HIV rapid diagnostic tests (RDTs) are now used widely in non-laboratory settings by non-laboratory-trained operators. Quality assurance programmes are essential in ensuring the quality of HIV RDT outcomes. However, there is no cost-effective means of supplying the many operators of RDTs with suitable quality assurance schemes. Therefore, it was examined whether photograph-based RDT results could be used and correctly interpreted in the non-laboratory setting. Further it was investigated if a single training session improved the interpretation skills of RDT operators. The photographs were interpreted, a 10-minute tutorial given and then a second interpretation session was held. It was established that the results could be read with accuracy. The participants (n = 75) with a range of skills interpreted results (>80% concordance with reference results) from a panel of 10 samples (three negative and seven positive) using four RDTs. Differences in accuracy of interpretation before and after the tutorial were marked in some cases. Training was more effective for improving the accurate interpretation of more complex results, e.g. results with faint test lines or for multiple test lines, and especially for improving interpretation skills of inexperienced participants. It was demonstrated that interpretation of RDTs was improved using photographed results allied to a 10-minute training session. It is anticipated that this method could be used for training but also for quality assessment of RDT operators without access to conventional quality assurance or training schemes requiring wet samples.

Severe immunodeficiency during primary human immunodeficiency virus (HIV) infection is unusual. Here, we characterized viral and immunological parameters in a subject presenting with Pneumocystis jirovecii pneumonia in the setting of prolonged primary HIV illness and delayed seroconversion. HIV antibody was only detected by enzyme-linked immunosorbent assay 12 months after presentation, and Western blot profiles remain indeterminate. Isolated virus was of R5 phenotype, exhibited poor viral fitness, but was otherwise unremarkable. Analysis of HIV antibody isotypes showed failure to mount a detectable HIV IgG response over nearly 2 years of infection, in particular IgG1- and IgG3-specific responses, despite normal responses to common infections and vaccines. Genetic analysis demonstrated homozygosity for part of an MHC haplotype containing susceptibility genes for common variable immunodeficiency (CVID) syndrome and other antibody deficiency disorders. Thus, a primary disorder of specific antibody production may explain exceptionally slow antibody development in an otherwise severe seroconversion illness. This highlights the need for multiparameter testing, in particular use of a fourth generation HIV test, for confirming HIV infection and underscores the importance of host factors in HIV pathogenesis.

Several clinical studies have shown that, relative to disease progression, HIV-1 isolates that are less fit are also less pathogenic. The aim of the present study was to investigate the relationship between viral fitness and control of viral load (VL) in acute and early HIV-1 infection. Samples were obtained from subjects participating in two clinical studies. In the PULSE study, antiretroviral therapy (ART) was initiated before, or no later than six months following seroconversion. Subjects then underwent multiple structured treatment interruptions (STIs). The PHAEDRA study enrolled and monitored a cohort of individuals with documented evidence of primary infection. The subset chosen were individuals identified no later than 12 months following seroconversion to HIV-1, who were not receiving ART. The relative fitness of primary isolates obtained from study participants was investigated ex vivo. Viral DNA production was quantified using a novel real time PCR assay. Following intermittent ART, the fitness of isolates obtained from 5 of 6 PULSE subjects decreased over time. In contrast, in the absence of ART the fitness of paired isolates obtained from 7 of 9 PHAEDRA subjects increased over time. However, viral fitness did not correlate with plasma VL. Most unexpected was the high relative fitness of isolates obtained at Baseline from PULSE subjects, before initiating ART. It is widely thought that the fitness of strains present during the acute phase is low relative to strains present during chronic HIV-1 infection, due to the bottleneck imposed upon transmission. The results of this study provide evidence that the relative fitness of strains present during acute HIV-1 infection may be higher than previously thought. Furthermore, that viral fitness may represent an important clinical parameter to be considered when deciding whether to initiate ART during early HIV-1 infection.

