RESUMEN
The administration of antiretrovirals (ARVs) for HIV pre-exposure prophylaxis (PrEP) is highly efficacious and may benefit from new long-acting (LA) drug delivery approaches. This paper describes a subcutaneous, reservoir-style implant for the LA delivery of tenofovir alafenamide (TAF) and documents the preclinical assessment of implant safety and pharmacokinetics (PK) in New Zealand White (NZW) rabbits (3 groups of n = 5), beagle dogs (2 groups of n = 6), and rhesus macaques (2 groups of n = 3). Placebo implants were placed in rabbits (n = 10) and dogs (n = 12). Implant parameters, including selection of the TAF form, choice of excipient, and PCL formulation were tuned to achieve targeted concentrations of the active anabolite of TAF, tenofovir diphosphate (TFV-DP), within peripheral blood mononuclear cells (PBMCs) and mucosal tissues relevant to HIV transmission. Sustained concentrations of TFV-DP in PBMCs over 100 fmol/106 cells were achieved in all animal species indicating that the implants effectively delivered TAF for 3-6 months. Unlike placebo implants without TAF, all active implants resulted in local adverse events (AEs) proximal to the implant ranging in severity from mild to moderate and included dermal inflammation and necrosis across all species. Despite these AEs, the implant performed as designed and achieved a constant drug release profile, supporting the continued development of this drug delivery platform.
RESUMEN
OBJECTIVES: To advance the initiative of ending the global epidemic, long-lasting HIV protection is needed through sustained release of antiretroviral drugs for months to years. We investigated in macaques the safety and efficacy of biodegradable polycaprolactone implants releasing tenofovir alafenamide for HIV pre-exposure prophylaxis (PrEP). METHODS: Implants were administered subcutaneously in the arm using a contraceptive trocar. Efficacy against vaginal simian-HIV (SHIV) infection was investigated in six pigtailed macaques that received two tenofovir alafenamide implants (0.35â mg/day), one in each arm, for a total release rate of tenofovir alafenamide at 0.7â mg/day. Macaques were exposed to SHIV twice weekly for 6â weeks. Statistical analyses were used to compare outcome with eight untreated controls. Histological assessments were performed on skin biopsies collected near implantation sites. RESULTS: Median (range) tenofovir diphosphate level in PBMCs was 1519 (1068-1898)â fmol/106 cells. All macaques with tenofovir alafenamide implants were protected against vaginal SHIV infection. In contrast, 7/8 controls were infected after a median of 4 SHIV exposures (Pâ=â0.0047). Histological assessment of tissues near tenofovir alafenamide implant sites showed inflammation and necrosis in 5/6 animals, which were not evident by visual inspection. CONCLUSIONS: We demonstrated complete protection against vaginal SHIV infection with two implants releasing a total of 0.7â mg of tenofovir alafenamide per day. We also identified tenofovir diphosphate concentrations in PBMCs associated with complete vaginal protection. Consistent with previous findings, we observed adverse local toxicity and necrosis near the tenofovir alafenamide implant site. Improved tenofovir alafenamide implants that are safe and maintain high efficacy have the potential to provide long-lasting protection against vaginal HIV infection.
Asunto(s)
Fármacos Anti-VIH , Infecciones por VIH , Virus de la Inmunodeficiencia de los Simios , Animales , Femenino , Infecciones por VIH/prevención & control , Infecciones por VIH/tratamiento farmacológico , Tenofovir/efectos adversos , Fármacos Anti-VIH/uso terapéutico , Macaca , Implantes Absorbibles , VIH , Necrosis/tratamiento farmacológico , Emtricitabina/uso terapéutico , Alanina/uso terapéuticoRESUMEN
In 2007, a previously uninfected kidney transplant recipient tested positive for human immunodeficiency virus type 1 (HIV) and hepatitis C virus (HCV) infection. Clinical information of the organ donor and the recipients was collected by medical record review. Sera from recipients and donor were tested for serologic and nucleic acid-based markers of HIV and HCV infection, and isolates were compared for genetic relatedness. Routine donor serologic screening for HIV and HCV infection was negative; the donor's only known risk factor for HIV was having sex with another man. Four organs (two kidneys, liver and heart) were transplanted to four recipients. Nucleic acid testing (NAT) of donor sera and posttransplant sera from all recipients were positive for HIV and HCV. HIV nucleotide sequences were indistinguishable between the donor and four recipients, and HCV subgenomic sequences clustered closely together. Two patients subsequently died and the transplanted organs failed in the other two patients. This is the first recognized cotransmission of HIV and HCV from an organ donor to transplant recipients. Routine posttransplant HIV and HCV serological testing and NAT of recipients of organs from donors with suspected risk factors should be considered as routine practice.
