HIV-1

Introduction: Vaccine design is mainly considered as a therapeutic strategy to elicit HIVspecific immunity. DNA vaccines encoding an antigen and also an adjuvant can induce an effective adaptive immunity. Due to having numerous roles in viral infection, the HIV-1 Nef protein is considered as an antigen candidate for development of therapeutic vaccines. A variety of adjuvants and delivery systems have been utilized to increase the potency of DNA vaccines against viral infections, such as heat shock proteins (HSPs) which possess chaperon activity and immunostimulatory properties. Methods: pEGFP mammalian expression vectors harboring nef, hsp27, hsp27-nef, hsp70 and hsp70-nef genes were prepared in large scale. Their concentration and purity were assessed by NanoDrop spectrophotometry. Human embryonic kidney 293T cells (HEK-293T) were grown in DMEM culture medium and transfected with these constructs using Lipofectamine 2000 transfection reagent. After 48 hours, their transfection efficiency was evaluated using fluorescent microscopy and flow cytometry. Results: The pEGFP-nef,-hsp27,-hsp27-nef,-hsp70 and-hsp70-nef constructs were successfully prepared in large scale and high purity. The results of cell transfection with each construct showed that the percentages of Nef

Although fishing communities (FCs) in Uganda are disproportionately affected by HIV-1 relative to the general population (GP), the transmission dynamics are not completely understood. We earlier found most HIV-1 transmissions to occur within FCs of Lake Victoria. Here, we test the hypothesis that HIV-1 transmission in FCs is isolated from networks in the GP. We used phylogeography to reconstruct the geospatial viral migration patterns in 8 FCs and 2 GP cohorts and a Bayesian phylogenetic inference in BEAST v1.8.4 to analyse the temporal dynamics of HIV-1 transmission. Subtype A1 (pol region) was most prevalent in the FCs (115, 45.1%) and GP (177, 50.4%). More recent HIV transmission pairs from FCs were found at a genetic distance (GD) <1.5% than in the GP (Fisher's exact test, p = 0.001). The mean time depth for pairs was shorter in FCs (5 months) than in the GP (4 years). Phylogeographic analysis showed strong support for viral migration from the GP to FCs without evidence of substantial viral dissemination to the GP. This suggests that FCs are a sink for, not a source of, virus strains from the GP. Targeted interventions in FCs should be extended to include the neighbouring GP for effective epidemic control. Human immunodeficiency virus type 1 (HIV-1) prevalence and incidence is higher among certain populations relative to other groups in Uganda. Among these, the fisher folk (FF) and female sex workers have the highest documented HIV-1 incidence rates 1. An earlier report showed that majority of new HIV-1 infections in key populations were likely to come from the fishing communities (FCs) 2 while a crosscountry analysis 3 among most-at-risk-populations in developing countries revealed that FF had the highest HIV-1 prevalence relative to other high-risk groups and the general population (GP). "Fishing communities" is a general term used in this study to refer to groups of persons living in villages that are located along the shores of Lake Victoria or on islands and who are largely dependent on the harvest or processing of fishery resources to meet their social and economic needs 1. In contrast, GP refers to people living mostly on the mainland or in towns adjacent to the FCs (approximately 10-40 kms) who do not derive their livelihood primarily from fishing-related activities 4. HIV prevalence in the FCs is very high; estimated at about 29% 5 and reaching as high as 40% in some communities 5. These figures significantly exceed the national average of 7.3% 6. Annual incidence rates of up to 6/100 person-years at risk (PYAR) 4 have been reported among high-risk individuals in the FCs, which is much higher than the national estimated rate of 1/100 PYAR 4. The high incidence rates have been attributed to risky sexual behaviour involving multiple partnerships, high alcohol consumption, low condom use, limited access to health services and transactional sex 7-9 .

Background: Identification of HIV-1 genomes responsible for establishing clinical infection in newly infected individuals is fundamental to prevention and pathogenesis research. Processing, storage, and transportation of the clinical samples required to perform these virologic assays in resource-limited settings requires challenging venipuncture and cold chain logistics. Here, we validate the use of dried-blood spots (DBS) as a simple and convenient alternative to collecting and storing frozen plasma.