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1.
Front Immunol ; 12: 723585, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34489974

RESUMO

Objectives: Our objective was to determine the antibody and cytokine profiles in different COVID-19 patients. Methods: COVID-19 patients with different clinical classifications were enrolled in this study. The level of IgG antibodies, IgA, IgM, IgE, and IgG subclasses targeting N and S proteins were tested using ELISA. Neutralizing antibody titers were determined by using a toxin neutralization assay (TNA) with live SARS-CoV-2. The concentrations of 8 cytokines, including IL-2, IL-4, IL-6, IL-10, CCL2, CXCL10, IFN-γ, and TNF-α, were measured using the Protein Sample Ella-Simple ELISA system. The differences in antibodies and cytokines between severe and moderate patients were compared by t-tests or Mann-Whitney tests. Results: A total of 79 COVID-19 patients, including 49 moderate patients and 30 severe patients, were enrolled. Compared with those in moderate patients, neutralizing antibody and IgG-S antibody titers in severe patients were significantly higher. The concentration of IgG-N antibody was significantly higher than that of IgG-S antibody in COVID-19 patients. There was a significant difference in the distribution of IgG subclass antibodies between moderate patients and severe patients. The positive ratio of anti-S protein IgG3 is significantly more than anti-N protein IgG3, while the anti-S protein IgG4 positive rate is significantly less than the anti-N protein IgG4 positive rate. IL-2 was lower in COVID-19 patients than in healthy individuals, while IL-4, IL-6, CCL2, IFN-γ, and TNF-α were higher in COVID-19 patients than in healthy individuals. IL-6 was significantly higher in severe patients than in moderate patients. The antibody level of anti-S protein was positively correlated with the titer of neutralizing antibody, but there was no relationship between cytokines and neutralizing antibody. Conclusions: Our findings show the severe COVID-19 patients' antibody levels were stronger than those of moderate patients, and a cytokine storm is associated with COVID-19 severity. There was a difference in immunoglobulin type between anti-S protein antibodies and anti-N protein antibodies in COVID-19 patients. And clarified the value of the profile in critical prevention.


Assuntos
Anticorpos Antivirais/sangue , COVID-19/imunologia , Citocinas/sangue , SARS-CoV-2/imunologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Anticorpos Neutralizantes/sangue , COVID-19/classificação , Proteínas do Nucleocapsídeo de Coronavírus/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Humanos , Imunoglobulina A/sangue , Imunoglobulina E/sangue , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Masculino , Pessoa de Meia-Idade , Índice de Gravidade de Doença , Glicoproteína da Espícula de Coronavírus/imunologia
2.
Arch Virol ; 165(3): 619-626, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31965315

RESUMO

Human pegivirus 2 (HPgV-2) is a recently recognized pegivirus of the family Flaviviridae. To investigate the epidemic features of HPgV-2 circulating in the human immunodeficiency virus (HIV)-infected population, we tested for antibodies and viral RNA of HPgV-2 and hepatitis C virus (HCV) with retrospective plasma samples collected from 771 HIV infections with multiple risk behaviors in Honghe Prefecture of Yunnan Province. A total of 195 subjects (25.29%) were seroreactive to HPgV-2, and 41 (5.32%) were RNA positive. Although the positive rate of HPgV-2 antibodies in HIV/HCV-coinfected individuals (27.69%) was significantly higher than that of HIV monoinfections (20.82%) (p = 0.036), this is the first report of HPgV-2 viremia in HIV-infected individuals without HCV infection and the presence of two HPgV-2 lineages in China. Our data indicate that HPgV-2 can also be transmitted sexually, which might be facilitated when combined with HCV infection, injecting drug use, and risky sexual behavior, which appear to have a synergistic effect on HPgV-2 infection. Phylogenetic analysis of 26 near-full-length genome sequences showed that the HPgV-2 strains in China are divided into two clusters.


Assuntos
Infecções por Flaviviridae/complicações , Infecções por Flaviviridae/epidemiologia , Flaviviridae/classificação , Infecções por HIV/complicações , Infecções por HIV/epidemiologia , Viremia , Anticorpos Antivirais/sangue , China/epidemiologia , Humanos , Incidência , Filogenia , Prevalência , RNA Viral/genética , RNA Viral/isolamento & purificação
3.
AIDS Res Hum Retroviruses ; 35(7): 679-683, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30924679

RESUMO

Jiangsu province has severe HIV-1 epidemic in China. Suqian which is located in north of the province has limited HIV epidemic information. Therefore, this study aimed to characterize the epidemic details in the area. A total of 196 plasma samples were collected from treated HIV-1-positive cases and viral RNA was extracted. Then HIV partial pol genes (nucleotide 2147-3462 by using HXB2 as calibrator) were amplified and sequenced. Finally, 84 partial pol genes were successfully obtained. The subtyping results indicate that multiple HIV-1 subtypes are circulating in Suqian district. Thereinto, CRF01_AE has been the dominant stains here and belonged to multiple lineages of CRF01_AE identified in China previously. Moreover, there is a high level of HIV drug resistance. All these results suggest HIV-1 epidemic in Suqian is rather complex and more measures must be performed for prevention and intervention in the area.


