Oxidative stress induced by HIV-1 reverse transcriptase modulates the enzyme’s performance in gene immunization

Abstract

quinone oxidoreductase (Nqo1) and heme oxygenase 1 (HO-1), indicating the induction of oxidative stress response. The capacity to induce oxidative stress and stress response appeared to be an intrinsic property of a vast variety of RTs: enzymatically active and inactivated, bearing mutations of drug resistance, following different routes of processing and presentation, expressed from viral or synthetic expression-optimized genes. The total ROS production induced by RT genes of the viral origin was found to be lower than that induced by the synthetic/expression-optimized or chimeric RT genes. However, the viral RT genes induced higher levels of ROS production and higher levels of HO-1 mRNA than the synthetic genes per unit of protein in the expressing cell. The capacity of RT genes to induce the oxidative stress and stress response was then correlated with their immunogenic performance. For this, RT genes were administered into BALB/c mice by intradermal injections followed by electroporation. Splenocytes of immunized mice were stimulated with the RT-derived and control antigens and antigen-specific proliferation was assessed by IFN-γ/IL-2 Fluorospot. RT variants generating high total ROS levels induced significantly stronger IFN-γ responses than the variants inducing lower total ROS, while high levels of ROS normalized per unit of protein in expressing cell were associated with a weak IFN-γ response. Poor gene immunogenicity was also associated with a high (per unit of protein) transcription of antioxidant response element (ARE) dependent phase II detoxifying enzyme genes, specifically HO-1. Thus, we have revealed a direct link between the propensity of the microbial proteins to induce oxidative stress and their immunogenicity.

Diallyl Sulfide: Potential Use in Novel Therapeutic Interventions in Alcohol, Drugs, and Disease Mediated Cellular Toxicity by Targeting Cytochrome P450 2E1

“…It appears that Tat mainly manipulates the glutathione system, a major cellular thiol molecule involved in the maintenance of cellular redox state. Similarly other viral proteins such as gp120, Vpr, Nef, and HIV reverse transcriptase [146–150] have been implicated in the alterations of oxidative stress indices in HIV infection and neuropathogenesis. Taken together these studies point out that HIV viral proteins have a propensity to induce oxidative stress in various tissue types through generation of ROS, inflammatory cytokines, and other secondary metabolites.…”

Nonstructural Protein 1 of Tick-Borne Encephalitis Virus Induces Oxidative Stress and Activates Antioxidant Defense by the Nrf2/ARE Pathway

“…The production of intracellular ROS was measured by epifluorescence of reporter dyes as described previously [21,22,28]. Two days after transfection the growth medium was removed and cells were incubated in DMEM containing 25 µ M dichlorofluorescein diacetate (DCFH 2 DA) or 25 µ M dihydroethidium (DHE) for 30 min.…”

“…cDNA was synthesized using Mint reverse transcriptase (Evrogen) and random hexamer primers (Evrogen). RT-qPCR was performed on an IQ5 Real-Time PCR Detection System (BioRad) using protocol described previously [21,28]. The primer and probe sequences were as follows: HO-1: 5′-CCAGCAACAAAGTGCAAGATTC-3′ (sense primer), 5′-TCACATGGCATAAAGCCCTACAG-3′ (antisense primer), and 5′- (Cy5)-TCTCCGATGGGTCCTTACACTCAGCTTTCT-(BHQ2)-3′ (probe); Nqo1: 5′-GTCATTCTCTGGCCAATTCAGAGT-3′ (sense primer), 5′-TTCCAGGATTTGAATTCGGG-3′ (antisense primer), and 5′-(CY5)-ACTGACATATAGCATGGGCACACTCCAGC-(BHQ2)-3′(probe); and β-actin: 5′-GATCATTGCTCCTCCTGAGC-3′ (sense primer), 5′-ACTCCTGCTTGCTGATCCAC-3′ (antisense primer), and 5′-(R6G)-CTCGCTGTCCACCTTCCAGCAGAT-(BHQ-1)-3′ (probe).…”

“…There is only controversial data on the implication of NS1 in TLR3 signaling [16,17,18]. In the case of other viruses, the induction of oxidative stress is often associated with viral nonstructural proteins, such as HIV Tat [19], Vpr [20], and RT [21], HCV core and NS5A [7,22,23] parvovirus H-1 NS1 [24], and the human parvovirus B19 NS1 protein [25]. In the current study we explore ability of TBEV NS1 to affect the redox state of the host cell.…”

Cardiovascular Disease and HIV: Pathophysiology, Treatment Considerations, and Nursing Implications

“…Among persons living with HIV, atherogenesis is accelerated because of a constant proinflammatory state that exists throughout the lifetime of the infected person, even during antiretroviral therapy. 8 Infection with HIV induces inflammatory oxidative stress, stimulates the production of reactive oxygen species, 17,18 and causes platelet dysfunction. 19 This proinflammatory environment combined with increased levels of reactive oxygen species contributes to plaque formation and accelerates the progression of atherosclerotic disease.…”