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Clinical and Experimental Immunology logoLink to Clinical and Experimental Immunology
. 1983 Dec;54(3):811–818.

Binding of mouse monoclonal antibodies to human leukaemic cells via the Fc receptor: a possible source of 'false positive' reactions in specificity screening.

S J Gadd, L K Ashman
PMCID: PMC1536158  PMID: 6580981

Abstract

Mouse monoclonal antibodies (MoAbs) of different classes and subclasses directed against antigens not expected to be present on human cells have been screened by indirect immunofluorescence using flow cytometry for binding to human non-lymphocytic leukaemic cells and normal peripheral blood leucocytes. Antibodies of the IgG2a and IgG3 subclass, but not of the IgG1, IgG2b or IgM class bound to the blast cell and monocyte populations in a peripheral blood mononuclear cell preparation from a patient with acute myelomonocytic leukaemia. F(ab')2 fragments of an anti-salmonella antibody of IgG2a subclass failed to bind, indicating that the results were not due to cross-reactivity with antigens of the cell membrane, thus implicating the Fc region in binding. Furthermore, binding was partly blocked by inclusion of 10% heat-inactivated normal rabbit serum in the assays. IgG2a bound to varying degrees to the leukaemic cell populations in seven of the nine non-lymphocytic leukaemic specimens tested, but no binding to normal peripheral blood mononuclear cells or granulocytes was detected. The results emphasize the importance of including appropriate controls when screening MoAbs for binding to various types of human cells.

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Selected References

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

  1. Alexander E. L., Sanders S. K. F(ab')2 reagents are not required if goat, rather than rabbit, antibodies are used to detect human surface immunoglobulin. J Immunol. 1977 Sep;119(3):1084–1088. [PubMed] [Google Scholar]
  2. Bennett J. M., Catovsky D., Daniel M. T., Flandrin G., Galton D. A., Gralnick H. R., Sultan C. Proposals for the classification of the acute leukaemias. French-American-British (FAB) co-operative group. Br J Haematol. 1976 Aug;33(4):451–458. doi: 10.1111/j.1365-2141.1976.tb03563.x. [DOI] [PubMed] [Google Scholar]
  3. Diamond B., Yelton D. E. A new Fc receptor on mouse macrophages binding IgG3. J Exp Med. 1981 Mar 1;153(3):514–519. doi: 10.1084/jem.153.3.514. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Ey P. L., Prowse S. J., Jenkin C. R. Isolation of pure IgG1, IgG2a and IgG2b immunoglobulins from mouse serum using protein A-sepharose. Immunochemistry. 1978 Jul;15(7):429–436. doi: 10.1016/0161-5890(78)90070-6. [DOI] [PubMed] [Google Scholar]
  5. Ezekowitz R. A., Bampton M., Gordon S. Macrophage activation selectively enhances expression of Fc receptors for IgG2a. J Exp Med. 1983 Feb 1;157(2):807–812. doi: 10.1084/jem.157.2.807. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ferrante A., Thong Y. H. Optimal conditions for simultaneous purification of mononuclear and polymorphonuclear leucocytes from human blood by the Hypaque-Ficoll method. J Immunol Methods. 1980;36(2):109–117. doi: 10.1016/0022-1759(80)90036-8. [DOI] [PubMed] [Google Scholar]
  7. Fleit H. B., Wright S. D., Unkeless J. C. Human neutrophil Fc gamma receptor distribution and structure. Proc Natl Acad Sci U S A. 1982 May;79(10):3275–3279. doi: 10.1073/pnas.79.10.3275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Johnson A. M., McNamara P. J., Neoh S. H., McDonald P. J., Zola H. Hybridomas secreting monoclonal antibody to Toxoplasma gondii. Aust J Exp Biol Med Sci. 1981 Jun;59(Pt 3):303–306. doi: 10.1038/icb.1981.23. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Lanier L. L., Warner N. L. Paraformaldehyde fixation of hematopoietic cells for quantitative flow cytometry (FACS) analysis. J Immunol Methods. 1981;47(1):25–30. doi: 10.1016/0022-1759(81)90253-2. [DOI] [PubMed] [Google Scholar]
  11. Maheu M., Baker M. A., Falk J. A., Taub R. N. Immunologic diagnosis and monitoring of human acute leukemias: a review. Am J Pathol. 1981 Apr;103(1):139–158. [PMC free article] [PubMed] [Google Scholar]
  12. O'Connor C. G., Ashman L. K. Application of the nitrocellulose transfer technique and alkaline phosphatase conjugated anti-immunoglobulin for determination of the specificity of monoclonal antibodies to protein mixtures. J Immunol Methods. 1982 Oct 29;54(2):267–271. doi: 10.1016/0022-1759(82)90068-0. [DOI] [PubMed] [Google Scholar]
  13. O'Keefe D., Ashman L. Peanut agglutinin: a marker for normal and leukaemic cells of the monocyte lineage. Clin Exp Immunol. 1982 May;48(2):329–338. [PMC free article] [PubMed] [Google Scholar]
  14. Unkeless J. C., Fleit H., Mellman I. S. Structural Aspects and Heterogeneity of Immunoglobulin Fc Receptors. Adv Immunol. 1981;31:247–270. doi: 10.1016/s0065-2776(08)60922-0. [DOI] [PubMed] [Google Scholar]

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