RESUMEN
BACKGROUND: Congenital rubella syndrome (CRS) case identification is challenging in older children since laboratory markers of congenital rubella virus (RUBV) infection do not persist beyond age 12 months. METHODS: We enrolled children with CRS born between 1998 and 2003 and compared their immune responses to RUBV with those of their mothers and a group of similarly aged children without CRS. Demographic data and sera were collected. Sera were tested for anti-RUBV immunoglobulin G (IgG), IgG avidity, and IgG response to the 3 viral structural proteins (E1, E2, and C), reflected by immunoblot fluorescent signals. RESULTS: We enrolled 32 children with CRS, 31 mothers, and 62 children without CRS. The immunoblot signal strength to C and the ratio of the C signal to the RUBV-specific IgG concentration were higher (P < .029 for both) and the ratio of the E1 signal to the RUBV-specific IgG concentration lower (P = .001) in children with CRS, compared with their mothers. Compared with children without CRS, children with CRS had more RUBV-specific IgG (P < .001), a stronger C signal (P < .001), and a stronger E2 signal (P ≤ .001). Two classification rules for children with versus children without CRS gave 100% specificity with >65% sensitivity. CONCLUSIONS: This study was the first to establish classification rules for identifying CRS in school-aged children, using laboratory biomarkers. These biomarkers should allow improved burden of disease estimates and monitoring of CRS control programs. Published by Oxford University Press on behalf of the Infectious Diseases Society of America 2014. This work is written by (a) US Government employee(s) and is in the public domain in the US.
Asunto(s)
Instituciones Académicas , Estudiantes , Síndrome de Rubéola Congénita/diagnóstico , Biomarcadores/sangre , Adolescente , Anticuerpos Antivirales , Afinidad de AnticuerposRESUMEN
CONTEXT: Childhood vaccination has reduced rubella disease to low levels in the United States, but outbreaks continue to occur. The largest outbreak in the past 5 years occurred in Nebraska in 1999. OBJECTIVES: To examine risk factors for disease, susceptibility of the risk population, role of vaccine failure, and the need for new vaccination strategies in response to the Nebraska rubella outbreak. DESIGN, SETTING, AND PATIENTS: Investigation of 83 confirmed rubella cases occurring in Douglas County, Nebraska, between March 23 and August 24, 1999; serosurvey of 413 pregnant women in the outbreak locale between October 1998 and March 1999 (prior to outbreak) and April and November 1999 (during and after outbreak). MAIN OUTCOME MEASURES: Case characteristics, compared with that of the general county population; area childhood rubella vaccination rates; and susceptibility among pregnant women before vs during and after the outbreak. RESULTS: All 83 rubella cases were unvaccinated or had unknown vaccination status and fell into 3 groups: (1) 52 (63%) were young adults (median age, 26 years), 83% of whom were born in Latin American countries where rubella vaccination was not routine. They were either employed in meatpacking plants or were their household contacts. Attack rates in the plants were high (14.4 per 1000 vs 0. 19 per 1000 for general county population); (2) 16 (19%), including 14 children (9 of whom were aged <12 months) and 2 parents, were US-born and non-Hispanic, who acquired the disease through contacts at 2 day care facilities (attack rate, 88.1 per 1000); and (3) 15 (18%) were young adults (median age, 22 years) whose major disease risk was residence in population-dense census tracts where meatpacking-related cases resided (R(2) = 0.343; P<.001); 87% of these persons were born in Latin America. Among pregnant women, susceptibility rates were 13% before the outbreak and 11% during and after the outbreak. Six (25%) of 24 susceptible women tested were seropositive for rubella IgM. Rubella vaccination rates were 90.2% for preschool children and 99.8% for school-aged children. CONCLUSIONS: A large rubella outbreak occurred among unvaccinated persons in a community with high immunity levels. Crowded working and living conditions facilitated transmission, but vaccine failure did not. Workplace vaccination could be considered to prevent similar outbreaks. JAMA. 2000;284:2733-2739.