Asunto(s)
Infecciones por VIH/transmisión , Hepatitis C/transmisión , Trasplante de Órganos/efectos adversos , Donantes de Tejidos , Ensayo de Inmunoadsorción Enzimática , Humanos , Factores de RiesgoRESUMEN
We present here a novel, distinct simian T-cell lymphotropic virus (STLV) found in a red-capped mangabey (Cercocebus torquatus) (CTO-NG409), wild-caught in Nigeria, that showed an HTLV-2-like Western blot (WB) seroreactivity. The complete genome (8920 bp) of CTO-NG409 STLV was related to but different from STLV-3/PHA-PH969 (13.5 %) and STLV-3/PPA-F3 (7.6 %), and STLV-3/CTO604 (11.3 %), found in Eritrean and Senegalese baboons, and red-capped mangabeys from Cameroon, respectively. Phylogenetic analysis of a conserved tax (180 bp) sequence and the env gene (1482 bp) confirmed the relatedness of STLV-3/CTO-NG409 to the STLV-3 subgroup. Molecular clock analysis of env estimated that STLV-3/CTO-NG409 diverged from East and West/Central African STLV-3s about 140,900+/-12,400 years ago, suggesting an ancient African origin of STLV-3. Since phylogenetic evidence suggests multiple interspecies transmissions of STLV-1 to humans, and given the antiquity and wide distribution of STLV-3 in Africa, a search for STLV-3 in human African populations with HTLV-2-like WB patterns is warranted.
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Cercocebus/virología , Infecciones por Deltaretrovirus/veterinaria , Enfermedades de los Monos/virología , Virus Linfotrópico T Tipo 3 de los Primates/clasificación , Animales , Animales Salvajes/virología , Anticuerpos Antivirales/sangre , Anticuerpos Antivirales/inmunología , Infecciones por Deltaretrovirus/virología , Evolución Molecular , Datos de Secuencia Molecular , Nigeria , Filogenia , Virus Linfotrópico T Tipo 3 de los Primates/genética , Virus Linfotrópico T Tipo 3 de los Primates/aislamiento & purificación , Análisis de Secuencia de ADNRESUMEN
Virtually all nonhuman primate species investigated thus far including prosimians, New World and Old World monkeys and apes all harbor distinct and species-specific clades of simian foamy virus (SFV). However, evidence supporting the existence of a human-specific foamy virus (FV) is not yet available. Early reports describing widespread infection of healthy and sick humans with FV could not be confirmed. In contrast, all FV infections documented in humans are of zoonotic origin and are identified in persons occupationally exposed to nonhuman primates. The introduction of SFV into humans raises several public health questions regarding disease outcomes and potential for human-to-human transmissibility. The available data from a very limited number of SFV-infected humans suggest that these infections are nonpathogenic and are not easily transmissible. Additional studies are needed to better define the prevalence and natural history of SFV in humans.
Asunto(s)
Infecciones por Retroviridae , Spumavirus/aislamiento & purificación , Animales , Anticuerpos Antivirales/sangre , Enfermedades de los Gatos/virología , Gatos , Preescolar , Haplorrinos , Humanos , Persona de Mediana Edad , Enfermedades de los Monos/virología , Filogenia , Infecciones por Retroviridae/diagnóstico , Infecciones por Retroviridae/epidemiología , Infecciones por Retroviridae/transmisión , Infecciones por Retroviridae/virología , Estudios Seroepidemiológicos , Enfermedades Virales de Transmisión Sexual/virología , Spumavirus/genética , Spumavirus/inmunología , Spumavirus/patogenicidad , Resultado del Tratamiento , ZoonosisRESUMEN
Resistance testing for treatment-naïve, recently HIV-infected persons is not currently recommended; its clinical value will depend on the prevalence of resistance-associated mutations among recently infected persons. To estimate this prevalence, specimens were collected during 1997-1999 in Seattle and Los Angeles from drug-naïve, recently HIV-infected persons. HIV-1 protease and reverse transcriptase (RT) RNA sequences were amplified from plasma by RT-polymerase chain reaction (RT-PCR), sequenced, and analysed. Of 69 patients, five (7%) had resistance-associated mutations: three (4%) had primary mutations associated with resistance to nucleoside reverse transcriptase inhibitors (NRTI) or non-nucleoside-RTIs, and three patients (4%) had secondary NRTI mutations. No primary mutation associated with resistance to protease inhibitors was observed. Mean age of the five persons with resistance-associated mutations (38 years) was higher than that of the 64 persons without resistance-associated mutations (31 years, P=0.04). The findings suggest that the prevalence of resistance-associated mutations among persons recently infected with HIV in these cities is low.
