Yang Han

Small Molecules Blocking the Entry of Severe Acute Respiratory Syndrome Coronavirus into Host Cells

ABSTRACT Severe acute respiratory syndrome coronavirus (SARS-CoV) is the pathogen of SARS, which caused a global panic in 2003. We describe here the screening of Chinese herbal medicine-based, novel small molecules that bind avidly with the surface spike protein of SARS-CoV and thus can interfere with the entry of the virus to its host cells. We achieved this by using a two-step screening method consisting of frontal affinity chromatography-mass spectrometry coupled with a viral infection assay based on a human immunodeficiency virus (HIV)-luc/SARS pseudotyped virus. Two small molecules, tetra-O-galloyl-β-d-glucose (TGG) and luteolin, were identified, whose anti-SARS-CoV activities were confirmed by using a wild-type SARS-CoV infection system. TGG exhibits prominent anti-SARS-CoV activity with a 50% effective concentration of 4.5 μM and a selective index of 240.0. The two-step screening method described here yielded several small molecules that can be used for developing new classes of anti-SARS-CoV drugs and is potentially useful for the high-throughput screening of drugs inhibiting the entry of HIV, hepatitis C virus, and other insidious viruses into their host cells.

CRF01_AE subtype is associated with X4 tropism and fast HIV progression in Chinese patients infected through sexual transmission.

Background:The molecular epidemiology of the HIV-1 CRF01_AE subtype as a risk factor for fast HIV-1 progression remains poorly understood. Methods:We analyzed HIV-1 tropism by utilizing samples from 201 treatment-naive patients in our multicenter cohort (12 research centers in different provinces of China). Tropism was determined by V3 loop sequencing. Data from 235 treatment-naive patients infected sexually (including aforementioned 201 patients) in this cohort with date of estimated seroconversion (EDS) were retrospectively evaluated. Median time from EDS to AIDS was analyzed by Kaplan–Meier curves. Hazard ratios were determined by Cox proportional model. Results:CRF01_AE subtype was predominant (46.0%), especially in the MSM group. Further analysis revealed that the proportion of X4 tropism was higher in the CRF01_AE subtype (45.5%) than in others (C/CRF07_BC/CRF08_BC, 4.3%; B, 6.1%; Pu200a<0.001). CRF01_AE subtype was associated with faster progression from EDS to AIDS (4.8 vs. 6.4 years, Pu200a=u200a0.018) compared with non-CRF01_AE subtypes. In a multivariate model, the adjusted hazard ratio (aHR) of CRF01_AE was 1.42 (95% confidence interval, CI 0.99–2.03, Pu200a=u200a0.057), independent of HIV-1 viral load; it was also associated with fast progression to advanced immunodeficiency (aHR, 1.81, 95% CI 1.03–3.18, Pu200a=u200a0.038). Conclusion:CRF01_AE, a predominant HIV-1 subtype in Chinese HIV-1 sexually infected patients, tends to be associated with fast progression to AIDS and advanced immunodeficiency, which might be ascribed to high proportion of X4 tropism. Further investigation of these risk factors may have significant implications to clinical practice and policy-making.

HIV infection: an independent risk factor of peripheral arterial disease.