Assuntos
Infecções por HIV/epidemiologia , Infecções por HIV/virologia , HIV-1/genética , Fármacos Anti-HIV/uso terapêutico , Sequência de Bases , China/epidemiologia , Farmacorresistência Viral/efeitos dos fármacos , Farmacorresistência Viral/genética , Feminino , Genes pol/genética , Genótipo , Infecções por HIV/tratamento farmacológico , Infecções por HIV/transmissão , HIV-1/classificação , HIV-1/efeitos dos fármacos , HIV-1/isolamento & purificação , Humanos , Masculino , Epidemiologia Molecular , Filogenia , RNA Viral/sangue , RNA Viral/genética
4.
Sci Rep ; 7(1): 6330, 2017 07 24.
Artigo em Inglês | MEDLINE | ID: mdl-28740095

RESUMO

As the most dominant HIV-1 strain in China, CRF01_AE needs to have its evolutionary and demographic history documented. In this study, we provide phylogenetic analysis of all CRF01_AE pol sequences identified in mainland China. CRF01_AE sequences were collected from the Los Alamos HIV Sequence Database and the local Chinese provincial centers of disease control and prevention. Phylogenetic trees were constructed to identify major epidemic clusters. Bayesian coalescent-based method was used to reconstruct the time scale and demographic history. There were 2965 CRF01_AE sequences from 24 Chinese provinces that were collected, and 5 major epidemic clusters containing 85% of the total CRF01_AE sequences were identified. Every cluster contains sequences from more than 10 provinces with 1 or 2 dominant transmission routes. One cluster arose in the 1990s and 4 clusters arose in the 2000s. Cluster I is in the decline stage, while the other clusters are in the stable stage. Obvious lineage can be observed among sequences from the same transmission route but not the same area. Two large clusters in high-level prevalence were found in MSM (Men who have sex with men), which highlighted that more emphasis should be placed on MSM for HIV control in mainland China.


Assuntos
Infecções por HIV/epidemiologia , HIV-1/classificação , Análise de Sequência de RNA/métodos , Produtos do Gene pol do Vírus da Imunodeficiência Humana/genética , Teorema de Bayes , China/epidemiologia , Evolução Molecular , Infecções por HIV/virologia , HIV-1/genética , Humanos , Masculino , Epidemiologia Molecular , Filogenia , Filogeografia
5.
AIDS Res Hum Retroviruses ; 33(10): 1065-1069, 2017 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-28569547

RESUMO

CRF07_BC was originally formed in Yunnan province of China in 1980s and spread quickly in injecting drug users (IDUs). In recent years, it has been introduced into men who have sex with men (MSM) and become the most dominant strain in China. In this study, we performed a comprehensively phylodynamic analysis of CRF07_BC sequences from China. All CRF07_BC sequences identified in China were retrieved from database. More sequences obtained in our laboratory were added to make the dataset more representative. A maximum-likelihood (ML) tree was constructed with PhyML3.0. Maximum clade credibility (MCC) tree and effective population size were predicted by using Markov Chains Monte Carlo sampling method with Beast software. A total of 610 CRF07_BC sequences coving 1,473 bp of the gag gene (from 817 to 2,289 according to HXB2 calculator) were included into the dataset. Three epidemic clusters were identified; two clusters comprised sequences from IDUs, while one cluster mainly contained sequences from MSMs. The time of the most recent common ancestor of clusters that composed of sequences from MSMs was estimated to be in 2000. Two rapid spreading waves of effective population size of CRF07_BC infections were identified in the skyline plot. The second wave coincided with the expanding of MSM cluster. The results indicated that the control of CRF07_BC infections in MSMs would help to decrease its epidemic in China.


