Attitudes of People in the UK with HIV Who Are Antiretroviral (ART) Na?ve to Starting ART at High CD4 Counts for Potential Health Benefit or to Prevent HIV Transmission

Objective

To assess if a strategy of early ART to prevent HIV transmission is acceptable to ART naïve people with HIV with high CD4 counts.

Design

ASTRA is a UK multicentre, cross sectional study of 3258 HIV outpatients in 2011/12. A self-completed questionnaire collected sociodemographic, behavioral and health data, and attitudes to ART; CD4 count was recorded from clinical records.

Methods

ART naïve participants with CD4 ≥350 cells/µL (n = 281) were asked to agree/disagree/undecided with the statements (i) I would want to start treatment now if this would slightly reduce my risk of getting a serious illness, and (ii) I would want to start treatment now if this would make me less infectious to a sexual partner, even if there was no benefit to my own health.

Results

Participants were 85% MSM, 76% white, 11% women. Of 281 participants, 49.5% and 45.2% agreed they would start ART for reasons (i) and (ii) respectively; 62.6% agreed with either (i) or (ii); 12.5% agreed with neither; 24.9% were uncertain. Factors independently associated (p<0.1) with agreement to (i) were: lower CD4, more recent HIV diagnosis, physical symptoms, not being depressed, greater financial hardship, and with agreement to (ii) were: being heterosexual, more recent HIV diagnosis, being sexually active.

Conclusions

A strategy of starting ART at high CD4 counts is likely to be acceptable to the majority of HIV-diagnosed individuals. Almost half with CD4 >350 would start ART to reduce infectiousness, even if treatment did not benefit their own health. However a significant minority would not like to start ART either for modest health benefit or to reduce infectivity. Any change in approach to ART initiation must take account of individual preferences. Transmission models of potential benefit of early ART should consider that ART uptake may be lower than that seen with low CD4 counts.     

Is the “3 by 5” Initiative the Best Approach to Tackling the HIV Pandemic?

BACKGROUND TO THE DEBATE: The World Health Organization (WHO) and its partners aim to treat 3 million people infected with HIV in poor and middle income countries with antiretroviral treatment by the end of 2005. The ambitious "3 by 5" initiative has had its supporters and its critics since its announcement in 2002.     

γ-Herpesvirus Load as Surrogate Marker of Early Death in HIV-1 Lymphoma Patients Submitted to High Dose Chemotherapy and Autologous Peripheral Blood Stem Cell Transplantation

Autologous stem cell transplantation (ASCT) is a feasible procedure for human immunodeficiency virus-1 (HIV-1) lymphoma patients, whose underlying disease and intrinsic HIV-1- and ASCT-associated immunodeficiency might increase the risk for γ-herpesvirus load persistence and/or reactivation. We evaluated this hypothesis by investigating the levels of Epstein-Barr virus (EBV)- and Kaposi sarcoma-associated herpesvirus (KSHV)-DNA levels in the peripheral blood of 22 HIV-1-associated lymphoma patients during ASCT, highlighting their relationship with γ-herpesvirus lymphoma status, immunological parameters, and clinical events. EBV-DNA was detected in the pre-treatment plasma and peripheral blood mononuclear cells (PBMCs) of 12 (median 12135 copies/mL) and 18 patients (median 417 copies/10⁶ PBMCs), respectively; the values in the two compartments were correlated (r = 0.77, p = 0.0001). Only EBV-positive lymphomas showed detectable levels of plasma EBV-DNA. After debulking chemotherapy, plasma EBV-DNA was associated with lymphoma chemosensitivity (p = 0.03) and a significant higher mortality risk by multivariate Cox analysis adjusted for EBV-lymphoma status (HR, 10.46, 95% CI, 1.11–98.32, p = 0.04). After infusion, EBV-DNA was detectable in five EBV-positive lymphoma patients who died within six months. KSHV-DNA load was positive in only one patient, who died from primary effusion lymphoma. Fluctuations in levels of KSHV-DNA reflected the patient’s therapy and evolution of his underlying lymphoma. Other γ-herpesvirus-associated malignancies, such as multicentric Castleman disease and Kaposi sarcoma, or end-organ complications after salvage treatment were not found. Overall, these findings suggest a prognostic and predictive value of EBV-DNA and KSHV-DNA, the monitoring of which could be a simple, complementary tool for the management of γ-herpesvirus-positive lymphomas in HIV-1 patients submitted to ASCT.     

