RESUMO
We compared the effectiveness of promoting bone healing between two teriparatide preparations for atypical femoral fracture (AFF). A total of 45 AFFs were included in this study, and we compared the duration of bone union. Teriparatide administered by daily injection enhanced bone union more than weekly administration in complete AFFs. INTRODUCTION: The efficacy of teriparatide for atypical femoral fracture (AFF) has been recently reported. Although two different teriparatide preparations can be used to treat osteoporosis in Japan, daily or weekly injection, all previous reports on the effectiveness of teriparatide for AFF only examined daily injection formulations. Therefore, we compared the promotion of bone healing between the two teriparatide preparations for AFF. METHODS: A total of 45 consecutive AFFs in 43 Japanese patients were included in this study. They received either a daily 20-µg teriparatide injection (daily group; n = 32) or a once-a-week 56.5-µg teriparatide injection (weekly group; n = 13). We compared the clinical background and duration of bone union between these two groups. RESULTS: When all patents were included, the fracture healing time was not significantly different between the two groups. Only patients with complete AFFs had significantly fewer daily bisphosphonate or denosumab injections than the weekly group (P < 0.05). The fracture healing time in the daily group (6.1 ± 4.1 months) was significantly shorter than that in the weekly group (10.1 ± 4.2 months) (P < 0.05). Even if the influence of bisphosphonate or denosumab usage was excluded, a similar significant difference was observed in the fracture healing time (P < 0.05). There was no significant difference between the two groups among patients with incomplete AFFs. CONCLUSIONS: Daily teriparatide injections enhance bone union more than weekly injections in complete AFF patients.
Assuntos
Conservadores da Densidade Óssea/administração & dosagem , Fraturas do Fêmur/tratamento farmacológico , Consolidação da Fratura/efeitos dos fármacos , Fraturas por Osteoporose/tratamento farmacológico , Teriparatida/administração & dosagem , Idoso , Idoso de 80 Anos ou mais , Conservadores da Densidade Óssea/uso terapêutico , Terapia Combinada , Esquema de Medicação , Feminino , Fraturas do Fêmur/fisiopatologia , Fraturas do Fêmur/cirurgia , Fixação Interna de Fraturas/métodos , Humanos , Injeções Subcutâneas , Masculino , Pessoa de Meia-Idade , Fraturas por Osteoporose/fisiopatologia , Fraturas por Osteoporose/cirurgia , Estudos Retrospectivos , Teriparatida/uso terapêuticoRESUMO
The ACTN3 R577X genotype has been found to associate with sprint/power phenotypes in all elite athlete cohorts investigated. This association has not been extensively studied in elite Asian athletes. The present study was undertaken to investigate the association between the ACTN3 R577X genotype and elite Japanese track and field athlete status. 299 elite Japanese track and field athletes (134 sprint/power athletes; 165 endurance/middle-power athletes) and 649 Japanese controls were genotyped for the ACTN3 R577X polymorphism. All athletes were of national or international level. Sprint/power athletes showed a higher frequency of RR + RX genotype than controls (111/134 [82.8%] vs. 478/649 [73.7%], P = 0.025 under the R-dominant model), while there was no significant difference between endurance/middle-power athletes and controls (126/165 [76.4%] vs. 478/649 [73.7%], P = 0.48 under the R-dominant model). Sprinters with the RR + RX genotype had significantly faster personal best times for the 100 m than those with XX genotype (10.42 ± 0.05 s vs. 10.64 ± 0.09 s, P = 0.042); no such association was found in the 400 m sprinters (47.02 ± 0.36 s vs. 47.56 ± 0.99 s, P = 0.62). ACTN3 R577X genotype is associated with sprint/power performance in elite Japanese track and field athletes, especially short sprint performance.
Assuntos
Actinina/genética , Povo Asiático/genética , Desempenho Atlético , Corrida , Atletismo , Feminino , Genótipo , Humanos , Japão , Masculino , CaminhadaRESUMO
Avian sarcoma virus 1 (ASV-1) was isolated from a spontaneous sarcoma of an adult chicken. It is a replication-defective virus that induces fibrosarcomas in young chickens and oncogenic transformation in cultured chick embryo fibroblasts. The ASV-1 genome has been cloned in the lambda gt WES.lambda B vector from closed circular viral DNA extracted from infected cells. The nucleotide sequence of the oncogene insert in the ASV-1 genome has been determined. It is virtually identical to the sequence of the oncogene crk recently discovered in the avian sarcoma virus CT10. ASV-1 and CT10 are two independent retrovirus isolates carrying the crk oncogene.