ABSTRACTThe development of a rapid and efficient system to identify human immunodeficiency virus type 1 (HIV-1)-infected individuals with broad and potent HIV-1-specific neutralizing antibody responses is an important step toward the discovery of critical neutralization targets for rational AIDS vaccine design. In this study, samples from HIV-1-infected volunteers from diverse epidemiological regions were screened for neutralization responses using pseudovirus panels composed of clades A, B, C, and D and circulating recombinant forms (CRFs). Initially, 463 serum and plasma samples from Australia, Rwanda, Uganda, the United Kingdom, and Zambia were screened to explore neutralization patterns and selection ranking algorithms. Samples were identified that neutralized representative isolates from at least four clade/CRF groups with titers above prespecified thresholds and ranked based on a weighted average of their log-transformed neutralization titers. Linear regression methods selected a five-pseudovirus subset, representing clades A, B, and C and one CRF01_AE, that could identify top-ranking samples with 50% inhibitory concentration (IC50) neutralization titers of ≥100 to multiple isolates within at least four clade groups. This reduced panel was then used to screen 1,234 new samples from the Ivory Coast, Kenya, South Africa, Thailand, and the United States, and 1% were identified as elite neutralizers. Elite activity is defined as the ability to neutralize, on average, more than one pseudovirus at an IC50titer of 300 within a clade group and across at least four clade groups. These elite neutralizers provide promising starting material for the isolation of broadly neutralizing monoclonal antibodies to assist in HIV-1 vaccine design.

Background: Elite non-progressors (plasma viral load <50 copies/ml while antiretroviral naive) constitute a tiny fraction of HIV-infected individuals. After 12 years follow-up of a cohort of 13 long-term non-progressors (LTNP) identified from 135 individuals with transfusion-acquired HIV infection, 5 remained LTNP after 23 to 26 years infection, but only 3 retained elite LTNP status. We examined the mechanisms that differentiated delayed progressors from LTNP in this cohort.Results: A survival advantage was conferred on 12 of 13 subjects, who had at least one host genetic factor (HLA, chemokine receptor or TLR polymorphisms) or viral attenuating factor (defective nef) associated with slow progression. However, antiviral immune responses differentiated the course of disease into and beyond the second decade of infection. A stable p24-specific proliferative response was associated with control of viraemia and retention of non-progressor status, but this p24 response was absent or declined in viraemic subjects. Strong Gag-dominant cytotoxic T lymphocyte (CTL) responses were identified in most LTNP, or Pol dominant-CTL in those with nef-defective HIV infection. CTL were associated with control of viraemia when combined with p24 proliferative responses. However, CTL did not prevent late disease progression. Individuals with sustained viral suppression had CTL recognising numerous Gag epitopes, while strong but restricted responses to one or two immunodominant epitopes was effective for some time, but failed to contain viraemia over the course of this study. Viral escape mutants at a HLA B27-restricted Gag-p24 epitope were detected in only 1 of 3 individuals, whereas declining or negative p24 proliferative responses occurred in all 3 concurrent with an increase in viraemia.Conclusion: Detectable viraemia at study entry was predictive of loss of LTNP status and/or disease progression in 6 of 8, and differentiated slow progressors from elite LTNP who retained potent virological control. Sustained immunological suppression of viraemia was independently associated with preserved p24 proliferative responses, regardless of the strength and breadth of the CTL response. A decline in this protective p24 response preceded or correlated with loss of non-progressor status and/or signs of disease progression.

Rapid antibody tests for the detection of human immunodeficiency virus (HIV) offer an effective means of providing a timely result of HIV serostatus to individuals. The increased use of rapid HIV antibody tests outside the laboratory has highlighted the need for new, cost-effective quality assurance methods to be developed for use in nonlaboratory-based and resource-limited settings. Photographed rapid HIV test results were used in a modified external quality assessment scheme to assess the interpretation proficiency and, therefore, to assess the feasibility of using this method as a basis for a quality assessment program for nonlaboratory-based testing. Participants (n = 148), both experienced and inexperienced in the performance and interpretation of rapid HIV testing, interpreted the photographed results of five rapid HIV assays. These were scored according to the degree of technical discordance. Error scores were grouped according to each participant's technical experience. The accuracy of interpretation for four of the five assays was between 80 and 97%, indicating that the photographed results of samples, including those difficult to read or borderline difficult to read, can be used to assess the proficiency of test operators in interpreting results. Participants had greater difficulty in interpreting samples of weak reactivity; this was consistent across the five assays. Experience played an important role in accurate interpretation, with experienced laboratory participants exhibiting greater proficiency (P < 0.05) in interpreting the results of three of the five rapid HIV assays. It was established that photographed results of rapid HIV assays could be interpreted with accuracy and demonstrated that prior experience resulted in a more accurate interpretation performance.