Asunto(s)
Brotes de Enfermedades , Hispánicos o Latinos/estadística & datos numéricos , Vacuna contra la Rubéola , Rubéola (Sarampión Alemán)/epidemiología , Vacunación/estadística & datos numéricos , Lugar de Trabajo , Adolescente , Adulto , Niño , Preescolar , Infecciones Comunitarias Adquiridas/epidemiología , Emigración e Inmigración , Femenino , Humanos , Lactante , Masculino , Nebraska/epidemiología , Embarazo , Factores de Riesgo , Rubéola (Sarampión Alemán)/prevención & control , Rubéola (Sarampión Alemán)/transmisión , Estudios Seroepidemiológicos , América del Sur , Lugar de Trabajo/estadística & datos numéricosRESUMEN
The association between human papillomavirus (HPV) DNA copy number and cervical disease was investigated. Viral DNA copy number for the most common high-risk HPV types in cervical cancer (types 16, 18, 31, and 45) was determined in cervical cytobrush specimens from 149 women with high-grade cervical intraepithelial neoplasias (CIN II-CIN III), 176 with low-grade CIN (CIN I), and 270 with normal cytology. Quantitative, PCR-based fluorescent assays for each of the HPV genotypes and for the beta-globin gene were used. The amount of cellular DNA increased significantly with increasing disease; thus, HPV was expressed as copies per microgram of cellular DNA. The assay had a dynamic range of >10(7), allowing documentation for the first time of the wide range of HPV copy numbers seen in clinical specimens. Median HPV DNA copy number varied by more than 10(4) among the viral types. HPV16 was present in the highest copy number; over 55% of HPV16-positive samples contained more than 10(8) copies/microgram. Median copy number for HPV16 showed dramatic increases with increasing epithelial abnormality, an effect not seen with the other HPV types. HPV16 increased from a median of 2.2 x 10(7) in patients with normal cytology, to 4.1 x 10(7) in CIN I patients, to 1.3 x 10(9) copies/microgram in CIN II-III patients. Even when stratified by cervical disease and viral type, the range of viral DNA copies per microgram of cellular DNA was quite large, precluding setting a clinically significant cutoff value for "high" copy numbers predictive of disease. This study suggests that the clinical usefulness of HPV quantitation requires reassessment and is assay dependent.
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ADN Viral/análisis , ADN Viral/genética , Papillomaviridae/genética , Papillomaviridae/aislamiento & purificación , Displasia del Cuello del Útero/virología , Neoplasias del Cuello Uterino/virología , Adolescente , Adulto , Anciano , Secuencia de Bases , Cuello del Útero/virología , Cartilla de ADN/genética , Sondas de ADN de HPV/genética , Femenino , Genotipo , Humanos , Persona de Mediana Edad , Papillomaviridae/clasificación , Infecciones por Papillomavirus/complicaciones , Infecciones por Papillomavirus/virología , Reacción en Cadena de la Polimerasa , Infecciones Tumorales por Virus/complicaciones , Infecciones Tumorales por Virus/virología , Neoplasias del Cuello Uterino/etiología , Neoplasias del Cuello Uterino/patología , Displasia del Cuello del Útero/etiología , Displasia del Cuello del Útero/patologíaRESUMEN
We conducted a case-control study of the association between SIL and HPV among whites (W), African Americans (AA), and Hispanics (H) in Harris County, Texas. Cases were identified at M.D. Anderson Cancer Center Colposcopy Clinic. Controls were identified among women obtaining routine Pap screening at two Harris County Health Department Clinics. HPV was detected by a PCR-based fluorescent assay. Dichotomous and polytomous logistic regression models were used to estimate adjusted odd ratios (AOR) and 95% confidence intervals (CI) for SIL among racial/ethnic groups and grade of disease. Prevalence of HPV infection was 64% in low grade SIL (LSIL), 84% in high grade SIL (HSIL), and 19% in controls. Risk of SIL was higher in H than in W and AA, AOR 29.5 (12.4-70.5), 15.3 (6.0-33.8), and 5.8 (2.6-12.6), respectively. Similarly, racial/ethnic differences were observed for both LSIL (AOR = 16.6, 7.7, and 4.3, respectively) and HSIL (AOR = 78.6, 34.6, and 14.2, respectively). Findings support the association between SIL and HPV and differences in the strength of the association with LSILs and HSILs. Data also suggest a higher risk for H and a lower risk for AA.