Assuntos
Evolução Molecular , Infecções por HIV/epidemiologia , HIV-1/genética , Minorias Sexuais e de Gênero , Sequência de Bases , China/epidemiologia , Genótipo , Infecções por HIV/virologia , HIV-1/classificação , Humanos , Masculino , Filogenia , RNA Viral/genética , Análise de Sequência de RNA
6.
AIDS Res Hum Retroviruses ; 33(6): 614-620, 2017 06.
Artigo em Inglês | MEDLINE | ID: mdl-28398773

RESUMO

Since the first identification of HIV-1 outbreak in Dehong, Yunnan province has been the epidemic center of HIV in China. Owing to the special geographic location and the frequent population mobility, Yunnan province contained complex HIV subtype distribution. Many new circulating recombinant forms (CRFs) and unique recombinant forms (URFs) have been found in recent years. In this study, a unique HIV-1 recombinant strain genome (YN10134) was characterized from an HIV-positive female in Yunnan, China. This virus genome had a complex intersubtype recombinant structure with eight breakpoints, composed of subtypes B and C. Although the sequence had a similar breakpoint with CRF07_BC in the start position in Env, the phylogenetic analysis showed that the segment was not originated from CRF07_BC. The identification of the URF indicated the severity of the HIV epidemic in Yunnan province and the urgent need for epidemiological surveillance of the new recombination.


Assuntos
Genótipo , Infecções por HIV/virologia , HIV-1/classificação , HIV-1/genética , Recombinação Genética , China , Feminino , HIV-1/isolamento & purificação , Humanos , Filogenia , Análise de Sequência de DNA
7.
AIDS Res Hum Retroviruses ; 33(1): 82-86, 2017 01.
Artigo em Inglês | MEDLINE | ID: mdl-27460636

RESUMO

Most HIV subtypes prevalent in China can be found in Shenzhen, including CRF07_BC, CRF01_AE, CRF08_BC, CRF55_01B, and subtype B. Multiple subtypes spreading in the same population always lead to the emergence of unique recombinant strains. Here, we report two unique recombinant forms (SZ44LS7251 and SZ95LS8027) of HIV-1 identified in a heterosexual population. Recombinant analyses were fulfilled based on the near full-length genomes. Both strains comprise subtypes B, C, and CRF01_AE. Phylogenetic analysis reveals that SZ44LS7251 is the second generation recombination originated from CRF55_01B andCRF07_BC, whereas SZ95LS8027 comprises CRF01_AE and CRF07_BC.The emergence of second generation recombination of HIV with complicated genomic structures supposed that high ratio of super infections or coinfections might happen in the Shenzhen area.


Assuntos
Genoma Viral , Infecções por HIV/virologia , HIV-1/classificação , HIV-1/genética , Recombinação Genética , Adulto , China , Análise por Conglomerados , Evolução Molecular , Feminino , Genótipo , HIV-1/isolamento & purificação , Heterossexualidade , Humanos , Masculino , Pessoa de Meia-Idade , Filogenia , Análise de Sequência de DNA
8.
BMC Infect Dis ; 16(1): 605, 2016 Oct 26.
Artigo em Inglês | MEDLINE | ID: mdl-27782811

RESUMO

BACKGROUND: The widespread use of antiretroviral therapies has led to considerable concerns about the prevalence of drug-resistant, as transmission of drug-resistant (TDR) strains poses a challenge for the control of the HIV-1 epidemic. METHODS: We conducted an epidemiological study enrolling treatment-naïve HIV-1-positive subjects at the Peking Union Medical College Hospital since 1991. Drug resistance was determined by submitting the sequences to the Stanford University Network HIV-1 database. RESULTS: Of 521 participants, 478 samples were amplified and sequenced successfully. HIV Transmitted drug resistance prevalence in China was determined to be 6.7 %. We did not find significant differences in the TDR rate by demographic characteristics. No significant time trend in the prevalence of overall TDR was observed (p > 0.05). CONCLUSIONS: We identified an intermediate prevalence of transmitted drug resistance (TDR), exhibiting a stable time trend. These findings enhance our understanding of HIV-1 drug resistance prevalence and time trend, and provide some guidelines for the comprehensive public health strategy of TDR prevention.


Assuntos
Fármacos Anti-HIV/uso terapêutico , Farmacorresistência Viral/efeitos dos fármacos , Infecções por HIV/virologia , HIV-1/efeitos dos fármacos , Adolescente , Adulto , Idoso , China/epidemiologia , Feminino , Infecções por HIV/epidemiologia , HIV-1/genética , Humanos , Masculino , Pessoa de Meia-Idade , Adulto Jovem
10.
PLoS One ; 11(3): e0149467, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26930645

