sepharose and Arthritis

To study the effects of recombinant tumor necrosis factor alpha (TNF-alpha), interleukin 1 beta (IL-1 beta) and interleukin 6 (IL-6) on proteoglycan metabolism of isolated chondrocytes.. Human articular cartilage cells were cultured in agarose gel. In these culture conditions, chondrocytes keep their phenotypic stability. They release cartilage specific proteoglycans into the surrounding artificial matrix. Proteoglycan synthesis was measured by the incorporation of 35sulfate (35S).. TNF-alpha and IL-1 beta depressed proteoglycan synthesis and induced proteoglycan degradation. The effects of both cytokines were additive, when used in submaximal doses. No mutual induction of TNF-alpha and IL-1 beta was shown, but both cytokines stimulated the chondrocytes to release IL-6, up to 100,000 pg/ml. Equal amounts of human recombinant IL-6 did not affect proteoglycan synthesis. IL-6 did not alter proteoglycan quality, nor did it modulate the IL-1 beta activities on proteoglycan metabolism.. These findings illustrate the role of IL-1 beta and TNF-alpha in cartilage degradation and suggest that the role of the large amounts of IL-6 released in response to IL-1 in chronic arthritis is not directly protective with regard to proteoglycan metabolism.

Topics: Arthritis; Cartilage, Articular; Cells, Cultured; Cytokines; Humans; Interleukin-1; Interleukin-6; Lipopolysaccharides; Microscopy, Electron; Proteoglycans; Recombinant Proteins; Sepharose; Tumor Necrosis Factor-alpha

Peripheral blood mononuclear cells (PBMC) from Rhesus monkeys previously immunized with bovine type II collagen to induce arthritis were cultured with the same antigen. Because the native protein is poorly soluble in culture medium a heating step is often used. The antigen in this form induced PBMC proliferation, but epitopes for the induction of antibody production and arthritis were lost. To keep the native protein intact it was coated on affigel beads. With the immobilized antigen specific antibody production could be induced.

Topics: Animals; Antibody Specificity; Arthritis; Autoantibodies; Collagen; Disease Models, Animal; Hot Temperature; Immunization; Lymphocytes; Macaca mulatta; Sepharose

An in vivo skin chamber method using lesions obtained by suction was evaluated. Neither dyspigmentation nor scarring were seen after 2 mth. The number of leucocytes accumulated in the collection chamber was correlated to the area of the lesion. Reproducibility remained essentially unchanged over an extended period and was 19% for one chamber and 13.6% for determinations with two chambers. No correlation was found between results obtained with the skin chamber technique and with chemotaxis or random migration as determined by an underagarose technique. When factors influencing in vivo and in vitro migration were studied, it was found that nonsteroidal anti-inflammatory drugs when given to arthritis patients or healthy volunteers inhibit leucocyte migration in vivo while in vitro migration was unchanged. Activated serum and LTB4 attracted leucocytes both in vivo and in vitro. The attraction by serum appeared at least partly to be caused by C5a. Polymorphonuclear leucocytes harvested from skin chambers were chemotactically deactivated and their bactericidal capacity reduced. The chemiluminescent response to formyl-methionyl-leucyl-phenylalanine and opsonized zymosan was increased. Exposed to zymosan activated serum, blood leucocytes showed a similar functional modification as leucocytes harvested from a skin chamber, and our findings suggest that the altered function of leucocytes in an inflammatory focus is largely the result of their exposure to chemotactic factors.

Topics: Anti-Inflammatory Agents; Arthritis; Bacteria; Cell Movement; Chemotaxis, Leukocyte; Humans; Inflammation; Kinetics; Leukocytes; Methods; Phagocytosis; Sepharose; Skin