Asunto(s)
Infecciones por VIH/virología , Inhibidores de la Proteasa del VIH/farmacología , VIH-1/efectos de los fármacos , Inhibidores de la Transcriptasa Inversa/farmacología , Adolescente , Adulto , Anciano , Farmacorresistencia Microbiana/genética , Femenino , Infecciones por VIH/tratamiento farmacológico , Proteasa del VIH/genética , Inhibidores de la Proteasa del VIH/uso terapéutico , Transcriptasa Inversa del VIH/genética , VIH-1/genética , Humanos , Los Angeles/epidemiología , Masculino , Persona de Mediana Edad , Mutación , Prevalencia , Inhibidores de la Transcriptasa Inversa/uso terapéutico , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Washingtón/epidemiologíaRESUMEN
Transmission of HIV-1 with reduced susceptibility to antiretroviral drugs raises public health concerns. Through surveillance of drug-resistant HIV-1 in 603 treatment-naive, recently diagnosed HIV-1-infected persons, we identified a distinct group of viruses that have mutations at codon 215 of the reverse transcriptase (RT) gene that are different from either the wild-type (WT) T or the zidovudine (AZT)-selected T215Y/F. These mutations included 215D/C/S and were found in 20 patients (3.3%). The 215D, 215C, and 215S mutations differ from 215Y by a 1-nt change compared with 2 nt for the WT T215 and likely represent revertants of 215Y. These viruses all were found to have WT susceptibility to AZT, and all replicated efficiently as WT HIV-1(T215). However, differences in fitness among HIV-1(215D), HIV-1(215C), and HIV-1(215S) were seen when RT backgrounds were changed, demonstrating a role of the RT background in the selection of these revertants. In vitro selection with AZT showed that HIV-1(215D) and HIV-1(215C) acquired 215Y more rapidly than did WT HIV-1(T215), likely reflecting the need for only 1-nt change to evolve to 215Y. Our study demonstrates that HIV-1 with unusual mutations at codon 215 replicate efficiently, have WT susceptibility, and are commonly found in treatment-naive persons. The increased ability for selecting resistance mutations defines this class of WT HIV-1 and highlights the higher potential of these viruses to compromise the efficacy of antiretroviral therapy.
Asunto(s)
Fármacos Anti-VIH/farmacología , Farmacorresistencia Viral/genética , Infecciones por VIH/virología , Transcriptasa Inversa del VIH/genética , VIH-1/enzimología , Inhibidores de la Transcriptasa Inversa/farmacología , Zidovudina/farmacología , Fármacos Anti-VIH/uso terapéutico , Secuencia de Bases , ADN Viral , Didanosina/farmacología , Didesoxinucleósidos/farmacología , Evolución Molecular , Infecciones por VIH/tratamiento farmacológico , VIH-1/efectos de los fármacos , VIH-1/genética , Humanos , Datos de Secuencia Molecular , Mutagénesis , Recombinación Genética , Inhibidores de la Transcriptasa Inversa/uso terapéutico , Estavudina/farmacología , Replicación Viral/genética , Zalcitabina/farmacología , Zidovudina/uso terapéuticoAsunto(s)
Etnicidad/genética , Frecuencia de los Genes , Genes MHC Clase II , Genes MHC Clase I , Antígenos de Histocompatibilidad Clase I/genética , Epítopos/genética , Variación Genética , Antígenos HLA-A/genética , Antígenos HLA-B/genética , Antígenos HLA-DQ/genética , Antígenos HLA-DR/genética , Humanos , Líbano , Linfocitos T/inmunología , Inmunología del TrasplanteRESUMEN
Nucleoside reverse-transcriptase (RT) inhibitors (NRTIs), including lamivudine (3TC) and zidovudine (Zdv), are being evaluated for the treatment of human T cell lymphotropic virus type 1 (HTLV-1)-associated disease. However, information on the susceptibility of HTLV-1 to these drugs is limited. The activity of 5 NRTIs on HTLV-1 RT was evaluated. IC(50) values for Zdv, zalcitabine (ddC), didanosine (ddI), 3TC, and stavudine (d4T) were determined, using an enzymatic assay, for 5 HTLV-1 isolates and for reference wild-type and NRTI-resistant human immunodeficiency virus type 1 (HIV-1). Both HTLV-1 and wild-type HIV-1 were equally susceptible to Zdv, ddC, ddI, and d4T. In contrast, high-level resistance to 3TC was found in all HTLV-1 isolates. The findings support the clinical use of Zdv, ddC, ddI, and d4T but not of 3TC for the antiretroviral treatment of HTLV-1-associated disease.