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Población Negra , Hispánicos o Latinos , Papillomaviridae , Infecciones por Papillomavirus/etnología , Infecciones Tumorales por Virus/etnología , Displasia del Cuello del Útero/etnología , Neoplasias del Cuello Uterino/etnología , Población Blanca , Adulto , Estudios de Casos y Controles , Femenino , Hispánicos o Latinos/estadística & datos numéricos , Humanos , Infecciones por Papillomavirus/diagnóstico , Prevalencia , Factores de Riesgo , Factores Socioeconómicos , Texas/epidemiología , Infecciones Tumorales por Virus/diagnóstico , Neoplasias del Cuello Uterino/diagnóstico , Displasia del Cuello del Útero/diagnósticoRESUMEN
BACKGROUND: The development of cervical carcinoma is influenced by multiple factors, including the presence of certain high risk types of human papillomavirus. The purpose of the current study was to investigate possible cooperating genetic changes by examining the expression of p53, p62 myc, and p21 ras in cervical biopsy specimens. METHODS: Three hundred and ninety-five cervical biopsy specimens representing normal through high grade cervical intraepithelial neoplasia (CIN) were screened by immunohistochemistry for expression of p53, p62myc, and p21ras. RESULTS: Neither the proportion of tissues staining positive for a given protein nor the staining patterns within the epithelial layers differed significantly among normal or CIN biopsy samples. However, grade specific nuclear staining of p21ras was found in the cells of 10 lesions that were classified as CIN I by histology. CONCLUSIONS: These results established the normal distribution and expression patterns of p53, p62myc, and p21ras within 395 cervical biopsy samples representing normal through CIN III histology. The expression of these proteins (e.g., staining intensity and layer of epithelium staining positive) is similar in normal tissues and those demonstrating all grades of CIN.
Asunto(s)
Carcinoma in Situ/genética , Proteína Oncogénica p21(ras)/análisis , Proteínas Proto-Oncogénicas c-myc/análisis , Proteína p53 Supresora de Tumor/análisis , Neoplasias del Cuello Uterino/genética , Femenino , Humanos , InmunohistoquímicaRESUMEN
We enrolled 85 patients with invasive cervical cancer and collected cervicovaginal lavage samples at each clinical visit for diagnosis, staging, treatment, and follow-up. Lavage samples were tested by L1 consensus polymerase chain reaction for human papillomavirus (HPV). Results were compared with HPV demonstrated in tumor tissue and the clinical status at time of sample collection. Sensitivity and specificity of the lavage for detection of tumor HPV, determined on the basis of results of tests on lavage samples collected prior to therapy, were found to be 56% and 76.9%, respectively. The proportion of lavage samples detecting tumor HPV decreased significantly with treatment, from 0.54 at diagnosis to 0.03 at complete response (P < .001). Local treatment failure was associated with increased detection of tumor HPV; however, no samples were positive prior to clinically detected treatment failure. These results suggest that cervicovaginal lavage is not an effective sampling method for epidemiological analysis of HPV in cervical tumors.
Asunto(s)
Papillomaviridae/aislamiento & purificación , Infecciones por Papillomavirus/virología , Infecciones Tumorales por Virus/virología , Neoplasias del Cuello Uterino/virología , Adulto , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Persona de Mediana Edad , Papillomaviridae/genética , Infecciones por Papillomavirus/fisiopatología , Población , Insuficiencia del Tratamiento , Infecciones Tumorales por Virus/fisiopatología , Neoplasias del Cuello Uterino/patología , Neoplasias del Cuello Uterino/fisiopatología , Neoplasias del Cuello Uterino/terapiaRESUMEN
A simple method for the detection of a number of human papillomavirus (HPV) genotypes associated with cervical cancer has been developed. The assay exploits the 5'-->3' exonucleolytic activity of Taq DNA polymerase to increase the signal from fluorescent dyes by releasing them from genotype-specific probes during PCR. The probes are oligonucleotides with a 5' reporter dye (6-carboxyfluorescein), a quencher dye (6-carboxy-tetramethyl-rhodamine), and a phosphate-blocked 3' end. In the intact probe, the proximity of the reporter and the quencher results in suppression of reporter fluorescence by Förster-type energy transfer (V. T. Förster. Ann. Phys. 2:55-75, 1948). If the probe is bound downstream of either primer during PCR, the 5'-->3' exonucleolytic activity of Taq polymerase degrades it, allowing the reporter to diffuse away from the quencher, which results in an increase in reporter fluorescence. The increased fluorescence is directly related to the amount of target DNA and can be detected with an automated fluorometer. Probes for the L1 region of the cervical-cancer-associated HPV types 16, 18, 31, 33, and 35 were synthesized and the assays were optimized. The most sensitive assay can detect as few as two copies of HPV DNA in human cervical specimens.