To the Editors: Highly active antiretroviral therapy (HAART) had significantly reduced the death rate resulted from HIV infection. However, HAART had also increased the risk of coronary heart disease in HIVinfected patients, especially in those on protease inhibitors (PIs). Whether HIV infection itself can promote the atherosclerosis is still controversial. Ankle brachial index (ABI) has been validated against lower extremity contrast angiography to determine its sensitivity, specificity, and accuracy as a diagnostic tool for lower extremity peripheral arterial disease, and the presence of a low ABI was predictive of total and cardiovascular mortality. Pulse wave velocity (PWV) is noninvasive parameter directly proportional to arterial wall stiffness. Our study aimed to determine whether the HIVinfected patients with or without receiving HAART are more likely to develop atherosclerosis in comparison to the general population, using ABI and PWV; and to assess the associated factors of peripheral arterial atherosclerosis. Our study had been approved by Peking Union Medical College Hospital Institutional Review Board. Eighty-two patients with HIV infection were divided into 2 groups: antiretroviral therapy (ART) na€ıve group comprising 41 antiretroviral-naive patients and HAARTtreated group comprising 41 patients on HAART for more than 12 months (34.76 17.1 months). Forty-three healthy people whose HIV screen tests were negative were enrolled as control subjects. Total cholesterol, triglyceride, lowdensity lipoprotein cholesterol (LDL-C), and high-density lipoprotein cholesterol (HDL-C) were determined by standard methods using Olympus AU5400 chemistry autoanalyzer (Olympus, Tokyo, Japan). CD4 lymphocyte count and CD8 lymphocyte count were measured by Epics XL flow cytometry (Beckman Coulter, Ramsey, MN). Plasma HIV RNA level was determined using a branched DNA assay (lower limit of quantification, 50 copies RNA/mL, version 3.0; Bayer Health Care, Leverkusen, Germany). ABI and PWV were determined using a pulse pressure analyzer (model BP-203RPE II; Nihon Colin, Komaki, Japan). The ABI was performed by measuring the systolic blood pressure from both brachial arteries and from both posterior tibial arteries; ABI was then calculated by the device as the ratio of systolic blood pressure in the leg to that in the arm on each side, and the average value was used for analysis. Pulse waves were recorded using sensors placed on both posterior tibial arteries. The time intervals required for the pulse waves to travel from the heart to both posterior tibial arteries were measured, and the distances between the heart and both posterior tibial arteries were estimated from the patient’s height. The PWV was calculated by dividing the distance by the time interval. For statistical analysis, between-group differences were compared by 2-sample t tests or Mann– Whitney tests (non-normal distribution) for continuous variables and by x analysis for categorical variables. Correlation was tested with Spearman rank order or Pearson correlation coefficient. Multiple linear regression analysis was used to test for independent associations between ABI and various factors. No significant statistical differences were observed among 3 groups considering age, sex, and family history of coronary heart disease or current smoking (Table 1). The ART-naive patients were more likely to be hypertensive and had lower level of total cholesterol and LDL-C than control subjects (P < 0.01). A lower HDL-C level was also noted in our ART-naive patients (P < 0.01). ART na€ıve HIV-infected patients have a lower ABI (P < 0.001) and a higher PWV (P = 0.010) when compared with control subjects. We included 41 ART-naive patients and 43 control subjects in a single group for linear regression analysis (R = 0.362; P = 0.001). We found that factors associated with reduced ABI were age (B = 0.03; 95% confidence interval: 0.000–0.005; P = 0.039) and HIV infection (B = 0.069; 95% confidence interval: 0.120 to 0.017; P = 0.01). That meant, after adjustment for other cardiovascular risk factors, HIV infection group was associated with a 0.069 lower ABI compared with control subjects. However, Pearson relation analysis shows not any association between ABI and common HIV infection parameters [CD4 lymphocyte count (r = 0.161; P = 0.314), CD8 lymphocyte count (r = 0.026; P = 0.871), CD4/CD8 ratio (r = 0.055; P = 0.731), and HIV RNA (r = 0.047; P = 0.770)] in patients with HIV. Patients in HAART-treated group mostly were treated with nonnucleoside reverse transcriptase inhibitors (NNRTIs, 40 of 41, 98%) and nucleoside reverse transcriptase inhibitors (NRTIs, 41 of 41, 100%). Only 7.3% (3 of 41) of the patients were on PIs–containing regimen. There were no differences in total cholesterol, triglyceride, LDL-C, HDL-C, and ABI between HAART-treated group and ART-naive group. Compared with ART-naive patients, the HAART-treated patients showed a lower body mass index, blood pressure, and PWV, as well as the HIV RNA level (P < 0.05). The idea that HIV-infected patients are likely to have an atherogenic lipid profile, such as high levels of total cholesterol, LDL-C, and triglyceride, has been reported by several studies. HAART is thought to be associated with these metabolic abnormalities. However, our ART-naive patients had lower total cholesterol and LDL-C levels, which is an atheroprotective lipid profile. This result supports the assumption that HAART may be contributory to dyslipidemia in HIV-infected patients. In this study, we proved that ART-naive patients have lower ABI compared with HIVuninfected control subjects, suggesting that HIV-infected patients are more likely to develop peripheral arterial disease. We also found in ART-naive HIV-infected patients having higher PWV level, which is also a marker of Supported by National Key Technologies R&D Program for the 11th Five-year Plan (2008ZX10001-006), Ministry of Health Clinical HIV/AIDS Research Grant (2007–2009), and Beijing Science and Technology Program Fund (D0906003040491).

Correlation between gag-specific CD8 T-cell responses, viral load, and CD4 count in HIV-1 infection is dependent on disease status.