In efforts to develop an effective vaccine, sterilizing immunity to primate lentiviruses has only been achieved by the use of live attenuated viruses carrying major deletions in nef and other accessory genes. Although live attenuated HIV vaccines are unlikely to be developed due to a myriad of safety concerns, opportunities exist to better understand the correlates of immune protection against HIV infection by studying rare cohorts of long-term survivors infected with attenuated, nef-deleted HIV strains such as the Sydney blood bank cohort (SBBC). Here, we review studies of viral evolution, pathogenicity, and immune responses to HIV infection in SBBC members. The studies show that potent, broadly neutralizing anti-HIV antibodies and robust CD8+ T-cell responses to HIV infection were not necessary for long-term control of HIV infection in a subset of SBBC members, and were not sufficient to prevent HIV sequence evolution, augmentation of pathogenicity and eventual progression of HIV infection in another subset. However, a persistent T-helper proliferative response to HIV p24 antigen was associated with long-term control of infection. Together, these results underscore the importance of the host in the eventual outcome of infection. Thus, whilst generating an effective antibody and CD8+ T-cell response are an essential component of vaccines aimed at preventing primary HIV infection, T-helper responses may be important in the generation of an effective therapeutic vaccine aimed at blunting chronic HIV infection.

Long-term survival of human immunodeficiency virus type 1 (HIV-1) infection has been noted in rare cohorts of individuals infected with nef-deleted virus. Enhanced macrophage tropism and cytopathicity contribute to pathogenicity of wild type HIV-1. To better understand the pathogenesis of nef-deleted HIV-1, we analyzed the replication capacity and macrophage cytopathicity of nef-deleted HIV-1 isolated sequentially from a long-term survivor during progression to AIDS (n=6 isolates). Compared with controls, all nef-deleted viruses replicated to low levels in peripheral blood mononu-clear cells and monocyte-derived macrophages (MDM). One nef-deleted virus that was isolated on the development of AIDS caused high levels of syncytia in MDM similar to control viruses, but five viruses isolated from earlier times prior to AIDS onset caused only minimal cytopathicity. Together, these results suggest that enhanced cytopathicity of nef-deleted HIV-1 for MDM can occur independently of replication capacity, and may contribute to the pathogenesis of nef-deleted HIV-1 infection.

The Sydney Blood Bank Cohort (SBBC) consists of eight blood transfusion recipients infected with nef-attenuated human immunodeficiency virus type 1 (HIV-1) acquired from a single donor. Here, we show that viral phenotypes and antibody responses differ considerably between individual cohort members, despite the single source of infection. Replication of isolated virus varied from barely detectable to similar to that of the wild-type virus, and virus isolated from five SBBC members showed coreceptor usage signatures unique to each individual. Higher viral loads and stronger neutralizing antibody responses were associated with better-replicating viral strains, and detectable viral replication was essential for the development of strong and sustained humoral immune responses. Despite the presence of strong neutralizing antibodies in a number of SBBC members, disease progression was not prevented, and each cohort member studied displayed a unique outcome of infection with nef-attenuated HIV-1.