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Bioensayo/métodos , ADN Viral/análisis , Papillomaviridae/genética , Colorantes Fluorescentes , Humanos , Datos de Secuencia Molecular , Sondas de Oligonucleótidos , Papillomaviridae/aislamiento & purificación , Sensibilidad y EspecificidadRESUMEN
It is not known whether DNA sequence variants of human papillomavirus type 16 (HPV-16) are distinct serotypes. To examine this question, the reactivities of women's sera from Zaire (n = 97) and Denmark (n = 123) were compared in IgG-specific ELISAs based on virus-like particles (VLPs) composed of the L1 major capsid protein derived from an HPV-16 variant common in central Africa (Z-1194) or one common in northern Europe (114K). These L1s differ in seven amino acids. There was a strong correlation between reactivity in the two assays for both sets of sera (correlation coefficients, 0.73 and 0.85 for Zairian and Danish sera, respectively). In only 1 serum was there evidence for a specific reaction to one but not the other VLP variant. The results support the conclusion that the virions of strains Z-1194 and 114K are serologically cross-reactive.
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Papillomaviridae/inmunología , Reacciones Cruzadas , Femenino , Humanos , Inmunoglobulina G/inmunología , Papillomaviridae/clasificación , Virión/inmunologíaRESUMEN
BACKGROUND: Cervical cancer remains an important public health problem, particularly for the urban minority population. To the authors' knowledge, determinants of cervical cancer survival have not been studied in this high risk population. METHODS: This study included all 158 women diagnosed and treated for invasive cervical cancer from January 1, 1986, through December 31, 1992, at the Grady Memorial Hospital and Clinics (Atlanta, GA). Medical records were abstracted to determine age at diagnosis, race, International Federation of Gynecology and Obstetrics (FIGO) clinical stage, treatment, and survival. Pathologic material was reviewed to confirm the diagnosis. RESULTS: Most patients (80%) were African American, and the stage distribution was similar for African American and white patients. Sixty-six (42%) had FIGO Stage I disease; 50%, Stage II or III; and 8%, Stage IV. Four-year actuarial survival differed significantly according to clinical stage (Ia = 94%, Ib = 79%, II = 39%, III = 26%, IV = 0%). Overall survival was lower for patients with glandular carcinomas than for those with squamous cell carcinomas (26% vs. 55%, P = 0.09). This difference was almost entirely due to increased mortality in patients with Stage Ib adenocarcinomas (53% vs. 88% for squamous cell carcinoma, Stage Ib, P = 0.03). CONCLUSIONS: The major prognostic markers for cervical cancer survival in this high risk patient population were clinical stage and histology, factors identical to those identified for other populations.
Asunto(s)
Salud Urbana , Neoplasias del Cuello Uterino/epidemiología , Negro o Afroamericano/estadística & datos numéricos , Femenino , Georgia/epidemiología , Humanos , Persona de Mediana Edad , Estadificación de Neoplasias , Programa de VERF , Tasa de Supervivencia , Neoplasias del Cuello Uterino/etnología , Neoplasias del Cuello Uterino/patología , Población Blanca/estadística & datos numéricosRESUMEN
The sequences of the capsid genes of a human papillomavirus type 16 (HPV 16) DNA and an HPV 31 DNA were determined. The HPV 16 DNA contained genes coding for the most variable HPV 16 capsid proteins yet identified (17 variable amino acids). Three of six coding changes in the HPV 31 DNA occurred at positions equivalent to ones where variable amino acids in HPV 16 have been observed. Variable amino acids in both viruses occurred predominantly in regions which showed amino acid variation when closely related types of HPV were compared; thus, most of the factors which determined the intratypic variation in the capsid proteins of the viruses described here were likely the same as those which determined differences between the capsid proteins of different HPV types.