Background: It still remains controversial which kind of relationships exist between HIV-1-specific CD8 T-cell responses and HIV RNA load or CD4 count over the course of the infection. This study was designed to investigate the role of HIV-specific CD8 responses in patients with different disease status. Methods: Three cohorts of patients were selected according to CD4 count levels: long-term nonprogressors (LTNPs, n = 19), asymptomatic progressors (CD4 counts between 500 and 350 cells/mm3, n = 14), and progressors (CD4 counts <350 cells/mm3, n = 23). Six of the LTNPs experiencing a quick loss of CD4 T-cells and another 6 LTNPs with stable CD4 counts were followed up. T-cell responses were studied using interferon (IFN) &ggr;-ELISpot assay against HIV p24 and 11 pools of HIV-Gag peptides. Results: No significant differences were found in Gag-specific CD8 responses among the 3 cohorts. However, inverse correlations were identified between CD8 responses and CD4 counts in asymptomatic progressors and between CD4 responses and viral loads in progressors. In addition, the sequential dynamics of CD8 responses in 6 LTNPs showed that with a quick loss of CD4 T-cells around the range of 500 to 300 cells/mm3, more vigorous CD8 responses were induced simultaneously, and plasma viremia was still kept relatively stable. Conclusions: These data suggest that the relationship between CD8 response and viral load or CD4 count is not universally consistent throughout the entire course of HIV-1 infection. Gag-specific CD8 responses may play differential roles in different stages of HIV-1 infection, and the maintenance of a threshold level of CD4 T-cells may contribute to mediate effective HIV-specific responses in natural control of HIV-1 infection.

Accelerating effect of human leukocyte antigen-Bw6 homozygosity on disease progression in Chinese HIV-1-infected patients.

Most HIV-1-infected individuals progress to AIDS within 8 to 10 years after seroconversion. Less than 5% of them, however, remain asymptomatic, although their CD4+ T-cell counts stay normal. In this study, our polymerase chain reaction sequence-specified primer (PCR-SSP) based human leukocyte antigen (HLA)-B genotyping of 28 typical progressors (TPs) and 15 long-term nonprogressors (LTNPs) revealed some evidence that an HLA-B locus polymorphism can influence the rate of disease progression in Chinese HIV-1-infected individuals: 12 of 28 TPs (43%) were HLA-Bw6 homozygotes. Only 1 of 15 LTNPs (6.7%) was homozygous for the polymorphism (P = 0.013), suggesting that HLA-Bw6 homozygosity is associated with accelerated disease progression. In contrast, 3 of 15 LTNPs (20%) were HLA-Bw4 homozygotes, whereas none of the 28 TPs were homozygotes (P = 0.037), supporting the conclusion that HLA-Bw4 homozygosity may have a protective role. Interestingly, the frequency of the HLA-B*15 allele was extremely high in the TP group (23.2%), which may be associated with faster disease progression in Chinese patients.

Prevalence of hepatitis B and C viruses in HIV-positive patients in China: a cross-sectional study.

Liver disease related to hepatitis B (HBV) and hepatitis C (HCV) may temper the success of antiretroviral therapy (ART) in China. Limited data exist on their prevalence in HIV‐positive Chinese. A multi‐centre, cross‐sectional study was carried out to determine the prevalence and disease characteristics of HBV and HCV co‐infection in HIV‐positive patients across 12 provinces.

Adiponectin and leptin levels in Chinese patients with HIV-related lipodystrophy: a 30-month prospective study.

The relationship of adipocytokine with the development of HIV-related lipodystrophy was investigated in a case-control study. Adipocytokine, lipid, and glycemic parameters were measured at every visit. Logistic regression analysis was used to assess the HIV-LD risk factors and the Spearman correlation coefficients test was used to assess the correlation between adiponectin with other metabolic variables. Most of the patients (96.3%) developed HIV-LD after month 12. Comparing the baseline adiponectin, the adiponectin concentration of the HIV-LD group rose by month 6 and began to decrease substantially by month 18; this reduction was maintained until month 30 (p < 0.05). Comparing the HIV-NLD group, the adiponectin concentration at months 18, 24, and 30 were significantly lower in the HIV-LD group. The leptin concentration of both the HIV-LD and HIV-NLD groups remained stable. Patients in the lower concentration of baseline adiponectin and greater adiponectin change rate at month 18 presented with increased odds ratio for HIV-LD. The adiponectin level had a correlation with serum triglycerides (r = -0.616, p < 0.0001), serum insulin concentration (r = -0.494, p = 0.001), and HDL-C (r = 0.673, p < 0.0001). The adiponectin concentration of HIV-LD began to decrease substantially by month 18. The lower baseline concentration of adiponectin and the greater change rate at month 18 were independent risk factors of HIV-LD. The adiponectin level had a correlation with serum triglycerides, serum insulin concentration, and HDL-C, suggesting that adiponectin may link the metabolic abnormalities and HIV-LD.

