The relative level of infection was determined by flow cytofluorimetry on an Epics 4XL Beckman Coulter instrument 48 h following the infection. Success AND INHIBITOR Building of pseudo HIV one particles and using them to infect numerous eukaryotic cell lines Efficiency of transduction of target cells with pseudo HIV one particles, and as a result the fluorescence degree within the resulting transgenic cells, may be the most significant parameter of a lentiviral program. This parameter depends upon the construction of pseudoviral particles as well as individual line of infected target cells. The transplaninhibitors human lymphoblastic cells Jurkat and CE M SS , Kasumi 1 , and mouse embryonic fibroblasts SC 1 have been applied as target cells. Two types of pseudo HIV 1 particles differing in coat proteins were obtained and subjected to study.
Particles on the to start with variety contain the HIV 1 coat protein selleck Vatalanib gp160 ; particles of the second type consist of the vesicular stomatitis virus G protein. Using particles of your 1st style resulted within a comparatively very low transduction efficiency and a weaker fluorescence signal from the contaminated cells . Inside the situation of pseudo HIV one particles carrying the VSV G protein, the share of infected cells and the degree of expression in the green fluorescent marker protein were substantially higher . In addition, the particles pseudo typed with the VSV G protein can be used to transfer marker genes to your cells with broad type and tissue specificity. This process permits one to execute the search for retroviruses affecting tissues besides blood.
For this reason, pseudo HIV 1 particles with the VSV G protein were the ones utilized in most experiments devoted towards the research with the properties of inhibitors of HIV 1 reverse transcriptase and integrase. Nucleoside inhibitors of HIV one reverse transcriptase Modified nucleosides selleck PA-824 and nucleotides have observed broad application while in the treatment of many different viral diseases, which include the HIV one infection . Their mechanism of action incorporates conversion of these compounds, in the cell, in to the corresponding nucleoside triphosphates, which act as terminating substrates for viral DNA and RN A polymerases. The integration of nucleotides to the growing chain of viral DNA RN A blocks viral replication and slows the spread on the infection. The 1st and most renowned anti HIV one agent of this class is 3? azido 3? deoxythymidine , which may inhibit viral replication even at a nano molar concentration.
The antiviral action of AZT was studied with respect to pseudo HIV 1 particles carrying the HIV 1 coat protein gp160 or the VSV protein G on their surface.