Skip to main content
NCBI home page
Clinical and Experimental Immunology logoLink to Clinical and Experimental Immunology
. 1993 Aug;93(2):279–285. doi: 10.1111/j.1365-2249.1993.tb07980.x

Antibodies to the protein core of the small cell lung cancer workshop antigen cluster-w4 and to the leucocyte workshop antigen CD24 recognize the same short protein sequence leucine-alanine-proline.

E Weber 1, H P Lehmann 1, A G Beck-Sickinger 1, E J Wawrzynczak 1, R Waibel 1, G Folkers 1, R A Stahel 1
PMCID: PMC1554848  PMID: 7688677

Abstract

We recently described the identity of the small cell lung cancer (SCLC) cluster-w4 antigen and the human B cell differentiation marker CD24, a glycosylphosphatidylinositol (GPI)-anchored, highly glycosylated surface molecule of only 31-35 amino acids [15]. The specificities of three anti-cluster-w4 and of eleven anti-CD24 MoAbs have been investigated with respect to their binding capacity to the protein core of cluster-w4/CD24 antigen. Four overlapping peptides spanning this protein core were synthesized. MoAbs shown to bind to two overlapping peptides by antibody binding inhibition using the cluster-w4/CD24-positive SCLC cell line SW2 and by direct peptide binding detected in an ELISA were investigated in more detail. To determine the exact epitopes recognized by these MoAbs, an epitope mapping assay using peptides synthesized onto polyethylene pins was established. The three anti-cluster-w4 MoAbs SWA11, SWA21 and SWA22 and the anti-CD24 MoAbs OKB2 and ALB9 recognized the same short leucine-alanine-proline (LAP) sequence in an area without potential glycosylation sites close to the GPI anchor of the protein core of the cluster-w4/CD24 antigen.

