1.Upregulation of cell adhesion molecules by endothelin-1 on human mesangial cells in vitro
Jingmei LIN ; Yipu CHEN ; Jin GAO
Chinese Journal of Nephrology 1994;0(02):-
Objective To study the expression of intercellular adhesion molecule-1 ( ICAM-1 ) and vascular cell adhesion molecule-l(VCAM-L) on human mesangial cells (HMC) after stimulation by endothelin-1 (ET-1). Methods The ex-pression of ICAM-1 and VCAM-1 was detected at mRNA and protein levels by using Northern bolt and cell ELISA respectively. Results There was basic mRNA and protein expression of ICAM-1 and VCAM-1 on HMC in the control groups without ET-1. ET-l(10-7mol)augmented the mRNA expression of ICAM-1 and VCAM-1 with the maximum at 4 -8 hr. Increased protein expression of ICAM-1 and VCAM-1 was followed thereafter in a time-depended manner. Conclusion The upregulated expression of ICAM-1 and VCAM-1 by ET-1 may have an important role in the development of glomerulonephritis.
2.Construction of recombinant influenza virus carrying human metapneumovirus epitopes
Yan Xiao LI ; Yipu LIN ; Mei KONG ; Liru GUO ; Ming ZOU ; Xu SU
Chinese Journal of Microbiology and Immunology 2019;39(1):6-11
Objective To construct and rescue recombinant influenza virus strains expressing hu-man metapneumovirus ( hMPV) epitopes. -ethods B cell, CTL and Th epitopes predicted by bioinformat-ics software were coupled together in different combinations. These different array genes were inserted into the NS1 gene of influenza virus strain A/PR/8/34 ( PR8 ) , respectively. Recombinant PR8 influenza virus vectors expressing different hMPV antigenic epitopes were rescued by reverse genetics using eight-plasmid system. Sequencing analysis was conducted to verify whether the rescued viruses carried the chimeric hMPV epitopes. Hemagglutination ( HA) titers, half tissue culture infection dose ( TCID50 ) and growth curves were detected. Results Interval sequences GPGPG and KK were introduced into hMPV epitope combinations to construct multi-epitope antigens (MEA). These MEA were inserted into the PR8 NS gene, respectively. Using 8 plasmid system, three recombinant influenza virus strains were rescued successfully. After cultured for three passages in Madin-Darby canine kidney ( MDCK) cells and one in eggs, these three recombinant strains could proliferate steadily. Whole genome sequencing verified that the three recombinant strains car-ried the chimeric MEA sequences, named as rFLU/hMPV/B, rFLU/hMPV/CTL-Th and rFLU/hMPV/B-Th. HA titers of the recombinant strains were 128, 128 and 256 using turkey erythrocyte, respectively. Their TCID50 were 107. 0/ml, 106. 8/ml and 107. 0/ml, respectively. Growth curve tests also verified that the recombinant strains could proliferate steadily in MDCK cells. Conclusions Three recombinant influenza vi-rus vector strains carrying the B cell, CTL and Th epitopes of hMPV were rescued successfully. This study lays the foundation for further evaluation of the immune effects of these recombinant viruses and their poten-tial application value in vaccine development.