Current HIV Research - Volume 11, Issue 4, 2013

Background: Highly Active Antiretroviral therapy (HAART) can effectively reduce the viral load to undetectable levels in most HIV-infected patients. However, some patients may still experience impaired immunologic response associated with increased risk of disease progression and death. Objective: The objective of this study was to assess the impact of the HIV DNA load on the immune alteration during successful HAART. Methods: 40 chronic HIV-infected adults initiating HAART were followed for 24 months. The CD4 count, HIV viral load, HIV DNA load, and levels of γ-cytokines IL-2, IL-7 and IL-21 were monitored at baseline (month 0), month 6, 12, 18 and 24 following HAART initiation. Results: The plasma viral load decreased significantly and remained below the detection limits after six months treatment. Likely the HIV DNA load decreased significantly in both cells during 12 months, was undetectable in CD14 monocytes after 18 months, but remained higher in CD3+ T cells during all the follow up. In addition, the HIV DNA load correlated positively between T cells and monocytes in 10 patients who maintained higher HIV DNA load in both cells during 12 months. The CD4 count, IL-2, and IL-21 levels increased significantly during 12 months, whereas IL-7 decreased significantly during 18 months, regardless of the HIV DNA load in T cells. Patients with CD4 count below 200/μl maintained higher HIV DNA load and showed lower increase in CD4 count compared to patients with CD4 count above 200/μl. In patients showing undetectable HIV DNA load in both cells, neither IL-2 nor IL-21 correlated with the CD4 count even after 24 months despite their partial restoration. Conclusion: These results suggest that the HIV DNA load could continuously hamper the CD4 restoration and γ-cytokines functional activities during HAART. This action seemed to be more severe in patients with pre-HAART CD4 count below 200/μl. The CD14 monocytes may contribute to this action as source of T cell infection both before and during HAART.

HIV infection results in the development of immunodeficiency mainly due to the apoptosis of infected and bystander CD4 cells. The aim of the study was to follow the mitochondrial dependent pathway of apoptosis, one of the suggested mechanisms of above process. The inner mitochondrial membrane potential (MMP), Adenosine-5'-triphosphate (ATP) generation, apoptosis and necrosis markers of peripheral mononuclear cells (PBMCs) were compared in HIV infected patients and HIV negative control group. The correlation of blood viral load, TNFα concentration, CD4 cells count and duration of ARV therapy was considered. Additionally, group of HIV infected ARV-naive patients was involved for the follow-up study and the effects of one year of ARV therapy on measured parameters were studied. PBMCs of HIV infected individuals (especially without ARV therapy) demonstrated lower MMP and ATP generation and higher percentage of apoptotic/necrotic PBMCs. Correlation between blood TNFα level and mitochondrial dysfunction was observed. The first months of ARV therapy resulted in most significant restoration of mitochondrial function and living PBMCs count. HIV infection and ARV therapy have significant impact on mitochondrial function and apoptosis of PBMCs. They are driven by abnormal mitochondrial function apoptosis of immune cells which seems to be the key element leading to immunosuppression, thus an early intervention in this process by therapy can be beneficial for symptomatology of HIV infected patients.

Two HIV-1 non B isolates, 98US_MSC5007 and 98US_MSC5016, which have been identified amongst the US Army personnel serving abroad, are known to have originated from other nations. Notwithstanding, they are categorized as American strains. This is because their countries of origin are unknown. American isolates are basically B subtype. 98US_MSC5007 belongs to Circulating Recombinant Form (CRF02_AG) while 98US_MSC5016 is of the C clade. Both sub-groups are recognized to have originated from African and Asian continents. It has become necessary to properly determine the countries of origin of microbes and viruses. This is because diversity and cross-subtyping have been found to mitigate the designing and development of vaccine and therapeutic interventions. The aim of this study therefore is to identify the countries of origin of the two American isolates found amongst US Army personnel serving abroad. A Digital Signal Processing-based Bioinformatics technique called Informational Spectrum Method (ISM) has been engaged. ISM entails translating the amino acids sequences of the protein into numerical sequences (signals) by means of one biological parameter (Amino Acids Scale). The signals are then processed using Discrete Fourier Transform (DFT) in order to uncover and present the embedded biological information as Informational Spectra (IS). Spectral Position of Maximum Binding Interaction (SPMBI) is used. Several approaches including Phylogeny have preliminarily been employed in the determination of evolutionary trends of organisms and viruses. SPMBI has preliminarily been used to re-establish the semblance and common originality that exist between human and Chimpanzee, evolutionary roadmaps in the Influenza and HIV viruses. The results disclosed that 98US_MSC5007 shared same semblance and originality with a Nigeria isolate (92NG083) while 98US_MSC5016 with the Zairian isolates (ELI, MAL, and Z2/CDC-34). These results appear to demonstrate that the American soldiers harboring these strains may have been infected by isolates from Nigeria and Zaire, respectively. This is because 98US_MSC5007 and the Nigerian isolate share SPMBI at position 44. Additionally, 98US_MSC5016, which has SPMBI at position 148, may have come from Zaire as it has similar SPMBI with the Zairian isolates at 150. SPMBI is a demonstration of Bio-functionality arising from maximum affinity by the proteins from different sources to a common protein. To help validate the findings, the experiment was further repeated using ISM-based Phylogenetic technique. The outcome appears not to be in complete accord with the results obtained in this study. It is therefore recommended that the countries in which these US Army personnel are deployed be identified and where the findings made and the locations of the Army personnel appropriately correlate, this novel procedure be engaged in the identification of the nations of origins of all other such HIV isolates across all clades and nations.

