Broad-Spectrum Antiviral Activity of an Ankyrin Repeat Protein on Viral Assembly against Chimeric NL4-3 Viruses Carrying Gag/PR Derived from Circulating Strains among Northern Thai Patients.

Certain proteins have demonstrated proficient human immunodeficiency virus (HIV-1) life cycle disturbance. Recently, the ankyrin repeat protein targeting the HIV-1 capsid, AnkGAG1D4, showed a negative effect on the viral assembly of the HIV-1NL4-3 laboratory strain. To extend its potential for future clinical application, the activity of AnkGAG1D4 in the inhibition of other HIV-1 circulating strains was evaluated. Chimeric NL4-3 viruses carrying patient-derived Gag/PR-coding regions were generated from 131 antiretroviral drug-naïve HIV-1 infected individuals in northern Thailand during 2001⁻2012. SupT1, a stable T-cell line expressing AnkGAG1D4 and ankyrin non-binding control (AnkA32D3), were challenged with these chimeric viruses. The p24CA sequences were analysed and classified using the K-means clustering method. Among all the classes of virus classified using the p24CA sequences, SupT1/AnkGAG1D4 demonstrated significantly lower levels of p24CA than SupT1/AnkA32D3, which was found to correlate with the syncytia formation. This result suggests that AnkGAG1D4 can significantly interfere with the chimeric viruses derived from patients with different sequences of the p24CA domain. It supports the possibility of ankyrin-based therapy as a broad alternative therapeutic molecule for HIV-1 gene therapy in the future.

Seroprevalence of antibodies to measles, mumps, and rubella, and serologic responses after vaccination among human immunodeficiency virus (HIV)-1 infected adults in Northern Thailand.

BACKGROUND: After the global implementation of national immunization programs for prevention of measles, mumps, and rubella (MMR), the prevalences of protective antibodies to these viruses are high in general population. However, there are limited data among human immunodeficiency virus (HIV)-1 infected individuals. This study aimed to determine the seroprevalence of antibodies to these viruses, and the serologic responses after vaccination among HIV-infected adults in Northern Thailand. METHODS: A cross-sectional study was conducted in 500 HIV-infected adults, aged 20-59 years, receiving combination antiretroviral therapy, CD4 cell count ≥200 cells/mm(3), and plasma HIV-1 RNA <50 copies/mL, and 132 HIV-uninfected controls, aged 20-59 years, at Chiang Mai University Hospital during July and August 2011. Prevalences of protective antibodies to these viruses as well as serologic responses after MMR vaccination in those without protective antibody to at least one of the three viruses were compared between groups. RESULTS: The prevalences of protective antibodies to measles, mumps, and rubella were 94.2, 55.0, and 84.6 % among HIV-infected adults, and 97.7, 67.5, and 89.4 % among HIV-uninfected controls, respectively. The prevalence of protective antibody to mumps was significantly lower in HIV-infected adults (p-value = 0.010). MMR vaccination was done in 249 HIV-infected and 46 HIV-uninfected controls; at week 8 to 12 after vaccination, the seroprotective rates against measles, mumps, and rubella in HIV-infected adults were 96.4, 70.7, and 98.0 %, respectively, whereas those in HIV-uninfected controls were 100, 87, and 100 %, respectively. No serious adverse effects were observed. CONCLUSIONS: In contrast to measles and rubella, the prevalence of protective antibody to mumps was low in both HIV-infected adults and HIV-uninfected controls in northern Thailand. The seroprotective rates after MMR vaccination in both groups were considerably high, except only for mumps. Therefore, MMR vaccination should be considered in all HIV-infected adults receiving antiretroviral therapy with undetectable plasma HIV-1 RNA and CD4 cell count ≥200 cells/mm(3). TRIAL REGISTRATION: ClinicalTrials.gov: NCT02724852 , registered on March 31, 2016.

Discontinuation of primary and secondary prophylaxis for opportunistic infections in HIV-infected patients who had CD4+ cell count <200 cells/mm(3) but undetectable plasma HIV-1 RNA: an open-label randomized controlled trial.

