A lavage method for dynamic intraarticular monitoring of animal joints in situ: quantification and release kinetics of histamine after selective synovial mast cell activation by diverse secretagogues.
Evidence implicating synovial mast cells in the initiation and perpetuation of arthritis has increased. We have developed a method of joint lavage to monitor dynamic intraarticular events in the intact animal. Lavage was done before and after immunologic (passive sensitization followed by intravenous specific antigen or intraarticular anti-rat immunoglobulin E [IgE] heteroantiserum) and nonimmunologic (intraarticular calcium ionophore A-23187; phorbol 12-myristate, 13-acetate; and compound 48/80) synovial mast cell activation. To quantify and analyze synovial mast cell mediator release kinetics in situ, we measured lavage fluid histamine. With all activation protocols except A-23187, histamine release was evident within 5 minutes after introduction of the stimulus. The quantitative and chronological similarities between immunologically induced and compound 48/80-induced synovial mast cell histamine release kinetics suggested that connective tissue type mast cells are an important source of inflammatory mediators in rat joints. We also measured joint lavage fluid histamine levels in rats immunized with an active sensitization protocol. Histamine levels were determined by the autoanalyzer method and were confirmed by using a commercially available radioimmunoassay that uses a monoclonal antibody against acetylated histamine. We found that in many of these animals, at the peak of the serum IgE response, joint lavage fluid histamine levels were very high even before challenge with specific antigen, and that this increase was not due to diffusion into the joint of abnormally elevated plasma histamine. These data suggested that synovial mast cells are preferentially activated in states of high serum IgE immune responses. We have used a simple, inexpensive, rapid lavage technique to generate the first data on histamine release kinetics after selective synovial mast cell activation in the intact animal. The technique can be adapted for investigation of release kinetics of a variety of other substances from activated synovial cells and can be used in other arthritis models.
Malone
D G
DG
Department of Medicine, University of Wisconsin.
Verbsky
J W
JW
Dolan
P W
PW
D000818:Animals; D000941:Antigens / Q000276:immunology; D000001:Calcimycin / Q000494:pharmacology; D002199:Capillary Permeability; D006636:Histamine Release; D007106:Immune Sera / Q000276:immunology; D007114:Immunization; D007073:Immunoglobulin E / Q000276:immunology; D007596:Joints; D008297:Male; D008407:Mast Cells / Q000378:metabolism* / Q000502:physiology; D008991:Monitoring, Physiologic / Q000379:methods*; D051381:Rats; D011919:Rats, Inbred Strains; D013583:Synovial Membrane / Q000098:blood supply / Q000166:cytology / Q000378:metabolism*; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D007507:Therapeutic Irrigation / Q000379:methods*; D003189:p-Methoxy-N-methylphenethylamine / Q000494:pharmacology
D000941:Antigens; D007106:Immune Sera; D007073:Immunoglobulin E; D000001:Calcimycin; D003189:p-Methoxy-N-methylphenethylamine; D013755:Tetradecanoylphorbol Acetate