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© 1992 British Society for Rheumatology

other

IL-1 SECRETING CELL ASSAY AND ITS APPLICATION TO CELLS FROM PATIENTS WITH RHEUMATOID ARTHRITIS

B. DULARAY, C. I. WESTACOTI and C. J. ELSON

Department of Pathology, University of Bristol

Correspondence to: Correspondence to Dr C. Elson, Department of Pathology, University of Bristol, Bristol BS8 1TD

The cells within a population that were secreting interleukin-1 (IL-1) were enumerated and visualized by an ELISA-SPOT assay. Initial experiments designed to validate the assay revealed that the number of IL-1ß spot forming cells was increased by exposing normal blood monocytes to LPS and that spot formation was prevented by incubating the cells with cycloheximide. Normal blood polymorphonuclear leucocytes (PMNs) produced IL-1{alpha} and IL-1ß in response to recombinant granulocyte monocyte colony stimulating factor (rhGMCSF) but not to cytochalasin B, calcium ionophore or LPS. Monocytes and PMN were isolated from the synovial fluid (SF) and blood of patients with rheumatoid arthritis (RA) and the ability of these cells to secrete IL-1{alpha} and IL-1ß compared. A higher proportion of SF derived monocytes were found to secrete IL-1{alpha} spontaneously compared to the corresponding blood cells. IL-1{alpha} secreting monocytes were not detected although high numbers of IL-1{alpha} secreting cells were found among cells isolated from rheumatoid synovium. By contrast SF PMNs did not produce IL-1{alpha} or IL-1ß whereas blood PMNs from some (3/8) RA patients produced IL-1{alpha} and/or IL-1ß. It is considered that the IL-1 ELISA-SPOT is a highly sensitive technique for detecting IL-1 secreting cells.

KEY WORDS: Synovial cells, Interleukin-i1{alpha}, Interleukin-1ß, Rheumatoid arthritis


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