Attempted Detection of Toxoplasma gondii Oocysts in Environmental Waters Using a Simple Approach to Evaluate the Potential for Waterborne Transmission in the Galápagos Islands,Ecuador

Toxoplasmosis is a health concern for wildlife and humans, particularly in island ecosystems. In the Galápagos Islands, exposure to Toxoplasma gondii has been found in marine avifauna on islands with and without domestic cats. To evaluate potential waterborne transmission of T. gondii, we attempted to use filtration and epifluorescent microscopy to detect autofluorescent T. gondii oocysts in fresh and estuarine surface water samples. T. gondii oocyst-like structures were microscopically visualized but were not confirmed by polymerase chain reaction and sequence analyses. Further research is needed to refine environmental pathogen screening techniques and to evaluate disease risk of waterborne zoonoses such as T. gondii for wildlife and humans, particularly in the Galápagos and other naive island ecosystems.     

Potential for the Anopheles gambiae Densonucleosis Virus To Act as an “Evolution-Proof” Biopesticide

“Evolution-proof” or “late-life-acting” insecticides (LLAIs) preferentially kill older adult mosquitoes and are of extreme interest to control vector-borne diseases such as malaria. We used quantitative PCR to assess whether the Anopheles gambiae densonucleosis virus (AgDNV) had potential as an LLAI. After infection, AgDNV titers increased modestly during larval development but replicated slower than the host cells, resulting in a significant decrease in the normalized virus titer during larval and pupal development. Normalized virus titers dramatically increased after adult emergence, peaking in 7- to 10-day-old adults. Unlike other DNVs, AgDNV does not significantly replicate in preadult mosquitoes but rather preferentially replicates in older adults. The natural dynamics of AgDNV make it ideal for expression of insect-specific toxin genes as a biological LLAI.Malaria infects several hundred million people and results in over one million deaths annually. Vector control is a major component of current malaria control strategies. However, the evolution of insecticide resistance by Anopheles mosquito vectors has severely hampered control efforts (5, 13, 14). Although novel chemistries are being explored, new insecticides will face similar problems with resistance evolution. Recently, late-life-acting insecticides (LLAIs) have been proposed as novel agents to control vector-borne diseases such as malaria (1, 10). LLAIs selectively kill the older mosquitoes responsible for the bulk of parasite transmission while allowing for reproduction of the younger age classes that contribute to the bulk of evolutionary fitness, i.e., there is an optimal window of time wherein mosquitoes live long enough to reproduce but not long enough to transmit pathogens (8, 9). Reproduction allows for relaxation of evolutionary pressures that select for resistance to the agent. If resistance alleles exert fitness costs, there are theoretical scenarios under which resistance is not expected to evolve, leading some to provocatively term LLAIs as “evolution-proof” (1, 10).LLAI do not have to be conventional chemical pesticides. Like all organisms, mosquitoes can be infected with pathogens of their own (including fungi, bacteria, or viruses) that can be exploited to shorten mosquito life span to control disease (6, 9-12). Some pathogens can be vertically transmitted (9, 11), which not only results in relaxed selection pressure but also allows for their transmission and spread into the vector population.Densonucleosis viruses (or densoviruses [DNVs]) are icosahedral, nonenveloped parvoviruses that have been identified from many invertebrate taxa, including multiple mosquito species (2, 11). Naturally occurring DNVs typically infect mosquitoes during the aquatic larval phase. Infection of young larvae (first or second instar) is generally lethal, resulting in virus amplification and release into the larval environment. Larvae infected at later time points (third or fourth instar) develop into infected adults that inoculate virus vertically and horizontally into the larval environment during oviposition, completing the virus life cycle (2, 11).The Aedes aegypti densovirus (AeDNV) is generally lethal to Ae. aegypti larvae in a dose-dependent manner, and high virus titers in larvae are observed both by quantitative PCR (qPCR) and by expression of foreign transgenes such as green fluorescent protein (GFP) (2, 8, 15). In contrast, the Anopheles gambiae densovirus (AgDNV) is not lethal to An. gambiae larvae (11). Using GFP-transducing virus and epifluorescence microscopy, we have also never seen GFP expression in larvae, pupae, or young adults; GFP is not observable until the adults are ∼1 week postemergence (J. L. Rasgon and X. Ren, unpublished observations) (Fig. ​(Fig.1).1). Based on these observations, we hypothesized that AgDNV has different replication kinetics in An. gambiae than AeDNV has in Ae. aegypti, remaining at relatively low titers in the immature life stages but replicating to high titers after adult emergence.Open in a separate windowFIG. 1.Epifluorescent image (dorsal view) of a 10-day-old adult An. gambiae female infected with GFP-expressing AgDNV (as described in reference 11). GFP is not visible in infected mosquitoes until 7 to 10 days postemergence.     

