RESUMEN
Spontaneous lymphomas occur at high frequency in NFS x V+ mice, strains congenic for ecotropic murine leukemia virus (MuLV) proviral genes and expressing virus at high titer. In the present study, a total of 703 NFS x V+ lymphomas were studied by histopathology, immunophenotypic analysis, immunoglobulin heavy chain or T cell receptor beta chain rearrangements, and somatic ecotropic MuLV integrations; 90% of the lymphomas tested were of B cell lineage. Low-grade tumors included small lymphocytic, follicular, and splenic marginal zone lymphomas, while high-grade tumors comprised diffuse large-cell (centroblastic and immunoblastic types), splenic marginal zone, and lymphoblastic lymphomas. Comparison of mice of similar genetic background except for presence (NFS x V+) or absence (NFS x V-) of functional ecotropic MuLV genomes showed that NFS x V-clonal lymphomas developed at about one-half the rate of those occurring in NFS x V+ mice, and most were low-grade B cell lymphomas with extended latent periods. In NFS x V+ mice, clonal outgrowth, defined by Ig gene rearrangements, was associated with acquisition of somatic ecotropic proviral integrations, suggesting that, although generation of B cell clones can be virus independent, ecotropic virus may act to increase the rate of generation of clones and speed their evolution to lymphoma. The mechanism remains undefined, because only rare rearrangements were detected in several cellular loci previously associated with MuLV insertional mutagenesis.
Asunto(s)
Virus de la Leucemia Murina/aislamiento & purificación , Linfoma de Células B/patología , Linfoma de Células B/virología , Animales , Southern Blotting , Reordenamiento Génico de la Cadena beta de los Receptores de Antígenos de los Linfocitos T , Genoma Viral , Cadenas Pesadas de Inmunoglobulina/genética , Inmunohistoquímica , Virus de la Leucemia Murina/genética , Linfoma de Células B/clasificación , Linfoma de Células B/genética , RatonesRESUMEN
Primary prostate epithelial and prostate adenocarcinoma cells cultured in serum-free medium grew for up to 10 passages before senescence. Cells from prostate adenocarcinoma of a 55-year-old patient without lymph node involvement were transfected with plasmids containing recombinant human papilloma virus HPV16 or HPV18 DNA and the selectable neomycin-resistance gene. After G-418 selection, cells underwent crisis, and surviving cells infected with retroviruses encoding the HPV18 E6/E7 genes (HPV-PAC1), transfected with a head-to-tail dimer of the complete HPV16 genome (HPV-PAC2), or transfected with HPV18 E6/E7 early genes (HPV-PAC3) were established. HPV-PAC1 and HPV-PAC2 cultures appeared morphologically similar to primary cultures even after 40 passages. However, HPV-PAC2 cultures had a clonal morphology. All lines were positive for cytokeratin 18, had acquired vimentin expression, and contained either HPV16 or HPV18 sequences integrated into host DNA. None was tumorigenic in nude mice or formed colonies in soft agar. These cells did not secrete prostate specific antigen nor respond to androgen although tamoxifen inhibited the growth of the cells. Immunohistochemistry showed no evidence of p53 overexpression. Further characterization of these cell lines and examination of their response to chemotherapeutic agents may provide relevant information for the study of hormone-independent PC.
Asunto(s)
Adenocarcinoma/virología , ADN Viral/genética , Papillomaviridae/genética , Neoplasias de la Próstata/virología , Adenocarcinoma/genética , Adenocarcinoma/patología , Andrógenos/farmacología , División Celular/efectos de los fármacos , Medios de Cultivo , Humanos , Masculino , Persona de Mediana Edad , Neoplasias de la Próstata/genética , Neoplasias de la Próstata/patología , Estimulación Química , Transfección , Células Tumorales CultivadasRESUMEN
To investigate clinical condyloma, abnormal cervical cytologic findings, and evidence of human papillomavirus infections, 89 adolescent girls were examined. Cellular DNAs extracted from exfoliated cervical cells were examined for human papillomavirus genomic sequences by Southern transfer hybridization using 32P-labeled human papillomavirus DNA probes. Human papillomavirus sequences were detected in 12 (13%) young women, abnormal cytologic specimens in 21 (24%), and vulvar condylomas in 12 (13%). The human papillomavirus types identified included HPV-6/11 (four instances), which is known to be associated with benign lesions, and HPV-16, -18, and -31 (eight instances) which are considered to have oncogenic potential. Two young women were infected with both HPV-16 and -31. Human papillomavirus sequences were found in 48% of the young women with abnormal cytologic findings and in 3% of patients with normal cytologic findings (P less than .0001). Condylomatous changes in the cervical smear were associated with the presence of HPV-6/11 and mild dysplasia with the presence of HPV-16, -18, and -31. The presence of vulvar condylomas correlated with condylomatous changes in the cervical smear and with the recovery of HPV-6/11 from the cervical epithelium. The results indicate that the prevalence of human papillomavirus infections in this population is high and that a majority of the infections are with viruses associated with lower genital tract malignancies.