Asunto(s)
Virus Linfotrópico T Tipo 1 Humano/efectos de los fármacos , Lamivudine/farmacología , ADN Polimerasa Dirigida por ARN/metabolismo , Inhibidores de la Transcriptasa Inversa/farmacología , Secuencia de Aminoácidos , Infecciones por Deltaretrovirus/virología , Farmacorresistencia Microbiana , Infecciones por VIH/virología , Transcriptasa Inversa del VIH/efectos de los fármacos , Transcriptasa Inversa del VIH/genética , VIH-1/efectos de los fármacos , VIH-1/enzimología , Virus Linfotrópico T Tipo 1 Humano/enzimología , Humanos , Pruebas de Sensibilidad Microbiana/métodos , Datos de Secuencia Molecular , ADN Polimerasa Dirigida por ARN/química , ADN Polimerasa Dirigida por ARN/genéticaRESUMEN
Treatment of HIV-1-infected persons with antiretroviral drugs including reverse transcriptase (RT) and protease inhibitors has significantly reduced the rate of HIV and AIDS-related morbidity and mortality. However, these treatments can select for drug-resistant viruses which are associated with poor virologic responses to the antiretroviral therapies and loss of clinical benefit. Drug resistance is conferred by single or several amino acid changes in the pol gene. These mutations can be classified as primary when they directly confer reduced drug susceptibility, or secondary when their influence is primarily on replication capabilities of resistant viruses. Both genotypic and phenotypic methods are used for drug resistance testing. Genotypic assays detect resistance-related mutations by sequence analysis or point mutations assays. Phenotypic testing measures drug susceptibility of patient-derived viruses in culture assays. Viruses can be conventionally isolated from peripheral blood lymphocytes, or generated more rapidly through recombination of plasma-derived RT/protease sequences and modified HIV-1 vectors. Phenotypic testing provides direct evidence of resistance, is easy to interpret, but is laborious and expensive. In contrast, genotypic testing provides indirect evidence of resistance, is relatively faster and cheaper, but some complex mutation patterns may be difficult to interpret. Non-culture based phenotypic assays that measure susceptibility of RT activity in plasma to RT inhibitors have been described recently, and provide new tools for rapid phenotypic testing. Resistance testing is currently recommended to help guide the choice of new regimens after treatment failure and for guiding therapy in pregnant women.
Asunto(s)
Fármacos Anti-VIH/farmacología , Infecciones por VIH/tratamiento farmacológico , Inhibidores de la Proteasa del VIH/farmacología , VIH-1/efectos de los fármacos , Inhibidores de la Transcriptasa Inversa/farmacología , Fármacos Anti-VIH/uso terapéutico , Farmacorresistencia Microbiana/genética , Genotipo , Infecciones por VIH/virología , Inhibidores de la Proteasa del VIH/uso terapéutico , VIH-1/genética , Humanos , Mutagénesis , Inhibidores de la Transcriptasa Inversa/uso terapéuticoRESUMEN
BACKGROUND: Nonhuman primates (NHPs) have been widely used in different porcine xenograft procedures inevitably resulting in exposure to porcine endogenous retrovirus (PERV). Surveillance for PERV infection in these NHPs may provide information on the risks of cross-species transmission of PERV, particularly for recipients of vascularized organ xenografts for whom data from human clinical trials is unavailable. METHODS: We tested 21 Old World and 2 New World primates exposed to a variety of porcine xenografts for evidence of PERV infection. These NHPs included six baboon recipients of pig hearts, six bonnet macaque recipients of transgenic pig skin grafts, and nine rhesus macaque and two capuchin recipients of encapsulated pig islet cells. Serologic screening for PERV antibody was done by a validated Western blot assay, and molecular detection of PERV sequences in peripheral blood mononuclear cells (PBMCs) and plasma was performed using sensitive polymerase chain reaction and reverse transcriptase-polymerase chain reaction assays, respectively. Spleen and lymph node tissues available from six bonnet macaques and three rhesus macaques were also tested for PERV sequences. RESULTS: All plasma samples were negative for PERV RNA suggesting the absence of viremia in these xenografted animals. Similarly, PERV sequences were not detectable in any PBMC and tissue samples, arguing for the lack of latent infection of these compartments. In addition, all plasma samples were negative for PERV antibodies. CONCLUSION: These data suggest the absence of PERV infection in all 23 NHPs despite exposure to vascularized porcine organs or tissue xenografts and the use of immunosuppressive therapies in some animals. These findings suggest that PERV is not easily transmitted to these NHP species through these types of xenografts.
