Subject(s)
Health Priorities , Neglected Diseases , Poverty , Uterine Cervical Neoplasms/mortality , Developing Countries , Early Detection of Cancer , Female , Humans , Tropical Medicine , Uterine Cervical Neoplasms/diagnosis , Uterine Cervical Neoplasms/prevention & control , Uterine Cervical Neoplasms/therapyABSTRACT
Protease inhibitor (PI)-based antiretroviral therapy (ART) can effectively suppress HIV-2 plasma load and increase CD4 counts; however, not all PIs are equally active against HIV-2, and few data exist to support second-line therapy decisions. To identify therapeutic options for HIV-2 patients failing ART, we evaluated the frequency of PI resistance-associated amino acid changes in HIV-2 sequences from a cohort of 43 Senegalese individuals receiving unboosted indinavir (n = 18 subjects)-, lopinavir/ritonavir (n = 4)-, or indinavir and then lopinavir/ritonavir (n = 21)-containing ART. Common protease substitutions included V10I, V47A, I54M, V71I, I82F, I84V, L90M, and L99F, and most patients harbored viruses containing multiple changes. Based on genotypic data, we constructed a panel of 15 site-directed mutants of HIV-2ROD9 containing single- or multiple-treatment-associated amino acid changes in the protease-encoding region of pol. We then quantified the susceptibilities of the mutants to the HIV-2 "active" PIs saquinavir, lopinavir, and darunavir using a single-cycle assay. Relative to wild-type HIV-2, the V47A mutant was resistant to lopinavir (6.3-fold increase in the mean 50% effective concentration [EC50]), the I54M variant was resistant to darunavir and lopinavir (6.2- and 2.7-fold increases, respectively), and the L90M mutant was resistant to saquinavir (3.6-fold increase). In addition, the triple mutant that included I54M plus I84V plus L90M was resistant to all three PIs (31-, 10-, and 3.8-fold increases in the mean EC50 for darunavir, saquinavir, and lopinavir, respectively). Taken together, our data demonstrate that PI-treated HIV-2 patients frequently harbor viruses that exhibit complex patterns of PI cross-resistance. These findings suggest that sequential PI-based regimens for HIV-2 treatment may be ineffective.
Subject(s)
Drug Resistance, Viral/drug effects , HIV Infections/drug therapy , HIV Protease Inhibitors/pharmacology , HIV Protease Inhibitors/therapeutic use , HIV-2/drug effects , Adult , Anti-HIV Agents/pharmacology , Anti-HIV Agents/therapeutic use , Cell Line , Female , Genotype , HIV Infections/virology , HIV Protease/drug effects , HIV Protease/genetics , HIV-2/enzymology , HIV-2/genetics , Humans , Male , Microbial Sensitivity Tests , Middle Aged , Molecular Sequence Data , Mutagenesis, Site-Directed , Mutation , Phylogeny , Senegal , Sequence Analysis, DNAABSTRACT
Spontaneous activity is expressed in many developing CNS structures and is crucial in correct network development. Previous work using [Ca(2+)](i) imaging showed that in the embryonic mouse hindbrain spontaneous activity is initiated by a driver population, the serotonergic neurons of the nascent raphe. Serotonergic neurons derived from former rhombomere 2 drive 90% of all hindbrain events at E11.5. We now demonstrate that the electrical correlate of individual events is a spontaneous depolarization, which originates at the rostral midline and drives events laterally. Midline events have both a rapid spike and a large plateau component, while events in lateral tissue comprise only a smaller amplitude plateau. Lateral cells have a large resting conductance and are highly coupled via neurobiotin-permeant gap junctions, while midline cells are significantly less gap junction-coupled and uniquely express a T-type Ca(2+) channel. We propose that the combination of low resting conductance and expression of T-type Ca(2+) current is permissive for midline neurons to acquire the initiator or driver phenotype, while cells without these features cannot drive activity. This demonstrates that expression of specific conductances contributes to the ability to drive spontaneous activity in a developing network.
