Identification and characterization of a 14 kDa immunosuppressive protein derived from IIB-MEL-J, a human melanoma cell line

• Published in: Cellular and molecular biology (Noisy-le-Grand, France) Vol. 42; no. 5; p. 779
• Main Authors: Morvillo, V, Bover, L, Mordoh, J
• Format: Journal Article
• Language: English
• Published: France 01-07-1996
• Subjects: Humans; Immune Tolerance; Immunosuppressive Agents - chemistry; Immunosuppressive Agents - isolation & purification; Immunosuppressive Agents - pharmacology; In Vitro Techniques; Interleukin-2 - pharmacology; Lymphocyte Activation - drug effects; Lymphocyte Culture Test, Mixed; Melanoma - chemistry; Melanoma - immunology; Molecular Weight; Neoplasm Proteins - chemistry; Neoplasm Proteins - immunology; Neoplasm Proteins - isolation & purification; Phytohemagglutinins - pharmacology; Solubility; Tumor Cells, Cultured
• ISSN: 0145-5680

The causes of decreased immune competence in melanoma patients as well as in other cancer patients are incompletely understood. The identification of the factor(s) responsible for this behaviour remains elusive. The present report demonstrates that an immunosuppressive activity (ISA), manifested in vitro as an inhibition of proliferation of human peripheral blood lymphocytes (PBL) induced both by phytohemagglutinin (PHA) or the cytokine IL-2, was exhibited by serum-free conditioned medium (SFCM) from the human melanoma cell line IIB-MEL-J, as well as by two other melanoma cell lines, IIB-MEL-LES and IIB-MEL-IAN, established in our laboratory. The ISA was found to be exerted by a protein, which co-eluted with serum albumin in anionic exchange Mono-Q, gel filtration chromatography and Blue Sepharose columns. It showed a molecular weight (Mw) of 14 kDa when separated from albumin traces by means of a Sephacryl S 200 column. It is not recognized by a pan-specific anti-transforming growth factor-beta (TGF-beta) antibody as determined by Western blots assays performed on the SFCM. The immunosuppressive factor (ISF) is secreted by IIB-MEL-J cell line in soluble from and in very scarce amounts, non-detectable by polyacrylamide gel electrophoresis (PAGE). This characteristic difficults its obtention in adequate quantities to sequentiation. Since this inhibitory factor may have a role in protecting melanoma tumors from attack by the host immune system, preparative isolation will be attempted. But we consider these results only as preliminary one's.