Assuntos
Vírus do Sarcoma Aviário/genética , Oncogenes , Sequência de Aminoácidos , Animais , Vírus do Sarcoma Aviário/isolamento & purificação , Sequência de Bases , Transformação Celular Neoplásica , Células Cultivadas , Embrião de Galinha , Galinhas , Clonagem Molecular , DNA Circular/genética , Genes Virais , Vetores Genéticos , Dados de Sequência Molecular , Proteína Oncogênica v-crk , Mapeamento por Restrição , Proteínas Oncogênicas de Retroviridae/genética , Homologia de Sequência do Ácido NucleicoRESUMO
An HIV-1 strain carrying a shorter form of the transmembrane glycoprotein (TM) with a mobility of 32 kD, named KB-1, was isolated from a Japanese male hemophiliac by coculture of his peripheral blood mononuclear cells (PBMCs) with MT-2 cells and adaption to TALL-1 cells. Another HIV-1 strain, named KB-2, was isolated from his seropositive spouse by coculture of her PBMCs with MT-2 cells. The KB-2 strain carried a TM of ordinary size, with a mobility of 41 kD. The KB-1 strain carrying a truncated form of the TM could replicate in MT-2, MT-4, TALL-1 and MOLT-4 cells. The KB-1 strain is a useful HIV-1 isolate for investigating the function of the cytoplasmic domain of the TM and the significance of the presence of an in-frame stop codon in HIV env gene.
Assuntos
Glicoproteínas/análise , Proteína gp41 do Envelope de HIV/análise , HIV-1/metabolismo , Proteínas do Envelope Viral/análise , Western Blotting , Feminino , HIV-1/isolamento & purificação , Hemofilia A/complicações , Hemofilia A/microbiologia , Humanos , Masculino , Peso MolecularRESUMO
The effect of 11 derivatives of 1-deoxynojirimycin (DNM) on the replication of HIV-1 was studied. Compared with DNM, seven of them showed remarkable inhibition of HIV-1-induced syncytium formation at significantly low concentrations which were not cytotoxic. Two derivatives were found to markedly reduce the infectious virus yields from cell lines chronically infected with HIV. Analysis of HIV-1 envelope glycoproteins showed that the derivatives induced modification of the processing of not only gp120/160 but also the transmembrane glycoprotein gp41. The modification of the processing of the transmembrane glycoprotein gp41 might play an important role in the inhibition of virus replication at a step after the binding of gp120 to CD4. The enhanced anti-HIV activity of DNM derivatives reported here could increase the possibility of non-toxic therapeutic intervention in HIV infections.
Assuntos
Glucosamina/análogos & derivados , HIV-1/efeitos dos fármacos , 1-Desoxinojirimicina , Western Blotting , Linhagem Celular , Células Gigantes/efeitos dos fármacos , Glucosamina/metabolismo , Glucosamina/farmacologia , Proteína gp120 do Envelope de HIV/metabolismo , Proteína gp41 do Envelope de HIV/metabolismo , HIV-1/fisiologia , Humanos , Processamento de Proteína Pós-Traducional/efeitos dos fármacos , Replicação Viral/efeitos dos fármacosRESUMO
OBJECTIVES: To determine the genetic variability of HIV-1 amongst infected Filipinos and to analyze phylogenetic relationships, temporal introductions and transmission dynamics of identified variants. METHODS: Polymerase chain reaction amplification and direct sequencing of a 204 base-pair fragment of the env C2-V3 region from uncultured peripheral blood mononuclear cells obtained from 51 HIV-1-positive Filipinos infected from 1987 to mid-1996. Evolutionary distance and phylogenetic relationships among the DNA sequences were estimated. RESULTS: The 51 Philippine strains were classified into five env V3 subtypes, namely subtype B (n = 37), subtype E (n = 8), subtype A (n = 3), subtype C (n = 2) and subtype D (n = 1). The overall env nucleotide divergence ranged from 11.7 to 32.2%. The nucleotide variation appeared to be random and no temporal ordering was observed. The variation of the sequences at the tip of the V3 loop was very broad. Subtypes B and C isolates did not show close genetic relationship to other Asian variants. Only three of the subtype E strains had close affinity to known Asian sequences. The majority (94%) of the subjects acquired the infection by sexual transmission. About two-thirds were presumably infected outside the Philippines, whereas the remaining were infected indigenously. Information was limited to allow segregation of the identified subtypes by mode of transmission or risk groups. CONCLUSION: Our findings demonstrate the presence of multiple genetic subtypes of HIV-1 in the Philippines. The apparent geographic range of previously reported genotypes in South and South-east Asia was extended and has obvious implications for env-based antiviral interventions.
