RESUMEN
A rapid method for identification of human immunodeficiency virus Type 1 (HIV-1) gag subtypes was developed based on restriction fragment length polymorphism (RFLP) analysis of 400 or 650 bp long polymerase chain reaction (PCR) fragments encompassing the start of the p17 (400 bp) and part of the p24 (650bp) regions. The consensus sequences of subtypes A-D, the only subtypes identified in South Africa, were analyzed to detect restriction endonucleases which generate unique patterns for each subtype. Four restriction endonucleases were identified: AluI, AccI, SwaI and XmnI. Digestion of a 400 bp fragment with AluI allowed identification of subtype C. Samples not identified were then reamplified, and a 650 bp fragment digested with AccI to identify subtype B, followed by SwaI and XmnI to distinguish between subtypes A and D. This strategy was applied to 87 samples previously subtyped by either sequence analysis of the gag p17 region (n = 33); or heteroduplex mobility assay (HMA) based on the env gene (n = 75); or both (n = 21). Out of the 87 samples, RFLP identified two samples as subtype A, 28 as subtype B, 56 as subtype C and one as a subtype D virus. No discrepancies were found between RFLP gag subtypes and gag sequence subtypes demonstrating the reliability of this method. There was also no discordance between gag RFLP subtypes and env HMA subtypes, suggesting that there were no recombinant viruses detected relating to the genomic regions analyzed. RFLP is an effective technique for the rapid screening in an HIV epidemic of limited diversity, such as in South Africa.
Asunto(s)
Genes gag , Infecciones por VIH/virología , VIH-1/clasificación , VIH-1/aislamiento & purificación , Polimorfismo de Longitud del Fragmento de Restricción , ADN Viral/aislamiento & purificación , VIH-1/genética , Humanos , Reacción en Cadena de la Polimerasa/métodos , Mapeo Restrictivo , SudáfricaRESUMEN
There are two families of viruses that contribute to lymphomagenesis in humans: herpesviruses and retroviruses. The two herpesviruses are the Epstein-Barr virus (EBV) and human herpesvirus-8 (HHV-8). EBV is an extremely well-characterized transforming agent: nine viral proteins contribute to transformation in vitro. In contrast, in vivo, the pattern of EBV gene expression varies with different types of malignancies. EBV is associated with endemic Burkitt's lymphoma, acquired immune deficiency syndrome (AIDS)-related lymphoma, post-transplantation lymphoproliferative disease, Hodgkin's disease (HD), and rare T-cell lymphomas. We have summarized studies on the different patterns of viral gene expression and signaling in different EBV-related malignancies, which have begun to reveal how EBV variably contributes to the malignant phenotype in different diseases. HHV-8 is associated with primary effusion lymphomas in patients with AIDS, and the rapidly accumulating information on this virus is summarized. Human T-cell leukemia virus-1 (HTLV-1) is a retrovirus which is the causative agent of adult T-cell leukemia/lymphoma (ATL). The specific mechanism of HTLV-1-mediated T-cell transformation is unclear, but the effects of HTLV-1 on interleukin-2 signaling are reviewed.
Asunto(s)
Herpesvirus Humano 4/patogenicidad , Herpesvirus Humano 8/patogenicidad , Virus Linfotrópico T Tipo 1 Humano/patogenicidad , Linfoma/virología , HumanosRESUMEN
Thirty-seven matched cerebrospinal fluid (CSF) and plasma samples from 34 human immunodeficiency virus type 1 (HIV-1)-infected patients with suspected meningitis were analyzed for levels of HIV-1 RNA and markers of inflammation. Patients with tuberculous (n = 9) or cryptococcal (n = 6) meningitis had the highest CSF virus loads, which in many cases exceeded the levels in plasma, compared with patients with meningococcal meningitis (n = 3), aseptic meningitis (n = 8), tuberculoma (n = 2), or AIDS dementia complex (n = 4) or with normal lumbar punctures (n = 3). CSF virus load correlated significantly with the number of infiltrating lymphocytes (r = .60, P < .001) but not with plasma virus load, the levels of beta2-microglobulin in the CSF, or the integrity of the blood-brain barrier. These data suggest significant intrathecal HIV-1 replication in patients with lymphocytic meningeal infections such as tuberculous and cryptococcal meningitis.
