Structural analysis of a human I-A homologue using a monoclonal antibody that recognizes an MB3-like specificity.

Abstract

Monoclonal antibody IVD12 was used to isolate and characterize a human Ia molecule present on B cells that generally display DR4 or DR5 phenotypes. The specificity of binding of IVD12 to human peripheral blood B cells from 75 normal individuals and 19 homozygous human lymphoblastoid B cell lines was identical to the supertypic specificity MB3 previously defined. Furthermore, IVD12-reactivity was shown to segregate with HLA in three informative families. In each family, individuals positive for IVD12 binding were also positive for DR4 or DR5. Using IVD12, a molecule has been isolated from the homozygous cell line PRIESS (DR4/4) and has been shown by amino acid sequence analysis to be homologous to the murine I-A and human HLA-DS molecules. These findings suggest that the MB3 specificity is found on a molecule encoded by loci distinct from those loci which encode HLA-DR molecules. This molecule represents the third family of HLA-D region molecules isolated from the cell line PRIESS. Both HLA-DR and HLA-SB molecules from this cell line were previously shown by amino acid sequence analysis to be I-E-like but distinct from one another. Collectively, these data provide evidence that the HLA-D region contains at least six loci encoding distinct alpha and beta chains for the HLA-SB, HLA-DR, and HLA-DS molecules.

L cells expressing DQ molecules of the DR3 and DR4 haplotypes: reactivity patterns with mAbs

“…(TA10 corresponds to DQ7 allele). The reactivity of the allele-specific anti-DQ mAbs C1, IVD12, PLM12, and GS200.1 on L transfectants was consistent with previous analyses (Knowles et al 1989;Griles et al 1983;Radka et al 1989). However, the DQw3-specific mAb PLM12 reacted with the DQAI*0301/ DQBI*0302 dimer but it did not react with the DQAI*OSO1/DQBI*0302 dimer, suggesting that this antibody recognizes the c~[~ complex and that the structural differences of the ~z chains may be responsible for the recognition pattern.…”

“…The mAbs used in this study were TAL-1B5 (Adams et al 1983), TU22 (Shaw et al 1985), HK19 (Shipp et al 1983), IIB3 (Koning et al 1985), Leu 10 (Brodsky 1984), BT3/4 (Corte et al 1981), GSP17.1 (Nelson and Joffe 1985), C1 (Knowles et al 1989), IVD12 (Griles et al 1983), PLM12 (Knowles et al 1989), TU39 (Shipp et al_ 1983), XD5.All (Radka et al 1984), WR18 (Pezzutto et al 1986), GS200.1 (Radka et al 1989), GS159.1 (Radka et al 1989), GS295-8B6 (S. Radka, personal communication), SFR16-PI.2 (Radka et al 1985), SFR20-DQM (Radka et al 1987), 10D2-7 (Radka et al 1987), LB3.1 (Gorga et al 1986), and B7/21 (Royston et al 1981;Watson et al 1983). …”

Coexpression of Susceptible and Resistant HLA Class II Transgenes in Murine Experimental Autoimmune Thyroiditis: DQ8 Molecules Downregulate DR3-Mediated Thyroiditis

“…Flow cytometric analysis of peripheral blood leukocytes was used to determine the HLA-DR3 and/or HLA-DQ8 expression. Monoclonal antibodies used were: L227, anti-DRB1 [19] and IVD12, anti-DQ [20]. The polymerase chain reaction technique was used to determine the homozygous or heterozygous presence of H2A g7 , based on the absence or presence of the neo marker for Ab 0 , respectively [16].…”

HLA-DQB1 polymorphism determines incidence, onset, and severity of collagen-induced arthritis in transgenic mice. Implications in human rheumatoid arthritis.

“…Lymphocytes were isolated from freshly obtained whole blood on a Ficoll-Hypaque gradient, and were washed in phosphate buffered saline containing 1% BSA and 0.1% sodium azide. The cells were incubated with mAb specific for HLA-DQ ␣ (IVD12; [47]), HLA-DQ6 ␤ (L227; [48] Assessment of CII-specific T cell proliferation. Mice were primed with 200 g of bovine CII emulsified 1:1 in CFA ( Mycobacterium tuberculosis H37 Ra; Difco Laboratories Inc., Detroit, MI), 100 g intradermally at the base of the tail, and 50 g in each of the hind footpads.…”