Construction and properties of new Lyt-congenic strains and anti-Lyt-2.2 and anti-Lyt-3.1 monoclonal antibodies |
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Authors: | Paul D Gottlieb Ann Marshak-Rothstein Karen Auditore-Hargreaves Dorothy B Berkoben Deborah A August Roger M Rosche James D Benedetto |
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Institution: | (1) Department of Biology and Center for Cancer Research, Massachusetts Institute of Technology, 02139 Cambridge, Massachusetts |
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Abstract: | Hybridomas producing mouse monoclonal IgM antibodies specific for Lyt-2.2 and Lyt-3.1 T-cell surface alloantigens have been constructed. Cytotoxic titers of ascites fluids were found to be 10–6 or greater and no lysis of thymocytes of congenic strains bearing the alternative allele was observed at the lowest dilutions tested (12). The anti-Lyt-2.2 monoclonal antibody (HO-2.2) specifically precipiated from extracts of Lyt-2.2-positive thymocytes molecular species indistinguishable from those precipitated by conventional anti-Lyt-2.2 sera. However, by immunoprecipitation criteria (though not by cytotoxicity), the anti-Lyt-3.1 antibody (HO-3.1) demonstrated some cross-reactivity with similar molecular species from Lyt-3.1-negative thymocytes.In addition, three new strains of mice differing from existing strains in the region of theLyt-2 and4Lyt-3 loci have been constructed. They are: C.C58-Lyt-2a, Lyt-3a and C.AKR-Lyt-2a, Lyt-3a, congenic with Balb/cAn and bearingLyt-2
a
andLyt-3
a
alleles of C58/J and AKR/J, respectively; and AKR.C-Lyt-2b, Lyt-3b, congenic with AKR/J and bearing theLyt-2
b
andLyt-3
b
alleles of Balb/cJ.Abbreviations used in this paper DMSO
dimethylsulfoxide
- NP40
Nonidet P-40 detergent
- SaCI
Staphylococcus aureus, Cowan I strain
- SDS
sodium dodecyl sulfate
- PAGE
polyacrylamide gel electrophoreses
- NP-NET buffer
0.15 M NaCl, 0.005 M EDTA, 0.05 M Tris, 0.02% sodium azide, pH 7.4, containing 0.5% or 0.05% NP40 as stated in text |
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Keywords: | |
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