RESUMO
Acceptable precision was achieved in a comparison study of the Abbott RealTime (RT) and Roche CAP/CTM-48 V2 HIV-1 assays, but viral load quantification was under- and overestimated, respectively, compared to the 2nd HIV-1 WHO International Standard. The same quantification patterns were observed for patient cohorts from Africa and the United States.
Assuntos
Infecções por HIV/diagnóstico , HIV-1/isolamento & purificação , Técnicas de Diagnóstico Molecular/métodos , Kit de Reagentes para Diagnóstico , Carga Viral/métodos , África , Infecções por HIV/virologia , Humanos , Estados UnidosRESUMO
BACKGROUND: To evaluate the prevalence of human papilloma virus (HPV) infection and types in the oral and cervix mucosa of treatment-naïve HIV-1-positive women with CD4 counts less than 300 cells per ml with no HPV-associated oral lesions. METHODS: Oral epithelium was harvested from the buccal mucosa and lateral borders of the tongue and cervical samples were collected from the endocervical area of 30 women, 22-64 years old. Cytobrush Plus cell collectors were used for sampling both anatomical areas. Genital pathology, obstetric and gynaecological history, co-morbid disease, hormone therapy, sexual behavior and smoking history were assessed via physical examination and clinical interviews. Special investigations included cervical Papanicolau smears, CD4 counts and HIV-1 viral loads. The linear array HPV test was used to determine HPV genotypes present in the specimens. RESULTS: Oral HPV were identified in 20% (n = 6) of the patients, of which two had infection with two HPV types. Genital HPV was found in 96.7% (n = 29) of the women, of which only 14 had cytological abnormalities on Papanicolau smear. Infection with multiple HPV types were present in 93.1% (n = 27) of the patients, with an average of four HPV types per individual. CONCLUSIONS: South African HIV-positive women with CD4 counts less than 300 cells per ml have a significant risk of cervical HPV strains and multiple strain infection of the cervix. The prevalence of HPV in normal oral mucosa was low but high-risk types were present. Limited correlation between oral HPV types and those identified in the cervical mucosa was found.
Assuntos
Infecções por HIV/complicações , Soropositividade para HIV/complicações , HIV-1/imunologia , Mucosa/virologia , Papillomaviridae/classificação , Infecções por Papillomavirus/virologia , Infecções Oportunistas Relacionadas com a AIDS/imunologia , Infecções Oportunistas Relacionadas com a AIDS/virologia , Adulto , Antirretrovirais/uso terapêutico , Contagem de Linfócito CD4 , Colo do Útero/virologia , DNA Viral/análise , Feminino , Infecções por HIV/tratamento farmacológico , Infecções por HIV/imunologia , Soropositividade para HIV/imunologia , Soropositividade para HIV/virologia , Humanos , Pessoa de Meia-Idade , Mucosa Bucal/virologia , Papillomaviridae/genética , Infecções por Papillomavirus/induzido quimicamente , Infecções por Papillomavirus/complicações , Prevalência , Fatores de Risco , África do Sul , Adulto JovemRESUMO
A safe and effective HIV-1 vaccine is urgently needed to control the worldwide AIDS epidemic. Traditional methods of vaccine development have been frustratingly slow, and it is becoming increasingly apparent that radical new approaches may be required. Computational and mathematical approaches, combined with evolutionary reasoning, may provide new insights for the design of an efficacious AIDS vaccine. Here, we used codon-based substitution models and maximum-likelihood (ML) methods to identify positively selected sites that are likely to be involved in the immune control of HIV-1. Analysis of subtypes B and C revealed widespread adaptive evolution. Positively selected amino acids were detected in all nine HIV-1 proteins, including Env. Of particular interest was the high level of positive selection within the C-terminal regions of the immediate-early regulatory proteins, Tat and Rev. Many of the amino acid replacements were associated with the emergence of novel (or alternative) myristylation and casein kinase II (CKII) phosphorylation sites. The impact of these changes on the conformation and antigenicity of Tat and Rev remains to be established. In rhesus macaques, a single CTL-associated amino substitution in Tat has been linked to escape from acute SIV infection. Understanding the relationship between host-driven positive selection and antigenic variation may lead to the development of novel vaccine strategies that preempt the escape process.