HIV-associated neurocognitive disorders (HAND) remain prevalent despite effective combined anti-retroviral therapy (cART). Cognitive function has been shown to inversely correlate with decreased synaptic and dendritic density. In this study, macaques inoculated with SIV were examined over a 3-month course of infection to characterize the appearance of the neuronal damage marker 14-3-3 protein in CSF and to determine whether CSF 14-3-3 levels directly reflected synaptic alterations. SIV-infected macaques with 14-3-3 in CSF had significantly lower levels of the presynaptic protein synaptophysin in cortical grey matter. Synaptophysin levels were inversely correlated with amount of SIV RNA in the CNS. In contrast, levels of 14-3-3 in CSF did not correspond with either alterations in levels of the postsynaptic protein PSD-95 or viral replication in the brain. These findings suggest that the appearance of 14-3-3 in CSF during asymptomatic infection reflects pre-synaptic damage in SIV-infected macaques and thus may serve as a marker of the early synaptic alterations that underlie HIV-induced neurocognitive impairment.

Transmitted drug resistance (TDR) is an important public health issue, because it may affect the outcome of antiretroviral treatment. The prevalence of human immunodeficiency virus type 1 (HIV-1) with TDR mutations defined according to the list of the World Health Organization was investigated among 53 therapy-naive persons with confirmed recent HIV-1 infection diagnosed in Poland, in the years 2008-2010. Proviral DNA was amplified, sequenced, and screened for the TDR mutations in the pol gene fragments coding for the whole protease and the initial 256 residues of the reverse transcriptase. The frequency of sequences with at least one TDR mutation was 11.3%. In four (7.5%) sequences at least one resistance mutation related to reverse transcriptase inhibitors was identified, and in further two (3.8%) sequences one mutation related to protease inhibitors' resistance was present. The moderate rate of TDR highlights the need for a continuous surveillance and resistance testing among treatment-naive individuals to optimize treatment effects within a country.

A portion of the gag gene cDNA for p24 protein from 30 Indian HIV-1 proviral DNA was amplified by PCR and sequenced. Phylogenetic analysis with reference samples of A1, A2, B, C, D, F1, F2, G, H, J, K, N and O subtypes revealed that 29 test samples aligned with subtype C reference strain while 1 matched with HIV-1 subtype A. Multiple alignment of predicted amino acid sequence of the Indian test samples and reference C subtype of HIV-1 samples from other countries indicated a molecular signature by way of rigid conservation of the amino acid ‘S’ at position 41 of the gag p24 protein in all Indian HIV-1 samples analyzed in this study as opposed to ‘T’in the same position in C subtype sequences from other parts of the world. A phylogenetic analysis and visualization of the resulting tree in radial position showed distinct clubbing of all Indian C subtypes and formation of a cluster when compared to C subtype sequences from other countries with a single Chinese sample as an exception which was found in the Indian cluster. The use of a portion of p24 gene sequence as tool for subtyping as well as phylogenetic grouping with special reference to its geographical location is discussed.

The present study assessed the phenotypic aspects of oral-cavity Candida albicans isolates from 300 HIV-1- positive patients, relating the most commonly investigated virulence factors (enzyme typing and germ-tube formation) to the most common morphotypes. The samples were seeded into specific media for isolation and subsequent identification using the automated Vitek 2 system. The following assays were performed for phenotypic characterization: morphotyping, germ-tube formation and enzyme typing. Out of 300 collected samples, 144 tested positive for yeasts of the Candida genus, 98 (32.7 %) of which were identified as C. albicans. The latter samples were attributed to seven different morphotypes; the three most common morphotypes were 7208 (49 %), 7308 (14.3 %) and 3208 (13.3 %). All of the C. albicans isolate samples formed germ tubes and produced the enzymes proteinase and phospholipase, with an activity classified as intermediate to high. Due to the identification of virulence factors among the analyzed samples, monitoring of HIV-1-positive patients colonized by different morphotypes must be established because these morphotypes are extremely pathogenic and can trigger severe fungal infections.

