Anti-Thy-1 antibody responses evoked by Thy-1 antigen expressed in transfected mouse mastocytoma cells and rat fibroblast.
The mouse genomic Thy-1.1 gene was isolated from a phage library constructed from AKR/J (Thy-1.1) mouse DNA. Partial nucleotide sequence analysis of the coding region showed that it has only a single nucleotide difference from the Thy-1.2 gene, namely that amino acid 89 reads CGA (Arg) in Thy-1.1 and CAA (Glu) in Thy-1.2, corresponding to the amino acid substitutions previously identified. It was subcloned into an SV-40 derived vector for transfection. Transient transfection into HeLa cells gave 2% positive staining by immunofluorescence. The gene in this vector was also co-transfected into L cells and mastocytoma cells (both of Thy-1.2 strain origin) together with the Agpt gene. L-cell clones selected for transformation proved almost negative for Thy-1.1 expression, and any positive clones gradually lost Thy-1.1 antigen expression in culture. On the contrary, all clones of mastocytoma transformants gave a high level of expression after more than 3 months in culture. The mastocytoma transformants were used to study the immunogenicity of Thy-1.1 molecules expressed on transfected cells. They evoked clear anti-Thy-1.1 plaque-forming cell (PFC) responses both in vivo and in vitro. The mastocytoma transformants also proved able to induce a T-dependent anti-Thy-1.1 antibody response in a cell transfer experiment. The immunogenicity of Thy-1.2 molecules on rat fibroblasts was also studied after transfection with a Thy-1.2 gene cosmid. Although Thy-1.2 expression was very low, these transfectants elicited a clear anti-Thy-1.2 PFC response from AKR spleen cells hyperimmunized against CBA thymocytes.
|Keywords||0 (Antibodies, Monoclonal), 0 (Antibodies, Neoplasm), 0 (Antigens, Surface), 0 (Antigens, Thy-1), 0 (Isoantibodies), 0 (anti-Thy antibody), Animals, Antibodies, Monoclonal/*immunology, Antibodies, Neoplasm/immunology, Antigens, Surface/*immunology, Antigens, Thy-1, Base Sequence, Fibroblasts/immunology, Genes, MHC Class II, Hela Cells, Hemolytic Plaque Technique, Isoantibodies/biosynthesis/*immunology, L Cells (Cell Line), Mast-Cell Sarcoma/immunology, Mice, Mice, Inbred AKR, Mice, Inbred CBA, Rats, Transfection|
Isobe, K., Fortunato, A., Giguere, V., Grosveld, F.G., & Mitchison, N.A.. (1985). Anti-Thy-1 antibody responses evoked by Thy-1 antigen expressed in transfected mouse mastocytoma cells and rat fibroblast.. Immunology: the journal of cells, molecules, system and technologies, 56, 505–512. Retrieved from http://hdl.handle.net/1765/2385