Comparison of genotypic resistance mutations in treatment-naive HIV type 1-infected patients in Korea and China.

Seventy-six treatment-naive human immunodeficiency virus (HIV)-1-infected patients were recruited from Korea and China to evaluate transmitted drug resistance (TDR). Although no major TDR was observed within the study population, some resistance-associated mutations in the reverse transcriptase region were observed (V118I 9.2%, V179D 7.9%). The frequencies of resistance-associated mutations in NNRTI (V179D) and PI minor mutations were higher in Korean patients compared with Chinese patients (13.6% vs. 0%, 45.5% vs. 12.5%, p < 0.05). Although unique clustering was observed in phylogenetic analyses according to geographic sources, cautious monitoring is recommended due to increasing TDR reports in this area where the population shares close geographic and cultural aspects.

Combination Antiretroviral Therapy Is Associated With Reduction in Liver Fibrosis Scores in HIV-1-Infected Subjects.

Abstract HIV increases the risk of liver disease as do two common coinfections, hepatitis B and C viruses (HBV and HCV). However, whether combination antiretroviral therapy (cART) reverses or exacerbates hepatic fibrosis remains unclear. This was an observational retrospective study. cART-naïve HIV-infected subjects without a history of substance abuse (including alcohol) had liver disease stage determined by aspartate aminotransferase-to-platelet ratio indices (APRIs) and fibrosis-4 (FIB-4) before and 24 and 48 weeks after cART. All the data were retrieved from previously established cohorts. Values before and after cART were compared using Wilcoxon test for paired samples. Regression analyses were used to determine factors associated with moderate-to-severe liver disease. Of the 1105 HIV-infected subjects, 120 were HBV coinfected and 64 were HCV coinfected. About 20% of HIV monoinfected participants had APRI and FIB-4 scores consistent with moderate-to-significant fibrosis compared to ∼36% of HIV–HBV coinfected and 67% to 77% of HIV–HCV coinfected participants. In adjusted analyses compared with HIV monoinfection, HBV coinfection was associated with 1.18-fold higher APRI (Pu200a<u200a0.001) and a 1.12-fold higher FIB-4 (Pu200a=u200a0.007) prior to cART; while HCV coinfection was associated with 1.94-fold higher APRI (Pu200a<u200a0.001) and a 1.43-fold higher FIB-4 (Pu200a<u200a0.001). After 48 weeks of cART, both fibrosis scores decreased in all subjects; however, HCV coinfection was still associated with higher fibrosis scores at week 48 compared to HIV monoinfection. cART was associated with improvement in hepatic fibrosis scores in the majority of HIV-hepatitis coinfected and HIV-monoinfected Chinese participants.

HIV-1 subtype B/B' and baseline drug resistance mutation are associated with virologic failure: a multicenter cohort study in China.

Background:Distribution of HIV-1 subtypes, transmitted drug resistance (TDR)/drug resistance mutation (DRM), and their impact on response to combination antiretroviral therapy remain poorly understood in China. Methods:We analyzed data from our multicenter cohort study with 444 antiretroviral-naive participants recruited between 2008 and 2010. HIV-1 subtype and tropism were determined by V3 sequencing, and TDR/DRM was determined by Pol sequencing. Virologic and immunologic responses were monitored over 96 weeks of follow-up. The initial combination antiretroviral therapy regimen for all patients was nevirapine + lamivudine + zidovudine or stavudine. Analysis 1 included patients who finished 96 weeks of follow-up (n = 379), and analysis 2 included all 444 patients. Results:Subtype B/B′ was associated with higher prevalence of TDR/DRM to nucleoside reverse transcriptase inhibitors and nonnucleoside reverse transcriptase inhibitors. Median time to HIV-1 suppression was 18 weeks in all 3 subtype groups. In Cox proportional models for viral suppression, neither viral tropism nor HIV-1 subtypes had any impact on viral suppression; however, subtypes CRF01_AE and C/CRF07_BC/CRF08_BC were associated with lower risk of virologic failure compared with subtype B/B′, with adjusted hazard ratio of 0.11 (P = 0.032) and 0.06 (P = 0.036), respectively in analysis 1, 0.42 (P = 0.047) and 0.22 (P = 0.008), respectively in analysis 2. This association was attenuated by adding DRM profiles to multivariate regression models. Neither subtype nor HIV-1 tropism affected immunologic response. Conclusions:HIV-1 subtype tended to be associated with virologic but not immunologic response; this effect could be ascribed to baseline DRM.