Assuntos
DNA Viral/genética , Genoma Viral , Infecções por HIV/virologia , HIV-1/genética , Adulto , Sequência de Aminoácidos , Criança , DNA Viral/análise , Feminino , Infecções por HIV/epidemiologia , HIV-1/isolamento & purificação , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Filipinas/epidemiologia , Filogenia , Reação em Cadeia da Polimerase , Alinhamento de Sequência , Análise de Sequência de DNARESUMO
The nef gene is considered to play a crucial role in the development of acquired immunodeficiency syndrome (AIDS). In this study, we analyzed the sequence of nef quasispecies obtained from replication-competent HIV-1 isolates from two Japanese hemophiliac patients infected with HIV-1. At least 10 nef clones were isolated at each time point and a total of 75 individual nef quasispecies were sequenced. We observed a gradual increase in genetic diversity of the nef gene over time. Among the various functional regions of Nef protein, myristoylation site and the central PXXP (SH3 ligand) motifs were well conserved. The scattered regions responsible for downregulation of CD4 and class I MHC were also conserved. These data suggest that these functions of Nef may be involved throughout the disease process.
Assuntos
Produtos do Gene nef/genética , Infecções por HIV/complicações , HIV-1/genética , Hemofilia A/complicações , Sequência de Aminoácidos , Antígenos CD4/metabolismo , Sequência Conservada , Progressão da Doença , Regulação para Baixo , Evolução Molecular , Produtos do Gene nef/química , Produtos do Gene nef/classificação , Produtos do Gene nef/metabolismo , Variação Genética , Infecções por HIV/virologia , HIV-1/metabolismo , Hemofilia A/metabolismo , Hemofilia A/virologia , Antígenos de Histocompatibilidade Classe I/metabolismo , Humanos , Dados de Sequência Molecular , Peptídeos/química , Peptídeos/genética , Filogenia , Domínios Proteicos Ricos em Prolina , Conformação Proteica , Análise de Sequência , Homologia de Sequência de Aminoácidos , Fatores de Tempo , Produtos do Gene nef do Vírus da Imunodeficiência HumanaRESUMO
The myeloid-monocytic cells ML-1, HL-60, THP-1, and U-937 were chronically infected (for > 2 years) with the lymphotropic human immunodeficiency virus type 1 (HIV-1) strain HTLV-IIIB. Reinfection experiments revealed that viruses obtained from chronically infected ML-1/HIV-1 and HL-60/HIV-1 cells showed a low infectivity if tested with uninfected ML-1 and HL-60 cells in contrast to virus preparations from chronically infected THP-1/HIV-1 and U-937/HIV-1 with their corresponding uninfected cell lines. Analyses of selected cell surface markers revealed a differential expression of CD4, CD8, CD11c, CD14, CD15, CD20, HLA-DR, and HLA-DQ in non- or chronically infected cells. In chronically infected cells, the steady-state levels for tumor necrosis factor-alpha, interleukin (IL)-1 beta, and granulocyte-macrophage colony-stimulating factor mRNA remained unchanged whereas the one for IL-6 dropped.
