Establishment of a consistent L929 bioassay system for TNF-alpha quantitation to evaluate the effect of lipopolysaccharide, phytomitogens and cytodifferentiation agents on cytotoxicity of TNF-alpha secreted by adherent human mononuclear cells.
TUMOR necrosis factor-alpha (TNF-alpha) plays an important role in the pathogenesis of rheumatoid arthritis. The present study was to evaluate the effects of lipopolysaccharide (LPS), phytomitogens and cytodifferentiation agents on cytotoxicity of TNF-alpha secreted by adherent human mononuclear cells (AMC). TNF-alpha cytotoxicity in LPS-treated, phytomitogen-treated, and cytodifferentiation agent-treated AMC supernatants were analyzed by the L929 bioassay system. Our results showed that LPS could induce homogeneous TNF-alpha production by AMC whereas, in addition to TNF-alpha, phytomitogens could also induce other TNF-like factors. Neither methotrexate, retinoic acid nor sodium butyrate can inhibit TNF-alpha cytotoxicity, while hexamethylene bisacetamide could not only inhibit TNF-alpha cytotoxicity but also TNF-alpha inducing ability of LPS to AMC.
Shiau
M Y
MY
Hung Kuang Institute of Technology, Chung Shan Medical University, Taiwan, Republic of China.
Chiou
H L
HL
Lee
Y L
YL
Kuo
T M
TM
Chang
Y H
YH
D000081:Acetamides / Q000494:pharmacology; D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D001681:Biological Assay / Q000379:methods*; D002087:Butyrates / Q000494:pharmacology; D002448:Cell Adhesion / Q000502:physiology; D002454:Cell Differentiation; D002460:Cell Line; D003601:Cytotoxicity Tests, Immunologic; D004306:Dose-Response Relationship, Immunologic; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology; D037102:Lectins / Q000494:pharmacology*; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D008070:Lipopolysaccharides / Q000494:pharmacology*; D008727:Methotrexate / Q000494:pharmacology; D051379:Mice; D011994:Recombinant Proteins / Q000378:metabolism; D014212:Tretinoin / Q000494:pharmacology; D014409:Tumor Necrosis Factor-alpha / Q000032:analysis* / Q000378:metabolism / Q000633:toxicity*
D000081:Acetamides; D000970:Antineoplastic Agents; D002087:Butyrates; D007166:Immunosuppressive Agents; D037102:Lectins; D008070:Lipopolysaccharides; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D014212:Tretinoin; C014026:hexamethylene bisacetamide; D008727:Methotrexate