Document Detail


A monoclonal mouse anti-human IL-2 receptor antibody (anti-Tac) will recognize molecules on the surface of Xenopus laevis immunocytes which specifically bind rIL-2 and are only slightly larger than the human Tac protein.
MedLine Citation:
PMID:  2354864     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
We have made visible the binding of a mouse monoclonal anti-human interleukin 2 (IL-2) receptor (anti-Tac) antibody on the surface of phytohemagglutinin (PHA)-stimulated Xenopus thymocytes using a colloidal gold-conjugated goat anti-mouse antibody and transmission electron microscopy. No binding was found when a different mouse monoclonal antibody (mAb) of the same isotype and subclass was tested, or when the anti-Tac antibody was omitted from the procedure. After metabolic radiolabeling of the IL-2 receptors with [35S]methionine using PHA-stimulated thymocytes of Xenopus laevis, the South African clawed toad, we show that a concentrated preparation of the mouse anti-human Tac antibody will immunoprecipitate a radiolabeled molecule just slightly larger than 55 kDa. Phorbol dibutyrate (PDB), an effective T cell mitogen, and cyclosporin A, an inhibitor of T cell mitogenesis in this species, are both capable of regulating the expression of this IL-2-binding molecule on Xenopus immunocytes. Here, we use the calcium ionophore A23187 to show that the relationship between IL-2 receptor expression and mitogenesis, which was previously established in X. laevis, is associated with a calcium ion flux. Flow cytometry is used for assaying alterations in epitope expression after binding the lectin-stimulated cells under test with a fluorescence (Fl*) conjugate of the anti-Tac antibody or a control mAb, which is either anti-DNP or anti-keyhole limpet hemocyanin (KLH) in specificity, but of the same mouse isotype and subclass as the anti-IL-2 receptor antibody.
Authors:
L N Ruben; L Langeberg; A Malley; R H Clothier; C Beadling; R Lee; S Shiigi
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Publication Detail:
Type:  Comparative Study; Journal Article; Research Support, U.S. Gov't, P.H.S.    
Journal Detail:
Title:  Immunology letters     Volume:  24     ISSN:  0165-2478     ISO Abbreviation:  Immunol. Lett.     Publication Date:  1990 May 
Date Detail:
Created Date:  1990-07-20     Completed Date:  1990-07-20     Revised Date:  2007-11-14    
Medline Journal Info:
Nlm Unique ID:  7910006     Medline TA:  Immunol Lett     Country:  NETHERLANDS    
Other Details:
Languages:  eng     Pagination:  117-25     Citation Subset:  IM    
Affiliation:
Department of Biology, Reed College, Portland, OR 97202-8199.
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MeSH Terms
Descriptor/Qualifier:
Animals
Antibodies, Monoclonal / immunology*
Antigens, CD4 / immunology*
Calcium / physiology
Interleukin-2 / metabolism
Lymphocyte Activation / drug effects
Mice
Phorbol 12,13-Dibutyrate / pharmacology
Receptors, Interleukin-2 / immunology*,  metabolism
Recombinant Proteins / metabolism
Signal Transduction
Species Specificity
T-Lymphocytes / drug effects,  immunology*
Xenopus laevis / immunology*
Grant Support
ID/Acronym/Agency:
AI-12846/AI/NIAID NIH HHS; RR 00163/RR/NCRR NIH HHS; RR 07168/RR/NCRR NIH HHS
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Antigens, CD4; 0/Interleukin-2; 0/Receptors, Interleukin-2; 0/Recombinant Proteins; 37558-16-0/Phorbol 12,13-Dibutyrate; 7440-70-2/Calcium

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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