RESUMO

OBJECTIVES: This study was designed to identify common HIV-1 mutation complexes affecting the slope of inhibition curve, and to propose a new parameter incorporating both the IC50 and the slope to evaluate phenotypic resistance. METHODS: Utilizing site-directed mutagenesis, we constructed 22 HIV-1 common mutation complexes. IC50 and slope of 10 representative approved drugs and a novel agent against these mutations were measured to determine the resistance phenotypes. The values of new parameter incorporating both the IC50 and the slope of the inhibition curve were calculated, and the correlations between parameters were assessed. RESULTS: Depending on the class of drug, there were intrinsic differences in how the resistance mutations affected the drug parameters. All of the mutations resulted in large increases in the IC50s of nucleoside reverse transcriptase inhibitors. The effects of the mutations on the slope were the most apparent when examining their effects on the inhibition of non-nucleoside reverse transcriptase inhibitors and protease inhibitors. For example, some mutations, such as V82A, had no effect on IC50, but reduced the slope. We proposed a new concept, termed IIPatoxic, on the basis of IC50, slope and the maximum limiting concentrations of the drug. The IIPatoxic values of 10 approved drugs and 1 novel agent were calculated, and were closely related to the IIPmax values (r > 0.95, p < 0.001). CONCLUSIONS: This study confirms that resistance mutations cannot be accurately assessed by IC50 alone, because it tends to underestimate the degree of resistance. The slope parameter is of very importance in the measurement of drug resistance and the effect can be applied to more complex patterns of resistance. This is the most apparent when testing the effects of the mutations on protease inhibitors activity. We also propose a new index, IIPatoxic, which incorporates both the IC50 and the slope. This new index could complement current IIP indices, thereby enabling predict the efficacy of pre-clinical drugs for which human pharmacokinetic is not available.


Assuntos
Fármacos Anti-HIV/farmacologia , Infecções por HIV/tratamento farmacológico , HIV-1/efeitos dos fármacos , HIV-1/genética , Mutação/efeitos dos fármacos , Linhagem Celular , Relação Dose-Resposta a Droga , Farmacorresistência Viral/efeitos dos fármacos , Infecções por HIV/virologia , Inibidores da Protease de HIV/farmacologia , Humanos , Concentração Inibidora 50 , Mutagênese Sítio-Dirigida , Inibidores da Transcriptase Reversa
11.
Curr HIV Res ; 14(2): 148-53, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26415701

RESUMO

INTRODUCTION: Multiple specific populations, including MSMs, IDUs, and FPDs, are involved in HIV epidemic in China. In the recent years, HIV transmission due to heterosexual transmission also contributed greatly to HIV epidemic in China. Very few studies have been fulfilled to characterize relationships of HIV-1 strains prevalent in different populations. In this study, the phylogenetic relationships of HIV-1 spreading in different populations were investigated. MATERIALS AND METHODS: HIV-1 sero-positive patients infected through different routes were enrolled into the study. Nested RT-PCR was used to amplify HIV gag and pol genes followed by sequencing. RESULTS: Multiple subtypes, including subtype B (52.1%), CRF01_AE (34.4%), CRF07_BC (6.3%), subtype C (4.2%), CRF02_AG (1.0%), CRF08_BC (1.0%) and unique recombination forms (1.0%) were identified. Phylogenetic analysis showed that strains from MSM, IDU, and FPDs grouped into clusters separately. However, strains identified in heterosexual transmitted population intermixed with all of other high risk populations. DISCUSSION AND CONCLUSION: The genetic data supposed that HIV-1 was spreading out of MSMs, IDUs, and FPDs through heterosexual transmission in Hebei, China. Urgent prevention and behavior intervention in the population will be necessary. Furthermore, the detailed sequence data will help the design of HIV-1 vaccines in China. Sequence Data: All of sequences have been deposited into the GenBank with the accession number: KJ820007-KJ820144.


Assuntos
Infecções por HIV/epidemiologia , Infecções por HIV/transmissão , HIV-1 , Heterossexualidade , Adolescente , Adulto , Idoso , Criança , Pré-Escolar , China/epidemiologia , Feminino , Genes gag/genética , Genes pol/genética , Genótipo , HIV-1/classificação , HIV-1/genética , HIV-1/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Epidemiologia Molecular , Filogenia , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Adulto Jovem
12.
BMC Infect Dis ; 15: 528, 2015 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-26572485