Full text

PDF
279

Selected References

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

  1. Abramson C. S., Kersey J. H., LeBien T. W. A monoclonal antibody (BA-1) reactive with cells of human B lymphocyte lineage. J Immunol. 1981 Jan;126(1):83–88. [PubMed] [Google Scholar]
  2. Alterman L. A., Crispe I. N., Kinnon C. Characterization of the murine heat-stable antigen: an hematolymphoid differentiation antigen defined by the J11d, M1/69 and B2A2 antibodies. Eur J Immunol. 1990 Jul;20(7):1597–1602. doi: 10.1002/eji.1830200728. [DOI] [PubMed] [Google Scholar]
  3. Barlow D. J., Edwards M. S., Thornton J. M. Continuous and discontinuous protein antigenic determinants. Nature. 1986 Aug 21;322(6081):747–748. doi: 10.1038/322747a0. [DOI] [PubMed] [Google Scholar]
  4. Cross G. A. Glycolipid anchoring of plasma membrane proteins. Annu Rev Cell Biol. 1990;6:1–39. doi: 10.1146/annurev.cb.06.110190.000245. [DOI] [PubMed] [Google Scholar]
  5. Derbyshire E. J., Henry R. V., Stahel R. A., Wawrzynczak E. J. Potent cytotoxic action of the immunotoxin SWA11-ricin A chain against human small cell lung cancer cell lines. Br J Cancer. 1992 Sep;66(3):444–451. doi: 10.1038/bjc.1992.294. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ferguson M. A., Williams A. F. Cell-surface anchoring of proteins via glycosyl-phosphatidylinositol structures. Annu Rev Biochem. 1988;57:285–320. doi: 10.1146/annurev.bi.57.070188.001441. [DOI] [PubMed] [Google Scholar]
  7. Fischer G. F., Majdic O., Gadd S., Knapp W. Signal transduction in lymphocytic and myeloid cells via CD24, a new member of phosphoinositol-anchored membrane molecules. J Immunol. 1990 Jan 15;144(2):638–641. [PubMed] [Google Scholar]
  8. Geysen H. M., Meloen R. H., Barteling S. J. Use of peptide synthesis to probe viral antigens for epitopes to a resolution of a single amino acid. Proc Natl Acad Sci U S A. 1984 Jul;81(13):3998–4002. doi: 10.1073/pnas.81.13.3998. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Geysen H. M., Rodda S. J., Mason T. J., Tribbick G., Schoofs P. G. Strategies for epitope analysis using peptide synthesis. J Immunol Methods. 1987 Sep 24;102(2):259–274. doi: 10.1016/0022-1759(87)90085-8. [DOI] [PubMed] [Google Scholar]
  10. Horsfall A. C., Hay F. C., Soltys A. J., Jones M. G. Epitope mapping. Immunol Today. 1991 Jul;12(7):211–213. doi: 10.1016/0167-5699(91)90029-S. [DOI] [PubMed] [Google Scholar]
  11. Hsu S. M., Jaffe E. S. Phenotypic expression of B-lymphocytes. 1. Identification with monoclonal antibodies in normal lymphoid tissues. Am J Pathol. 1984 Mar;114(3):387–395. [PMC free article] [PubMed] [Google Scholar]
  12. Jackson D., Waibel R., Weber E., Bell J., Stahel R. A. CD24, a signal-transducing molecule expressed on human B cells, is a major surface antigen on small cell lung carcinomas. Cancer Res. 1992 Oct 1;52(19):5264–5270. [PubMed] [Google Scholar]
  13. Kadmon G., Eckert M., Sammar M., Schachner M., Altevogt P. Nectadrin, the heat-stable antigen, is a cell adhesion molecule. J Cell Biol. 1992 Sep;118(5):1245–1258. doi: 10.1083/jcb.118.5.1245. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Kay R., Rosten P. M., Humphries R. K. CD24, a signal transducer modulating B cell activation responses, is a very short peptide with a glycosyl phosphatidylinositol membrane anchor. J Immunol. 1991 Aug 15;147(4):1412–1416. [PubMed] [Google Scholar]
  15. Kemshead J. T., Fritschy J., Asser U., Sutherland R., Greaves M. F. Monoclonal antibodies defining markers with apparent selectivity for particular haemopoietic cell types may also detect antigens on cells of neural crest origin. Hybridoma. 1982;1(2):109–123. doi: 10.1089/hyb.1.1982.1.109. [DOI] [PubMed] [Google Scholar]
  16. Knapp W., Majdic O., Bettelheim P., Liszka K., Aberer W., Stingl G. Typing of leukemic cells with monoclonal antibodies. Ann N Y Acad Sci. 1983;420:251–260. doi: 10.1111/j.1749-6632.1983.tb22210.x. [DOI] [PubMed] [Google Scholar]
  17. Larkin M., Knapp W., Stoll M. S., Mehmet H., Feizi T. Monoclonal antibodies VIB-E3, IB5 and HB9 to the leucocyte/epithelial antigen CD24 resemble BA-1 in recognizing sialic acid-dependent epitope(s). Evidence that VIB-E3 recognizes NeuAc alpha 2-6GalNAc and NeuAc alpha 2-6Gal sequences. Clin Exp Immunol. 1991 Sep;85(3):536–541. doi: 10.1111/j.1365-2249.1991.tb05763.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. May R. D., Finkelman F. D., Wheeler H. T., Uhr J. W., Vitetta E. S. Evaluation of ricin A chain-containing immunotoxins directed against different epitopes on the delta-chain of cell surface-associated IgD on murine B cells. J Immunol. 1990 May 1;144(9):3637–3642. [PubMed] [Google Scholar]
