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. 1980 May;77(5):2899–2903. doi: 10.1073/pnas.77.5.2899

Identification and characterization of rabbit-mouse hybridomas secreting rabbit immunoglobulin chains.

M L Yarmush, F T Gates 3rd, D R Weisfogel, T J Kindt
PMCID: PMC349513  PMID: 6771762

Abstract

Standard cell-fusion techniques have been used to generate hybrid cells from rabbit spleen cells and mouse myeloma cell lines. The hybrids were selected for secretion of rabbit immunoglobulin. Detailed allotype analyses were carried out for 189 cell lines found to be immunoglobulin positive: 1 produced an intact immunoglobulin molecule with antibody activity, 143 produced rabbit light (L) chains, 36 produced rabbit heavy (H) chains, and 9 cell lines gave negative results in tests for group a and b allotypes. Fusions with a nonproducing murine myeloma cell line (SP-2) yielded only L chain-secreting hybrids, whereas 27% of hybrids resulting from fusion with an L chain-producing line (PU) secreted rabbit H chains paired with mouse L chains. Several of the hybridomas have maintained the ability to produce rabbit immunoglobulin chains in culture for almost 1 year and can be propagated as ascites tumors in athymic (nude) mice. Cytogenetic analyses have detected no rabbit chromosomes in the stable hybrid cell lines.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Buttin G., LeGuern G., Phalente L., Lin E. C., Medrano L., Cazenave P. A. Production of hybrid lines secreting monoclonal anti-idiotypic antibodies by cell fusion on membrane filters. Curr Top Microbiol Immunol. 1978;81:27–36. doi: 10.1007/978-3-642-67448-8_4. [DOI] [PubMed] [Google Scholar]
  2. Coffino P., Scharff M. D. Rate of somatic mutation in immunoglobulin production by mouse myeloma cells. Proc Natl Acad Sci U S A. 1971 Jan;68(1):219–223. doi: 10.1073/pnas.68.1.219. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Collins J. J., Black P. H., Strosberg A. D., Haber E., Bloch K. J. Transformation by simian virus 40 of spleen cells from a hyperimmune rabbit: evidence for synthesis of immunoglobulin by the transformed cells. Proc Natl Acad Sci U S A. 1974 Feb;71(2):260–262. doi: 10.1073/pnas.71.2.260. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Engvall E., Perlmann P. Enzyme-linked immunosorbent assay, Elisa. 3. Quantitation of specific antibodies by enzyme-labeled anti-immunoglobulin in antigen-coated tubes. J Immunol. 1972 Jul;109(1):129–135. [PubMed] [Google Scholar]
  5. Farnsworth V., Goodfliesh R., Rodkey S., Hood L. Immunoglobulin allotypes of rabbit kappa chains: polymorphism of a control mechanism regulating closely linked duplicated genes? Proc Natl Acad Sci U S A. 1976 Apr;73(4):1293–1296. doi: 10.1073/pnas.73.4.1293. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gefter M. L., Margulies D. H., Scharff M. D. A simple method for polyethylene glycol-promoted hybridization of mouse myeloma cells. Somatic Cell Genet. 1977 Mar;3(2):231–236. doi: 10.1007/BF01551818. [DOI] [PubMed] [Google Scholar]
  7. Gottlieb A. B., Seide R. K., Kindt T. J. Quantitative determination of allotypic and idiotypic markers with antisera coupled to N-hydroxysuccinimide-activated sepharose. J Immunol. 1975 Jan;114(1 Pt 1):51–54. [PubMed] [Google Scholar]
  8. Handmaker S. D. Hybridization of eukaryotic cells. Annu Rev Microbiol. 1973;27:189–204. doi: 10.1146/annurev.mi.27.100173.001201. [DOI] [PubMed] [Google Scholar]