Assuntos
Antígenos CD/análise , HIV-1/fisiologia , Fator Estimulador de Colônias de Granulócitos e Macrófagos/genética , HIV-1/genética , Antígenos HLA-DQ/análise , Antígenos HLA-DR/análise , Humanos , Interleucina-1/genética , Interleucina-6/genética , Leucemia Promielocítica Aguda , Monócitos , RNA Mensageiro/análise , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Viral/genética , RNA Viral/isolamento & purificação , Células Tumorais Cultivadas , Fator de Necrose Tumoral alfa/genéticaRESUMO
Previously it has been reported that cocultivation of human immunodeficiency virus type 1 (HIV-1)-infected cells with uninfected cells results in formation of multinuclear giant cells, generated via an interaction of gp120 on the surface of infected cells with CD4 on the uninfected cells. Formation of multinuclear giant cells as occurring in the presence of normal fetal calf serum was not observed when HIV-infected MOLT-4 or MOLT-3 cells (chronically infected with HTLV-IIIB) and uninfected cells were cocultured in both serum-free medium and fibrinogen-depleted serum. Addition of sera (human and rabbit) as well as of fibrinogen (human and bovine), fibronectin (human), and alpha-globulin (human), but not of albumin, transferrin or gamma-globulin to serum-free medium caused formation of multinuclear giant cells. In contrast, HIV production from MOLT-3 cells proceeds also in the absence of serum. In control experiments it was established that the cells maintained at reduced serum concentration, or in serum-free medium without or with fibrinogen are viable even though displaying a lower metabolic rate (ATP formation and DNA synthesis). From these findings we conclude that serum components (e.g., fibrinogen, fibronectin, and alpha-globulin) are absolutely required for syncytium formation but are not essential for virus release.
Assuntos
Fenômenos Fisiológicos Sanguíneos , Células Gigantes/microbiologia , HIV-1/fisiologia , Linfócitos T/microbiologia , Trifosfato de Adenosina/metabolismo , Sequência de Aminoácidos , Divisão Celular , Linhagem Celular , Humanos , Dados de Sequência Molecular , Linfócitos T/patologia , Replicação ViralRESUMO
Two hundred forty nucleotides from the pre-membrane gene region of 12 Japanese encephalitis virus (JEV) strains isolated from three different regions of Malaysia from 1993 to 1994 were sequenced and compared with each other and with the JEV strains from different geographic areas in Asia. These 12 Malaysian isolates were classified into two genotypes. The four JEV strains isolated from Sarawak in 1994 and the four JEV strains isolated from Sepang, Selangor in 1993 were classified into one genotype that included earlier isolated strains from Malaysia (JE-827 from Sarawak in 1968 and WTP/70/22 from Kuala Lumpur in 1970). The four JEV strains from Ipoh, Perak in 1994 were classified into another genotype that included JEV strains isolated from northern Thailand and Cambodia. In an earlier report, 10 JEV strains from Sabak Bernam, Selangor in 1992 were classified into the largest genotype that included strains isolated in temperate regions such as Japan, China, and Taiwan. The data indicate that at least three genotypes of JEV have been circulating in Malaysia.
Assuntos
DNA Viral/química , Vírus da Encefalite Japonesa (Espécie)/genética , RNA Viral/genética , Sequência de Aminoácidos , Animais , Sequência de Bases , Culicidae/virologia , Primers do DNA/química , Vírus da Encefalite Japonesa (Espécie)/classificação , Genótipo , Humanos , Insetos Vetores/virologia , Malásia , Dados de Sequência Molecular , Análise de Sequência de DNA , SuínosRESUMO
Human anti-HIV antibody-coated magnetic beads and magnetic particle concentrators were used to eliminate HIV antigens and the infected cells in vitro. Fifty micrograms/ml of the antibody coated on 30 mg/ml of tosyl-activated beads, and 10 min of reaction time between the antigens and the immunobeads were sufficient to eliminate the virus antigens from phosphate buffered saline, serum and peripheral blood. HIV-infected cells were eliminated in vitro, but not completely. This method will be useful in eliminating viral antigens and infected cells from clinical material such as blood and blood products.