Asunto(s)
Infecciones por VIH/líquido cefalorraquídeo , VIH-1/aislamiento & purificación , Meningitis/líquido cefalorraquídeo , Meningitis/virología , ARN Viral/aislamiento & purificación , Complejo SIDA Demencia/sangre , Complejo SIDA Demencia/líquido cefalorraquídeo , Complejo SIDA Demencia/inmunología , Barrera Hematoencefálica , Recuento de Linfocito CD4 , Criptococosis/sangre , Criptococosis/líquido cefalorraquídeo , Criptococosis/inmunología , Proteína p24 del Núcleo del VIH/análisis , Infecciones por VIH/sangre , Infecciones por VIH/inmunología , VIH-1/genética , Humanos , Recuento de Leucocitos , Linfocitos/inmunología , Meningitis/sangre , Meningitis Aséptica/sangre , Meningitis Aséptica/líquido cefalorraquídeo , Meningitis Aséptica/inmunología , Meningitis Fúngica/sangre , Meningitis Fúngica/líquido cefalorraquídeo , Meningitis Fúngica/inmunología , Meningitis Meningocócica/sangre , Meningitis Meningocócica/líquido cefalorraquídeo , Meningitis Meningocócica/inmunología , ARN Viral/análisis , Tuberculosis Meníngea/sangre , Tuberculosis Meníngea/líquido cefalorraquídeo , Tuberculosis Meníngea/inmunología , Carga Viral , Microglobulina beta-2/análisis , Microglobulina beta-2/metabolismoRESUMEN
BACKGROUND: A simple, inexpensive serological assay is required for the early determination of HIV infection status among infants born to HIV-1-seropositive women in developing countries. OBJECTIVES: To evaluate the use of a commercially available capture enzyme immunoassay (EIA), the MUREX*ICE HIV-1.O.2, for the early identification of seroreverting, uninfected infants. STUDY DESIGN: Infants with a clearly defined HIV-1 infection status, as determined by polymerase chain reaction results and/or seroreactivity at 18 months, were tested for antibodies to HIV. The time to seroreversion using the capture EIA was compared with the results obtained using an indirect assay, the GENELAVIA MIXT EIA. RESULTS: Seroreverting infants were identified earlier with the capture than the indirect EIA; all of the uninfected infants were seronegative at 12 months with the capture EIA while 100% seroreversion was only seen at 18 months with the indirect EIA. CONCLUSIONS: In general, the capture EIA identified seroreverting infants 3-6 months earlier than the indirect EIA. However, caution must be exercised in interpreting seroreactivity in a breast-fed population where HIV infection may occur in a child who has previously seroreverted.
Asunto(s)
Infecciones por VIH/inmunología , VIH-1/aislamiento & purificación , Técnicas para Inmunoenzimas , África/epidemiología , Estudios de Evaluación como Asunto , Femenino , Anticuerpos Anti-VIH/inmunología , Infecciones por VIH/diagnóstico , Infecciones por VIH/virología , VIH-1/inmunología , Humanos , LactanteRESUMEN
OBJECTIVE: To describe the endogenous cytokine profile of HIV-1-infected lymph nodes (LN) and to identify the phenotype of individual cells expressing intracytoplasmic cytokines. DESIGN AND METHODS: Whole LN biopsies were collected from three HIV-seronegative controls and four HIV-1-positive individuals with persistent generalized lymphadenopathy. Three had established infection (Centers for Disease Control and Prevention 1993 criteria, stages A2, C1 and C3) and one was undergoing seroconversion illness. A combination of three methods was used to assess the impact of HIV-1 on LN architecture and endogenous cytokine expression. Immunocytochemistry was used to locate follicular dendritic cells (FDC), interdigitating cells and T and B cells. Reverse transcriptase-polymerase chain reaction was used to assess mRNA for interleukin (IL)-1 beta, IL-2, IL-4, IL-6, IL-10, tumour necrosis factor (TNF)-alpha and interferon (IFN)-gamma in collagenase-digested LN cells. Three-colour flow cytometry was used to identify intracytoplasmic cytokine expression within cell subsets. RESULTS: Germinal center (GC) hyperplasia was observed in LN from two patients with established HIV-1 infection, and the third, coinfected with Mycobacterium tuberculosis, showed extensive necrosis. In the patient undergoing seroconversion, there was an extensive FDC network within the expanded and confluent GC which covered expansive areas of the LN. There was varied expression of IL-1 beta, IL-4, IL-6, IL-10 and TNF-alpha mRNA from the four HIV-1-infected LN and the patient undergoing seroconversion showed evidence for a mixed cytokine profile, which also included IL-2 and IFN-gamma. Flow cytometry revealed intracytoplasmic IL-1 beta protein restricted to cells expressing CD19, CD21 and CD38 antigens. CONCLUSIONS: Cytokines were detected in freshly isolated HIV-1-infected LN cells without requiring an exogenous stimulus. Seroconversion was associated with an expanded FDC network within enlarged GC, bounded by defined mantle zones containing B cells. There was diverse cytokine mRNA expression and IL-1 beta protein was restricted to cells expressing B-cell-associated antigens.