Asunto(s)
Cápside/genética , Papillomaviridae/genética , Secuencia de Bases , ADN Viral , Femenino , Genes Virales , Variación Genética , Humanos , Datos de Secuencia Molecular , Sistemas de Lectura AbiertaRESUMEN
To determine if human papillomavirus (HPV) in the primary tumor was associated with disease-free survival of stage Ib cervical cancer patients, archival tissues from 47 patients were analyzed for HPV DNA by in situ hybridization (ISH) and polymerase chain reaction. HPV integration was determined by ISH signal pattern. Laboratory data were correlated with clinical parameters and disease-free survival. Kaplan-Meier estimates of 4-year disease-free survival were 56% in women with HPV detected in the primary tumor by ISH and 100% for women in whom HPV was not detected (P = .02). Four-year disease-free survival was 39% for patients with integrated HPV in the primary tumor (P = .005 vs. HPV-negative tumors and .05 vs. HPV episomal or episomal/integrated). HPV detection and integration state was not associated with any other clinical variable. Detection of integrated HPV DNA in the primary tumor was strongly associated with treatment failure.
Asunto(s)
ADN de Neoplasias/análisis , ADN Viral/análisis , Papillomaviridae/aislamiento & purificación , Neoplasias del Cuello Uterino/patología , Neoplasias del Cuello Uterino/virología , Adulto , Anciano , Anciano de 80 o más Años , Supervivencia sin Enfermedad , Femenino , Humanos , Hibridación in Situ , Metástasis Linfática , Persona de Mediana Edad , Estadificación de Neoplasias , Papillomaviridae/genética , Reacción en Cadena de la Polimerasa , Estudios Retrospectivos , Factores de Tiempo , Ultrasonografía , Neoplasias del Cuello Uterino/diagnóstico por imagen , Neoplasias del Cuello Uterino/radioterapia , Integración ViralRESUMEN
The crystal structure of the P1/Mahoney poliovirus empty capsid has been determined at 2.9 A resolution. The empty capsids differ from mature virions in that they lack the viral RNA and have yet to undergo a stabilizing maturation cleavage of VP0 to yield the mature capsid proteins VP4 and VP2. The outer surface and the bulk of the protein shell are very similar to those of the mature virion. The major differences between the 2 structures are focused in a network formed by the N-terminal extensions of the capsid proteins on the inner surface of the shell. In the empty capsids, the entire N-terminal extension of VP1, as well as portions corresponding to VP4 and the N-terminal extension of VP2, are disordered, and many stabilizing interactions that are present in the mature virion are missing. In the empty capsid, the VP0 scissile bond is located some 20 A away from the positions in the mature virion of the termini generated by VP0 cleavage. The scissile bond is located on the rim of a trefoil-shaped depression in the inner surface of the shell that is highly reminiscent of an RNA binding site in bean pod mottle virus. The structure suggests plausible (and ultimately testable) models for the initiation of encapsidation, for the RNA-dependent autocatalytic cleavage of VP0, and for the role of the cleavage in establishing the ordered N-terminal network and in generating stable virions.
Asunto(s)
Cápside/química , Poliovirus/química , Cristalización , Cristalografía por Rayos X , Modelos Moleculares , Poliovirus/fisiología , Pliegue de Proteína , ARN Viral/químicaRESUMEN
This study sought to determine risk factors for genital infection with papillomavirus (HPV) in Panamanian women 20-49 years old. Subjects were randomly selected from Herrera and Panama provinces (cervical cancer incidence 79 and 25/100,000, respectively). Participants were interviewed to determine sexual behavior. Cervicovaginal lavage specimens were obtained to test for HPV DNA by commercial dot blot hybridization. HPV-16/18 DNA was detected significantly more frequently (5%) in Panama than Herrera (2%) samples (P = .002). Clearly, infection with high-risk HPV types alone cannot account for the differences in cervical cancer incidence between the two populations. HPV-16/18 detection decreased with increasing years of sexual experience among all women in Panama and among women with multiple partners in Herrera. However, HPV-16/18 detection did not change with sexual experience among monogamous women in Herrera. Thus, the epidemiology of HPV is complex and reflects both virus- and population-specific factors.