Assuntos
Mapeamento Cromossômico , Variação Genética , Genoma Viral , HIV-1/genética , Filogenia , Seleção Genética , Vacinas/genética , Sequência de Aminoácidos , Bases de Dados Genéticas , Epitopos/genética , Genes rev/genética , Genes tat/genética , Funções Verossimilhança , Modelos Genéticos , Dados de Sequência Molecular , Alinhamento de SequênciaRESUMO
South Africa has one of the fastest growing HIV-1 epidemics worldwide, consisting mostly of subtype C. However, HIV-1 subtype B and subtype D viruses were isolated in the beginning of the epidemic in the early 1980s. This study describes the amplification, cloning, and near full-length genome sequencing of four HIV-1 subtype D primary strains, isolated from 1984 to 1986 in Cape Town, in what seems to have been a small restricted subtype D epidemic in the country.
Assuntos
Surtos de Doenças , Genoma Viral , Infecções por HIV/epidemiologia , HIV-1/classificação , Análise de Sequência de DNA , Infecções por HIV/virologia , HIV-1/genética , Humanos , Masculino , Dados de Sequência Molecular , Filogenia , África do Sul/epidemiologiaRESUMO
By the end of 2012, more than 6.1 million people were infected with HIV-1 in South Africa. Subtype C was responsible for the majority of these infections and more than 300 near full-length genomes (NFLGs) have been published. Currently very few non-subtype C isolates have been identified and characterized within the country, particularly full genome non-C isolates. Seven patients from the Tygerberg Virology (TV) cohort were previously identified as possible non-C subtypes and were selected for further analyses. RNA was isolated from five individuals (TV047, TV096, TV101, TV218, and TV546) and DNA from TV016 and TV1057. The NFLGs of these samples were amplified in overlapping fragments and sequenced. Online subtyping tools REGA version 3 and jpHMM were used to screen for subtypes and recombinants. Maximum likelihood (ML) phylogenetic analysis (phyML) was used to infer subtypes and SimPlot was used to confirm possible intersubtype recombinants. We identified three subtype B (TV016, TV047, and TV1057) isolates, one subtype A1 (TV096), one subtype G (TV546), one unique AD (TV101), and one unique AC (TV218) recombinant form. This is the first NFLG of subtype G that has been described in South Africa. The subtype B sequences described also increased the NFLG subtype B sequences in Africa from three to six. There is a need for more NFLG sequences, as partial HIV-1 sequences may underrepresent viral recombinant forms. It is also necessary to continue monitoring the evolution and spread of HIV-1 in South Africa, because understanding viral diversity may play an important role in HIV-1 prevention strategies.
Assuntos
Genoma Viral , Genótipo , Infecções por HIV/virologia , HIV-1/classificação , HIV-1/genética , Análise de Sequência de DNA , Adulto , Análise por Conglomerados , Feminino , HIV-1/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Filogenia , Recombinação Genética , Homologia de Sequência , África do SulRESUMO
Understanding the origin, distribution, and evolving dominance of HIV-1 subtype C strains is an important component in the design and evaluation of a globally effective AIDS vaccine. To better understand subtype C viruses, we constructed complete molecular clones of primary, CCR-5-using isolates from South Africa and analyzed the molecular phylogenies of these clones using best fitting evolutionary substitution models. Analyses were performed on three full-length sequences, and on the individual genes. All clones were nonrecombinant, and although two of three had open reading frames and intact splice sites, they were not infectious. At the genomic level, the models demonstrated the increasing variability of subtype C in South Africa. At the subgenomic level, they revealed marked differences in the evolutionary patterns of individual genes, a finding that suggests that the genes are under different selective pressures and constraints. These data underscore the dynamic nature of the subtype C epidemic and emphasize the need for continuous monitoring of local strains.
