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Cell lines derived from human small cell carcinoma of the lung express high levels of a surface polypeptide termed the cluster-w4 antigen, which was previously identified as a potential target for toxin-based immunotherapy of lung cancer. We have cloned a complementary DNA encoding the cluster-w4 antigen from COS-1 fibroblasts transfected with a SW2 small cell carcinoma library, by panning with a mixture of the cluster-w4-specific monoclonal antibodies SWA11, SWA21, and SWA22. The sequence of the cluster-w4 complementary DNA encodes an unusually short (80-amino acid) protein identical to that recently reported for the leukocyte activation molecule CD24 except for a single valine-alanine substitution due to a single-base polymorphism within the region of the gene coding for the extracellular domain. Biochemical analyses of the cloned cluster-w4 antigen confirmed both the presence of the phosphatidylinositol tail and the extensive glycosylation reported for the CD24 molecule. Furthermore, the cloned cluster-w4 antigen expressed on COS cells was shown to react with a comprehensive panel of CD24-specific monoclonal antibodies, as assessed by indirect immunofluorescence staining. Northern blot hybridization indicated the presence of several transcript sizes for the cluster-w4 antigen that were greatly overexpressed in small cell carcinoma cell lines, compared with normal hemopoietic cells and CD24-positive cell lines. Southern blot hybridization of restriction digests of genomic DNA identified a complex pattern of bands consistent with either a complex gene structure containing many exons or the presence of a family of closely related genes.

Type

Journal article

Journal

Cancer research

Publication Date

10/1992

Volume

52

Pages

5264 - 5270

Addresses

Molecular Immunology Group, John Radcliffe Hospital, Oxford, UK.

Keywords

B-Lymphocytes, Animals, Mice, Inbred BALB C, Humans, Mice, Carcinoma, Small Cell, Lung Neoplasms, Alanine, Valine, Membrane Glycoproteins, DNA, Neoplasm, Antigens, CD, Antibodies, Monoclonal, Antigens, Neoplasm, Antigens, Surface, Cloning, Molecular, Signal Transduction, Amino Acid Sequence, Base Sequence, Sequence Homology, Molecular Weight, Molecular Sequence Data, CD24 Antigen