RESUMO

BACKGROUND: In this study, the prevalence of HIV-1 CRF01_AE intrasubtype recombinants in China is estimated and their contributions to the epidemic are explored. METHODS: Available HIV-1 complete genomes of CRF01_AE were retrieved from the HIV database. The two alignments were evaluated with RDP3. Recombinants were defined as cases in which the recombination signal was supported by at least 3 methods with P-values of ≤0.05 after Bonferroni correction for multiple comparisons implemented in RDP3. Phylogenetic analysis was performed to further investigate the role of intrasubtype recombinants in epidemics. RESULTS: Here, 124 out of the 339 sequences from around the world (36.6 %) showed significant evidence of recombination. Here, 84 of these recombinants were from China, accounting for 54.9 % of local total sequences (84 out of 153). The results indicated non-negligible levels of intrasubtype recombination. Subsequent phylogenetic analysis indicated that a considerable proportion of CRF01_AE strains in China originated from circulating intrasubtype recombinant forms. Three large, well-supported intrasubtype recombinants clusters were identified here. Through a survey of risk factors and sampling cities and provinces, cluster I and cluster II were found to be prevalent primarily among men who have sex with men in major northern cities. Cluster III was prevalent among heterosexuals and intravenous drug users in southern and southwestern provinces. CONCLUSIONS: The current work highlighted the remarkable prevalence of intrasubtype recombination within the CRF01_AE epidemic and emphasized the value of intrasubtype recombinants, which came to circulate in the same manner as intersubtype recombinants.


Assuntos
Infecções por HIV/virologia , HIV-1/genética , China/epidemiologia , Usuários de Drogas , Infecções por HIV/epidemiologia , HIV-1/patogenicidade , Heterossexualidade , Humanos , Masculino , Filogenia , Recombinação Genética
13.
Virol J ; 12: 187, 2015 Nov 14.
Artigo em Inglês | MEDLINE | ID: mdl-26578099

RESUMO

BACKGROUND: Highly active antiretroviral therapy (HAART) is recommended to control the infection of HIV-1. HIV-1 drug resistance becomes an obstacle to HAART due to the accumulation of specific mutations in the RT coding region. The development of resistance mutations may be more complex than previously thought. METHODS: We followed two HIV-1 infectors from a HIV-1 drug resistance surveillance cohort in Henan province and evaluated CD4+ T-cell number and viral load thereafter at ten time-periods and characterized their reverse transcriptase-associated mutation patterns at each time point. Then we constructed the recombinant virus strains with these mutation patterns to mimick the viruses and test the phenotypic resistance caused by the mutation patterns on TZM-b1 cells. RESULTS: CD4+ T-cell number initially increased and then decreased rapidly, while viral load decreased and then dropped sharply during initial antiretroviral treatment. The number of mutations and the combination patterns of mutations increased over time. According to the phenotypic resistance performed by recombinant virus strains, VirusT215Y/V179E/Y181C/H221Y exhibited high levels of resistance to EFV (5.57-fold), and T215Y/V179E-containing virus increased 20.20-fold in AZT resistance (p < 0.01). VirusT215Y/V179E/Y181C increased markedly in EFV resistance (p < 0.01). The IC50 for VirusT215Y/V179E/H221Y was similar to that for VirusT215Y/V179E/Y181C. VirusT215Y/K103N/Y181C/H221Y induced a dramatic IC50 increase of all the four agents (Efavirenz EFV, Zidovudine AZT, Lamivudine 3TC, and Stavudine d4T) (p < 0.01). As for VirusT215Y/K103N/Y181C, only the IC50 of EFV was significantly increased. T215Y/K103N resulted in a 26.36-fold increase in EFV (p < 0.01). T215Y/K103N/H221Y significantly increased the resistance to AZT and 3TC. The IC50 of EFV with T215Y/V179E was lower than with T215Y/K103N (F = 93.10, P < 0.0001). With T215Y/V179E, Y181C significantly increase in EFV resistance, while the interaction between 181 and 221 in EFV was not statistically significant (F = 1.20, P = 0.3052). With T215Y/K103N, neither H221Y nor Y181C showed a significant increase in EFV resistance, but the interaction between 181 and 221 was statistically significant (F = 38.12, P = 0.0003). CONCLUSIONS: Data in this study suggests that pathways of viral evolution toward drug resistance appear to proceed through distinct steps and at different rates. Phenotypic resistance using recombinant virus strains with different combination of mutation patterns reveals that interactions among mutations may provide information on the impact of these mutations on drug resistance. All the result provides reference to optimize clinical treatment schedule.