Assuntos
HIV-1/classificação , Síndrome da Imunodeficiência Adquirida/virologia , Animais , Sequência de Bases , Células COS , Evolução Molecular , HIV-1/genética , Humanos , Dados de Sequência Molecular , Filogenia , África do SulRESUMO
Human immunodeficiency virus type 1 (HIV-1) subtype C has become the major etiological agent in the global and especially African epidemic. To gain better understanding of the genetic diversity and rapid transmission of HIV-1 subtype C, we have characterized the complete 5' long terminal repeat (LTR) region along with the regulatory genes tat and rev as well as the accessory gene nef of 14 South African HIV-1 subtype C isolates. Phylogenetic analysis revealed a subtype C 5' LTR cluster, as well as subclustering of our nef sequences with various subtype C strains separate from the India and China subclusters. At least 3 NF-kappaB sites were present in the 5' LTR of most isolates and 13 isolates had the subtype C-specific Rev truncation. Some length variation in exon 2 and the absence of a critical cysteine were found in Tat. Residue variation in the myristoylation signal and motifs involved in CD4 and MHC-I downregulation was recorded in our nef gene sequences.
Assuntos
Genes nef , Genes rev , Genes tat , Infecções por HIV/virologia , Repetição Terminal Longa de HIV , HIV-1/genética , África Austral/epidemiologia , Sequência de Aminoácidos , Infecções por HIV/epidemiologia , HIV-1/classificação , Humanos , Dados de Sequência Molecular , Alinhamento de Sequência , Homologia de Sequência de AminoácidosAssuntos
Síndrome da Imunodeficiência Adquirida/epidemiologia , Surtos de Doenças/prevenção & controle , Infecções por HIV/epidemiologia , Infecções por HIV/virologia , Vacinas contra a AIDS/uso terapêutico , Síndrome da Imunodeficiência Adquirida/prevenção & controle , Adolescente , Adulto , África/epidemiologia , Criança , Pré-Escolar , Saúde Global , HIV-1/classificação , HIV-1/patogenicidade , HIV-2/patogenicidade , Humanos , Incidência , Lactente , Recém-Nascido , Pessoa de Meia-Idade , Vigilância da População , Prevalência , PesquisaRESUMO
We analyzed the HIV-1 pol gene from patients in Cape Town to determine the genetic diversity of HIV-1 in the region and to assess the baseline HIV-1 resistance level of treatment-naive patients. Plasma was collected prior to the national antiretroviral therapy (ART) program. RNA was extracted, followed by RT-PCR and automated DNA sequencing of the viral protease (PR) and reverse transcriptase (RT) coding region. Genotyping was done through phylogenetic analysis. The sequences were inspected for resistance-associated mutations against PR and RT inhibitors. A total of 140 pol sequences were analyzed, of which 133 (95%) belong to HIV-1 subtype C, five (3.6%) were subtype B, and one each was subtype G and CRF02_AG. Five sequences (3.6%) had resistance-associated mutations. These include three (2.1%) NNRTI mutations. With the progression of the national ART program, it is important to monitor the resistance profile of naive and treatment-experienced patients.
Assuntos
Fármacos Anti-HIV/farmacologia , Infecções por HIV/virologia , HIV-1/efeitos dos fármacos , HIV-1/genética , Antirretrovirais , Farmacorresistência Viral/genética , Feminino , Genes pol/genética , Variação Genética , Protease de HIV/genética , Transcriptase Reversa do HIV/genética , Humanos , Masculino , Dados de Sequência Molecular , Mutação , Fases de Leitura Aberta/genética , Filogenia , Inibidores de Proteases/farmacologia , Inibidores da Transcriptase Reversa/farmacologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , África do SulRESUMO
South Africa has the highest number of HIV-1-infected individuals in the world, with HIV-1 subtype C prevailing. However, HIV-1 subtype C accessory genes are rarely characterized in the country. These genes are important for establishing viral pathogenesis. The Vif protein has been shown to counteract the antiretroviral activity of APOBEC3G/F cytidine deaminases. In this study an additional 50 HIV-1 vif sequences are characterized. These include 48 HIV-1 subtype C and 2 HIV-1 subtype B sequences. Highly conserved HIV-1 subtype C motifs are outlined. The previously identified RLRR (90-93) motif does not seem to be conserved among our newly analyzed sequences. Conserved motifs can be useful for developing new vaccine strategies or antiretroviral drugs.