  19. May R. D., Wheeler H. T., Finkelman F. D., Uhr J. W., Vitetta E. S. Intracellular routing rather than cross-linking or rate of internalization determines the potency of immunotoxins directed against different epitopes of sIgD on murine B cells. Cell Immunol. 1991 Jul;135(2):490–500. doi: 10.1016/0008-8749(91)90292-j. [DOI] [PubMed] [Google Scholar]
  20. Melink G. B., LeBien T. W. Construction of an antigenic map for human B-cell precursors. J Clin Immunol. 1983 Jul;3(3):260–267. doi: 10.1007/BF00915350. [DOI] [PubMed] [Google Scholar]
  21. Pirruccello S. J., LeBien T. W. Monoclonal antibody BA-1 recognizes a novel human leukocyte cell surface sialoglycoprotein complex. J Immunol. 1985 Jun;134(6):3962–3968. [PubMed] [Google Scholar]
  22. Press O. W., Martin P. J., Thorpe P. E., Vitetta E. S. Ricin A-chain containing immunotoxins directed against different epitopes on the CD2 molecule differ in their ability to kill normal and malignant T cells. J Immunol. 1988 Dec 15;141(12):4410–4417. [PubMed] [Google Scholar]
  23. Rock K. L., Reiser H., Bamezai A., McGrew J., Benacerraf B. The LY-6 locus: a multigene family encoding phosphatidylinositol-anchored membrane proteins concerned with T-cell activation. Immunol Rev. 1989 Oct;111:195–224. doi: 10.1111/j.1600-065x.1989.tb00547.x. [DOI] [PubMed] [Google Scholar]
  24. Rougon G., Alterman L. A., Dennis K., Guo X. J., Kinnon C. The murine heat-stable antigen: a differentiation antigen expressed in both the hematolymphoid and neural cell lineages. Eur J Immunol. 1991 Jun;21(6):1397–1402. doi: 10.1002/eji.1830210611. [DOI] [PubMed] [Google Scholar]
  25. Smith A., Waibel R., Westera G., Martin A., Zimmerman A. T., Stahel R. A. Immunolocalisation and imaging of small cell cancer xenografts by the IgG2a monoclonal antibody SWA11. Br J Cancer. 1989 Feb;59(2):174–178. doi: 10.1038/bjc.1989.36. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Stefanová I., Horejsí V., Ansotegui I. J., Knapp W., Stockinger H. GPI-anchored cell-surface molecules complexed to protein tyrosine kinases. Science. 1991 Nov 15;254(5034):1016–1019. doi: 10.1126/science.1719635. [DOI] [PubMed] [Google Scholar]
  27. Waibel R., O'Hara C. J., Stahel R. A. Monoclonal antibodies defining the cluster-5A small cell lung carcinoma antigen. Br J Cancer Suppl. 1991 Jun;14:29–32. [PMC free article] [PubMed] [Google Scholar]
  28. Wang C. Y., Azzo W., Al-Katib A., Chiorazzi N., Knowles D. M., 2nd Preparation and characterization of monoclonal antibodies recognizing three distinct differentiation antigens (BL1, BL2, BL3) on human B lymphocytes. J Immunol. 1984 Aug;133(2):684–691. [PubMed] [Google Scholar]
  29. Wawrzynczak E. J., Derbyshire E. J., Henry R. V., Parnell G. D., Smith A., Waibel R., Stahel R. A. Selective cytotoxic effects of a ricin A chain immunotoxin made with the monoclonal antibody SWA11 recognising a human small cell lung cancer antigen. Br J Cancer. 1990 Sep;62(3):410–414. doi: 10.1038/bjc.1990.308. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wawrzynczak E. J., Zangemeister-Wittke U., Waibel R., Henry R. V., Parnell G. D., Cumber A. J., Jones M., Stahel R. A. Molecular and biological properties of an abrin A chain immunotoxin designed for therapy of human small cell lung cancer. Br J Cancer. 1992 Aug;66(2):361–366. doi: 10.1038/bjc.1992.271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Wenger R. H., Ayane M., Bose R., Köhler G., Nielsen P. J. The genes for a mouse hematopoietic differentiation marker called the heat-stable antigen. Eur J Immunol. 1991 Apr;21(4):1039–1046. doi: 10.1002/eji.1830210427. [DOI] [PubMed] [Google Scholar]
  32. Wilson J. E. The use of monoclonal antibodies and limited proteolysis in elucidation of structure-function relationships in proteins. Methods Biochem Anal. 1991;35:207–250. doi: 10.1002/9780470110560.ch4. [DOI] [PubMed] [Google Scholar]
  33. Xia M. Q., Tone M., Packman L., Hale G., Waldmann H. Characterization of the CAMPATH-1 (CDw52) antigen: biochemical analysis and cDNA cloning reveal an unusually small peptide backbone. Eur J Immunol. 1991 Jul;21(7):1677–1684. doi: 10.1002/eji.1830210714. [DOI] [PubMed] [Google Scholar]
  34. Zangemeister-Wittke U., Lehmann H. P., Waibel R., Wawrzynczak E. J., Stahel R. A. Action of a CD24-specific deglycosylated ricin-A-chain immunotoxin in conventional and novel models of small-cell-lung-cancer xenograft. Int J Cancer. 1993 Feb 1;53(3):521–528. doi: 10.1002/ijc.2910530327. [DOI] [PubMed] [Google Scholar]

Articles from Clinical and Experimental Immunology are provided here courtesy of British Society for Immunology

RESOURCES