Assuntos
Separação Celular/métodos , Antígenos HIV/isolamento & purificação , HIV/isolamento & purificação , HIV/ultraestrutura , Antígenos HIV/sangue , Humanos , Técnicas Imunoenzimáticas , Magnetismo , MicroesferasRESUMO
Immunofluorescence assays (IFA) that simultaneously distinguish between antibodies against closely related human immunodeficiency virus type 1 (HIV-1) and type 2 (HIV-2) infections have not been readily available. Serum specimens from 95 HIV-1-infected, 26 HIV-2-infected and 3 HIV-1/HIV-2 dually infected individuals and 106 seronegative blood donors were evaluated for the ability to serologically discriminate HIV-1 and HIV-2 infections by means of IFA employing three types of cells whose morphology varied within one field of microscopy. Mixtures of HIV-1-infected, HIV-2-infected and uninfected cells were used in the present study. In consequence, all serum specimens from individuals infected with HIV were confirmed to contain antibodies to HIV-1 and/or HIV-2. None of the sera from the blood donors were positive. Serum specimens from HIV-1-infected or HIV-2-infected individuals were diagnosed as single infection with HIV-1 (85/95) and HIV-2 (22/26), respectively, by this new assay. Although another 14 (10/95 and 4/26) were shown to be seropositive for both HIV-1-infected and HIV-2-infected cells, these results suggest that this assay is potentially simple and useful for screening and confirming both HIV-1 and HIV-2 infections simultaneously.
Assuntos
Imunofluorescência , Anticorpos Anti-HIV/sangue , Soropositividade para HIV/diagnóstico , HIV-1/imunologia , HIV-2/imunologia , Linhagem Celular , Reações Cruzadas , Soronegatividade para HIV/imunologia , Soropositividade para HIV/imunologia , Soropositividade para HIV/virologia , Células HeLa , Humanos , Valor Preditivo dos TestesRESUMO
An indirect immunofluorescence method was developed for the assay of antibodies to lymphadenopathy-associated virus (LAV) which is known to be associated with the aetiology of the acquired immunodeficiency syndrome (AIDS). Samples of serum or plasma from 1353 healthy volunteer blood donors, 53 homosexual males, 29 patients who had received multiple blood transfusions and 163 haemophiliacs in Japan were tested for antibody to LAV. Results showed that 47 (29%) of the haemophiliacs, who had been treated largely with factor VIII or IX produced in the USA, were anti-LAV antibody positive, whereas all other subjects were anti-LAV antibody negative. The incidence of antibodies to Adult T-cell leukaemia virus or Human T-cell leukaemia virus I (ATLV or HTLV-I) in these subjects was high.
Assuntos
Síndrome da Imunodeficiência Adquirida/epidemiologia , Síndrome da Imunodeficiência Adquirida/imunologia , Síndrome da Imunodeficiência Adquirida/microbiologia , Anticorpos Antivirais/análise , Doadores de Sangue , Transfusão de Sangue , Deltaretrovirus/imunologia , Métodos Epidemiológicos , Imunofluorescência , Hemofilia A/imunologia , Homossexualidade , Humanos , Japão , Retroviridae/imunologiaRESUMO
To analyse the appearance of AZT-resistant HIV in HIV carriers after AZT treatment and compare the mutations responsible for resistance employing cloned HIV DNA derived from provirus and free virions in plasma, serial blood specimens were taken before and after AZT treatment. RNA in virions in plasma, proviral DNA and RNA from virus isolates by coculture of PBMCs of HIV carriers and healthy blood donors were cloned and sequenced. DNA clones were compared for their nucleotide sequences responsible for AZT resistance. AZT resistance was acquired as early as 2 months after the start of the treatment and follow-up study was performed for 16 months of the treatment. Population of DNA clones was different according to the origin of the DNA or RNA, which indicated that the provirus population in PBMC was different from that in virions in plasma. These data demonstrated the possibility of selective activation of provirus or activation of provirus in organs other than peripheral blood, although the number of cases was small.
