Development and evaluation of a MAb-based ELISA for detection of Chlamydophila pneumoniae infection with variable domain 2 and 3 of the major outer membrane protein / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>This paper aims to develop a monoclonal antibodies (MAbs)- based ELISA for detecting Chlamydophila pneumoniae (C. pneumoniae) antigens in humans with the variable domains (VD) 2 and 3 of the major outer membrane protein (MOMPVD2-VD3) and to assess its sensitivity and specificity by comparing with a widely used MAb that is able to recognize the elementary bodies of C. pneumoniae.</p><p><b>METHODS</b>MOMPVD2-VD3 were overexpressed in Escherichia coli and purified by affinity chromatography. Mice were immunized with the recombinant antigen, and hybridomas secreting MAbs were screened. Three stable hybridomas clones were selected and named 5D6, 7G3, and 8C9. The MAbs-based ELISA was scrutinized for species-specific recognition with a number of human throat swab samples from Group I (156 patients with typical respiratory illness clinically confirmed before) and Group II (57 healthy donors).</p><p><b>RESULTS</b>In Group I, 55 positive cases were detected by anti-EB MAb-based ELISA, 51 cases were positive by MAbs 5D6-based ELISA, and 33 and 38 cases were positive by MAb 8C9 and 7G3-based ELISA respectively. Of the 57 samples from Group II "healthy donors", 5 were positive and 52 were negative with both anti-EB and 5D6-based tests, while 2 and 3 positive cases were identified by the other two MAb-based ELISAs respectively.</p><p><b>CONCLUSION</b>The novel MOMPVD2-VD3 MAb-based assay may have higher specificity than the anti-EB MAb, which may possibly be used as an alternative tool for the diagnosis of C. pneumoniae infection.</p>

Apoptosis induced by NNAMB, a novel polyamine conjugate, in human erythroleukemia K562 cells and its mechanism / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the apoptosis-inducing effects of NNAMB, a novel polyamine conjugate, in erythroleukemia K562 cells and its molecular mechanism.</p><p><b>METHODS</b>Cell viability was assessed by MTT assay and trypan blue dye exclusion method. The cell morphology was observed by fluorescence microscopy. The cell cycle distribution, apoptosis and mitochondrial membrane potential were measured by flow cytometry. The expression of caspase-3, -8, -9, cytochrome c in the K562 cells was detected by Western blot.</p><p><b>RESULTS</b>NNAMB inhibited the proliferation of K562 cells. The cells treated with NNAMB showed a typical apoptotic morphology, Sub-G1 peak and loss of mitochondrial membrane potential. Western blot assay showed that NNAMB increased the expression of caspase-3, -9, cytochrome c but not caspase-8 in a dose-and time-dependent manner.</p><p><b>CONCLUSION</b>NNAMB induces apoptosis via mitochondrial pathway in K562 cells.</p>

Adriamycin enhances anti-human DR5 monoclonal antibody (mDRA-6) induced HL-60 cells apoptosis / 中华血液学杂志

<p><b>OBJECTIVE</b>To investigate synergistic killing effect of anti-human DR5 (death receptor 5 of TRAIL) monoclonal antibody (mDRA-6) and adriamycin(Adr) on HL-60 cells.</p><p><b>METHODS</b>mDRA-6 was prepared by immunizing BALB/c mice with DR5 protein. DR5 expression on Adr-treated HL-60 cells was detected by flow cytometry. Morphologic changes of HL-60 cells were observed under fluorescence microscope. Cytotoxic and apoptotic effects of mDRA-6 and Adr on HL-60 cells were measured by MTT analysis. DNA fragmentation was detected by agarose gel electrophoresis.</p><p><b>RESULTS</b>Adr induce DR5 expression on HL-60 cells. Cell budding, chromatin condensation and apoptotic body formation were observed in HL-60 cells treated by mDRA-6 and Adr. Death and apoptosis of these cells and DNA ladder were exhibited on agarose gel electrophoresis.</p><p><b>CONCLUSION</b>mDRA-6 and Adr have synergistic killing effect on HL-60 cells.</p>