Due to the fact that a definite treatment for AIDS disease has not been discovered so far, antiretroviral drugs are relatively significant in controlling the disease. In this study, Lamivudine- which is an old and effective anti-AIDS drug- was connected to PEGylated chitosan nanoparticle in order to produce a new and greater version of Lamivudine. First, physicochemical studies such as HNMR, FTIR spectroscopy and CHN analysis were performed to ensure the proper synthesis. Following that, Lamivudine-PEGylated Chitosan (LPC) drug was evaluated in terms of its inhibitory capability in producing HIV virions and its cytotoxicity in different doses. HIV virions were created by transfection of psPAX2, PmzNL4-3 and pMD2.G plasmids into HEK293T cell line. Also, assessment of the P24 amounts of cell supernatant was performed using ELISA method. Among the different doses of LPC drug (0.1, 1, 10 and 100μM), it was found that 0.1μM was the most effective and least toxic dose compared to Lamivudine in the same dose. Results of this study indicate that LPC drug has the ability to inhibit HIV virus replication in a more significant way in comparison to the old drug. Besides, the drug has a low cytotoxicity and is effective with a lower dose.

The case of an HIV-positive treatment-naive male with toxic shock syndrome (TSS) is presented herein. The course of TSS was favorable; however, the patient had extremely high plasma levels of MCP-1 and CD38 and HLA-DR expression on CD8+ T cells during the acute illness. Furthermore, the numbers of CD8+ T cells were reduced and CD4+ T cells remained stable during acute illness in comparison to baseline values. MCP-1 and HLA-DR gradually decreased, but they were still elevated after a month, whereas the number of circulating CD8+ T cells increased more than fivefold. CD38 expression remained stable during this period. A further decrease in CD38, HLA-DR and MCP-1 was noted five months after the initiation of antiretroviral therapy.

Intestinal infection causing diarrheal disease is a dominant contributor to high morbidity and mortality in developing countries. This intervention study aimed to assess the response of specific anti-microbial and anti-retroviral therapy (ART) on enteropathogens identified in HIV/AIDS adult subjects from northern India. Seventy five ART naive (group 1) and seventy five ART adherent (group 2) HIV/AIDS adult subjects with diarrhea were enrolled. Stool samples from all subjects were examined for enteropathogens by wet mount, staining methods, culture and ELISA. Subjects with enteropathogens were started on specific therapy as per National AIDS Control Organisation, Government of India's guidelines. Follow-up stool samples were examined after 2-4 weeks of completion of therapy for persistence/clearing of enteropathogens. CD4+ T lymphocyte count was done for all subjects. At enrollment, group 1 had 26.13% bacterial, 57.66% parasitic & 16.22% fungal pathogens while group 2 had 11.9%, 69.05% & 19.05% pathogens, respectively. Parasitic diarrhea was more common than bacterial diarrhea. The coccidian parasites (Cryptosporidium spp. & Isospora belli) were the common parasites identified. Clearance of enteric pathogens was significant after specific anti-microbial therapy (p = 0.0001). Persistence of enteropathogens was seen primarily for coccidian parasites. Clearance of enteropathogens after specific therapy and the diagnostic yield of stool specimens were influenced by the CD4+ counts. Immune competence coupled with specific anti-microbial therapy displays the best response against enteric pathogens.

Clinical data on antiretroviral effectiveness in women are limited, especially long-term data, because women are usually underrepresented in clinical trials. This sub-analysis of a large European non-comparative, retrospective, observational cohort study evaluated gender differences in long-term outcomes in antiretroviral-experienced adult patients with HIV-1 infection switched to an ATV/r-based regimen between October 2004 and March 2007. Data were extracted from 3 European HIV databases every 6 months (maximum follow-up 5 years). Time to virological failure (VF), defined as two consecutive HIV-1 RNA≥50 c/mL or one HIV-1 RNA≥50 c/mL followed by treatment discontinuation (TD), and time to TD were analyzed using the Kaplan-Meier method. Associations of gender with VF and TD were analyzed using multivariate Cox proportional models. Safety and tolerability were evaluated. In total, 1294 patients (336 women, 958 men) were analyzed. No gender differences in time to VF were observed; at 3 years, the probability of not having VF was 0.59 (95%CI: 0.52, 0.65) and 0.63 (95%CI: 0.59, 0.67) for women and men, respectively. In multivariate analyses, women had a higher risk of TD than men (hazard ratio [HR], 1.54; 95%CI: 1.28, 1.85) but no increased risk of VF (HR, 1.06; 95%CI: 0.85, 1.33). Safety and tolerability were comparable between genders. In a clinical setting, long-term efficacy and safety outcomes of ATV/r-based regimens were similar by gender. Women had a higher risk of TD but no increased risk of VF. ATV/r is an effective and well-tolerated therapeutic option for treatment-experienced men and women with HIV-1 infection.