BACKGROUND: CD44 is a cell surface adhesion molecule which has been implicated in various biologic functions as lymphocyte homing and activation, cellular migration and extracellular matrix adhesion. Over-expression of CD44v8- 10 has been found in several cancers and is considered to be associated with tumor progression and metastasis. Recently, a novel molecular method, CD44v8- 10/CD44v10 competitive reverse transcription-polymerase chain reaction(RT-PCR) has been developed for detecting cancer cells over-expressing CD44v8-10. METHODS: We analyzed from benign and malignant pleural effusion and ascites by CD44 competitive RT-PCR and compared to the conventional cytology. RESULTS: The CD44 competitive RT-PCR analysis showed that all the 24 samples associated with benign disease presented a predominant expression of the CD44v10 transcript (v8-10/v10 ratio: 0.126-0.948), whereas 6 of 7 malignant pleural samples associated with cytology positive cancer expressed the CD44v8-10 transcript (v8-10/v10 ratio > 1.00). CONCLUSION: These results indicate that CD44 competitive RT-PCR assay is a useful and adjunct to cytological examination in cancer diagnosis, especially in detecting exfoliated cancer cells in pleural effusion.
Adult ; Aged ; Aged, 80 and over ; Antigens, CD44/analysis* ; Ascites/pathology* ; Ascites/immunology* ; Base Sequence ; Carcinoma, Non-Small-Cell Lung/pathology ; Carcinoma, Non-Small-Cell Lung/immunology ; Comparative Study ; Female ; Gastrointestinal Neoplasms/pathology ; Gastrointestinal Neoplasms/immunology ; Human ; Lung Neoplasms/pathology ; Lung Neoplasms/chemistry ; Male ; Middle Age ; Molecular Sequence Data ; Pleural Effusion, Malignant/pathology* ; Pleural Effusion, Malignant/chemistry* ; Reverse Transcriptase Polymerase Chain Reaction* ; Sensitivity and Specificity ; Support, Non-U.S. Gov't

OBJECTIVETo detect Mycobacterium tuberculosis RD1-encoded antigens-specific, interferon-gamma (INF-gamma)-secreting T cells in pleural effusions, ascites, and cerebrospinal fluid.

METHODThe early secretory antigenic target-6 (ESAT-6) and culture filtrate protein-10 (CFP-10) peptides-specific T cells in peripheral blood mononuclear cell (MC), ascites MC, pleural effusions MC, and cerebrospinal fluid MC were detected using enzyme-linked immunospot assay (ELISPOT) for INF-gamma.

RESULTSESAT-6 or CFP-10 peptides-specific, INF-gamma-secreting T cells were detected in peripheral blood, ascites, pleural effusions, and cerebrospinal fluid, which marked the presence of tuberculosis infection. Patients with positive ELISPOT results of INF-gamma-release assay were all diagnosed as active tuberculosis. Spot forming cells in ascites and pleural effusions were much higher than those in peripheral blood (up to 6.4 and 31.9 times).

CONCLUSIONDetection of RD1-encoded antigens-specific, INF-gamma-secreting T cells in pleural effusions, ascites, and cerebrospinal fluid provides a new way to diagnose tuberculosis infection.

Antigens, Bacterial ; genetics ; Ascites ; metabolism ; Bacterial Proteins ; Humans ; Interferon-gamma ; cerebrospinal fluid ; metabolism ; Leukocytes, Mononuclear ; Mycobacterium tuberculosis ; Peptides ; Pleural Effusion ; immunology ; Recombinant Proteins ; T-Lymphocytes ; metabolism ; Tuberculosis, Pulmonary ; diagnosis