Asunto(s)
Antígenos CD/análisis , Citocinas/inmunología , Infecciones por VIH/inmunología , VIH-1/inmunología , Ganglios Linfáticos/inmunología , Subgrupos de Linfocitos T/inmunología , Adulto , Citocinas/genética , Citoplasma/inmunología , Expresión Génica , Infecciones por VIH/patología , Humanos , Inmunohistoquímica , Ganglios Linfáticos/patología , Masculino , Persona de Mediana Edad , Coloración y Etiquetado , Subgrupos de Linfocitos T/clasificaciónRESUMEN
BACKGROUND: Both human immunodeficiency virus type 1 (HIV-1) and African viral haemorrhagic fever (VHF) viruses cause similar symptoms in acutely infected individuals and must be included in the differential diagnosis in areas where HIV-1 and VHF viruses both occur. OBJECTIVES: To determine the cause of an acute illness in a patient at risk of exposure to both HIV-1 and African VHF viruses. RESULTS: Serological examination revealed the presence of high levels of the p24 core antigen of HIV-1 in the absence of antibodies to HIV-1 in a specimen collected during the acute stage of the infection. On follow-up, the antigen enzyme-linked immunosorbent assay (ELISA) became negative while the antibody ELISA and confirmatory Western blot for HIV-1 became positive. CONCLUSIONS: Acute HIV seroconversion illness may have protean manifestations and, in the more severe forms, may cause diagnostic dilemmas, particularly in regions where African VHFs occur.
RESUMEN
A 3-cm peripheral nerve gap in a rabbit hind-limb model was repaired by using a polyglycolic acid (PGA) conduit filled with a gelatin/Schwann-cell suspension. Postoperative nerve function after 16 weeks, as measured by isometric twitch and tetanic muscle strengths, and nerve conduction velocities, failed to demonstrate a statistically significant difference, compared to a control group in which the nerve gap was reconstructed by using a PGA conduit filled with gelatin only. The 3-cm gap in the described model may not have been long enough to show a significant difference between the two groups. Alternatively, the transferred cultured Schwann cells may have been functionally inactive.
