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Purification from a human lung cancer cell line of a water soluble molecule mediating leukocyte adherence inhibition for patients with lung cancer
Author(s) -
Thomson D. M. P.,
Sutherland Mary,
Durko Margaret,
Dubois Tony,
Scanzano Rosemarie,
Labateya Nabil,
Shenouda George
Publication year - 1985
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.2910350603
Subject(s) - lung cancer , monoclonal antibody , cancer , affinity chromatography , melanoma , antibody , microbiology and biotechnology , cancer research , lung , cancer cell , cell culture , antigen , medicine , biology , immunology , pathology , biochemistry , genetics , enzyme
Soluble lung tumor activity as determined by LAI 2 was enriched by physiocochemical methods from chemically ‐ defined spent medium of a lung cancer cell line (NCI‐H69). To identify the polypeptide carrying the antigenic determinant, splenic lymphocytes of BALB/c mice were immunized with the enriched isolate and hybridized with mouse plasmacytoma cells. Eight hybrids were cloned successfully and produced MAbs that immunoprecipitated principally a single chain of M r 40,000 (p40) as well as minor chains of M r 25,000 (p25) and M r 13,000 (p13) which were probably degradation products of p40. On 2D gels, p40 was composed of 7 spots with a p1 of 6.3 to 7.6, which was not altered by neuraminidase digestion. Affinity chromatography with MAb anti‐p40 absorbed p40 and LAI activity. The bound and recovered fraction was enriched for p40 and LAI activity. Affinity‐purified p40 also contained the previously identified p25 and p13 as well as a M r 32,000 peptide (p32). MAb anti‐p40 was directed to a common framework determinant on p40 since MAb anti‐p40 bound to cancer cells from other organs. The comparatively lung cancer organ‐specific determinant recognized by leukocytes from lung cancer patients was not recognized by the MAb. Affinity‐purified p40 triggered LAI for leukocytes from patients with lung cancer but not for leukocytes from control subjects or patients with colon cancer or malignant melanoma in rigorous blind testing. Crossreactivity was observed with leukocytes from patients with breast cancer. LAI activity of affinity‐purified p40 seems unlikely to result from an unidentified impurity. Thus a p40 molecule has been purified that is expressed on the membranes of lung cancer cells and triggers immunologically‐mediated LAI.