Assuntos
Fármacos Anti-HIV/farmacologia , Farmacorresistência Viral , Infecções por HIV/virologia , Transcriptase Reversa do HIV/genética , HIV-1/enzimologia , HIV-1/genética , Mutação de Sentido Incorreto , Adulto , Contagem de Linfócito CD4 , China , Evolução Molecular , HIV-1/efeitos dos fármacos , HIV-1/isolamento & purificação , Humanos , Estudos Longitudinais , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Carga Viral
15.
Int J Infect Dis ; 37: 86-92, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-26117343

RESUMO

OBJECTIVE: This study utilized genotypic and in-house recombinant virus phenotypic assays to examine HIV-1 variant susceptibility to antiretroviral (ARV) drugs; comparisons were made between the analyses. METHODS: A nested PCR was employed to amplify the HIV-1 gag-pol gene, which comprised the entire PR gene (codons 1-99) and the former RT gene (codons 1-312). Genetic resistance was determined by submitting the sequences to the Stanford University Network HIV-1 Database. Phenotypic susceptibilities to six ARV drugs were measured using a high-throughput, multi-cycle, recombinant virus phenotypic assay. Results were expressed in terms of the IC50 (half maximal inhibitory concentration) and fold-change values. The relationship between phenotypic drug resistance and genetic polymorphisms was determined. RESULTS: Nineteen fragment sequences for which recombinant viruses were successfully constructed were translated and compared with the consensus B sequences in the Stanford University Network HIV-1 Database. No recognizable genotypic resistance-associated mutations were noted, except in one sample. Each homologous replication-competent recombinant viral fold-change in the presence of six ARV drugs used widely in China was measured. According to the clinical and statistical criteria, 16 of the 19 samples were susceptible to the six drugs tested. The majority of phenotypic and genotypic results obtained were in agreement, with a concordance rate of 97.4%. Both phenotypic and genotypic assays suggested that sample HN2009001 was resistant to all drugs tested. All phenotypic and genotypic results obtained regarding the susceptibility of the 19 recombinant viruses to nucleoside reverse transcriptase inhibitors (NRTIs) were in agreement. With regard to the genotypic results for the non-nucleoside reverse transcriptase inhibitors (NNRTIs), 7.9% (3/38) were inconsistent with the phenotypic results. CONCLUSIONS: The in-house recombinant virus phenotypic assay was able to provide a straightforward quantitative assessment of resistance. In most cases, the genotypic and novel phenotypic assays yielded similar results. The disparity in HIV-1 susceptibility indicates a need to further investigate the clinical outcomes of antiretroviral therapy in certain individuals.


Assuntos
Fármacos Anti-HIV/farmacologia , HIV-1/efeitos dos fármacos , Síndrome de Imunodeficiência Adquirida/virologia , Farmacorresistência Viral , Feminino , Genótipo , HIV-1/genética , HIV-1/isolamento & purificação , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Mutação , Fenótipo , Polimorfismo Genético , Inibidores da Transcriptase Reversa/farmacologia
16.
PLoS One ; 10(5): e0127696, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26018591

RESUMO

BACKGROUND: The complex epidemic and significant diversity of HIV-1 strains in China pose serious challenges for surveillance and diagnostic assays, vaccine development and clinical management. There is a lack of HIV-1 isolates in current canonical HIV-1 subtype panels that can represent HIV-1 diversity in China; an HIV-1 subtype panel for China is urgently needed. METHODS: Blood samples were collected from HIV-1 infected patients participating in the drug-resistance surveillance program in China. The samples were isolated, cultured and stored as neat culture supernatant. The HIV-1 isolates were fully characterized. The panel was used to compare 2 viral load assays and 2 p24 assays as the examples of how this panel could be used. RESULTS: An HIV-1 subtype panel for China composed of 30 HIV-1 primary strains of four subtypes (B [including Thai-B], CRF01_AE, CRF07_BC and G) was established. The samples were isolated and cultured to a high-titer (10(6)-10(9) copies/ml)/high-volume (40 ml). The HIV-1 isolates were fully characterized by the final viral load, p24 concentration, gag-pol and envC2V3 sequencing, co-receptor prediction, determination of the four amino acids at the tip of the env V3-loop, glycosylation sites in the V3 loop and the drug-resistance mutations. The comparison of two p24 assays and two viral load assays on the isolates illustrated how this panel may be used for the evaluation of diagnostic assay performance. The Pearson value between p24 assays were 0.938. The viral load results showed excellent concordance and agreement for samples of Thai-B, but lower correlations for samples of CRF01_AE. CONCLUSION: The current panel of 30 HIV-1 isolates served as a basis for the development of a comprehensive panel of fully characterized viral isolates, which could reflect the current dynamic and complex HIV-1 epidemic in China. This panel will be available to support HIV-1 research, assay evaluation, vaccine and drug development.


