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jem Home » 1993 Archive » 1 February » 177 (2): 351
Article

Production and characterization of an Mls-1-specific monoclonal antibody.

N Mohan, D Mottershead, M Subramanyam, U Beutner, B T Huber
N Mohan
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D Mottershead
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M Subramanyam
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U Beutner
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B T Huber
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DOI: 10.1084/jem.177.2.351 | Published February 1, 1993
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Abstract

Superantigens (SAGs) represent a new class of antigens, characterized as T cell receptor (TCR) V beta-reactive elements. Bacterial toxins constitute the major group of exogenous SAGs, while the mouse mammary tumor virus (MMTV)-encoded Mls molecules represent the endogenous SAGs. Mls-1 is the prototype of the latter SAGs, because it elicits a very potent T cell stimulatory response in vitro in unprimed T cells expressing the TCR V beta 6 or 8.1 chains. In vivo, Mls-1 causes deletion of immature T cells bearing the V beta 6, 7, 8.1, or 9 chains. Although Mls-1 was functionally discovered > 20 yr ago, it has not been possible to raise antibodies against this molecule. We have previously cloned and sequenced the Mtv-7 sag gene, which encodes Mls-1. Sequence comparisons with other MMTV sag genes suggested that the polymorphic 3' end encodes the TCR V beta specificity of these SAGs. We have, therefore, immunized hamsters with a 14-amino acid peptide from the deduced COOH-terminal sequence of the Mtv-7 sag gene. We describe here the production of a monoclonal antibody (mAb), 3B12, which is peptide specific and reacts with a recombinant baculovirus product of Mtv-7 sag. This mAb blocks Mls-1-specific T cell recognition and detects the Mls-1 protein on the surface of the B cell hybridoma LBB.A, but not on LBB.11, which is an Mtv-7 loss variant of LBB.A. Transfection of the Mtv-7 sag gene into LBB.11 renders this cell functionally Mls-1+ as well as positive for 3B12 binding, confirming the specificity of this mAb. It is well documented that B cells and CD8+ T cells express T cell stimulatory Mls-1 determinants, and we show here that this functional profile correlates with the expression of MMTV-specific mRNA. However, primary lymphocytes derived from Mls-1+ mice do not stain with 3B12, even after in vitro activation with mitogens or phorbol ester.

© 1993 Rockefeller University Press
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Production and characterization of an Mls-1-specific monoclonal antibody.
N Mohan, D Mottershead, M Subramanyam, U Beutner, B T Huber
Journal of Experimental Medicine Feb 1993, 177 (2) 351-358; DOI: 10.1084/jem.177.2.351

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The Journal of Experimental Medicine: 215 (4)

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April 2, 2018
Volume 215, No. 4

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