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KMID : 0357319930280030229
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Journal of the Korean Society for Microbiology
1993 Volume.28 No. 3 p.229 ~ p.237
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Detection of Human CTLA-4 by using Anti-petide Antibody
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정용훈/Bo Dupont
조양자/임석원/박장환/최 용
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Abstract
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CTLA-4 was first isolated as a gene by reverse genetics using a cDNA subtractive hybridization followed by serial differential hybridization screenings. CTLA-4 belongs to lymphocyte activation antigens which express transiently on T cell surdface
when
it is activated. The nature of thisa molecule has made it difficult ot develop anti-CTLA-4 specific antibody and to characterize protein counter part of this molecule. Here we describe a method to produce anti-human CTLA-4 antibody by immunizing
with
synthetic peptide coupled Keyhole Limpet hemocyanin. In western blot 3 antipeptide antibodies showed titers between 1:800 & 1:1,600. These 3 different anti-poptide antibodies recognized 34kD annd as huan CTLA-4 protein in western blot. It was
found
that
humnaNK and CD8+ T cell clone, LOH and NBL77 could produce CTLA-4. Polyclonal mitogenic combinations, PHA, PMA+ionomycin, SEB, anti-CD2 antibodies 9.1+9.6, and MLC with allogenic EBV transformed B cell, could iduce CTLA-4 expression in peripheral
blood
lymphocyte which peaked on day 3 by PHA and anti CD2 stimulation and on day 5 by PMA+ionomycin and SEB stimulation. And IL-2 was able to increase CTLA-4 induction in MLC.
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KEYWORD
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