Asunto(s)
Enfermedades de los Genitales Femeninos/epidemiología , Papillomaviridae , Infecciones por Papillomavirus/epidemiología , Infecciones Tumorales por Virus/epidemiología , Neoplasias del Cuello Uterino/epidemiología , Adulto , Factores de Edad , ADN Viral/análisis , Femenino , Enfermedades de los Genitales Femeninos/patología , Enfermedades de los Genitales Femeninos/virología , Humanos , Persona de Mediana Edad , Panamá/epidemiología , Papillomaviridae/aislamiento & purificación , Infecciones por Papillomavirus/patología , Distribución Aleatoria , Factores de Riesgo , Salud Rural/estadística & datos numéricos , Conducta Sexual , Infecciones Tumorales por Virus/patología , Salud Urbana/estadística & datos numéricos , Neoplasias del Cuello Uterino/patología , Neoplasias del Cuello Uterino/virologíaRESUMEN
Cericovaginal lavage samples from 124 human immunodeficiency virus type 1 (HIV-1)-seropositive and 126 HIV-1-seronegative women were collected monthly for 8 months and tested for human papillomavirus (HPV) DNA. The estimated prevalence of HPV was 42.8% in HIV-1-seropositive and 13.4% in -seronegative women (P < .001). There was no significant difference in HPV DNA detection in HIV-1-seropositive women with CD4 cell counts of < 300/mm3 (50% HPV-positive), 300-499/mm3 (36.4% HPV-positive), or > or = to 500/mm3 (40.5% HIV-positive). However, HIV-1-seropositive women who were more immunocompromised, as indicated by lower CD4 cell counts, were more likely to shed HPV persistently. The quantity of HPV DNA detected in cervicovaginal lavage samples was similar in HIV-1-seropositive and -seronegative women. This study further defined the characteristics of HPV infections in HIV-1-infected women.
Asunto(s)
Seropositividad para VIH/complicaciones , Papillomaviridae/aislamiento & purificación , Infecciones por Papillomavirus/virología , Infecciones Tumorales por Virus/virología , Cuello del Útero/virología , Estudios de Cohortes , ADN Viral/aislamiento & purificación , Femenino , Seronegatividad para VIH , Humanos , Estudios Longitudinales , Papillomaviridae/genética , Infecciones por Papillomavirus/complicaciones , Infecciones por Papillomavirus/epidemiología , Prevalencia , Estudios Prospectivos , Infecciones Tumorales por Virus/complicaciones , Infecciones Tumorales por Virus/epidemiología , Vagina/virologíaRESUMEN
Human papillomaviruses (HPVs) are associated with at least 80% of cervical carcinomas and are classified as high-risk or low-risk based on whether or not they are commonly found in cervical cancers. The high-risk HPVs have early gene products (E6 and E7) that immortalize human keratinocytes and are at least partially responsible for causing cervical carcinoma. E6 and E7 from the high-risk viruses interact strongly with the tumor suppressors p53 and Rb; those from the low-risk HPVs do not. Transformation involves a multi-step process and requires additional factors besides high-risk HPV infection. High-risk HPVs are capable of immortalizing primary human keratinocytes in tissue culture, but such cells become transformed only after certain chromosomal changes take place, possibly having to do with oncogene activation. The DNA of high-risk HPVs is frequently (if not always) integrated into the genome of cancer cells; it is normally episomal in premalignant lesions. Integration disrupts the E2 and E5 genes and viral gene regulation. Cells containing integrated viral DNA show excessively high levels of E6 and E7. While there is some conflicting evidence, it appears that the p53 and Rb tumor-suppressor genes are more frequently mutated in HPV-negative tumors than they are in HPV-positive tumors, suggesting that for tumor formation to proceed the p53 and Rb proteins must be inactivated either by interaction with the viral proteins or by mutation. The presence of an activated oncogene in a cell lacking functional p53 or Rb may then be sufficient to cause tumor progression.
