Clearance of HIV infection by selective elimination of host cells capable of producing HIV

The RNA genome of the human immunodeficiency virus (HIV) is reverse-transcribed into DNA and integrated into the host genome, resulting in latent infections that are difficult to clear. Here we show an approach to eradicate HIV infections by selective elimination of host cells harboring replication-competent HIV (SECH), which includes viral reactivation, induction of cell death, inhibition of autophagy and the blocking of new infections. Viral reactivation triggers cell death specifically in HIV-1-infected T cells, which is promoted by agents that induce apoptosis and inhibit autophagy. SECH treatments can clear HIV-1 in >50% mice reconstituted with a human immune system, as demonstrated by the lack of viral rebound after withdrawal of treatments, and by adoptive transfer of treated lymphocytes into uninfected humanized mice. Moreover, SECH clears HIV-1 in blood samples from HIV-1-infected patients. Our results suggest a strategy to eradicate HIV infections by selectively eliminating host cells capable of producing HIV.


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nature research | reporting summary
October 2018

Life sciences study design
All studies must disclose on these points even when the disclosure is negative. , the potential toxicity of drug treatment and the efficacy of treatment, at least 15 mice were employed in the experimental groups, which we have found to be sufficient to produce a statistically reliable results. The number of independent experiments was indicated in each figure legend.
No data were excluded for data analyses.
For each experiment, the number of biological independent animals/samples/patients is reported in the figure legend. A total of 8 sets of experiments were performed to establish HIV-1 infections and treatments using HSC-Hu mice, including 3 sets of experiments to test the reconstitution of NSG-SGM3 mice with human CD34+ stem cells and the establishment of HIV-1 infections in these mice, 2 sets of experiments to determine the dose of drugs that could be safely used in C57BL/6 mice and NSG-SGM3-derived HSC-Hu mice. Findings from in vitro cell culture studies were successfully replicated at least 3 independent experiments.
Age-and sex-matched animals were randomly assigned to the experimental groups.
The investigators were not blinded to experimental group assignments since they need to administrate specific ART or SECH treatments and assess the mice daily according to the regimen procedure. Note that full information on the approval of the study protocol must also be provided in the manuscript.

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Recruitment
Ethics oversight Note that full information on the approval of the study protocol must also be provided in the manuscript.
The validation for all antibodies for Western blot and flow cytometry to detect intracellular proteins and cell surface makers have been validated by the suppliers and published on their web pages. These antibodies for flow cytometry were further tested by staining cells with known makers and confirmed by flow cytometry before analyzing experimental samples. Human PBMCs with or without infection by HIV-1 were used as positive and negative controls for staining with PE-anti-HIV-1 p24 to confirm the specificity in the detection of HIV-1 p24 (Supplementary Figure 6a).
TZM-bl Cell line was obtained from the NIH AIDS Reagent Program (Cat# 8129).
TZM-bl Cells were further confirmed by flow cytometry to determine the expression of CD4, CXCR4 and CCR5. The cell line was further tested using HIV-1 standards with known titers to confirm its validity in quantitatively detecting HIV-1.
TZM-bl cells was obtained from NIH AIDS Reagent Program and tested to be negative for mycoplasma.
No commonly misidentified lines were used.
NSG-SGM3 mice were purchased from the Jackson Laboratory. Within 3 days after birth from the mating of NSG-SGM3 mice, newborn male and female mice were implanted intrahepatically with CD34+ human stem cells (AllCells) to generate a humanized mouse model. During treatments, mice were monitored daily for body weight, food consumption, activity and any other discomfort signs related to treatment. At the end of experiment, histological analysis by H&E staining was carried out in the major vital organs (brain, liver, lung and kidney).
No wild animals were used in this study.
No field-collected samples were used in this study.
Experiments were performed according to federal and institutional guideline, and with the approval of the Institutional Animal Care and Use Committee of the Houston Methodist Research Institute.
Buffy coats of anonymous healthy donors were purchased from the Gulf Coast Blood Center. The investigators were blinds to any covariate.
HIV-1+ patients of all age, gender and ethnic background were included. Ten ART-naive patients who were HIV-1+ but never received ART treatments were recruited from The University of Texas Health Science Center at Houston. Ten ART-experience samples from the outpatients undergoing ART treatments were collected at the Houston Methodist Hospital, Houston, Texas.
ART-naive patients were recruited with written informed consents and the approval of the Institutional Review Boards of University of Texas Health Science Center at Houston and Houston Methodist Research Institute. Experiments with ART-naïve patients were performed according to federal and institutional guidelines For ART-experienced patients, specimens of ART-treated, HIV-1+ patients from Houston Methodist Hospital Research Institute (HMRI) Biorepository were included. Experiments with de-identified samples from ART-treated patients were performed according to federal and institutional guidelines with the approval of the Institutional Review Board of the Houston Methodist Research Institute. For ART experienced patients, the informed consents for the de-identified biospecimens in Figure 7 from HMRI Biorepository were waived according to the federal regulation 45 CFR 46.116.
Samples used in this study were de-identified and assigned with unique identification number before provided to the laboratory for research. This number is used to label the samples that are stored and/or distributed, and also is used to for sample information and/or related clinical data. The identification system is password-protected and is only accessible by the PI and assigned research staff for the study. The protocols for this study were approved and oversaw by The University of Texas at Houston and the Houston Methodist Hospital Research Institute.