BACKGROUND:The Sydney blood bank cohort (SBBC) of long-term survivors consists of multiple individuals infected with attenuated, nef-deleted variants of human immunodeficiency virus type 1 (HIV-1) acquired from a single source. Long-term prospective studies have demonstrated that the SBBC now comprises slow progressors (SP) as well as long-term nonprogressors (LTNP). Convergent evolution of nef sequences in SBBC SP and LTNP indicates the in vivo pathogenicity of HIV-1 in SBBC members is dictated by factors other than nef. To better understand mechanisms underlying the pathogenicity of nef-deleted HIV-1, we examined the phenotype and env sequence diversity of sequentially isolated viruses (n = 2) from 3 SBBC members.RESULTS:The viruses characterized here were isolated from two SP spanning a three or six year period during progressive HIV-1 infection (subjects D36 and C98, respectively) and from a LTNP spanning a two year period during asymptomatic, nonprogressive infection (subject C18). Both isolates from D36 were R5X4 phenotype and, compared to control HIV-1 strains, replicated to low levels in peripheral blood mononuclear cells (PBMC). In contrast, both isolates from C98 and C18 were CCR5-restricted. Both viruses isolated from C98 replicated to barely detectable levels in PBMC, whereas both viruses isolated from C18 replicated to low levels, similar to those isolated from D36. Analysis of env by V1V2 and V3 heteroduplex tracking assay, V1V2 length polymorphisms, sequencing and phylogenetic analysis showed distinct intra- and inter-patient env evolution.CONCLUSION:Independent evolution of env despite convergent evolution of nef may contribute to the in vivo pathogenicity of nef-deleted HIV-1 in SBBC members, which may not necessarily be associated with changes in replication capacity or viral coreceptor specificity.

Movement of HIV testing to point of care (POC) has been facilitated by the availability of simple/rapid anti-HIV immunoassays. These assays have been used at health centres in the United States, Canada and at voluntary counselling and testing centres in developing countries. While this approach has revolutionised HIV screening in some areas, there are ethical, legal and quality assurance considerations that must be addressed before POC HIV testing is universally accepted.

The authors studied the transcriptional activity of blood- and cerebrospinal fluid (CSF)-derived nef/long-terminal repeat (LTR) sequences isolated from a slow progressor infected with nef-deleted human immunodeficiency virus type 1 (HIV-1) who developed HIV-associated dementia (HIVD). The transcriptional activity of CSF-derived nef/LTR clones isolated during HIVD was up to 4.5-fold higher than blood-derived clones isolated before and during HIVD when tested under basal, phorbol 12-myristate 13-acetate-(PMA−), and Tat-activated conditions, and was associated with the presence of duplicated nuclear factor (NF)-κB and specificity factor-1 (Sp-1) binding sites coupled with a truncated nef sequence, increased replication capacity, and high CSF viral load. Thus, nef and LTR mutations that augment transcription may contribute to neuropathogenesis of nef-deleted HIV-1.

We studied the evolution and compartmentalization of nef/ long terminal repeat ( nef/ LTR) - deleted human immunodeficiency virus type 1 (HIV-1) from a long-term survivor who developed HIV-associated dementia (HIVD). Analysis of sequential blood-derived HIV-1 isolated before and during HIVD revealed a persistent R5X4 phenotype and a progressive loss of nef/ LTR sequence; in contrast, HIV-1 present in cerebrospinal fluid during HIVD had an R5 phenotype, distinct nef/ LTR sequence of unique deletions and additional nuclear factor - kappaB sites and specificity factor - 1 sites, and enhanced transcriptional activity, compared with the blood-derived isolates. Thus, nef/LTR-deleted HIV-1 strains may undergo compartmentalized evolution in long-term survivors and cause neurologic disease.

Virions produced after HIV-1 infection of HTLV-I transformed cells have an expanded tropism that has been attributed to the pressence of HTLV-I glycoproteins in the envelope. This report now directly identifies these phenotypically mixed virions by immunogold labelling electron microscopy. Furthermore we estimate there are 2% of these in cell-free supernatant, which represents up to 1 × 107 particles/ml from an in vitro infection. HTLV-1 envelope labelling was localised to a single region, suggesting a defined event in packaging of foreign envelope proteins into HIV-1 virus particles.