Asunto(s)
Cebidae/virología , Trasplante de Células/efectos adversos , Cercopithecidae/virología , Trasplante de Órganos/efectos adversos , Infecciones por Retroviridae/transmisión , Enfermedades de los Porcinos/transmisión , Trasplante Heterólogo/efectos adversos , Animales , Cebus , Quimera , Islotes Pancreáticos/citología , Macaca , Papio , ARN Viral/análisis , Retroviridae/genética , Retroviridae/inmunología , Trasplante de Piel/efectos adversos , Porcinos/genética , Porcinos/virologíaRESUMEN
OBJECTIVES: Information on early HIV-1 infection has come primarily from studies of persons infected with subtype B in North America and Europe; much less is known about other subtypes. The purpose of the present study was to compare the virologic and immunologic parameters following seroconversion among recently-infected persons infected with either of two different HIV-1 subtypes. METHOD: A prospective cohort study was carried out at methadone treatment clinics administered by the Bangkok Metropolitan Administration, Thailand. A total of 130 HIV-1-infected seroconverters (103 with HIV-1 subtype E and 27 with subtype B) were included in the study. The main outcome measures were serial HIV-1 RNA viral load, natural killer cell percentage, CD4 and CD8 lymphocyte counts since seroconversion. RESULTS: The demographic and behavioral characteristics of persons with either subtype were similar. Median RNA viral levels at the earliest time within 3 months of seroconversion were more than three times higher for persons infected with subtype E than subtype B (63 100 versus 18 050 copies/ml, P = 0.001). However, this difference decreased over time such that viral loads were similar at 12, 18, and 24 months following seroconversion. The CD4 and CD8 lymphocyte counts were similar in infections with either subtype during the entire period up to 24 months post-seroconversion. CONCLUSIONS: Higher viral loads associated with subtype E may result from inter-subtype biological differences; however, the epidemiological dynamics of transmission in Bangkok may have also contributed to this phenomenon.
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Infecciones por VIH/inmunología , Infecciones por VIH/virología , VIH-1 , Adulto , Recuento de Linfocito CD4 , Linfocitos T CD8-positivos , Femenino , Infecciones por VIH/epidemiología , Seropositividad para VIH , VIH-1/clasificación , Humanos , Masculino , Estudios Prospectivos , ARN Viral/sangre , Tailandia/epidemiología , Carga ViralRESUMEN
The identification of endogenous avian leukosis virus (ALV) and endogenous avian retrovirus (EAV) in chick cell-derived measles and mumps vaccines in current use has raised concern about transmission of these retroviruses to vaccine recipients. We used serologic and molecular methods to analyze specimens from 206 recipients of measles, mumps, and rubella (MMR) vaccine for evidence of infection with ALV and EAV. A Western blot assay for detecting antibodies to endogenous ALV was developed and validated. All serum samples were negative for antibodies to endogenous ALV by Western blot analysis. Peripheral blood lymphocyte samples from 100 vaccinees were further tested by polymerase chain reaction for both ALV and EAV proviral sequences; all were negative. Matching serum samples were tested by reverse transcriptase polymerase chain reaction for ALV and EAV RNA, and all 100 samples were negative, providing no evidence of viremia. These findings do not indicate the presence of either ALV or EAV infection in MMR vaccine recipients and provide support for current immunization policies.