Assuntos
Infecções por HIV/virologia , HIV-1/isolamento & purificação , China , Feminino , Infecções por HIV/sangue , Humanos , Masculino , Filogenia , Carga Viral/métodos
17.
AIDS Res Hum Retroviruses ; 31(5): 559-63, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25748656

RESUMO

Multiple subtypes were found to be epidemic in the Shenzhen men who have sex with men (MSM) population, which always predicts the emergence of a unique recombinant. In 2012, CRF55_01B was first reported, which later was proven to have originated in MSM in Shenzhen city. In this study, we reported a unique recombinant form (URF) of HIV-1 identified in a man who has had sex with men in Shenzhen city. The strain showed a genomic schematic map similar to CRF55_01B with subtype C segments inserted in the gag and pol genes. The full-length genome was amplified in two halves with 1-kb overlap regions. The PCR products were cloned and sequenced. A recombination detection program showed that two subtype C fragments and two subtype B fragments were inserted into the CRF01_AE backbone genome in the gag and pol regions. In the phylogenetic tree, the subtype C fragments clustered with CRF07_BC variants and the other segments grouped with CRF55_01B strains except for one segment that clustered with CRF01_AE. Similar breakpoints between our strain and CRF65_cpx were also observed. The data suggested that the URF strain might be the recombinant form of CRF55_01B, CRF01_AE, and CRF07_BC. This is the first report of a third generation of recombination of HIV-1 that originated from CRF55_01B in China. The identification of the URF indicated the severity of the HIV epidemic in Shenzhen MSM and the urgent need for epidemiological surveillance of the new recombination.


Assuntos
Infecções por HIV/virologia , HIV-1/classificação , HIV-1/genética , RNA Viral/genética , China/epidemiologia , Análise por Conglomerados , Ordem dos Genes , Genes Virais , Genoma Viral , Infecções por HIV/epidemiologia , HIV-1/isolamento & purificação , Homossexualidade Masculina , Humanos , Masculino , Dados de Sequência Molecular , Mutagênese Insercional , Filogenia , Recombinação Genética , Análise de Sequência de DNA , Homologia de Sequência do Ácido Nucleico , Adulto Jovem
18.
AIDS Res Hum Retroviruses ; 31(5): 479-87, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-25560398

RESUMO

The Liangshan prefecture in Sichuan province is an area in China severely affected by the HIV epidemic, with intravenous drug use (IDU) as the main risk factor. No reports on HIV subtypes prevalent in IDUs in Liangshan prefecture could be found. In this study, we have characterized the genotypes of HIV-1 in the IDU population in Liangshan prefecture and further determined the phylogenetic relationship of the CRF07_BC strains to HIV-1 sequences from the other regions of China, including Xinjiang and Yunnan provinces, to explore the pattern and possible diffusion pathway of HIV-1 in these regions. HIV-1-seropositive drug-naive IDUs identified in Liangshan prefecture, Sichuan province were enrolled in 2009. Full-length gag and pol genes were amplified by reverse transcription and nested PCR and then sequenced. All of the sequences were subtyped. Phylogenetic trees were constructed using the neighbor-joining and maximum likelihood methods. Divergence times were estimated using a Bayesian molecular clock approach. CRF07_BC was found to be the predominant strain in IDUs in Liangshan prefecture (95.5%). The CRF07_BC strains from Liangshan prefecture were found to be intermixed with those from Yunnan province in phylogenetic trees. The CRF07_BC sequences from Xinjiang province can be grouped into several clusters, suggesting that the expansion of the CRF07_BC epidemic in Xinjiang province was the result of a local epidemic driven by multiple independent introductions in the late 1990s. Only low-level drug-resistant viruses were found in the IDU population. CRF07_BC strains from Liangshan prefecture were more similar to those from Yunnan province than those from Xinjiang province. This finding will contribute to our understanding of the distribution, the evolution, and the potential source of CRF07_BC founder strains, and will also provide useful information for the development of strategies to prevent transmission.


Assuntos
Genótipo , Infecções por HIV/epidemiologia , Infecções por HIV/virologia , HIV-1/classificação , HIV-1/genética , Abuso de Substâncias por Via Intravenosa/complicações , Adolescente , Adulto , Criança , Pré-Escolar , China/epidemiologia , Análise por Conglomerados , Transmissão de Doença Infecciosa , Usuários de Drogas , Feminino , Infecções por HIV/transmissão , HIV-1/isolamento & purificação , Humanos , Masculino , Pessoa de Meia-Idade , Epidemiologia Molecular , Dados de Sequência Molecular , Filogenia , Reação em Cadeia da Polimerase , Análise de Sequência de DNA , Homologia de Sequência , Adulto Jovem , Produtos do Gene gag do Vírus da Imunodeficiência Humana/genética , Produtos do Gene pol do Vírus da Imunodeficiência Humana/genética
19.
Zhonghua Liu Xing Bing Xue Za Zhi ; 36(9): 988-93, 2015 Sep.
Artigo em Chinês | MEDLINE | ID: mdl-26814868

