OBJECTIVE: Present study addresses the issue whether apoptosis and necrosis increases the antigenicity of proteins recognized by antinuclear antibodies. MATERIAL AND METHODS: HEp-2 cells were cultured in standard conditions; apoptosis was induced by camptothecin and necrosis by mercuric chloride. Protein antigenicity of cell extracts was tested onto nitrocellulose membranes and probed with positive or negative sera for antinuclear antibodies by a luminescent-dot-ELISA system. RESULTS: Apoptotic changes in HEp-2 cells appeared by 24 hours of camptothecin exposure, meanwhile the necrotic features become visible earlier. Luminescence was significantly superior in ANA positive sera than in ANA negative controls. Antinuclear antibody sera recognized better the antigens from the apoptotic and necrotic cells than controls without chemical treatments. CONCLUSIONS: Apoptosis and necrosis increase the ANA binding by better availability of intracellular antigens, or by disclosing cryptic epitopes.
OBJECTIVE: Present study addresses the issue whether apoptosis and necrosis increases the antigenicity of proteins recognized by antinuclear antibodies. MATERIAL AND METHODS:HEp-2 cells were cultured in standard conditions; apoptosis was induced by camptothecin and necrosis by mercuric chloride. Protein antigenicity of cell extracts was tested onto nitrocellulose membranes and probed with positive or negative sera for antinuclear antibodies by a luminescent-dot-ELISA system. RESULTS: Apoptotic changes in HEp-2 cells appeared by 24 hours of camptothecin exposure, meanwhile the necrotic features become visible earlier. Luminescence was significantly superior in ANA positive sera than in ANA negative controls. Antinuclear antibody sera recognized better the antigens from the apoptotic and necrotic cells than controls without chemical treatments. CONCLUSIONS: Apoptosis and necrosis increase the ANA binding by better availability of intracellular antigens, or by disclosing cryptic epitopes.
Authors: Christopher W Pohlmeyer; Ching Shang; Pei Han; Zhi-Hua Cui; Randall M Jones; Astrid S Clarke; Bernard P Murray; David A Lopez; David W Newstrom; M David Inzunza; Franziska G Matzkies; Kevin S Currie; Julie A Di Paolo Journal: BMC Rheumatol Date: 2021-03-30