Two Australian HIV-1 isolates, derived from patient blood (HIV MBC200 ) and cerebrospinal fluid (HIV MBC925 ), were characterized after in vitro culture in peripheral blood mononuclear cells (PBMC). Although virus replication was similar, as measured by cell-free reverse transcriptase activity, only one of the two isolates (HIV-1 MCB200 ) consistently induced cell syncytia and depleted the PBMC population of CD4+ cells by cell killing. A novel technique, devised for rapidly obtaining high-quality viral sequence data and the full-length genomic sequence of these two isolates, is presented. Analysis of the predicted sequence of the viral Env proteins provides correlates of the observed phenotypes. Phylogenetic analysis derived using near full-length sequence of these and other HIV-1 subtype B genomic sequences (including two other Australian isolates) shows a star-shaped phylogeny with each member having a similar genetic diversity. These data expand the database of genomic sequence available from well-characterized primary clinical isolates of HIV-1 using a novel rapid technique.

AIDS Research and Human RetrovirusesVol. 14, No. 9 Sequence Note: Genomic Sequence of HIV Type 1 from Four Members of the Sydney Blood Bank Cohort of Long-Term NonprogressorsROBERT OELRICHS, ANNA TSYKIN, DAVID RHODES, AJANTHA SOLOMON, ANNE ELLETT, DALE McPHEE, and NICHOLAS DEACONROBERT OELRICHS, ANNA TSYKIN, DAVID RHODES, AJANTHA SOLOMON, ANNE ELLETT, DALE McPHEE, and NICHOLAS DEACONPublished Online:15 Mar 2009https://doi.org/10.1089/aid.1998.14.811AboutSectionsPDF/EPUB Permissions & CitationsPermissionsDownload CitationsTrack CitationsAdd to favorites Back To Publication ShareShare onFacebookTwitterLinked InRedditEmail FiguresReferencesRelatedDetailsCited byHIV-1 Rev-RRE functional activity in primary isolates is highly dependent on minimal context-dependent changes in Rev1 November 2022 | Scientific Reports, Vol. 12, No. 1HIV-1 Hijacking of Host ATPases and GTPases That Control Protein Trafficking8 July 2021 | Frontiers in Cell and Developmental Biology, Vol. 9Validation of Variant Assembly Using HAPHPIPE with Next-Generation Sequence Data from Viruses14 July 2020 | Viruses, Vol. 12, No. 7Sequence and Functional Variation in the HIV-1 Rev Regulatory AxisCurrent HIV Research, Vol. 18, No. 2Structure of HIV-2 Nef Reveals Features Distinct from HIV-1 Involved in Immune RegulationiScience, Vol. 23, No. 1CD4 downregulation by the HIV-1 protein Nef reveals distinct roles for the γ1 and γ2 subunits of the AP-1 complex in protein traffickingJournal of Cell Science, Vol. 130, No. 2HIV-1 Nef: Taking Control of Protein Trafficking3 June 2016 | Traffic, Vol. 17, No. 9HIV-1 Nef Sequence and Functional Compartmentalization in the Gut Is Not Due to Differential Cytotoxic T Lymphocyte Selective Pressure13 September 2013 | PLoS ONE, Vol. 8, No. 9Viral Genetic Determinants of Nonprogressive HIV Type 1 Subtype C Infection in Antiretroviral Drug-Naive Children D.B. Tzitzivacos, C.T. Tiemessen, W.S. Stevens, and M.A. Papathanasopoulos29 November 2009 | AIDS Research and Human Retroviruses, Vol. 25, No. 11Simultaneous assessment of CD4 and MHC-I downregulation by Nef primary isolates in the context of infectionJournal of Virological Methods, Vol. 161, No. 2Protein structure and oligomerization are important for the formation of export-competent HIV-1 Rev-RRE complexes2 January 2009 | Protein Science, Vol. 