Asunto(s)
Infecciones Tumorales por Virus/epidemiología , Adolescente , Cuello del Útero/patología , Condiloma Acuminado/epidemiología , Condiloma Acuminado/patología , ADN Viral/análisis , Femenino , Humanos , Maryland , Papillomaviridae/análisis , Infecciones Tumorales por Virus/patología , Frotis VaginalRESUMEN
Southern transfer analysis for human papillomavirus genomic sequences was conducted on 152 vulvar and vaginal tissue specimens obtained from 86 patients. Histopathologic diagnoses included condyloma acuminatum, intraepithelial neoplasia, and invasive cancer. In six patients, lesions of more than one pathologic type were identified. Vaginal lesions constituted less than 5% of tissues examined. Distribution of lesions was as follows: condyloma, 93 lesions from 57 patients; intraepithelial neoplasia, 47 lesions from 29 patients; and invasive carcinoma, 12 lesions from six patients. Seventy-five percent of the patients were white. The mean age of the patients increased from 25 years for condyloma to 38 years for vulvar intraepithelial neoplasia III to 56 years for invasive cancer. A viral diagnosis was made in 81% of condylomas, 84% of vulvar intraepithelial neoplasia III, and 58% of invasive carcinomas. Distribution of viral types differed markedly for the various histopathologies. Types 6/11 accounted for 77% of condylomas and 0% of vulvar intraepithelial neoplasia III. Type 16 was recovered from 12% of condylomas and 81% of vulvar intraepithelial neoplasia III. Type 18 was identified in a small proportion in both categories; type 31 was seen in a few vulvar intraepithelial neoplasia III lesions. In invasive carcinomas, type 16 was the predominantly identified virus. Papillomavirus type 16 emerges as the dominant oncogenic virus in vulvar neoplasms. Its presence in a large percentage of condylomas raises the issue of an "atypical condyloma" as a precursor of neoplasia.
Asunto(s)
Papillomaviridae/aislamiento & purificación , Neoplasias de la Vulva/microbiología , Adulto , Condiloma Acuminado/microbiología , Condiloma Acuminado/patología , Femenino , Humanos , Persona de Mediana Edad , Invasividad Neoplásica , Papillomaviridae/clasificación , Neoplasias de la Vulva/patologíaRESUMEN
DNA of human papillomavirus (HPV) type 45, a new HPV type 18-related papillomavirus of the genital tract, was cloned from a recurrent cervical lesion displaying mild to moderate dysplasia with koilocytosis. HPV-45 DNA was identified in paraffin sections of biopsies of both the initial and recurrent lesions of the patient, taken 7 months apart. HPV-45 DNA hybridized efficiently to that of many different HPV types under low and moderate stringency conditions (Tm - 37 degrees C to Tm - 25 degrees C) but with only HPV-18 DNA under high stringency conditions (Tm - 17 degrees C). HPV-45 DNA was distinguished from HPV-18 DNA by (i) differences in restriction enzyme digest patterns, (ii) lack of hybridization at Tm - 17 degrees C between HPV-18 and some fragments of HPV-45, (iii) a value of 25% in liquid reassociation kinetics between HPV-18 and HPV-45 and (iv) differences in intensities of hybridization with selected tissue DNAs. The prevalence of HPV-45 infection in the genital tract was low. In tests of over 600 tissue DNAs from female genital tract lesions, HPV-45 sequences were detected in three additional tissues, one each of invasive cervical carcinoma, condyloma, and normal cervical epithelium. HPV-45 is a newly recognized papillomavirus which rarely infects the genital tract and is associated with lesions across a wide histological spectrum.