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Virus de la Leucosis Aviar/aislamiento & purificación , Retrovirus Endógenos/aislamiento & purificación , Vacuna contra el Sarampión-Parotiditis-Rubéola/efectos adversos , Infecciones por Retroviridae/transmisión , Animales , Western Blotting , Pollos , Niño , Humanos , Reacción en Cadena de la Polimerasa de Transcriptasa InversaRESUMEN
Previous findings of low levels of reverse transcriptase (RT) activity in chick cell-derived measles and mumps vaccines showed this activity to be associated with virus particles containing RNA of both subgroup E endogenous avian leukosis viruses (ALV-E) and endogenous avian viruses (EAV). These particles originate from chicken embryonic fibroblast (CEF) substrates used for propagating vaccine strains. To better characterize vaccine-associated ALV-E, we examined the endogenous ALV proviruses (ev loci) present in a White Leghorn CEF substrate pool by restriction fragment length polymorphism. Five ev loci were detected, ev-1, ev-3, ev-6, ev-18, andev-19. Both ev-18 and ev-19 can express infectious ALV-E, while ev-1, ev-3, and ev-6 are defective. We analyzed the full-length sequence of ev-1 and identified an adenosine insertion within the pol RT-beta region at position 5026, which results in a truncated RT-beta and integrase. We defined the 1,692-bp deletion in the gag-pol region of ev-3, and we found that in ev-6, sequences from the 5' long terminal repeat to the 5' pol region were absent. Based on the sequences of the ev loci, RT-PCR assays were developed to examine expression of ALV-E particles (EV) in CEF supernatants. Both ev-1- and ev-3-like RNA sequences were identified, as well as two other RNA sequences with intact pol regions, presumably of ev-18 and ev-19 origin. Inoculation of susceptible quail fibroblasts with CEF culture supernatants from both 5-azacytidine-induced and noninduced CEF led to ALV infection, confirming the presence of infectious ALV-E. Our data demonstrate that both defective and nondefective ev loci can be present in CEF vaccine substrates and suggest that both ev classes may contribute to the ALV present in vaccines.
Asunto(s)
Virus de la Leucosis Aviar/aislamiento & purificación , Fibroblastos/virología , Vacuna Antisarampión/biosíntesis , Vacuna contra la Parotiditis/biosíntesis , Animales , Virus de la Leucosis Aviar/genética , Virus de la Leucosis Aviar/fisiología , Azacitidina/farmacología , Southern Blotting , Embrión de Pollo , Medios de Cultivo Condicionados , Virus Defectuosos/genética , Virus Defectuosos/aislamiento & purificación , Virus Defectuosos/fisiología , Contaminación de Medicamentos , Fibroblastos/efectos de los fármacos , Genoma , Datos de Secuencia Molecular , Polimorfismo de Longitud del Fragmento de Restricción , Provirus/genética , Provirus/aislamiento & purificación , Codorniz/virología , ARN Viral/análisis , ARN Viral/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Análisis de Secuencia de ADNRESUMEN
Simian type D retrovirus (SRV) is enzootic in many populations of Asian monkeys of the genus Macaca and is associated with immunodeficiency diseases. However, the zoonotic potential of this agent has not been well defined. Screening for antibodies to SRV was performed as part of an ongoing study looking for evidence of infection with simian retroviruses among persons occupationally exposed to nonhuman primates (NHPs). Of 231 persons tested, 2 (0.9%) were found to be strongly seropositive, showing reactivity against multiple SRV antigens representing gag, pol, and env gene products by Western immunoblotting. Persistent long-standing seropositivity, as well as neutralizing antibody specific to SRV type 2, was documented in one individual (subject 1), while waning antibody with eventual seroreversion was observed in a second (subject 2). Repeated attempts to detect SRV by isolation in tissue culture and by using sensitive PCR assays for amplification of two SRV gene regions (gag and pol) were negative. Both individuals remain apparently healthy. We were also unable to transmit this seropositivity to an SRV-negative macaque by using inoculation of whole blood from subject 1. The results of this study provide evidence that occupational exposure to NHPs may increase the risk of infection with SRV and underscore the importance of both occupational safety practices and efforts to eliminate this virus from established macaque colonies.
Asunto(s)
Enfermedades de los Monos/transmisión , Exposición Profesional , Infecciones por Retroviridae/transmisión , Retrovirus de los Simios/aislamiento & purificación , Infecciones Tumorales por Virus/transmisión , Zoonosis , Animales , Anticuerpos Antivirales/sangre , ADN Viral/sangre , Humanos , Macaca mulatta , Enfermedades de los Monos/virología , Pruebas de Neutralización , Reacción en Cadena de la Polimerasa , Infecciones por Retroviridae/virología , Retrovirus de los Simios/genética , Retrovirus de los Simios/inmunología , Infecciones Tumorales por Virus/virologíaRESUMEN
Plasma viral load from 71 HIV-1-infected neonates was measured by using Amp-RT, an ultrasensitive quantitative reverse transcriptase (RT) assay and by nucleic acid sequence-based amplification (NASBA), an RNA-based quantitative assay. Results were then compared with those obtained from detection of proviral DNA in peripheral blood mononuclear cells (PBMCs) by polymerase chain reaction (PCR) using Turnbull analysis. At 5 days of life, 50% of neonates were positive by Amp-RT, 30% were NASBA positive, and 20% were DNA-PCR positive. Through the first 12 days of life, Amp-RT was more sensitive than either NASBA or DNA-PCR in detecting HIV-1 infection. Amp-RT values correlated well with NASBA RNA values, with an overall Pearson's r = 0.63 (95% confidence interval [CI], 0.40-0.78). In proportional hazards analysis of infants aged 14 to 61 days (N = 31), a one-log increase in RNA-based viral load was associated with a > fivefold risk of disease progression when using the U.S. Centers for Disease Control and Prevention (CDC) clinical Category C (CDC-C) or death as an endpoint (p =.014). Kaplan-Meier analysis of these data found that RNA viral loads were able to predict disease progression using CDC-C/death as an endpoint (p = .013). Early quantitative viral load measurements may assist clinicians in diagnosing HIV-1 infection, stratifying risk of disease progression, and implementing a treatment plan using highly active antiretroviral therapy for infants within the first few weeks of life.