Asunto(s)
Transformación Celular Viral , Genes Supresores de Tumor , Papillomaviridae/fisiología , Proteínas Proto-Oncogénicas/fisiología , Secuencia de Bases , ADN , Humanos , Datos de Secuencia Molecular , Oncogenes , Papillomaviridae/genética , Factor de Crecimiento Transformador beta/fisiología , Proteínas Virales/fisiologíaRESUMEN
We have investigated the diversity of a hypervariable segment of the human papillomavirus type 16 (HPV-16) genome among 301 virus isolates that were collected from 25 different ethnic groups and geographic locations. Altogether, we distinguished 48 different variants that had diversified from one another along five phylogenetic branches. Variants from two of these branches were nearly completely confined to Africa. Variants from a third branch were the only variants identified in Europeans but occurred at lower frequency in all other ethnic groups. A fourth branch was specific for Japanese and Chinese isolates. A small fraction of all isolates from Asia and from indigenous as well as immigrant populations in the Americas formed a fifth branch. Important patterns of HPV-16 phylogeny suggested coevolution of the virus with people of the three major human races, namely, Africans, Caucasians, and East Asians. But several minor patterns are indicative of smaller bottlenecks of viral evolution and spread, which may correlate with the migration of ethnic groups in prehistoric times. The colonization of the Americas by Europeans and Africans is reflected in the composition of their HPV-16 variants. We discuss arguments that today's HPV-16 genomes represent a degree of diversity that evolved over a large time span, probably exceeding 200,000 years, from a precursor genome that may have originated in Africa. The identification of molecular variants is a powerful epidemiological and phylogenetic tool for revealing the ancient spread of papillomaviruses, whose trace through the world has not yet been completely lost.
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Papillomaviridae/genética , Infecciones por Papillomavirus/historia , Infecciones Tumorales por Virus/historia , África , Asia , Secuencia de Bases , Europa (Continente) , Femenino , Frecuencia de los Genes , Historia Antigua , Humanos , India/etnología , Indígenas Norteamericanos , Masculino , Datos de Secuencia Molecular , Infecciones por Papillomavirus/genética , Infecciones Tumorales por Virus/genética , Neoplasias del Cuello Uterino/microbiologíaRESUMEN
Compared with other laboratory techniques, the polymerase chain reaction (PCR) is a simple, rapid, sensitive method for detecting human papillomavirus (HPV) DNA in cervical samples. However, since many cervical samples contain multiple HPV types, we decided to investigate whether PCR results from such samples accurately reflected the relative amounts of each HPV type present. Theoretical calculations of product accumulation when multiple DNAs with different amplification efficiencies are present in the same sample were done. In addition a set of samples in which cloned HPV DNAs were mixed in varying proportions prior to PCR was tested. Finally, four clinical samples containing multiple HPV types by hybridization assays were subjected to PCR, using two different primer sets. Each of these lines of investigation showed that selective amplification of one HPV DNA over another can occur when mixed HPV types are present. This effect may lead to inaccurate information regarding both types and relative amounts of HPV DNAs in samples containing multiple HPV types. A protocol to avoid this problem is described.
