Abstract
The existence of an antisense Open Reading Frame (ORF) that encodes a putative AntiSense Protein (ASP) on the proviral genome of Human Immunodeficiency Virus type 1 (HIV-1) was a source of debate for 30 years. During the last years, some progresses have been made to characterize the cellular immune response against ASP in HIV-1 seropositive patients. However, no tools were available for the detection of antibodies to ASP in the plasma of HIV-1-infected patients during the natural course of the infection. The aim of our study was to develop a Luciferase Immuno-Precipitation System (LIPS) to monitor the quantitative detection of ASP-specific antibodies in the plasma of HIV-1-infected patients [antiretroviral therapy (ART) naive-patients, patients under ART and HIV-1 controllers], patients who discontinued antiretroviral drugs (ARV). We further used this approach to delineate the epitopes of ASP targeted by antibodies. Antibodies directed against ASP were detected in 3 out of 19 patients who discontinued ARV (15%) and in 1 out of 10 ART-naive patients (10%), but were neither detected in HIV-1 infected patients under ART nor in HIV-1 controllers. Individual variations in levels of ASP-specific antibodies were detected overtime. Both the conserved prolin-rich motif and the core 60–189 region of ASP were found to be essential for antibody recognition in the four patients tested positive for anti-ASP antibodies, who were all untreated at the time of sampling. Moreover, for two of these patients, increased levels of ASP-specific antibodies were observed concomitantly to viremia declines. Overall, our method may represent a useful tool to detect a humoral response to ASP in HIV-1-infected patients, which allowed us to confirm the expression of ASP during the course of HIV-1 infection. Further studies will be needed to fully characterize the humoral response to ASP in HIV-1-infected patients.
Highlights
The existence of a conserved Open Reading Frame (ORF) located on the antisense strand of the proviral genome of the Human Immunodeficiency Virus type 1 (HIV-1) was first suggested in 1988 (Miller, 1988)
To detect anti-AntiSense Protein (ASP) antibodies in the plasma of HIV-1 infected patients, we used a codon-optimized ASP derived from pNL4-3 fused to the nano-luciferase protein
Using Luciferase Immuno-Precipitation System (LIPS) assay, we evaluated the presence of specific anti-ASP antibodies in the plasma samples of HIV-1-infected patients
Summary
The existence of a conserved Open Reading Frame (ORF) located on the antisense strand of the proviral genome of the Human Immunodeficiency Virus type 1 (HIV-1) was first suggested in 1988 (Miller, 1988). The same year, our group demonstrated the existence of Human-Tleukemia Virus Type 1 (HTLV-1) B-ZIP Factor (HBZ), another antisense retroviral protein (Gaudray et al, 2002) Following this discovery, we and others have investigated the antisense transcriptional activity of HIV-1 (Michael et al, 1994; Landry et al, 2007; Laverdure et al, 2012; Arpin-André et al, 2014) and studied ASP both in vitro and ex vivo. Vaquero conducted in 1995 previously revealed by western blot an in vitro translated ASP after immunoprecipitation by several serum samples from HIV-1-infected individuals, suggesting the existence of antibodies targeting ASP in vivo (Vanhée-Brossollet et al, 1995). These promising results have never been reproduced nor confirmed by any other study. In this study, we provided evidence of the existence of antibodies directed against ASP in vivo, we gave insights into the targeted epitopes of ASP, and, for the first time, we described the humoral response against ASP in the plasma of HIV-1-infected individuals
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