A new system for parallel drug screening against multiple-resistant HIV mutants based on lentiviral self-inactivating (SIN) vectors and multi-colour analyses

Abstract

BackgroundDespite progress in the development of combined antiretroviral therapies (cART), HIV infection remains a significant challenge for human health. Current problems of cART include multi-drug-resistant virus variants, long-term toxicity and enormous treatment costs. Therefore, the identification of novel effective drugs is urgently needed.MethodsWe developed a straightforward screening approach for simultaneously evaluating the sensitivity of multiple HIV gag-pol mutants to antiviral drugs in one assay. Our technique is based on multi-colour lentiviral self-inactivating (SIN) LeGO vector technology.ResultsWe demonstrated the successful use of this approach for screening compounds against up to four HIV gag-pol variants (wild-type and three mutants) simultaneously. Importantly, the technique was adapted to Biosafety Level 1 conditions by utilising ecotropic pseudotypes. This allowed upscaling to a large-scale screening protocol exploited by pharmaceutical companies in a successful proof-of-concept experiment.ConclusionsThe technology developed here facilitates fast screening for anti-HIV activity of individual agents from large compound libraries. Although drugs targeting gag-pol variants were used here, our approach permits screening compounds that target several different, key cellular and viral functions of the HIV life-cycle. The modular principle of the method also allows the easy exchange of various mutations in HIV sequences. In conclusion, the methodology presented here provides a valuable new approach for the identification of novel anti-HIV drugs.

Intestinal Parasitosis in Relation to CD4+T Cells Levels and Anemia among HAART Initiated and HAART Naive Pediatric HIV Patients in a Model ART Center in Addis Ababa, Ethiopia

“…On the other hand, the 27.8% prevalence of IPs detected among patients on HAART in this study is higher than that reported in Brazil (24%) and Dessie (17.6%) [ 19 , 24 ] but lower than from studies done in Indonesia (50%), in selected ART centers of Adama, Afar and Dire-Dawa (48%), Jimma (39.6%) and Addis Ababa (34.3%), [ 11 , 23 , 25 , 26 ]. This lower prevalence might be due to better laboratory test supported follow-up and better awareness of the patients themselves in adopting prevention and treatment measures against intestinal parasites in our study area plus the combined effect of anti-helminthes and HAART may better decrease occurrence of IPs.…”

“…The prevalence of Strongyloides stercoralis is higher in non-HAART (6.9%) than patients on HAART (2.5%) in the present study which is comparable with a study done in Nigeria (6.39%) [ 6 ]. But higher than studies done in Malaysia (0%), China (0%) and Dessie (0.7%) [ 17 , 18 , 24 ]. The possible reasons might be due to geographical differences, comparatively small sample size in this study and age differences i.e.…”

Fucoidans as Potential Inhibitors of HIV-1

“…A model system based on lentiviral vectors was used to study antiviral activity of fucoidans [16,22]. Pseudo-HIV-1 particles are recombinant lentiviruses based on HIV-1.…”

“…DNA of the lentiviral vector containing the marker eGFP gene and the plasmids directing the synthesis of the proteins that are required for the formation of pseudo-HIV-1 particles were introduced into HEK293 cells via calcium phosphate transfection. The pseudo-HIV-1 particles were collected 24 h following transfection with a 12 h interval [16]. …”

“…Limitations in working with infectious viruses greatly complicate the search for and testing of antiviral drugs. Using the system designed at the Engelhardt Institute of Molecular Biology of the Russian Academy of Sciences, it is possible to investigate the activity of potential inhibitors that prevent the interaction of simple and complex retroviruses with the primary surface receptors of the cell thus preventing the infection of cells [15,16]. The aim of this work was to study the antiviral activity of a number of natural fucoidans and choose the most active fucoidans samples to be studied further as potential candidates for antiviral drug development.…”