Identification of a Human Cancer Related Organ-Specific Neoantigen

• Published in: MICROBIOLOGY and IMMUNOLOGY Vol. 37; no. 2; pp. 119 - 128
• Main Authors: Ilantzis, Christian, Thomson, David M.P., Michaelidou, Alice, Benchimol, Sarita, Stanners, Clifford P.
• Format: Journal Article
• Language: English
• Published: Tokyo Blackwell Publishing Ltd 01.01.1993; Center For Academic Publications Japan; Center for Academic Publications Japan
• Subjects: Amino Acid Sequence; Antigens, Neoplasm - chemistry; Antigens, Neoplasm - genetics; Antigens, Neoplasm - isolation & purification; Bacterial Proteins; Base Sequence; Biological and medical sciences; Blotting, Western; Cloning, Molecular; Colonic Neoplasms - immunology; Colonic Neoplasms - microbiology; DNA, Bacterial - chemistry; Host-tumor relations. Immunology. Biological markers; Humans; Leukocyte adherence inhibition; Leukocyte Adherence Inhibition Test; Medical sciences; Molecular Sequence Data; Mycoplasma; Mycoplasma - genetics; Mycoplasma - isolation & purification; Oligonucleotides - chemistry; Polymerase Chain Reaction; RNA, Bacterial - analysis; Transcription, Genetic; Tumor antigen; Tumor Cells, Cultured; Tumors; Human; Carcinoma; Tumor associated antigen; Gene product; Mycoplasmataceae; Mollicutes; Polymerase chain reaction; Mycoplasmatales; Complementary DNA; Immunogenicity; Mycoplasma hyorhinis; Bacteria; Tumor; Organ specificity; Colon; Neoantigen
• ISSN: 0385-5600; 1348-0421
• DOI: 10.1111/j.1348-0421.1993.tb03188.x

Human cancers express organ-specific neoantigens (OSNs) which elicit specific cellular immune responses in the cancer patient, as demonstrated by leukocyte adherence inhibition (LAI), an in vitro immune response assay. A purified protein of MW 40, 000 (p40) exhibiting OSN (colon specific) activity was cleaved into specific peptide fragments and their partial amino acid sequences determined. This information was used in the polymerase chain reaction (PCR) to obtain a 992bp cDNA clone (PCR-992) from a human colon adenocarcinoma cell line (LS-180). By comparison of the predicted amino acid sequence of PCR-992 with the known sequence of p40 peptides, PCR-992 was shown to correspond to almost the entire coding region of p40. Nucleotide sequence analysis suggested that the protein was mycoplasmal in origin due to its high A+T content (76%) and the presence of five in frame TGA termination codons; at least two of the latter are actually read as tryptophan, a known feature of mycoplasma translation. We have confirmed this origin by direct isolation of a contaminating mycoplasma species from the LS-180 cell line and demonstration that it could be hybridized with the PCR-992 probe. Northern and PCR analysis of RNA preparations from the contaminated LS-180 cell line showed that p40 was part of the high affinity transport system operon of Mycoplasma hyorhinis (Dudler et al, EMBO J., 7: 3963-3970, 1988). Total protein lysates of Mycoplasma hyorhinis cultivated without animal cells could elicit positive LAI responses when incubated with cancer patient leukocytes but not with normal patient leukocytes. The organ-specific nature of the response was, however, not observed indicating that host cell-mycoplasmal interactions may play a role in determining the organ-specific nature of p40 seen with the LAI. The significance of these findings will be discussed in the context of previous thinking regarding the origin of OSNs.