines the more acidic beta chain as the product of the DR2 beta a sequence, and the more basic chain as the product of the DR2 beta b sequence. The LDR2a transfectants present antigen efficiently to M.leprae-specific T cell clones and are capable of presenting synthetic peptide, 65-kD recombinant mycobacterial antigen and M.leprae. Of the DR2Dw2-restricted T cell clones we have tested, all use the DR2 beta a chain as their restriction element. Inhibition studies with mAbs demonstrate the dependence of presentation by the transfectant on class II and CD4, while mAbs against LFA-1, which substantially inhibit presentation by B-lymphoblastoid cell lines, do not inhibit transfectant presentation.

BT - The Journal of experimental medicine C1 - http://www.ncbi.nlm.nih.gov/pubmed/3128633?dopt=Abstract DA - 1988 Apr 01 DO - 10.1084/jem.167.4.1442 IS - 4 J2 - J. Exp. Med. LA - eng N2 -

We have used DNA-mediated gene transfer to express HLA class II molecules in mouse L cells for serological, biochemical, and functional analysis. cDNA clones encoding the DR2 beta a and DR2 beta b products of the DR2Dw2 haplotype were subcloned into a mouse Moloney leukemia virus-based expression vector (pJ4) and transfected separately into mouse L cells together with a HLA-DR alpha/pJ4 construct. These transfectants have allowed differential analysis of the two DR2 beta products in a manner normally prohibited by the concomitant expression seen in B cells. Two-dimensional SDS-PAGE analysis of the transfectants defines the more acidic beta chain as the product of the DR2 beta a sequence, and the more basic chain as the product of the DR2 beta b sequence. The LDR2a transfectants present antigen efficiently to M.leprae-specific T cell clones and are capable of presenting synthetic peptide, 65-kD recombinant mycobacterial antigen and M.leprae. Of the DR2Dw2-restricted T cell clones we have tested, all use the DR2 beta a chain as their restriction element. Inhibition studies with mAbs demonstrate the dependence of presentation by the transfectant on class II and CD4, while mAbs against LFA-1, which substantially inhibit presentation by B-lymphoblastoid cell lines, do not inhibit transfectant presentation.

PY - 1988 SP - 1442 EP - 58 T2 - The Journal of experimental medicine TI - Analysis of HLA-DR glycoproteins by DNA-mediated gene transfer. Definition of DR2 beta gene products and antigen presentation to T cell clones from leprosy patients. VL - 167 SN - 0022-1007 ER -