Characterization of three mouse monoclonal antibodies that react with high-molecular-mass glycopeptides isolated from F9 mouse teratocarcinoma cells.
Three IgM mouse monoclonal antibodies, NL-9, Thy-22, and HL-5, which were produced primarily against human hematopoietic cells, were tested for their reactivity with various mouse cell lines and were found to react predominantly with mouse embryonal carcinoma cells. Thy-22 reacted with 2-cell-stage mouse embryos, whereas the other two antibodies were not reactive at this stage. All three antibodies, however, reacted with 8-cell-stage embryos. At the blastocyst stage, Thy-22 reacted with the entire surface of the trophectoderm cells, whereas the reactivity of NL-9 and HL-5 was weaker and was polarized on the mural trophectoderm. Immunohistological examination of 6th-day mouse embryos using anti-complement immunofluorescence demonstrated that the embryonic ectoderm was positive for all three antibodies: the reaction of NL-9 and Thy-22 was uniformly distributed over these cells, whereas HL-5 predominantly stained the luminal aspects of the cells lining the proamniotic cavity. Visceral-endoderm cells and trophoblastic cells were positive with all three monoclonal antibodies, whereas the parietal endoderm, extraembryonic ectoderm, and ectoplacental cone were negative. In 19th-day fetuses and adult tissues, certain epithelial cells were stained by these three antibodies. The biochemical nature of the antigens detected was also investigated. Farr's assay showed that both NL-9 and Thy-22 precipitated approximately 10% of the high-molecular-mass glycopeptides isolated from F9 cells, while HL-5 reacted with about 5% of these glycopeptides. The reactivity of the three antibodies against the glycopeptides was completely inhibited by the presence of X-hapten-conjugated silica.(ABSTRACT TRUNCATED AT 250 WORDS)
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification*; D000918:Antibody Specificity; D000954:Antigens, Surface / Q000276:immunology; D001667:Binding, Competitive; D002241:Carbohydrates / Q000494:pharmacology; D002460:Cell Line; D004622:Embryo, Mammalian / Q000276:immunology; D000939:Epitopes / Q000032:analysis; D005333:Fetus / Q000276:immunology; D006020:Glycopeptides / Q000032:analysis / Q000276:immunology*; D006412:Hematopoietic Stem Cells / Q000276:immunology; D051379:Mice; D008970:Molecular Weight; D013724:Teratoma / Q000032:analysis* / Q000276:immunology
D000911:Antibodies, Monoclonal; D000954:Antigens, Surface; D002241:Carbohydrates; D000939:Epitopes; D006020:Glycopeptides