RESUMO

OBJECTIVE: To analyze genetic characteristics of HIV isolated from men who have sex with men (MSM) in Beijing and predict the epidemic trend in this population. METHODS: All of the HIV gene sequences in our laboratory obtained from MSM in Beijing were used, which were aligned with all of the HIV gene sequences from MSM and other populations in China downloaded from Los Alamos HIV Database. Phylogenetic trees were constructed by using software PhyML 3.0, based on which the relationships of prevalent HIV strains between Beijing MSM and other populations in China were further explored. The evolution rate, the time of most recent common ancestor (tMRCA) , the epidemic parameters, the reproductive number (R0) were calculated by using software BEAST to predict HIV evolution and epidemic characteristics. RESULTS: Multiple HIV subtypes, including subtype B, CRF01_AE and CRF07_BC, were found to be prevalent among MSM in Beijing. In ML tree constructed based on strains from the whole country, three clusters including B-1, CRF01_AE-1, and CRF01_AE-2 were found among the MSM in Beijing (accounting for 40%) . At least three independent introduction of B1 cluster strains into Beijing MSM were found, which were at March 1991 (July 1984-February 1997) , January 1994 (January 1989-January 1998) , April 1991 (August 1984-January 1996) . For CRF01_AE strains, two clusters including CRF01_AE-1 and CRF01_AE-2 were introduced into the population at December 2000 (March 1998-January 2003) and December 2001 (January 2000-July 2003) respectively. The population epidemiology of HIV in Beijing MSM was reconstructed based on sequences. The CRF01_AE-1 cluster spread more quickly than the other two clusters, and the evolution rate was higher. CONCLUSION: Multiple HIV subtypes were found prevalent among MSM in Beijing. Although subtype B strain was introduced into Beijing MSM earlier than CRF01_AE strain, CRF01_AE strain increased more quickly than subtype B strain. More research and control of the CRF01_AE prevalence will be helpful for prevention and control of HIV epidemic in MSM in Beijing.


Assuntos
Infecções por HIV/diagnóstico , Infecções por HIV/virologia , HIV-1/genética , Homossexualidade Masculina/estatística & dados numéricos , China/epidemiologia , Epidemias , Infecções por HIV/epidemiologia , HIV-1/isolamento & purificação , Humanos , Masculino , Epidemiologia Molecular , Filogenia , Prevalência
20.
PLoS One ; 9(9): e107349, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25203725

RESUMO

BACKGROUND: Recombination is one of the major mechanisms underlying the generation of HIV-1 variability. Currently 61 circulating recombinant forms of HIV-1 have been identified. With the development of recombination detection techniques and accumulation of HIV-1 reference stains, more accurate mosaic structures of circulating recombinant forms (CRFs), like CRF04 and CRF06, have undergone repeated analysis and upgrades. Such revisions may also be necessary for other CRFs. Unlike previous studies, whose results are based primarily on a single recombination detection program, the current study was based on multiple recombination analysis, which may have produced more impartial results. METHODS: Representative references of 3 categories of intersubtype recombinants were selected, including BC recombinants (CRF07 and CRF08), BG recombinants (CRF23 and CRF24), and BF recombinants (CRF38 and CRF44). They were reanalyzed in detail using both the jumping profile hidden Markov model and RDP3. RESULTS: The results indicate that revisions and upgrades are very necessary and the entire re-analysis suggested 2 types of revision: (i) length of inserted fragments; and (ii) number of inserted fragments. The reanalysis also indicated that determination of small regions of about 200 bases or fewer should be performed with more caution. CONCLUSION: Results indicated that the involvement of multiple recombination detection programs is very necessary. Additionally, results suggested two major challenges, one involving the difficulty of accurately determining the locations of breakpoints and the second involving identification of small regions of about 200 bases or fewer with greater caution. Both indicate the complexity of HIV-1 recombination. The resolution would depend critically on development of a recombination analysis algorithm, accumulation of HIV-1 stains, and a higher sequencing quality. With the changes in recombination pattern, phylogenetic relationships of some CRFs may also change. All these results may be critical to understand the role of recombination in a complex and dynamic HIV evolution.


Assuntos
HIV-1/genética , Recombinação Genética/genética , Genoma Viral/genética , Dados de Sequência Molecular , Filogenia , Análise de Sequência de DNA/métodos
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