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A matrix approach to human class II histocompatibility antigens: Reactions of four monoclonal antibodies with the products of nine haplotypes

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Abstract

Three putative HLA-DC-specific monoclonal antibodies, Genox 3.53, BT3/4 and anti-Leu-10, and the HLA-DR-specific antibody, L243, were compared. Their interactions with molecules from homozygous cell lines expressing DR types 1 through 9 were studied. Indirect radioimmunoassays on 29 cell lines demonstrated that Genox 3.53 reactivity correlated with DR1, 2, 6; BT3/4 reactivity correlated with DR 1, 2, 4, 6, 8; and anti-Leu-10 reactivity correlated with DR1, 2, 4, 5, 6, 8, and 9. In addition, one of six DR3-positive cells and three DR7, DRw10-positive cells reacted with anti-Leu-10 and one of two DR9-positive cells reacted with BT3/4. Binding studies with soluble antigen and competitive radioimmunoassays demonstrated that all three antibodies reacted with the DC1 molecule. Preincubation with BT3/4 blocked anti-Leu-10 binding; Genox 3.53 and L243 did not. Genox 3.53 and L243 were only blocked by themselves. Serial immuno-precipitation showed anti-Leu-10 reacted with non-HLA-DR molecules from cells expressing DR types 1–6, 8 and 9. However, the molecules precipitated by anti-Leu-10 were characteristic class II major histocompatibility complex (MHC) molecules. Their α and β chains were of lower apparent molecular weight than the DR chains in all haplotypes. They also comigrated with the DC1 molecule precipitated by Genox 3.53. Serial immuno-precipitation also showed that anti-Leu-10 removed all Genox 3.53 reactive molecules from cell lysates, but Genox 3.53 removed only a subset of anti-Leu-10 reactive molecules. These studies show Genox 3.53, BT3/4, and anti-Leu-10 react exclusively with class II MHC molecules that are not HLA-DR, and most likely define different polymorphisms of DC molecules, the human equivalent of mouse I-A products.

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Abbreviations

BSA:

bovine serum albumin

PBS:

phosphate-buffered saline, pH 7.4

RIA:

radioimmunoassays

125I-RAM:

125I-labeled F(ab′)2 of rabbit anti-mouse IgG

NP40:

Nonidet P40 OVA-LB, 0.1% ovalbumin/0.5% NP40, 10mM Tris pH 7.3, 1MM M9Cl2 0.5% phenyl methyl sulfonyl fluoride

SDS-PAGE:

sodium dodecyl sulfate-polyacrylamide gel electrophoresis

MHC:

major histocompatibility complex

KD:

kilodaltons

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  1. Becton Dickinson Monoclonal Center, 2375 Garcia Avenue, 94043, Mountain View, California

    Frances M. Brodsky

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Brodsky, F.M. A matrix approach to human class II histocompatibility antigens: Reactions of four monoclonal antibodies with the products of nine haplotypes. Immunogenetics 19, 179–194 (1984). https://doi.org/10.1007/BF00364762

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  • DOI: https://doi.org/10.1007/BF00364762

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