Abstract The CDC recommends discontinuing opportunistic infections (OIs) prophylaxis in HIV-infected patients who have CD4+ cell count >200 cells/mm(3) after receiving combination antiretroviral therapy (cART). A prospective randomized controlled trial was conducted at Chiang Mai University Hospital from June 1, 2009 to January 31, 2012 in 74 adult HIV-infected patients who had received cART and had CD4+ cell count <200 cells/mm(3) but plasma HIV-1 RNA<50 copies/ml. Forty-three patients (58.1%) were male and the mean age was 41.8±8.1 years; 68 (91.9%) and 59 (79.7%) patients were receiving co-trimoxazole and antifungal prophylaxis, respectively. The median CD4+ cell counts at enrollment were 142 (IQR 108, 161) and 158 (IQR 141, 176) cells/mm(3) among patients who discontinued and continued OIs prophylaxis, respectively (p value=0.041). One of 37 patients (2.7%) in the discontinuation group developed Pneumocystis jiroveci pneumonia, giving the incidence rate of 1.57/1000 person-months. None of the 37 patients in the continuation group developed OIs. The difference in the prevention rates of OIs between groups was -2.7% (95% CI -7.9, 2.5). In conclusion, in the setting where plasma HIV-RNA measurement is available, e.g., Asia-Pacific region, discontinuation of prophylaxis is considerably safe in HIV-infected patients receiving cART with undetectable plasma HIV-RNA but incomplete immune recovery.

Resistance-associated mutations after initial antiretroviral treatment failure in a large cohort of patients infected with HIV-1 subtype CRF01_AE.

The nucleoside reverse transcriptase inhibitors (NRTIs), zidovudine (AZT) and stavudine (d4T) are thymidine analog drugs recommended as first-line antiretroviral therapy in HIV-1-naive patients. Two thymidine analog mutation (TAM) pathways, TAM-1 and TAM-2, confer high levels of resistance with mutations in the viral RT. The relative prevalence of TAM pathways and their associations with other NRTI resistance mutations acquired under the pressure of drug treatment in a large cohort of 1,876 patients infected with HIV-1 CRF01_AE attending the Infectious Disease Clinic, Chiang Mai University Hospital, Chiang Mai, Thailand, were studied. From 117 patients infected with HIV-1 CRF01_AE who had plasma HIV-1 RNA of ≥500 copies/mL, 69 patients had at least one TAM. The most common mutation associated with NRTI resistance was M184V/I (89.9%). The TAM-2 (89.9%) pathway occurred approximately two times more frequently than the TAM-1 (43.5%) pathway. The presence of TAM and the TAM-1 pathway was significantly more frequent in the AZT- than the d4T-receiving group ((OR, 2.89; 95% CI, 1.12-7.46; P< 0.05) and (OR, 3.33; 95% CI, 1.19-9.37; P< 0.05), respectively). In conclusion, the TAM-2 pathway was selected more frequently than the TAM-1 pathway by thymidine analog drugs in HIV-1 CRF01_AE-infected patients, while the TAM-1 pathway occurred more frequently than the TAM-2 pathway in such patients with AZT-based treatment. Routine monitoring of plasma HIV-1 RNA may result in less exposure to failing regimens and reduce the opportunity for TAMs to accumulate. However, the low frequency of the TAM-1 pathway in our cohort data suggests that these patients should respond well to second-line regimens containing a ritonavir-boosted protease inhibitor.

Current molecular epidemiology and recombination of HIV type 1 subtypes in northern Thailand.

HIV subtype characterization is an important tool to monitor the genetic variation of the HIV epidemic. This study investigated the current HIV subtype distribution and recombination among the northern Thai population. An in-house genotypic assay of HIV protease and reverse transcriptase genes was performed on 420 plasma specimens from HIV-infected patients residing in several northern Thai provinces. HIV subtyping was determined by phylogenetic analysis. Three hundred and ninety-eight sequences (94.8%) were identified as CRF01_AE with the genetic distance of 1.848 ± 0.957% and 12 (2.9%) as subtype B with the genetic distance of 4.186 ± 0.849%. In addition, two sequences (0.5%) of HIV subtype C were found, suggesting that these patients were either immigrants from another country or were infected through heterosexual contact with HIV-infected subjects from another country. Bootscan analysis showed that there were eight (1.9%) unique recombinant forms (URFs) consisting of a recombinant of CRF01_AE with subtype B or subtype C. The information from this study is useful for prevention programs to halt the onward transmission of a particular HIV outbreak. However, characterization of the full genome of these CRF01_AE/B and CRF01_AE/C intersubtype recombinants, and also subtype C, is required for confirmation and elucidation.

Impact of the frequency of plasma HIV-1 RNA monitoring on the outcome of antiretroviral therapy.

