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. 1985 Nov;56(3):505–512.

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

K Isobe, A Fortunato, V Giguere, F Grosveld, N A Mitchison
PMCID: PMC1453741  PMID: 2867029

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.

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Selected References

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  1. Campbell D. G., Williams A. F., Bayley P. M., Reid K. B. Structural similarities between Thy-1 antigen from rat brain and immunoglobulin. Nature. 1979 Nov 15;282(5736):341–342. doi: 10.1038/282341a0. [DOI] [PubMed] [Google Scholar]
  2. Clark E. A., Lake P., Favila-Castillo L. Modulation of Thy-1 alloantibody responses: donor cell-associated H-2 inhibition and augmentation without recipient Ir gene control. J Immunol. 1981 Nov;127(5):2135–2140. [PubMed] [Google Scholar]
  3. Cohen F. E., Novotný J., Sternberg M. J., Campbell D. G., Williams A. F. Analysis of structural similarities between brain Thy-1 antigen and immunoglobulin domains. Evidence for an evolutionary relationship and a hypothesis for its functional significance. Biochem J. 1981 Apr 1;195(1):31–40. doi: 10.1042/bj1950031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Evans G. A., Ingraham H. A., Lewis K., Cunningham K., Seki T., Moriuchi T., Chang H. C., Silver J., Hyman R. Expression of the Thy-1 glycoprotein gene by DNA-mediated gene transfer. Proc Natl Acad Sci U S A. 1984 Sep;81(17):5532–5536. doi: 10.1073/pnas.81.17.5532. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fuji H., Zaleski M., Milgrom F. Immune response to alloantigens of thymus studied in mice with plaque assay. J Immunol. 1971 Jan;106(1):56–64. [PubMed] [Google Scholar]
  6. Giguére V., Isobe K., Grosveld F. Structure of the murine Thy-1 gene. EMBO J. 1985 Aug;4(8):2017–2024. doi: 10.1002/j.1460-2075.1985.tb03886.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Grosveld F. G., Lund T., Murray E. J., Mellor A. L., Dahl H. H., Flavell R. A. The construction of cosmid libraries which can be used to transform eukaryotic cells. Nucleic Acids Res. 1982 Nov 11;10(21):6715–6732. doi: 10.1093/nar/10.21.6715. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Grosveld G. C., de Boer E., Shewmaker C. K., Flavell R. A. DNA sequences necessary for transcription of the rabbit beta-globin gene in vivo. Nature. 1982 Jan 14;295(5845):120–126. doi: 10.1038/295120a0. [DOI] [PubMed] [Google Scholar]
  9. Isobe K. I., Nakashima I., Nagase F., Kato N., Mizoguchi K., Kawashima K., Lake P. Cellular mechanism of primary anti-Thy-1 antibody responses in vitro induced by uniquely immunogenic thymocyte antigens. J Immunol. 1984 Mar;132(3):1100–1105. [PubMed] [Google Scholar]
  10. Isobe K., Nakashima I., Nagase F., Yoshida T., Ando K., Inagi R., Kato N., Ohashi K. Genetic controls of T cell-independent Thy-1 alloantibody responses. Immunogenetics. 1984;20(3):331–340. doi: 10.1007/BF00364214. [DOI] [PubMed] [Google Scholar]
  11. Isobe K., Nakashima I., Nagase N., Nagase F., Lake P., Kato N., Mizoguchi K., Saito M., Kido N., Kawashima K. Intact and subcellular form of thymocytes of rats are exceptionally immunogenic in mice for inducing anti-Thy-1 T cell-independent class 2 antibody responses. Immunol Lett. 1983;7(2):111–115. doi: 10.1016/0165-2478(83)90043-3. [DOI] [PubMed] [Google Scholar]