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Regeneración Nerviosa/fisiología , Nervio Peroneo/cirugía , Células de Schwann/trasplante , Animales , Células Cultivadas , Miembro Posterior , Contracción Isométrica/fisiología , Músculo Esquelético/inervación , Músculo Esquelético/fisiología , Conducción Nerviosa/fisiología , Nervio Peroneo/fisiología , Ácido Poliglicólico , ConejosAsunto(s)
VIH-1/fisiología , Trasplante de Células Madre Hematopoyéticas , Transfusión de Linfocitos , Linfocitos/inmunología , Linfocitos/virología , Animales , Secuencia de Bases , Gonadotropina Coriónica Humana de Subunidad beta/biosíntesis , Gonadotropina Coriónica Humana de Subunidad beta/genética , ADN , Cartilla de ADN , Sangre Fetal , Humanos , Ganglios Linfáticos , Datos de Secuencia Molecular , Oligonucleótidos Antisentido , Papio , Reacción en Cadena de la Polimerasa , Bazo , Replicación ViralRESUMEN
Several previous studies reported various immunosuppressive effects of hyperbaric oxygen on nonspecific and specific cell-mediated reactions. A highly immunogenic skin allograft mouse model was used to evaluate the clinical relevance of the previously described immunosuppressive effects of hyperbaric oxygen. A 1.5 x 2.0-cm full-thickness skin allograft was cross-grafted between paired immunohistoincompatible mouse strains (N = 40, C57BL/6 and BALB/c female mice) that were randomly assigned to four groups receiving (1) no treatment (controls), (2) cyclosporine 1 mg per kilogram intraperitoneally daily, (3) cyclosporine plus a low-dose hyperbaric oxygen treatment (two treatments per day, once a week), and (4) cyclosporine plus a high-dose hyperbaric oxygen treatment (two treatments per day, three times a week) following surgery (N = 32). Allograft samples were taken from each group at day 9 after cross-grafting (N = 8). Skin allograft rejection was significantly delayed in all treatment groups compared to controls. No difference was found between animals who received cyclosporine only and the combined treatment regimen including low-dose hyperbaric oxygen. High-dose hyperbaric oxygen treatment in combination with cyclosporine substantially prolonged skin allograft survival compared to other treatments. These findings were histologically confirmed. We conclude that hyperbaric oxygen treatment as an adjunct to standard immunosuppressive therapy may only be advantageous if frequently applied.
Asunto(s)
Ciclosporina/farmacología , Rechazo de Injerto/prevención & control , Prueba de Histocompatibilidad , Oxigenoterapia Hiperbárica , Inmunosupresores/farmacología , Trasplante de Piel/patología , Animales , Terapia Combinada , Femenino , Rechazo de Injerto/patología , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Trasplante HomólogoRESUMEN
The effect of hyperbaric oxygen (HBO) as an immunosuppressive agent was evaluated by using a highly immunogenic skin allograft mouse model. Immune-histoincompatible female C57BL/6 and BALB/c mice (N = 30) were randomly assigned to three groups receiving no treatment (control group), low dose HBO treatment (two treatments once a week), and intermediate HBO treatment (two treatments 3 times/wk) 1 wk before and 2 wk after transplantation of a 1.5 x 2 cm full thickness skin allograft from the back. Rejection was observed a Day 7 and was completed 14 days after surgery in controls. Low dose and intermediate HBO treatment delayed skin allograft rejection, which was histologically confirmed.
Asunto(s)
Rechazo de Injerto/prevención & control , Oxigenoterapia Hiperbárica , Animales , Femenino , Inmunocompetencia , Ratones , Ratones Endogámicos BALB C , Ratones Endogámicos C57BL , Distribución Aleatoria , Colgajos QuirúrgicosRESUMEN
Three rapid enzyme immunobinding assays, Abbott's TestPack HIV-1/HIV-2, Clonatec's Rapid HIV1/HIV2 AB and the DuPont HIVCHEK 1 + 2, were evaluated using a panel of 20 selected sera with Western blot-proven reactivity to at least one envelope glycoprotein of HIV-1. The Abbott assay had the highest sensitivity and detected 18 of the 20 sera with no indeterminates (i.e. specimens with high background coloration which interferes with interpretation of the assay). Both the Clonatec and DuPont rapid assays correctly identified 12 sera as HIV-1 antibody-positive; the former produced 5 false-negatives and 3 indeterminates and the latter 7 false-negatives and 1 indeterminate. The majority of the sera used in the evaluation showed evidence of early sero-conversion as many had low absorbance ratios in a screening enzyme immunosorbent assay. Although they represent only a small portion of clinical specimens, they point to the need for careful evaluation of new methodologies, using appropriate, well-characterised sera, before such techniques are accepted for general use.