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Papillomaviridae/clasificación , Reacción en Cadena de la Polimerasa/métodos , Secuencia de Bases , Cartilla de ADN , Femenino , Genotipo , Humanos , Datos de Secuencia Molecular , Papillomaviridae/genética , Infecciones por Papillomavirus/microbiología , Reproducibilidad de los Resultados , Infecciones Tumorales por Virus/microbiología , Frotis VaginalRESUMEN
Increasing evidence indicates that infection with genital types of human papillomavirus (HPV) can occur prior to the onset of sexual activity, possibly by perinatal transmission. Evidence is also accumulating that women infected with the human immunodeficiency virus (HIV) more frequently express HPV. We conducted this study to measure HPV prevalence in HIV-seropositive and -seronegative women in Kinshasa, Zaire and in their children. We collected cervico-vaginal lavage specimens from 80 mothers (52 HIV-seropositive and 28 HIV-seronegative at the time of delivery) and oropharyngeal and perineal specimens from their 81 3-year old children (21 HIV-seropositive and 60-seronegative). We used the ViraPap and ViraType assay to test specimens for HPV DNA by the dot-blot technique. Detection of HPV in the mother was highly associated with HIV: 20 HIV-seropositive women and one seronegative woman had HPV DNA. Ten children had HPV DNA. However, detection of HPV in the children was not associated with the mothers' HPV or HIV status or with the child's own HIV status. These findings document a substantial prevalence in young children of HPV DNA types that are linked to genital-tract neoplasia in adults, but do not specifically support a hypothesis of mother-to-child transmission of genital HPV types.
Asunto(s)
Infecciones por VIH/complicaciones , Papillomaviridae/patogenicidad , Infecciones Tumorales por Virus/transmisión , ADN Viral/análisis , República Democrática del Congo , Femenino , Infecciones por VIH/epidemiología , Humanos , Madres , Infecciones Tumorales por Virus/epidemiología , Enfermedades del Cuello del Útero/microbiologíaRESUMEN
Epidemiologic studies show an association between infection with human immunodeficiency virus type 1 (HIV-1) and human papillomavirus (HPV) associated anogenital disease. To investigate possible molecular mechanisms of HIV-1 modulation of HPV expression, we studied the effect of an HIV-1 regulatory protein, tat1, on gene expression directed by the upstream regulatory region (URR) of HPV type 16 (HPV 16). HPV 16 URR-directed chloramphenicol acetyltransferase (CAT) expression driven by the native HPV 16 promoter (P97) was increased in the presence of tat1 alone. Tat1 also reversed E2-mediated repression of P97-directed CAT expression. E2 mediated CAT expression with URR constructs containing the SV40 promoter was enhanced when tat1 and E2 were cotransfected. Using a cervical carcinoma cell line (SiHa), E2 enhancement of URR-directed gene expression was elevated in the presence of extracellular tat1 or during cocultivation with HIV-1-infected cells. These results show HIV can modulate HPV gene expression in cell culture and that the increased rate of HPV-associated cervical disease in asymptomatic HIV-seropositive women may result from HPV-HIV molecular interactions.
Asunto(s)
Proteínas de Unión al ADN , Productos del Gen tat/genética , VIH-1/genética , Papillomaviridae/genética , Infecciones Oportunistas Relacionadas con el SIDA/complicaciones , Línea Celular , Femenino , Regulación Viral de la Expresión Génica/efectos de los fármacos , Productos del Gen tat/farmacología , Infecciones por VIH/complicaciones , VIH-1/patogenicidad , Humanos , Proteínas Oncogénicas Virales/genética , Papillomaviridae/efectos de los fármacos , Papillomaviridae/patogenicidad , Transcripción Genética/efectos de los fármacos , Activación Transcripcional , Transfección , Infecciones Tumorales por Virus/complicaciones , Neoplasias del Cuello Uterino/complicaciones , Productos del Gen tat del Virus de la Inmunodeficiencia HumanaRESUMEN
Human papillomavirus (HPV) DNA was found in cervicovaginal lavage fluids from 9 of 11 human immunodeficiency virus type 1 (HIV-1)-seropositive female prostitutes with cervical intraepithelial neoplasia (CIN) in Kinshasa, Zaire. Since 7 yielded complex nucleic acid hybridization results consistent with mixed HPV infections, limited sequencing of HPV DNA was used to identify the HPVs present. Three of HPV 16 and 1 each of HPV 18, 31, 33, and 56 and ME180-HPV were identified by sequencing in 8 samples. Each of these genotypes has been found in specimens from HIV-1-seronegative women with CIN. Some DNAs had nucleic acid and amino acid sequence variations relative to the reference HPVs, but the variants were closely related to variants that have been found in HIV-1-seronegative women. Variant amino acids were found predominantly at three positions in one 40-amino-acid segment of the L1 open reading frame sequenced. The predominant HPV 16 variant observed has been found rarely in other countries.