  9. Kimball E. S., Wolf B. Regulation of allotype expression in heterozygous rabbits. III. Concomitant modulation and concomitant suppression oa a2 and a3 allotypes on individual peripheral blood lymphocytes. Eur J Immunol. 1977 Dec;7(12):898–906. doi: 10.1002/eji.1830071215. [DOI] [PubMed] [Google Scholar]
  10. Kindt T. J. Rabbit immunoglobulin allotypes: structure, immunology, and genetics. Adv Immunol. 1975;21:35–86. doi: 10.1016/s0065-2776(08)60218-7. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Krause R. M. The search for antibodies with molecular uniformity. Adv Immunol. 1970;12:1–56. doi: 10.1016/s0065-2776(08)60167-4. [DOI] [PubMed] [Google Scholar]
  12. Köhler G., Howe S. C., Milstein C. Fusion between immunoglobulin-secreting and nonsecreting myeloma cell lines. Eur J Immunol. 1976 Apr;6(4):292–295. doi: 10.1002/eji.1830060411. [DOI] [PubMed] [Google Scholar]
  13. Köhler G., Milstein C. Continuous cultures of fused cells secreting antibody of predefined specificity. Nature. 1975 Aug 7;256(5517):495–497. doi: 10.1038/256495a0. [DOI] [PubMed] [Google Scholar]
  14. LITTLEFIELD J. W. SELECTION OF HYBRIDS FROM MATINGS OF FIBROBLASTS IN VITRO AND THEIR PRESUMED RECOMBINANTS. Science. 1964 Aug 14;145(3633):709–710. doi: 10.1126/science.145.3633.709. [DOI] [PubMed] [Google Scholar]
  15. Margulies D. H., Kuehl W. M., Scharff M. D. Somatic cell hybridization of mouse myeloma cells. Cell. 1976 Jul;8(3):405–415. doi: 10.1016/0092-8674(76)90153-7. [DOI] [PubMed] [Google Scholar]
  16. Minna J. D., Coon H. G. Human times mouse hybrid cells segregating mouse chromosomes and isozymes. Nature. 1974 Nov 29;252(5482):401–404. doi: 10.1038/252401a0. [DOI] [PubMed] [Google Scholar]
  17. Ruddle F. H., Creagan R. P. Parasexual approaches to the genetics of man. Annu Rev Genet. 1975;9:407–486. doi: 10.1146/annurev.ge.09.120175.002203. [DOI] [PubMed] [Google Scholar]
  18. Schall D., Rechsteiner M. Kinetics of human chromosome loss from 3T3-human hybrid cells. Somatic Cell Genet. 1978 Nov;4(6):661–676. doi: 10.1007/BF01543157. [DOI] [PubMed] [Google Scholar]
  19. Schwaber J., Cohen E. P. Pattern of immunoglobulin synthesis and assembly in a human-mouse somatic cell hybrid clone. Proc Natl Acad Sci U S A. 1974 Jun;71(6):2203–2207. doi: 10.1073/pnas.71.6.2203. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Seabright M. A rapid banding technique for human chromosomes. Lancet. 1971 Oct 30;2(7731):971–972. doi: 10.1016/s0140-6736(71)90287-x. [DOI] [PubMed] [Google Scholar]
  21. Shulman M., Wilde C. D., Köhler G. A better cell line for making hybridomas secreting specific antibodies. Nature. 1978 Nov 16;276(5685):269–270. doi: 10.1038/276269a0. [DOI] [PubMed] [Google Scholar]
  22. Siebert G. R., Harris J. F., Gefter M. L. Regulation of immunoglobulin biosynthesis in the murine plasmacytoma MOPC 315. J Immunol. 1978 Nov;121(5):1808–1818. [PubMed] [Google Scholar]
  23. Sumner A. T. A simple technique for demonstrating centromeric heterochromatin. Exp Cell Res. 1972 Nov;75(1):304–306. doi: 10.1016/0014-4827(72)90558-7. [DOI] [PubMed] [Google Scholar]
  24. Yarmush M. L., Sogn J. A., Kindt T. J. Latent allotypes: a window to a genetic enigma. Ann Immunol (Paris) 1979 Mar-Apr;130(2):143–156. [PubMed] [Google Scholar]
  25. Yelton D. E., Diamond B. A., Kwan S. P., Scharff M. D. Fusion of mouse myeloma and spleen cells. Curr Top Microbiol Immunol. 1978;81:1–7. doi: 10.1007/978-3-642-67448-8_1. [DOI] [PubMed] [Google Scholar]

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