Study of Different Copper (I) Catalysts for the “Click Chemistry” Approach to Carbanucleosides

We compare herein the scope of three copper (I) catalysts on the synthesis of various 1,4-disubstitued-1,2,3-triazolo-carbanucleosides through a microwave (and thermic) assisted Huisgen 1,3-dipolar cycloaddition. The tetrakis(acetonitrile)copper hexafluorophosphate ([Cu(CH ₃ CN)₄]PF ₆ ), the imidazoline(mesythyl)copper bromide (Imes)CuBr, and the copper/copper sulfate Cu(0)/CuSO ₄ (II) mixture have been chosen for this study. Their influence in a catalytic amount will be analyzed according to the substituent of the alkyne, the solvent, or the heating method.     

CD4 Counts at Entry to HIV Care in Mexico for Patients under the “Universal Antiretroviral Treatment Program for the Uninsured Population,” 2007–2014

In Mexico, public health services have provided universal access to antiretroviral therapy (ART) since 2004. For individuals receiving HIV care in public healthcare facilities, the data are limited regarding CD4 T-lymphocyte counts (CD4e) at the time of entry into care. Relevant population-based estimates of CD4e are needed to inform strategies to maximize the impact of Mexico’s national ART program, and may be applicable to other countries implementing universal HIV treatment programs. For this study, we retrospectively analyzed the CD4e of persons living with HIV and receiving care at state public health facilities from 2007 to 2014, comparing CD4e by demographic characteristics and the marginalization index of the state where treatment was provided, and assessing trends in CD4e over time. Our sample included 66,947 individuals who entered into HIV care between 2007 and 2014, of whom 79% were male. During the study period, the male-to-female ratio increased from 3.0 to 4.3, reflecting the country''s HIV epidemic; the median age at entry decreased from 34 years to 32 years. Overall, 48.6% of individuals entered care with a CD4≤200 cells/μl, ranging from 42.2% in states with a very low marginalization index to 52.8% in states with a high marginalization index, and from 38.9% among individuals aged 18–29 to 56.5% among those older than 50. The adjusted geometric mean (95% confidence interval) CD4e increased among males from 135 (131,142) cells/μl in 2007 to 148 (143,155) cells/μl in 2014 (p-value<0.0001); no change was observed among women, with a geometric mean of 178 (171,186) and 171 (165,183) in 2007 and 2014, respectively. There have been important gains in access to HIV care and treatment; however, late entry into care remains an important barrier in achieving optimal outcomes of ART in Mexico. The geographic, socioeconomic, and demographic differences observed reflect important inequities in timely access to HIV prevention, care, and treatment services, and highlight the need to develop contextual and culturally appropriate prevention and HIV testing strategies and linkage programs.     

High Producer Haplotype (CAG) of -863C/A, -308G/A and -238G/A Polymorphisms in the Promoter Region of TNF-α Gene Associate with Enhanced Apoptosis of Lymphocytes in HIV-1 Subtype C Infected Individuals from North India

Introduction

The natural history of HIV-1 infection and its progression towards AIDS vary considerably among individuals. Host genetic factors may be one of the possible reasons for variable HIV-1 disease progression. Single nucleotide polymorphisms (SNPs) in the promoter region of TNF-α gene can influence its production. The aim of the present study was to determine the association of functional TNF-α SNPs and its associated parameters related to apoptosis that may influence the rate of HIV-1 disease progression.