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Anticuerpos Anti-VIH/sangre , Técnicas para Inmunoenzimas/normas , Estudios de Evaluación como Asunto , HumanosRESUMEN
A sero-epidemiological surveillance study to monitor the prevalence of HIV-1 infection in Johannesburg, South Africa, was commenced in February 1988. The population selected for study were attenders at clinics for sexually transmitted diseases (STD) and at family planning (FP) clinics. In the 12 months of the study 6631 sera were tested. Of the STD attenders, 15 of 1224 black females (1.2%) and 21 of 2482 black males (0.8%) were positive. Of the 449 white males tested 49 were homosexual, amongst whom 10 (20.4%) were positive; in the heterosexual white male group 4 of 400 (1.0%) were positive. Of the FP clinic attenders, 4 of 1459 black females (0.3%) were positive. 68 of the 6631 sera tested were indeterminate for infection. No attenders were positive for HIV-2 infection. These data confirmed the entry of HIV infection into the black population in South Africa.
Asunto(s)
Seropositividad para VIH/epidemiología , VIH-1 , Instituciones de Atención Ambulatoria , Donantes de Sangre , Femenino , Seropositividad para VIH/etnología , Humanos , Masculino , Vigilancia de la Población , Sudáfrica/epidemiologíaRESUMEN
An HIV1 antibody screening assay, the "ImmunoComb" HIV1 kit which uses a dip-stick methodology, was evaluated using a panel of selected sera. The ImmunoComb detected 144 of 147 (98%) Western-blot-confirmed positive sera. However, when the ImmunoComb was compared with a recombinant EIA capable of detecting early seroconversion, only 144 of 156 (92.3%) recombinant EIA or Western-blot-positive sera were ImmunoComb-positive. Thus, while the kit is suitable for serological surveys and prevalence studies, especially in areas where skilled laboratory staff and sophisticated equipment are unavailable, the low sensitivity precludes its use in blood banks and transfusion services.
Asunto(s)
Síndrome de Inmunodeficiencia Adquirida/diagnóstico , Anticuerpos Anti-VIH/análisis , VIH-1/inmunología , Juego de Reactivos para Diagnóstico , Ensayo de Inmunoadsorción Enzimática , Estudios de Evaluación como Asunto , Humanos , Pruebas InmunológicasRESUMEN
An epidemiological composite of the human immunodeficiency virus (HIV) epidemic in South Africa was constructed from reported acquired immunodeficiency syndrome (AIDS) figures, blood donor seroprevalence data and a family of active surveillance studies in the Johannesburg area. Of great concern is the extensive and continuing silent spread of HIV in the urban black population, manifested by doubling times in male and female sexually transmitted disease (STD) clinic attenders of 10.67 and 9.78 months respectively, a doubling time in female family planning (FP) attenders of 6.55 months, and HIV infection rates of 1:56, 1:37 and 1:91, respectively. These data suggest a spread of infection far more extensive than the relatively lower AIDS figures. However, seroprevalence figures in municipal employees of mainly rural origin were considerably lower, only 1:1,250 in black men and none in women, but regular movements between urban and rural areas could well introduce and amplify infection in rural areas. A complete epidemiological picture of HIV can only be attained by employing both the above approach of the family of surveys complemented by a nationwide seroprevalence study to assess HIV penetration into the general 'background' population. Educational interventions should now be particularly targeted to the major risk groups in the urban black heterosexual community.
Asunto(s)
Síndrome de Inmunodeficiencia Adquirida/epidemiología , Seroprevalencia de VIH , Negro o Afroamericano , Población Negra , Femenino , Homosexualidad , Humanos , Masculino , Conducta Sexual , Sudáfrica/epidemiología , Población BlancaRESUMEN
The significance of indeterminate Western blotting results was assessed by determining their distribution in attenders of sexually transmitted diseases (STD) and family planning (FP) clinics. As expected, it was found that the former had a highly significantly greater prevalence of HIV infection. Indeterminate results were not found more frequently in STD attenders refuting any association with sexual promiscuity. They were, however, found significantly more frequently amongst black compared to white subjects, but no difference was found between males and females. Indeterminate results appear to be a feature of black African sera and what the significance is, still remains to be elucidated.