Pharmacological Effects Of Camptothecin On Lymphocytes In Vitro

AIM:To investigate the effects of camptothecin (CPT) on lymphocyte behaviors, especially its underlying mechanisms, in order to establish immunological base on which newly immunosuppressive reagent be developed.METHODS:1.The lymphocytes were isolated under sterile condition from lymph nodes of BALB/c mice and were stimulated by polyclonal activators after treated with CPT, concanavalin (ConA) or phorbol 12, 13-dibutyrate (PDB) combined with Ion. Expression level of CD69 activation of T cells was evaluated by Fluorescence conjugated monoclonal antibodies labeling. In addition, proliferation of lymphocytes was detected by CFDA-SE labeling.2.The lymphocytes were isolated from lymph nodes of BALB/c mice and were stimulated with ConA or PDB combined with Ion after treated with CPT.Distribution of cell cycle was analyzed by propidium iodide (PI) staining associated with flow cytometry. Cell apoptosis was analyzed by AnnexinV-APC/CFSE double stainging, flow cytometry and by method of DNA Ladder.3.Jurkat cells were treated with CPT. AnnexinV-FITC/ PI double stainig was used to detect early stage of apoptosis and PI staining, to analyse cell cycle. Jurkat cells were stained by Annexin V-PE/DiOC₆(3) to detect changes of (mitochondrial membrane pontential) △Ψ_m and by NAO, of mitochondrial mass.