Transcriptomic crosstalk between viral and host factors drives aberrant homeostasis of T-cell proliferation and cell death in HIV-infected immunological non-responders.

BACKGROUND: Immunological non-responders (INRs) among people living with HIV have inherently higher mortality and morbidity rates. The underlying immunological mechanisms whereby failure of immune reconstitution occurs in INRs require elucidation. METHOD: HIV-1 DNA and HIV-1 cell-associated RNA (CA-HIV RNA) quantifications were conducted via RT-qPCR. Transcriptome sequencing (RNA-seq), bioinformatics, and biological verifications were performed to discern the crosstalk between host and viral factors. Flow cytometry was employed to analyze cellular activation, proliferation, and death. RESULTS: HIV-1 DNA and CA-HIV RNA levels were observed to be significantly higher in INRs compared to immunological responders (IRs). Evaluation of CD4/CD8 ratios showed a significantly negative correlation with HIV-1 DNA in IRs, but not in INRs. Bioinformatics analyses and biological verifications showed IRF7/INF-α regulated antiviral response was intensified in INRs. PBMCs of INRs expressed significantly more HIV integrase-mRNA (p31) than IRs. Resting (CD4+CD69- T-cells) and activated (CD4+CD69+ T-cells) HIV-1 reservoir harboring cells were significantly higher in INRs, with the co-occurrence of significantly higher cellular proliferation and cell death in CD4+ T-cells of INRs. CONCLUSION: In INRs, the systematic crosstalk between the HIV-1 reservoir and host cells tends to maintain a persistent antiviral response-associated inflammatory environment, which drives aberrant cellular activation, proliferation, and death of CD4+ T-cells.

Differences in drug resistance of HIV-1 genotypes in CSF and plasma and analysis of related factors.

The emergence of HIV drug resistance seriously affects the quality of life of patients. However, there has been no extensive study of CSF resistance. The aim of this study is to evaluate common HIV-1 resistance in CSF and compare it with resistance in matched plasma, and analyse the influencing factors of cerebrospinal fluid drug resistance. The matched CSF and plasma samples of 62 HIV-1 patients were tested at one study site in China (Chongqing; 2019-2022). HIV genotyping and drug resistance was evaluated using the Stanford v8.7 algorithm. The diagnosis and treatment data and basic information were collected from the clinical case system, and the influencing factors of drug resistance mutations in CSF was obtained by variance analysis. CSF and matched plasma HIV-1 subtypes were confirmed in 62 patients, and the most frequent recombinant form was CRF07-BC (64.5%). Thirteen patients (21.0%) were detected with drug-resistant mutations, and the sites were consistent in both CSF and matched plasma. The drug-resistant ratios of untreated patients and treated patients were 5/51 (9.8%) and 8/11 (72.7%), respectively. The type with the highest mutation frequency was NNRTI, and no mutation was found in INSTI. Multivariate analysis indicated that ARV treatment was associated with CSF resistance (P < 0.001). The subtypes and drug resistance mutation sites are consistent in CSF and matched plasma samples of HIV-1 patients, and there is a correlation between ARV treatment and possible drug resistance, especially in CSF reservoirs. These findings highlight the concern about CSF drug resistance in HIV patients.

Pretreatment drug resistance in people living with HIV: A large retrospective cohort study in Chongqing, China.

OBJECTIVES: The emergence of pretreatment drug resistance (PDR) caused by increased usage of antiretroviral therapy (ART) represents a significant challenge to HIV management. In this study, we evaluated the prevalence of PDR in people living with HIV (PLWH) in Chongqing, China. METHODS: We retrospectively collected the data of 1110 ART-naïve PLWH in Chongqing from January 1, 2018 to June 30, 2021. HIV-1 genotypes and drug resistance were analyzed using the HIV-1 pol sequence. Risk factors associated with PDR were evaluated via the logistic regression model. RESULTS: Nine genotypes were detected among 1110 participants, with CRF07_BC (55.68%) being the dominant genotype, followed by CRF01_AE (21.44%), CRF08_BC (14.14%), and other genotypes (8.74%). Of all the participants, 24.14% exhibited drug resistance mutations (DRMs). The predominant DRMs for non-nucleoside reverse transcriptase inhibitors (NNRTIs) and nucleoside reverse transcriptase inhibitors (NRTIs) were V179D/E/A/DIN (13.60%) and M184V/I (1.44%), respectively, whereas only two major DRMs (M46L and I54L) were identified for protease inhibitors (PIs). The total prevalence of PDR was 10.54%, with 2.43%, 7.66%, and 1.71% participants exhibiting PDR to NRTIs, NNRTIs, and PIs, respectively. Furthermore, female PLWH, delays in ART initiation, and the CRF08_BC genotype were associated with a higher risk of PDR. CONCLUSIONS: Our study provides the first large cohort data on the prevalence of PDR in Chongqing, China. HIV-1 genotypes are diverse and complex, with a moderate level of PDR, which does not reach the threshold for the initiation of a public health response. Nevertheless, continuous surveillance of PDR is both useful and advisable.

Multicenter evaluation of Xpert HIV-1 viral load assay for HIV quantification in China.

New approaches to increase HIV-1 testing and HIV-1 viral load (VL) monitoring are needed for people living with HIV (PLHIV) in China. The Xpert HIV-1 VL assay was prequalified by the World Health Organization in 2017 but has not been evaluated in China. A multicenter evaluation was conducted to assess the accuracy of the Cepheid Xpert HIV-1 VL assay compared to the Abbott RealTime HIV-1 assay in China. Overall agreement was seen in 558 of 562 specimens (99.29%) with a κ value of 0.962. Pearson's coefficient between the two assays was 0.943. Analyzed by the Bland-Altman method, the mean bias was -0.54 log10 copies/mL, and 94.05% results fell within the 95% confidence limit of agreement (-1.248 to 0.168 log10 copies/mL). The coefficient of variation of the Cepheid Xpert HIV-1 VL assay ranged from 0.61% to 1.55%, as determined by testing eight positive plasma specimens with three different lots on different days. Due to its simplicity, random-access, rapid turnaround time, and accuracy, the Xpert HIV-1 VL assay can be used in local hospitals and clinics that bear the burden of identifying and treating HIV patients in China.

Target-induced aptamer release strategy based on electrochemical detection of staphylococcal enterotoxin B using GNPs-ZrO2-Chits film.

A novel electrochemical aptasensor was developed for ultrasensitive detection of staphylococcal enterotoxin B (SEB) by combining signal amplification and target-induced aptamer release strategy. A gold electrode was modified with a nanocomposite made of gold nanoparticles reduced in situ, zirconia nanoparticles, and chitosan. The SEB aptamer was hybridized by anchoring the capture probe on the modified gold electrode surface through AuS binding. In the presence of SEB, the capture probe-aptamer duplex was compelled to open, releasing the aptamer from the electrode. The resulting single-strand capture probe was hybridized with a biotinylated detection probe and labeled with streptavidin-horseradish peroxidase, producing an ultrasensitive enzyme-catalyzed electrochemical signal. Under optimal conditions, the amperometric responses were proportional to the SEB concentrations ranging from 2 to 512ngmL(-1), with a detection limit of as low as 0.24ngmL(-1) (S/N=3). The aptasensor exhibited good stability, outstanding reproducibility, and high selectivity. The as-prepared aptasensor was used to analyze SEB in human serum specimens, and validated through enzyme-linked immunosorbent assay method. Analytical results suggest that the developed assay is a promising alternative approach for detecting SEB in clinical diagnosis.