17, No. 3Nef interference with HIV-1–specific CTL antiviral activity is epitope specificBlood, Vol. 108, No. 10Evasion of cytotoxic T lymphocytes is a functional constraint maintaining HIV-1 Nef expressionEuropean Journal of Immunology, Vol. 35, No. 11Full-Length Genome Characterization of HIV Type 1 Subtype C Isolates from Two Slow-Progressing Perinatally Infected Siblings in South Africa M. A. Papathanasopoulos, T. Patience, T. M. Meyers, F. E. McCutchan, and L. Morris5 July 2004 | AIDS Research and Human Retroviruses, Vol. 19, No. 11Broadly Increased Sensitivity to Cytotoxic T Lymphocytes Resulting from Nef Epitope Escape MutationsThe Journal of Immunology, Vol. 171, No. 8Will we be able to 'spot' an effective HIV-1 vaccine?Trends in Immunology, Vol. 24, No. 2A Human Immunodeficiency Virus Type 1 Isolate from an Infected Person Homozygous for CCR5Δ32 Exhibits Dual Tropism by Infecting Macrophages and MT2 Cells via CXCR4Journal of Virology, Vol. 76, No. 7Nef-Mediated Resistance of Human Immunodeficiency Virus Type 1 to Antiviral Cytotoxic T LymphocytesJournal of Virology, Vol. 76, No. 4Situación actual en el desarrollo de una vacuna frente al virus de la inmunodeficiencia humanaEnfermedades Infecciosas y Microbiología Clínica, Vol. 20, No. 10An examination of signs of disease progression in survivors of the Sydney Blood Bank Cohort (SBBC)Journal of Clinical Virology, Vol. 22, No. 3Unusual Polymorphisms in Human Immunodeficiency Virus Type 1 Associated with Nonprogressive InfectionJournal of Virology, Vol. 74, No. 9Rapid full-length genomic sequencing of two cytopathically heterogeneous Australian primary HIV-1 isolatesJournal of Biomedical Science, Vol. 7, No. 2The Explosive Human Immunodeficiency Virus Type 1 Epidemic among Injecting Drug Users of Kathmandu, Nepal, Is Caused by a Subtype C Virus of Restricted Genetic DiversityJournal of Virology, Vol. 74, No. 3Identification of a New Recipient in the Sydney Blood Bank Cohort: A Long-Term HIV Type 1-Infected Seroindeterminate Individual David Rhodes, Ajantha Solomon, Wayne Bolton, Jeanette Wood, John Sullivan, Jenny Learmont, and Nicholas Deacon5 July 2004 | AIDS Research and Human Retroviruses, Vol. 15, No. 16AIDS vaccine research in Asia: needs and opportunitiesAIDS, Vol. 13, No. 11Immunologic and Virologic Status after 14 to 18 Years of Infection with an Attenuated Strain of HIV-1 — A Report from the Sydney Blood Bank CohortNew England Journal of Medicine, Vol. 340, No. 22Full-Length Genomic Sequence of an HIV Type 1 Subtype G from Kinshasa Robert B. Oelrichs, Anne-Mieke Vandamme, Kristel Van Laethem, Zeger Debyser, Francine E. Mccutchan, and Nicholas J. Deacon5 July 2004 | AIDS Research and Human Retroviruses, Vol. 15, No. 6Sequence Note: A Novel Subtype A/G/J Recombinant Full-Length HIV Type 1 Genome from Burkina Faso ROBERT B. OELRICHS, CASSY WORKMAN, TIINA LAUKKANEN, FRANCINE E. McCUTCHAN, and NICHOLAS J. DEACON15 March 2009 | AIDS Research and Human Retroviruses, Vol. 14, No. 16 Volume 14Issue 9Jun 1998 To cite this article:ROBERT OELRICHS, ANNA TSYKIN, DAVID RHODES, AJANTHA SOLOMON, ANNE ELLETT, DALE McPHEE, and NICHOLAS DEACON.Sequence Note: Genomic Sequence of HIV Type 1 from Four Members of the Sydney Blood Bank Cohort of Long-Term Nonprogressors.AIDS Research and Human Retroviruses.Jun 1998.811-814.http://doi.org/10.1089/aid.1998.14.811Published in Volume: 14 Issue 9: March 15, 2009PDF download