High tumour contamination of leukaphereses in patients with small cell carcinoma of the lung: a comparison of immunocytochemistry and RT-PCR

In small-cell lung carcinoma (SCLC) tumour cell contamination of leukaphereses is unknown. The present study was performed to define appropriate markers for reverse transcriptase polymerase chain reaction (RT-PCR), then to assess the contamination rate of leukaphereses and corresponding bone marrow...

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Bibliographic Details
Published in:British journal of cancer 2001-11, Vol.85 (11), p.1713-1721
Main Authors: PEREY, L, BENHATTAR, J, PETERS, R, JAUNIN, P, LEYVRAZ, S
Format: Article
Language:English
Subjects:
Adult
Aged
Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy
Antigens, Neoplasm - analysis
Antigens, Neoplasm - genetics
Biological and medical sciences
Blood. Blood and plasma substitutes. Blood products. Blood cells. Blood typing. Plasmapheresis. Apheresis
Bone Marrow - chemistry
Bone Marrow - pathology
Carcinoembryonic Antigen - analysis
Carcinoembryonic Antigen - genetics
Carcinoma, Small Cell - blood
Carcinoma, Small Cell - therapy
Cell Adhesion Molecules - analysis
Cell Adhesion Molecules - genetics
Chromogranins - analysis
Chromogranins - genetics
Epithelial Cell Adhesion Molecule
Female
Gastric Mucins - analysis
Gastric Mucins - genetics
Humans
Immunohistochemistry
Keratins - analysis
Keratins - genetics
Leukapheresis
Lung Neoplasms - blood
Lung Neoplasms - therapy
Male
Medical sciences
Middle Aged
Neoplasm Proteins
Neoplastic Cells, Circulating - metabolism
Neoplastic Cells, Circulating - pathology
Neural Cell Adhesion Molecules - analysis
Neural Cell Adhesion Molecules - genetics
Phosphopyruvate Hydratase - analysis
Phosphopyruvate Hydratase - genetics
Proteins - analysis
Proteins - genetics
Proteoglycans
Regular
Reverse Transcriptase Polymerase Chain Reaction
RNA, Neoplasm - genetics
RNA, Neoplasm - metabolism
Synaptophysin - analysis
Synaptophysin - genetics
Transfusions. Complications. Transfusion reactions. Cell and gene therapy
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Summary:In small-cell lung carcinoma (SCLC) tumour cell contamination of leukaphereses is unknown. The present study was performed to define appropriate markers for reverse transcriptase polymerase chain reaction (RT-PCR), then to assess the contamination rate of leukaphereses and corresponding bone marrow samples. Immunocytochemistry (ICC) and RT-PCR methods were also compared. Among the 33 patients included, analyses were performed in 16 who had multiple leukaphereses and 17 who had only bone marrow. Leukapheresis products and bone marrow were analysed by ICC using several specific monoclonal antibodies against neural-cell adhesion molecule (N-CAM), epithelial glycoprotein (EGP-40) and cytokeratins (CK). Samples were also analyzed by RT-PCR for expression for N-CAM, synaptophysin, neuron-specific enolase, chromogranin, cytokeratin-18/-19, CEA, EGP-40, apomucin type 1 (MUC-1) and human endothelial cell-specific molecule (ESM-1). Using ICC staining, contaminating tumour cells were detected in 34% of leukaphereses (27% in patients with limited disease and 43% in those with extensive disease). N-CAM was the most reliable marker for detection of contamination. For RT-PCR, CK-19 and CEA were the only appropriate markers. Positive signal rate in leukaphereses increased to 78% (89% for patients with limited disease and 67% for extensive disease). In bone marrow, both techniques were in agreement whereas in leukaphereses, RT-PCR was better than ICC. A high rate of tumour cell contamination was demonstrated not only in bone marrow but also in leukaphereses from SCLC patients. The most appropriate technique was RT-PCR mainly in patients with limited disease.
ISSN:0007-0920
1532-1827
DOI:10.1054/bjoc.2001.2177