Asunto(s)
ADN Viral/sangre , Infecciones por VIH/congénito , Infecciones por VIH/diagnóstico , Transcriptasa Inversa del VIH/sangre , Enfermedades del Recién Nacido/diagnóstico , ARN Viral/sangre , Negro o Afroamericano , Fármacos Anti-VIH/uso terapéutico , Terapia Antirretroviral Altamente Activa , Peso al Nacer , Centers for Disease Control and Prevention, U.S. , Demografía , Progresión de la Enfermedad , Femenino , Infecciones por VIH/sangre , Infecciones por VIH/tratamiento farmacológico , Infecciones por VIH/virología , Transcriptasa Inversa del VIH/metabolismo , VIH-1/enzimología , VIH-1/genética , VIH-1/aislamiento & purificación , VIH-1/fisiología , Humanos , Lactante , Recién Nacido , Enfermedades del Recién Nacido/sangre , Enfermedades del Recién Nacido/tratamiento farmacológico , Enfermedades del Recién Nacido/virología , Recien Nacido Prematuro , Masculino , Reacción en Cadena de la Polimerasa , Pronóstico , Modelos de Riesgos Proporcionales , Sensibilidad y Especificidad , Tasa de Supervivencia , Estados Unidos , Carga ViralRESUMEN
In view of the concern over potential infection hazards in the use of porcine tissues and organs for xenotransplantation to humans, we investigated the diversity of porcine endogenous retrovirus (PERV) genomes in the DNA of domestic pigs and related species. In addition to the three known envelope subgroups of infectious gamma retroviruses (PERV-A, -B, and -C), classed together here as PERV group gamma 1, four novel groups of gamma retrovirus (gamma 2 to gamma 5) and four novel groups of beta retrovirus (beta 1 to beta 4) genomes were detected in pig DNA using generic and specific PCR primers. PCR quantification indicated that the retroviral genome copy number in the Landrace x Duroc F(1) hybrid pig ranged from 2 (beta 2 and gamma 5) to approximately 50 (gamma 1). The gamma 1, gamma 2, and beta 4 genomes were transcribed into RNA in adult kidney tissue. Apart from gamma 1, the retroviral genomes are not known to be infectious, and sequencing of a small number of amplified genome fragments revealed stop codons in putative open reading frames in several cases. Analysis of DNA from wild boar and other species of Old World pigs (Suidae) and New World peccaries (Tayassuidae) showed that one retrovirus group, beta 2, was common to all species tested, while the others were present among all Old World species but absent from New World species. The PERV-C subgroup of gamma1 genomes segregated among domestic pigs and were absent from two African species (red river hog and warthog). Thus domestic swine and their phylogenetic relatives harbor multiple groups of hitherto undescribed PERV genomes.
Asunto(s)
Retrovirus Endógenos/clasificación , Retrovirus Endógenos/genética , Genoma Viral , Porcinos/virología , Animales , ADN Viral/análisis , ADN Viral/genética , Datos de Secuencia Molecular , Reacción en Cadena de la Polimerasa/métodos , ARN Viral/genética , ARN Viral/metabolismoRESUMEN
Since 1984, unheated porcine clotting factor VIII (Hyate:C) has been used to treat severe bleeding episodes in persons with hemophilia who have antibodies to human clotting factor. We document the presence of porcine endogenous retrovirus (PERV) in plasma samples of pigs and in clinical lots of Hyate:C. Both gag and pol PERV RNA sequences were detected by reverse-transcriptase (RT) polymerase chain reaction in 13 of 13 lots of Hyate:C tested. Among 10 of these lots, RT activity also was detected, which confirms the presence of retroviral particles. To assess the transmission of PERV to Hyate:C recipients, we tested serum specimens from 88 recipients of Hyate:C and 23 noninfused control subjects for anti-PERV antibodies by using a Western blot assay. None of the samples was positive. Our data document that PERV particles are a common contaminant of Hyate:C products and suggest that the risk of PERV transmission from these percutaneous exposures is very low.