Assuntos
Fármacos Anti-HIV/uso terapêutico , Portador Sadio , Infecções por HIV/virologia , HIV-1/efeitos dos fármacos , Zidovudina/uso terapêutico , DNA Viral/análise , Resistência Microbiana a Medicamentos , Seguimentos , Infecções por HIV/tratamento farmacológico , HIV-1/genética , Humanos , RNA Viral/análiseRESUMO
We investigated the prevalence of human immunodeficiency viruses-1 and 2 (HIV-1 and HIV-2), human T-lymphotropic virus type I and II, hepatitis B virus (HBV), hepatitis C virus (HCV) and hepatitis D virus among intravenous drug users (IVDU) in Hiroshima, Japan, where little is known about their present levels. From June to December 1993, serum samples were collected from 47 IVDU and 98 alcoholics in Hiroshima, Japan, and examined for markers of virus infection. The prevalence of antibody to HCV (anti-HCV) and/or HCV-RNA was significantly higher in IVDU than alcoholics (74.5% vs 20.4%, 44.7% vs 10.2% respectively, P < 0.001). In contrast, the prevalence of antibody to hepatitis B surface antigen and/or core antigen (anti-HBs and/or anti-HBc) showed no significant difference between the 2 groups (57.4% vs 66.3%). HIV-1 infection was found in one (2.1%) IVDU and genome analysis indicated that it was subtype B according to Myers' classification. Thus, an extremely low level of HIV infection and a high level of HCV infection was found in IVDU. Careful follow-up of this group is thought to be needed to minimize an outbreak of HIV-1 infection in Japan.
Assuntos
Alcoolismo/complicações , Infecções por Deltaretrovirus/complicações , Infecções por HIV/complicações , Hepatite Viral Humana/complicações , Abuso de Substâncias por Via Intravenosa/complicações , Adulto , Alcoolismo/virologia , Sequência de Aminoácidos , Western Blotting , DNA Viral/análise , Deltaretrovirus/imunologia , Anticorpos Antideltaretrovirus/análise , Feminino , HIV/genética , Anticorpos Anti-HIV/análise , Anticorpos Anti-Hepatite C/análise , Humanos , Japão , Masculino , Pessoa de Meia-Idade , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Prevalência , RNA Viral/análiseRESUMO
HIV spread in South and South-East Asia is most alarming, and genetic variability of HIV-1 is an important consideration in vaccine development. In this study, we examined the third variable (V3) region of env gene of HIV-1 variants prevalent in Thailand, Malaysia, India, and the Philippines. By phylogenetic tree analyses, an HIV-1 variant from an injecting drug user (IDU) in Thailand belonged to subtype B, and HIV-1 variants from 2 IDUs in Malaysia were classified into 2 subtypes, B and E. One HIV-1 variant from a male homosexual in the Philippines belonged to subtype B. Out of 8 HIV-1 variants from sexually transmitted disease patients in India, 7 belonged to subtype C, and one to subtype A. Although the total number of individuals examined in this study was limited, 4 HIV-1 subtypes were found in South and South-East Asia and large international movements of HIV-1-infected individuals in this region could induce global dissemination of these HIV-1 variants.
Assuntos
Variação Genética , Infecções por HIV/virologia , HIV-1/genética , Sequência de Aminoácidos , Feminino , Genes Virais/genética , Infecções por HIV/epidemiologia , HIV-1/classificação , Homossexualidade Masculina , Humanos , Índia/epidemiologia , Malásia/epidemiologia , Masculino , Dados de Sequência Molecular , Filipinas/epidemiologia , Filogenia , Homologia de Sequência do Ácido Nucleico , Trabalho Sexual , Tailândia/epidemiologiaRESUMO
In this study, we examined the difference in susceptibility to anti-HIV activity of the CC-chemokines (RANTES, MIP-1 alpha and MIP-1 beta) among HIV-1 isolates and analysed its relation with phenotype (syncytium inducibility) and V3 domain of gp120 of the HIV-1 isolates. Of 11 cases tested in endogenous assay, at a concentration of 200 ng/ml, RANTES, MIP-1 alpha, and MIP-1 beta showed more than 80% suppression of HIV-1 replication in 10, 8, and 7 cases, respectively. HIV-1 isolates sensitive to more than one CC-chemokine showed non-syncytium-inducing phenotype, whereas HIV-1 isolates resistant to all of the 3 CC-chemokines showed syncytium-inducing phenotype. HIV-1 isolates resistant to all of the 3 CC-chemokines contained more positively charged amino acid residues in the V3 domain of the gp120. These results indicated that utilization of the CC-chemokine receptors as co-receptors for virus entry could vary among HIV-1 isolates.