  12. Lake P. Antibody response induced in vitro to the cell-surface alloantigen Thy-1. Nature. 1976 Jul 22;262(5566):297–298. doi: 10.1038/262297a0. [DOI] [PubMed] [Google Scholar]
  13. Lake P., Clark E. A., Khorshidi M., Sunshine G. H. Production and characterization of cytotoxic Thy-1 antibody-secreting hybrid cell lines. Detection of T cell subsets. Eur J Immunol. 1979 Nov;9(11):875–886. doi: 10.1002/eji.1830091109. [DOI] [PubMed] [Google Scholar]
  14. Lake P., Douglas T. C. Recognition and genetic control of helper determinants for cell surface antigen Thy-1. Nature. 1978 Sep 21;275(5677):220–222. doi: 10.1038/275220a0. [DOI] [PubMed] [Google Scholar]
  15. Lake P., Mitchison N. A. Associative control of the immune response to cell surface antigens. Immunol Commun. 1976;5(9):795–805. doi: 10.3109/08820137609047620. [DOI] [PubMed] [Google Scholar]
  16. Lake P., Mitchison N. A. Regulatory mechanisms in the immune response to cell-surface antigens. Cold Spring Harb Symp Quant Biol. 1977;41(Pt 2):589–595. doi: 10.1101/sqb.1977.041.01.068. [DOI] [PubMed] [Google Scholar]
  17. Marshak-Rothstein A., Fink P., Gridley T., Raulet D. H., Bevan M. J., Gefter M. L. Properties and applications of monoclonal antibodies directed against determinants of the Thy-1 locus. J Immunol. 1979 Jun;122(6):2491–2497. [PubMed] [Google Scholar]
  18. Maxam A. M., Gilbert W. A new method for sequencing DNA. Proc Natl Acad Sci U S A. 1977 Feb;74(2):560–564. doi: 10.1073/pnas.74.2.560. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mitchison N. A. Linked help in the cytotoxic T-cell response revealed by adoptive transfer. Transplant Proc. 1983 Dec;15(4):2121–2124. [PubMed] [Google Scholar]
  20. Peterson P. A., Cunningham B. A., Berggård I., Edelman G. M. 2 -Microglobulin--a free immunoglobulin domain. Proc Natl Acad Sci U S A. 1972 Jul;69(7):1697–1701. doi: 10.1073/pnas.69.7.1697. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. REIF A. E., ALLEN J. M. THE AKR THYMIC ANTIGEN AND ITS DISTRIBUTION IN LEUKEMIAS AND NERVOUS TISSUES. J Exp Med. 1964 Sep 1;120:413–433. doi: 10.1084/jem.120.3.413. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Scherer G., Telford J., Baldari C., Pirrotta V. Isolation of cloned genes differentially expressed at early and late stages of Drosophila embryonic development. Dev Biol. 1981 Sep;86(2):438–447. doi: 10.1016/0012-1606(81)90202-5. [DOI] [PubMed] [Google Scholar]
  23. Wigler M., Sweet R., Sim G. K., Wold B., Pellicer A., Lacy E., Maniatis T., Silverstein S., Axel R. Transformation of mammalian cells with genes from procaryotes and eucaryotes. Cell. 1979 Apr;16(4):777–785. doi: 10.1016/0092-8674(79)90093-x. [DOI] [PubMed] [Google Scholar]
  24. Williams A. F., Gagnon J. Neuronal cell Thy-1 glycoprotein: homology with immunoglobulin. Science. 1982 May 14;216(4547):696–703. doi: 10.1126/science.6177036. [DOI] [PubMed] [Google Scholar]
  25. Zaleski M. B., Gorzynski T. Genetic and cellular requirements for primary antibody immune response to Thy-1 antigens in mice. J Immunol. 1979 May;122(5):2074–2076. [PubMed] [Google Scholar]
  26. Zaleski M., Klein J. Genetic control of the immune response to Thy-1 antigens. Immunol Rev. 1978;38:120–162. doi: 10.1111/j.1600-065x.1978.tb00386.x. [DOI] [PubMed] [Google Scholar]

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