Asunto(s)
Contaminación de Medicamentos , Retrovirus Endógenos/aislamiento & purificación , Factor VIII/efectos adversos , Hemofilia A/terapia , Infecciones por Retroviridae/transmisión , Porcinos/virología , Animales , Anticuerpos Antivirales/sangre , Retrovirus Endógenos/clasificación , Retrovirus Endógenos/genética , Retrovirus Endógenos/inmunología , Hemofilia A/virología , Humanos , Datos de Secuencia Molecular , Plasma/virología , ARN Viral/análisis , ARN Viral/sangre , Infecciones por Retroviridae/veterinaria , Infecciones por Retroviridae/virología , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Enfermedades de los Porcinos/virologíaRESUMEN
Porcine xenografts may offer a solution to the shortage of human donor allografts. However, all pigs contain the porcine endogenous retrovirus (PERV), raising concerns regarding the transmission of PERV and the possible development of disease in xenotransplant recipients. We evaluated 11 antiretroviral drugs licensed for human immunodeficiency virus type 1 (HIV-1) therapy for their activities against PERV to assess their potential for clinical use. Fifty and 90% inhibitory concentrations (IC(50)s and IC(90)s, respectively) of five nucleoside reverse transcriptase inhibitors (RTIs) were determined enzymatically for PERV and for wild-type (WT) and RTI-resistant HIV-1 reference isolates. In a comparison of IC(50)s, the susceptibilities of PERV RT to lamivudine, stavudine, didanosine, zalcitabine, and zidovudine were reduced >20-fold, 26-fold, 6-fold, 4-fold, and 3-fold, respectively, compared to those of WT HIV-1. PERV was also resistant to nevirapine. Tissue culture-based, single-round infection assays using replication-competent virus confirmed the relative sensitivity of PERV to zidovudine and its resistance to all other RTIs. A Gag polyprotein-processing inhibition assay was developed and used to assess the activities of protease inhibitors against PERV. No inhibition of PERV protease was seen with saquinavir, ritonavir, indinavir, nelfinavir, or amprenavir at concentrations >200-fold the IC(50)s for WT HIV-1. Thus, following screening of many antiretroviral agents, our findings support only the potential clinical use of zidovudine.
Asunto(s)
Antivirales/farmacología , Retrovirus Endógenos/efectos de los fármacos , Inhibidores de Proteasas/farmacología , Inhibidores de la Transcriptasa Inversa/farmacología , Secuencia de Aminoácidos , Animales , Farmacorresistencia Microbiana , Resistencia a Múltiples Medicamentos , Retrovirus Endógenos/enzimología , Retrovirus Endógenos/fisiología , Endopeptidasas/metabolismo , Transcriptasa Inversa del VIH/genética , VIH-1/efectos de los fármacos , VIH-1/enzimología , Humanos , Pruebas de Sensibilidad Microbiana/métodos , Datos de Secuencia Molecular , ADN Polimerasa Dirigida por ARN/química , ADN Polimerasa Dirigida por ARN/genética , ADN Polimerasa Dirigida por ARN/metabolismo , Porcinos , Cultivo de VirusRESUMEN
A group of three rhesus macaques were inoculated with SIV isolated from a human (SIVhu) accidentally exposed and infected with SIVsm. Extensive sequence analyses of SIVhu obtained from the human and macaques following infection indicated the presence of truncated nef. Not only did nef fail to repair itself in vivo postinfection (p.i.), but instead, further mutations added additional stop codons with increasing time p.i. Infection of these animals was associated with minimal acute viral replication, followed by undetectable plasma viral loads and only intermittent PCR detection up to 5 years p.i. The three SIVhu infected and three control monkeys were then challenged with the heterologous highly pathogenic SHIV89.6p. All three controls became infected and showed rapid declines in peripheral CD4(+) lymphocytes, disease, and death at 10 and 32 weeks p.i., respectively. In contrast, all three animals previously infected with SIVhu are healthy and exhibit stable CD4(+) lymphocyte levels and undetectable plasma viral loads at >20 months post-SHIV89. 6p challenge. Only transient, low levels of SHIV replication were noted in these animals. Whereas responses to SIVgag/pol were noted, no evidence for SIV/SHIV envelope cross-reactivity was detected by antibody or CTL analyses, suggesting that the protective immune mechanisms to the heterologous challenge isolate were most likely not directed to envelope but rather to other viral determinants.