Establishment of an immunoassay system for monocyte chemotacticand activating factor (MCAF) and its clinical application

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The results of the project is summarized as follows.1. We established MCAF-producing cell line by transfecting MCAF cDNA into a mouse myeloma cell line and purified MCAF from the culture supernatants of this cell line. We observed that the preadministration of purified recombinant MCAF could prevent either normal or granulocytopenic mice from lethal infection caused by pseudomonas aeruginosa o r Salmonella typhymurium.2. We established a specific and sensitive ELASA against human MCAF.Using this ELASA, we observed that the pleural fluid levels of MCAF increased in patients with malignant pleurisy upon the administration of an immunomodulator, OK-432. In addition, most human glioblastoma cell lines could produce MCAF in response to IL-1 or TNFalpha, suggesting a potential role of MCAF in tumor immunity.3. We also examined the role of MCAF-related molecule, interleukin-8 (IL-8) in the acute inflammation by administering a neutralizing antibody against IL-8. these experiments demonstrated that IL-8 plays an essential role in neutrophil infiltration and neutrophi-dependent tissue injuries in lipopolysaccharide-induced dermatitis and lung reperfusion injury.4. The determination of urinary IL-8 lavels using ELISA revealed that urinary IL-8 levels increased in the urines of patients with urinary tract infections and those with several types of glomerulonephritis such as lupus nephritis, acute glomerulonephritis, and IgA nephropathy with acute exacerbations.These results imply that the determination of urinary IL-8 level can be used for diagnosing and monitoring several diseases in urinary tracts.In addition, we explored the molecular biological aspects of IL-8 gene activation and established that NF_<-k>B site was essentially involved in the transcription of IL-8 gene in every cell line that we examined.