Assuntos
Produtos do Gene vif/genética , Infecções por HIV/virologia , Motivos de Aminoácidos/genética , Sequência de Aminoácidos , Feminino , HIV-1/genética , Humanos , Masculino , Dados de Sequência Molecular , Alinhamento de Sequência , África do SulRESUMO
BACKGROUND: High-risk human papilloma viruses (HPV) are reported to be significant independent risk factors for oral squamous cell carcinoma (OSCC). The prevalence of HPV in OSCC in a South African population sample was evaluated comparing three different HPV detection methods. METHODS: Tumour and adjacent morphologically normal oral mucosa of 59 resections of primary OSCC were evaluated for the presence of HPV using real-time polymerase chain reaction (PCR), conventional in situ hybridization (ISH), and a signal amplification ISH technique (Dako GenPoint). RESULTS: HPV18 DNA was detected in seven cases using real-time PCR. No positivity was found with the other two ISH techniques. CONCLUSIONS: We support the view that HPV is probably unimportant in the pathogenesis of OSCC and hypothesize HPV detection techniques as the main reason for the positive results in many studies. Real-time PCR was confirmed as the most sensitive technique, but researchers are urged to incorporate strict internal controls when using this detection method.
Assuntos
Carcinoma de Células Escamosas/virologia , Neoplasias Bucais/virologia , Papillomaviridae/isolamento & purificação , Adulto , Idoso , DNA Viral/análise , Contaminação de Equipamentos , Feminino , Técnicas de Preparação Histocitológica , Papillomavirus Humano 18/genética , Papillomavirus Humano 18/isolamento & purificação , Humanos , Hibridização In Situ/métodos , Masculino , Pessoa de Meia-Idade , Mucosa Bucal/virologia , Infecções por Papillomavirus/diagnóstico , Reação em Cadeia da Polimerase , Fatores de Risco , Simbiose/fisiologia , Infecções Tumorais por Vírus/diagnósticoRESUMO
The efficacy of cellular immune responses elicited by HIV vaccines is dependent on their strength, durability and antigenic breadth. The regulatory proteins are abundantly expressed early in the viral life cycle and CTL recognition may bring about early killing of infected cells. We synthesised DNA vaccine constructs that encode consensus HIV-1 subtype C Tat, Rev and Nef proteins. Proteins carrying inactivating mutations were tested for functional activity and highly expressing, inactive Tat, Rev and Nef mutants were identified and their reading frames fused into a TatRevNef cassette. Single- and polygene Tat, Rev and/or Nef constructs were immunogenic in BALB/c mice. These constructs may serve to increase the antigenic breadth for an HIV-1 vaccine that is relevant for sub-Saharan Africa.
Assuntos
Vacinas contra a AIDS/genética , Sequência Consenso/imunologia , Genes nef/genética , Genes rev/genética , Genes tat/genética , HIV-1/genética , Vacinas de DNA/genética , Vacinas contra a AIDS/imunologia , Animais , Linhagem Celular , Feminino , Genes nef/imunologia , Genes rev/imunologia , Genes tat/imunologia , HIV-1/imunologia , Células HeLa , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Mutação , Vacinas de DNA/classificação , Vacinas de DNA/imunologia , Vacinas de Produtos Inativados/genética , Vacinas de Produtos Inativados/imunologiaRESUMO
CXC chemokine ligand 12 (CXCL12), or stromal cell-derived factor 1 (SDF1), is the only known natural ligand for the HIV-1 coreceptor, CXC chemokine receptor 4 (CXCR4). A single nucleotide polymorphism (SNP) in the CXCL12 gene (SDF1-3'A) has been associated with disease progression to AIDS in some studies, but not others. Mutations in the CXCR4 gene are generally rare and have not been implicated in HIV-1/AIDS pathogenesis. This study analyzed the SDF1-3'A SNP and performed mutation screening for polymorphic markers in the CXCR4 gene to determine the presence or absence of significant associations with susceptibility to HIV-1 infection. The study consisted of 257 HIV-1-seropositive patients and 113 HIV-1-seronegative controls representing a sub-Saharan African population belonging to the Xhosa ethnic group of South Africa. The SDF1-3'A SNP was associated with an increased risk for HIV-1 infection (P = 0.0319) whereas no significant association was observed between the occurrence of the SDF1-3'A SNP and increased or decreased plasma levels of CXCL12. Comprehensive mutation analysis of the CXCR4 gene confirmed a high degree of genetic conservation within the coding region of this ancient population.
