Monoclonal antibodies to HLA-DP-transfected mouse L cells.

Abstract

Mouse L cells transfected with human HLA-DP (DPw4) a and f3 genes were used to make monoclonal antibodies in C3H mice. A polymorphic antibody, DPl1. Alloantisera, mainly produced by fetal maternal stimulation, were the original basis for the definition of the HLA-DR and -DQ region products and their polymorphism. It is clear that most of the serologically detected variation is in the DR and DQ 83 chains. The DP specificities were identified by cellular typing techniques and there is, so far, no corresponding serological identification. One monoclonal antibody, IRL1, has been identified, which appears to react preferentially to DPw2 and DPw3 (4). Monoclonal antibodies that are monomorphic have made a major contribution to the biochemical analysis of the HLA-D region products (5, 6), but useful polymorphic antibodies have been harder to obtain (7). A wider range of polymorphic HLA-D monoclonal antibodies, particularly for the definition of HLA-DP types, would be extremely useful both for serological typing and for biochemical and functional studies.The production by transfection of mouse L cells expressing a single human HLA-D region product, in this case HLA-DP (8), provides the basis for a novel approach to making anti-HLA class II monoclonal antibodies. If an HLA-DPexpressing L-cell transfectant is used to immunize C3H mice, the strain from which L cells were derived, then the only product the mice should react to, apart from L-cell-specific determinants, is HLA-DP. This principle is an extension of that used to make human-specific antisera (9) and monoclonal antibodies (10) by immunizing mice with human-mouse somatic cell hybrids containing a single human chromosome. In this paper, we describe the recognition of polymorphic specificities on L-cell DP transfectants and the use of these transfectants to make a polymorphic monoclonal anti-DP antibody with, surprisingly, specificity for the a chain of DPw4 and to an apparently lesser extent, DPw2.

First report on the antibody verification of HLA-DR, HLA-DQ and HLA-DP epitopes recorded in the HLA Epitope Registry

“…Although there are several reports about DPA-reactive antibodies in sera from transplant patients [65,[73][74][75], there is at present insufficient documentation which DPA epitopes are specifically verified.…”

Southern blot analysis of HLA-DP gene polymorphisms in Caucasoid rheumatoid arthritis (RA) patients and controls

“…Despite considerable efforts to DP type Caucasoids using PLT (Shaw et al 1981, Pawelec et al 1982, Moen et al 1983, Wank and Schendel 1984, Morling et al 1984, the lack of alternative methods is probably responsible for the paucity of information on DP allele frequencies in non-Caucasoid populations, or within disease groups. However, transfection of DPA1 and DPB1 genes into mouse L cells , and the subsequent immunization of these transfectants into mice has produced new serological DPtyping reagents (Heyes et al 1986). Similarly, Southern blot analysis or genotyping of genomic DNA with DPA and DPB gene probes has shown strong correlations between restriction fragment length polymorphisms (RFLPs) and specific DP alleles (Hyldig-Nielsen et al 1987, Bodmer et al 1987, Mitsuishi et al 1987, Maeda et al 1988.…”

Detection of HLA‐DP serological allodeterminants by the use of radioiodinated DP molecules

“…The Butterfield allospecificity may be the same detected by the mAb DPll.l on mouse cells transfected either with DPw2 or DPw4 genes [25]. The DP1l.l determinant has been shown to reside on the a subunit.…”