BACKGROUND: Current guidelines for HIV management recommend monitoring plasma HIV-1 RNA level every 3-6 months in patients on a stable antiretroviral regimen. However, cost is the major obstacle to follow the guidelines in resource-limited settings. OBJECTIVE: This study aimed to compare the outcome of antiretroviral therapy among HIV-infected patients on a stable regimen who had plasma HIV-1 RNA monitoring once vs. twice yearly. METHODS: A retrospective cohort study was conducted among HIV-infected patients receiving antiretroviral therapy since 2002 at Chiang Mai University Hospital, Thailand. We evaluated the incidence of virological failure and number of reverse transcriptase (RT) mutations between groups. RESULTS: Of 551 patients on a stable antiretroviral regimen, 405 (73.5%) and 146 (26.5%) patients had plasma HIV-1 RNA measurement once and twice yearly, respectively. Forty-seven of 405 patients (11.6%) in once-yearly group and 15 of 146 patients (10.3%) in twice-yearly group developed virological failure, giving the incidence rate of 2.03/100 and 1.95/100 person-years, respectively. The probability of virological failure did not differ between groups (p=0.897, log-rank test). The number of RT mutations was not statistically different between groups (all p-values>0.05). The predicting factors for virological failure from a multivariate analysis were adherence rate <95% and baseline CD4 cell count <50 cells/mm3 but not the frequency of HIV-1 RNA monitoring. CONCLUSIONS: The incidence of virological failure and the number of RT mutations were not different between groups. Therefore, in resource-limited settings, the recommendation to perform plasma HIV-1 RNA measurement once yearly in patients on a stable antiretroviral regimen is justified.

Impact of HIV-1 viral load on genotypic characteristics among patients failing non-nucleoside reverse trancriptase inhibitor-based first-line regimens in Northern Thailand.

Widespread use of antiretroviral drugs has significantly increased drug resistance. In the resource limited countries, delayed detection of drug resistance may lead to accumulation of drug resistance mutations. We investigated the genotypic drug resistance mutation patterns in HIV-infected patients with various levels of plasma HIV RNA levels. Fifty-nine HIV-infected patients with antiviral therapy failure were recruited. Genotypic assays of HIV-1 protease and reverse transcriptase genes were analyzed. There was a significant difference in CD4 cell counts and percentage of CD4 (p < 0.05) between groups of patients with high and low viral load, who failed first-line non nucleoside reverse transcriptase inhibitor-based regimens. In addition, patients with HIV-1 viral load > or = 4 log10 have a significantly higher likelihood of being infected with HIV-1 containing 3 to 5 resistance-associated mutations than those with HIV-1 viral load < 4 log10. Thus, delayed detection of increased HIV-1 viral load and antiretroviral drug-resistance may lead to accumulation of drug-resistant mutations and decreased CD4 cell count and percentage.

Long-term CD4 cell count recovery among Thai naive HIV-infected patients initiating HAART at low CD4 cell count.

CD4 T cell recovery after highly active antiretroviral therapy (HAART) has been reported mostly from developed countries. A retrospective cohort study was conducted among naïve HIV-infected patients initiating HAART between July 1, 2001 and December 31, 2004 at Chiang Mai University, Thailand. We evaluated the CD4 cell count recovery over 4 years among patients initiated HAART at low (CD4 count 51-200 cells/mm(3)) and very low (CD4 count < or = 50 cells/mm(3)) CD4 counts. Of 287 patients, 153 and 134 had low and very low baseline CD4 count, respectively. There were 126 men (43.9%), and the mean age was 34.2 +/- 7.9 years. The median baseline CD4 count was 50 cells/mm(3) (IQR 25, 104). GPO-VIR (a combination of lamivudine, stavudine, and nevirapine) was the most common prescribed HAART (262 patients, 91.3%). Overall, the mean CD4 count significantly increased 108 cells/mm(3) in the first 6 months after HAART initiation and continued to increase up to 4 years, but in the lesser extent. The overall slope of CD4 count was not significantly different between groups. (p = 0.052) The median time to achieve CD4 count of > or = 200 cells/mm(3) was 6 and 18 months in those with low and very low baseline CD4 count, respectively (p<0.001). By 4 years, 19.9% of patients achieved CD4 count of > or = 500 cells/mm(3). The earlier HAART is initiated among patients with low and very low baseline CD4 count, the sooner the patients will achieve adequate immune status to prevent morbidity and mortality from opportunistic infections.