Assuntos
População Negra , Quimiocinas CXC/genética , Variação Genética , Infecções por HIV/genética , Polimorfismo de Nucleotídeo Único , Receptores CXCR4/genética , Quimiocina CXCL12 , Genótipo , Infecções por HIV/etnologia , Infecções por HIV/fisiopatologia , HIV-1/patogenicidade , Humanos , Fatores de Risco , África do Sul/epidemiologiaRESUMO
MOTIVATION: Genetic analysis of HIV-1 is important not only for vaccine development, but also to guide treatment strategies, track the emergence of new viral variants and ensure that diagnostic assays are contemporary and fully optimized. However, most genotyping methods are laborious and complex, and involve the use of multiple software applications. Here, we describe the development of an automated genotyping system that can be easily applied to HIV-1 and other rapidly evolving viral pathogens. RESULTS: The new REGA subtyping tool, developed using Java programming and PERL scripts, combines phylogenetic analyses with boot-scanning methods for the genetic subtyping of full-length and subgenomic fragments of HIV-1. When used to investigate the subtype of previously published reference datasets that were analysed using manual phylogenetic methods, the automated method correctly identified 97.5-100% of non-recombinant and circulating recombinant forms of HIV-1, including 108 full-length, 108 gag and 221 env sequences downloaded from the Los Alamos database.
Assuntos
Algoritmos , Inteligência Artificial , Mapeamento Cromossômico/métodos , DNA Viral/genética , HIV-1/genética , Reconhecimento Automatizado de Padrão/métodos , Análise de Sequência de DNA/métodos , Sequência de Bases , DNA Viral/análise , Genótipo , Dados de Sequência Molecular , Filogenia , Alinhamento de Sequência/métodos , SoftwareRESUMO
Human immunodeficiency virus type 1 (HIV-1) subtype C infections are on the rise in Sub-Saharan Africa and Asia. Therefore, there is a need to develop an HIV vaccine capable of eliciting broadly reactive immune responses against members of this subtype. We show here that modified HIV envelope (env) DNA vaccines derived from the South African subtype C TV1 strain are able to prime for humoral responses in rabbits and rhesus macaques. Priming rabbits with DNA plasmids encoding V2-deleted TV1 gp140 (gp140TV1DeltaV2), followed by boosting with oligomeric protein (o-gp140TV1DeltaV2) in MF59 adjuvant, elicited higher titers of env-binding and autologous neutralizing antibodies than priming with DNA vaccines encoding the full-length TV1 env (gp160) or the intact TV1 gp140. Immunization with V2-deleted subtype B SF162 env and V2-deleted TV1 env together using a multivalent vaccine approach induced high titers of oligomeric env-binding antibodies and autologous neutralizing antibodies against both the subtypes B and C vaccine strains, HIV-1 SF162 and TV1, respectively. Low-level neutralizing activity against the heterologous South African subtype C TV2 strain, as well as a small subset of viruses in a panel of 13 heterologous primary isolates, was observed in some rabbits immunized with the V2-deleted vaccines. Immunization of rhesus macaques with the V2-deleted TV1 DNA prime/protein boost also elicited high titers of env-binding antibodies and moderate titers of autologous TV1 neutralizing antibodies. The pilot-scale production of the various TV1 DNA vaccine constructs and env proteins described here should provide an initial platform upon which to improve the immunogenicity of these subtype C HIV envelope vaccines.
