Anti-Thy-1 antibody responses evoked by Thy-1 antigen expressed in transfected mouse mastocytoma cells and rat fibroblast.

Abstract

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.

• Authors: K Isobe, A Fortunato, V Giguere, F Grosveld, N A Mitchison
• Journal: Immunology (Immunology) Vol. 56 Issue 3 Pg. 505-12 (Nov 1985) ISSN: 0019-2805 [Print] England
• PMID: 2867029 (Publication Type: Journal Article)

Chemical References

• Antibodies, Monoclonal
• Antibodies, Neoplasm
• Antigens, Surface
• Isoantibodies
• Thy-1 Antigens
• anti-Thy antibody

Topics

• Animals
• Antibodies, Monoclonal (immunology)
• Antibodies, Neoplasm (immunology)
• Antigens, Surface (immunology)
• Base Sequence
• Fibroblasts (immunology)
• Genes, MHC Class II
• HeLa Cells
• Hemolytic Plaque Technique
• Isoantibodies (biosynthesis, immunology)
• L Cells
• Mast-Cell Sarcoma (immunology)
• Mice
• Mice, Inbred AKR
• Mice, Inbred CBA
• Rats
• Thy-1 Antigens
• Transfection