Methods

Therapy naive, 100 HIV slow progressors (SPs), 100 HIV fast progressors (FPs), 50 HIV exposed but seronegative individuals (ESNs) and 260 healthy controls from same ethnic origin were recruited. Genotyping of TNF-α variants (−863C/A, -308G/A and -238G/A) was done using PCR-RFLP. CD4 counts were determined by flow cytometry. Plasma viral load was estimated by COBAS AMPLICOR HIV-1 monitor test. Plasma TNF-α concentration was estimated by Human CBA Th1/Th2 cytokine kit. The lymphocyte mitochondrial membrane potential was measured by JC-1 dye by flow cytometry.

Results

Genotype and allele frequency of TNF-α -238G/A and -863C/A was not significantly different in HIV-1-infected patients when compared to controls, while that of TNF-α -308G/A variant (high TNF-α producer) was significantly higher in FPs compared to SPs (p<0.01, OR = 3.43). Haplotype analyses also showed that carriers of high TNF-α producing haplotype CAG was significantly more common among FPs compared to SPs (p<0.01, OR = 3). The circulating TNF-α levels in blood also correlated well with genotypes. The lymphocyte mitochondrial membrane potential of FPs having CAG haplotype was significantly low as compared to wild type (CGG) haplotype (417±22 vs 571±28, p<0.01).

Conclusion

High producer haplotype, CAG of TNF-α gene associates with enhanced apoptosis of lymphocytes in HIV-1 infected individuals, hence faster progression to AIDS. However, further functional studies are needed to confirm this association and this knowledge may help clinicians to better understand the disease outcome.     

The Lyt phenotype of the T cells in an antitumor adoptive transfer differs for “parent to F1” and “parent to parent” combinations

The T-cell subset mediating tumor graft neutralization was characterized in a methylcholanthrene (MC) tumor system. Lyt 1⁺ cells were critical for the successful prevention of outgrowth of the tumor cells in graft neutralization assays with irradiated recipients. Elimination of Lyt 1⁺ cells abolished the outgrowth inhibitory effect exerted by T-cell-enriched populations derived from syngeneic or semisyngeneic mice immunized with the H-2-carrying MC-induced M-A tumor. In accordance, lymphocyte populations containing 98% Lyt 1⁺ cells derived from M-A-immune mice, mediated a complete transplantation immunity against this tumor. When the immune T cells admixed to the tumor inoculum were syngeneic to the recipient (i.e., A-derived cells were transferred to A recipients, or F₁ to F₁), elimination of the Lyt 2⁺ cells did not influence the potential to inhibit tumor outgrowth. The presence of Lyt 1⁺2^? cells were thus necessary, and sufficient, in the syngenic combination. A reduction of the graft-inhibiting potential occurred after elimination of Lyt 2⁺ cells from the A-derived M-A immune T-cell population when the recipients were semisyngeneic (i.e., (A/Sn X A.SW)F₁, (A/Sn X CBA)F₁, or (A/Sn X C57Bl/6)F₁). Consequently, only in the semisyngeneic, but not in the syngeneic combinations, was the transfer of Lyt 2⁺ cells necessary for optimal graft inhibition. It can be concluded that the genotypic relation between the donor and the recipient influences the prerequisites of the tumor cell neutralization.     

Novel Approach to the Ligation of Single-Stranded DNA Fragments by T4 DNA Ligase—DNA Mobile Multiple-Restriction Fragments: “UNI-LINKERS” for Cloning of Genes

Abstract

A group of uniquely designed single-stranded oligodeoxyribo-nucleotides that form hairpin loops were synthesized. These oligonucleotides can be ligated to other synthetic single-stranded fragents differing in length and design without the need for external annealing templates. A novel approach to building a limitless variety of mobile multiple-restriction DNA fragments termed “uni-linkers” and which are open only at one end, is described. The latter were used as “multi-stage” linkers in cloning experiments. The cloning of the cDNA for a carrot proline-rich protein, DC-5, into a plant vector is presented as an example. Other possible applications are also discussed.