Assuntos
Vacinas contra a AIDS/imunologia , Produtos do Gene env/imunologia , Anticorpos Anti-HIV/sangue , Infecções por HIV/imunologia , HIV-1/imunologia , Vacinação , Vacinas contra a AIDS/administração & dosagem , Sequência de Aminoácidos , Animais , Avaliação Pré-Clínica de Medicamentos , Deleção de Genes , Produtos do Gene env/genética , Proteína gp160 do Envelope de HIV/genética , Proteína gp160 do Envelope de HIV/imunologia , Infecções por HIV/sangue , Imunização Secundária , Injeções Intramusculares , Macaca mulatta , Dados de Sequência Molecular , Mutação , Testes de Neutralização , Coelhos , Alinhamento de Sequência , África do Sul , Vacinas de DNA/administração & dosagem , Vacinas de DNA/imunologia , Produtos do Gene env do Vírus da Imunodeficiência HumanaRESUMO
OBJECTIVES: To evaluate the willingness of medical doctors working at a tertiary hospital to participate in HIV vaccine trials, their perceptions of patients' willingness to participate, and the major reasons underlying these views. DESIGN: A self-administered, anonymous postal survey conducted in two rounds between May and July 2001. SETTING: A tertiary care hospital in the Western Cape. SUBJECTS: All medical doctors listed on the hospital's staff directory. OUTCOME MEASURES: Willingness to participate in, and to recruit patients into, HIV vaccine trials, and the reasons for this. RESULTS AND CONCLUSIONS: Of the 289 individuals surveyed, 80% stated either that they would not be willing to participate in HIV vaccine trials or that they were unsure about their participation. Meanwhile, 37% stated that they would be willing to recruit patients into vaccine trials. The most common concerns with trial participation were the possibility of vaccine-induced infection and the possibility of testing positive for antibodies to HIV. The surprisingly low level of willingness to participate in trials in this sample of medical professionals highlights the importance of preparatory work to overcome substantial barriers to HIV vaccine trial participation.
Assuntos
Vacinas contra a AIDS/uso terapêutico , Ensaios Clínicos como Assunto/estatística & dados numéricos , Infecções por HIV/prevenção & controle , Participação do Paciente/estatística & dados numéricos , Seleção de Pessoal/estatística & dados numéricos , Médicos/estatística & dados numéricos , Adulto , Atitude do Pessoal de Saúde , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Seleção de Pacientes , África do SulRESUMO
Human herpesvirus 8 (HHV-8) has been identified as the causative agent for all forms of Kaposi's sarcoma and is also associated with the development of body cavity-based B-cell lymphomas and multicentric Castleman's disease. HHV-8 genomes are now classified into five major subtypes (A-E) that reflect sequence heterogeneity in the highly variable open reading frame (ORF) K1. To identify HHV-8 subtypes associated with different forms of Kaposi's sarcoma, we compared the ORF 26 and ORF-K1 gene sequences from South African patients with the prototype strains of the major subtypes, as well as published sequences from other African strains. DNA prepared from Kaposi's sarcoma biopsies and/or peripheral blood lymphocytes were available from 14 patients with postrenal transplant (iatrogenic) Kaposi's sarcoma, six patients with the African endemic form, and one patient with AIDS-related body cavity-based B-cell lymphoma. We identified a B2 subtype in six patients, four of whom also had a novel B5 type ORF 26 polymorphism. Two patients had B2 type patterns for both the ORF 26 and ORF-K1 genes. The ORF-K1 subtype A5 was identified in samples from three patients with a B3/C2 type polymorphism in the ORF 26 gene. A novel ORF-K1 B variant strain was identified in a patient with African endemic Kaposi's sarcoma, who also had a B3/C2 class ORF 26 pattern. In 58.3% of iatrogenic Kaposi's sarcoma patients, a B5-type ORF 26 gene sequence pattern was identified. No association was found among particular subtypes, geographical origin of patients, or clinical presentation.
Assuntos
Variação Genética , Herpesvirus Humano 8/classificação , Herpesvirus Humano 8/genética , Sarcoma de Kaposi/epidemiologia , Sarcoma de Kaposi/virologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Sequência de Aminoácidos , DNA Viral/análise , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Fases de Leitura Aberta/genética , Filogenia , Análise de Sequência de DNA , África do Sul/epidemiologia , Proteínas Virais/genéticaRESUMO
The objective of the study was to assess the genotypic and phenotypic properties of 18 viral strains from human immunodeficiency virus-1 (HIV-1) positive patients and to identify subtype C isolates for vaccine design strategies. All the isolates were non-syncytium-inducing (NSI) in both the primary and MT-2 cell cultures. The amino acid charge of the V3 loop correlated with the NSI phenotype of the strains. The V3 competitive peptide enzyme immunoassay and DNA sequencing of the partial gp120 region gave concordant results on the 15 subtype C strains, whereas the three B genotypes gave a positive to B, a nonreactive to B, and a dual reaction to the B-D peptides, respectively. Sixteen of the isolates used only CCR5 as coreceptor whereas two isolates made use of additional coreceptors including CXCR4. In summary, all our subtype C isolates are NSI phenotypically and almost all of them use CCR5 exclusively as their coreceptor.