Assuntos
Quimiocinas/farmacologia , HIV-1/efeitos dos fármacos , Linfócitos T CD4-Positivos/virologia , Quimiocina CCL4 , Quimiocina CCL5/farmacologia , HIV-1/isolamento & purificação , Humanos , Proteínas Inflamatórias de Macrófagos/farmacologia , Replicação Viral/efeitos dos fármacosRESUMO
The discordant cases of seronegative, but culture and proviral HIV-2 DNA positive were found in Mumbai, India. This was corroborated by the successful isolation of HIV-2-RNA in culture medium, HIV-2 cDNA sequence determination and the detection of the antigen. The sequence of the isolated HIV-2 genomic RNA does not seem to be altered to the extent that the change will alter antibody binding. Furthermore, antibody from the same individual (even at 8 months from initial sampling) from whom HIV-2 was isolated did not react with the antigen of this strain. Those evidences imply that extremely low or non-production of the antibody may be due to suboptimal immune stimulation due to extremely slow HIV-2 replication. This low virus-load may be responsible for the negative antibody results in the HIV-2 carriers.
PIP: This paper describes the characteristics of HIV-2 seropositive and seronegative cases in Mumbai, India, and characterizes the differences between HIV-1 and HIV-2. More than 200 outpatients considered to be at high risk of HIV infection were screened for HIV-1 and HIV-2 antibody and proviral DNA. The study found 11 cases that were discordant for antibody test and HIV proviral DNA (i.e., negative for anti-HIV but positive for HIV-2 proviral DNA). The presence of this provirus was further corroborated by the detection of HIV-2 RNA in the culture medium upon HIV isolation, HIV-2 cDNA sequencing, and antigen detection. The sequence of the isolated HIV-2 genomic RNA did not seem to be altered to the extent that the change would affect antibody binding. Moreover, antibody from the same person in whom HIV-2 was detected did not react with the antigen of this strain even 8 months after the initial sampling. These findings indicate that extremely low production or non-production of the antibody may be brought about by suboptimal immune stimulation due to very low HIV-2 replication speed. This low virus load may account for the negative antibody results in the HIV-2 carriers in India.
Assuntos
Portador Sadio/veterinária , Soronegatividade para HIV , HIV-2 , Linhagem Celular , DNA Viral/isolamento & purificação , Técnica Indireta de Fluorescência para Anticorpo , Soropositividade para HIV/virologia , Humanos , Índia/epidemiologia , Reação em Cadeia da Polimerase , Linfócitos T/virologia , Carga ViralRESUMO
The inhibitory effect of CD8+ T-cells from HIV-infected or HIV-seronegative individuals on HIV replication in the naturally-infected CD4+ T-cells in vitro was examined. Not only autologous CD8+ T-cells from HIV-infected individuals but also allogeneic CD8+ T-cells from HIV-seronegative individuals prevented or delayed HIV replication, even in transwell cocultures using a semi-permeable 0.45 micron filter. The level of the inhibitory effect of allogeneic CD8+ T-cells from the HIV-seronegative individuals on the HIV replication was varied among CD4+ T-cells obtained from HIV-infected individuals used. The results suggested that CD8+ T-cells from HIV-seronegative individuals as well as HIV-infected individuals could produce some cytokine(s) which suppress HIV replication in vitro. The sensitivity to the cytokine(s) might be variable among HIV strains, depending on differences in the nucleotide sequence of different HIV-1 strains. Further studies of control of HIV replication by CD8+ anti-HIV cytokine(s) should provide new strategies for the therapy of HIV infection.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Infecções por HIV/imunologia , Soronegatividade para HIV/imunologia , HIV-1/imunologia , Linfócitos T CD8-Positivos/citologia , Células Cultivadas , Proteína do Núcleo p24 do HIV/análise , Infecções por HIV/sangue , HIV-1/fisiologia , Humanos , Replicação ViralRESUMO
Isolation of Japanese encephalitis virus (JEV) from mosquitoes in Sabak Bernam, Selangor, Malaysia, was attempted. An aliquot of homogenate from each pool of mosquitoes, 50 per tube, was inoculated into Aedes albopictus clone C6/36 cells for virus isolation. Each cell culture was tested for the presence of viral antigen by immunoperoxidase staining using an anti-JEV polyclonal antibody. Out of 4 Culex sitiens mosquito pools, 2 pools were positive for JEV by cell culture. Presence of JEV genome in the cell cultures for Cx. sitiens was confirmed by using reverse transcriptase-polymerase chain reaction and JEV-specific primers. This is the first report on the isolation of JEV from Cx. sitiens.