Asunto(s)
Papillomaviridae/genética , Enfermedades del Cuello del Útero/microbiología , Mapeo Cromosómico , Enzimas de Restricción del ADN , Femenino , Humanos , Hibridación de Ácido Nucleico , Papillomaviridae/clasificación , Homología de Secuencia de Ácido NucleicoRESUMEN
Human papillomavirus deoxyribonucleic acid (DNA) was identified by Southern blot hybridization in 21 of 24 patients with multicentric anogenital lesions and in 46 of 61 individual lesions. Type 6/11 was present in nine patients, type 16 in one, an undetermined type in one, and more than one type in ten patients. Mixed types were present in eight of 46 virus-positive individual lesions. Abnormal mitotic figures were found in 16, 87, and 75% of lesions associated with type 6/11, type 16, and mixed types, respectively. Colposcopic presentation or location of lesions was not predictive of viral types. The relatively high rate of mixed human papillomavirus types in multicentric lesions and in single lesions, and the lack of absolute correlation between viral types and abnormal mitotic figures, suggest that lesions should be removed to prevent viral transmission and possible progression to carcinoma.
Asunto(s)
Neoplasias de los Genitales Femeninos/microbiología , Mitosis , Lesiones Precancerosas/microbiología , Infecciones Tumorales por Virus/microbiología , Biopsia , Cuello del Útero/microbiología , Cuello del Útero/patología , Colposcopía , Condiloma Acuminado/diagnóstico , Condiloma Acuminado/microbiología , Condiloma Acuminado/patología , ADN Viral/análisis , Epitelio/microbiología , Epitelio/patología , Femenino , Neoplasias de los Genitales Femeninos/diagnóstico , Neoplasias de los Genitales Femeninos/patología , Humanos , Papillomaviridae/análisis , Lesiones Precancerosas/diagnóstico , Lesiones Precancerosas/patología , Infecciones Tumorales por Virus/diagnóstico , Infecciones Tumorales por Virus/patologíaRESUMEN
Tissues from two cases of bowenoid papulosis of the vulva with coexistent invasive squamous cell carcinoma were evaluated for the presence of human papillomaviruses (HPVs) and for nuclear DNA content. In both cases, HPV type 16 and nuclear aneuploidy were found in bowenoid papulosis as well as in invasive carcinoma. Patterns of hybridization suggested that the viral genome was integrated into the cellular genome in both bowenoid papulosis tissues as well as in invasive carcinoma tissues. These observations suggest that lesions designated as bowenoid papulosis may have invasive cancer potential. The term vulvar intraepithelial neoplasia seems to be more appropriate for HPV-16-containing aneuploid, intraepithelial lesions of the vulva.
Asunto(s)
Enfermedad de Bowen/microbiología , Carcinoma de Células Escamosas/complicaciones , Carcinoma de Células Escamosas/microbiología , Papillomaviridae/genética , Neoplasias Cutáneas/microbiología , Neoplasias de la Vulva/complicaciones , Adulto , Anciano , Anciano de 80 o más Años , Aneuploidia , Enfermedad de Bowen/complicaciones , ADN de Neoplasias/genética , ADN Viral/genética , Epitelio/microbiología , Femenino , Genes Virales , Humanos , Hibridación Genética , Neoplasias Cutáneas/complicacionesRESUMEN
An infant boy born of a mother who had condylomata (genital warts) during pregnancy and at delivery developed recurrent conjunctival papillomas and papillomas on the soft palate and the false vocal cords. A conjunctival lesion was first noticed by the mother when the infant was 4 months old and was excised and histologically diagnosed as a papilloma when he was 11 months old. The DNA sequences of genital tract human papillomavirus type 6 (HPV-6) were identified in conjunctival papilloma tissue by Southern transfer hybridization of tissue DNA extracted from a lesion excised at 29 months of age as well as by in situ hybridization of paraffin sections of the diagnostic biopsy specimen obtained at 11 months of age. It is probable that the infant acquired conjunctival infection from the mother, very likely during passage through the infected birth canal.
Asunto(s)
Neoplasias de la Conjuntiva/microbiología , Genitales Femeninos/microbiología , Intercambio Materno-Fetal , Papiloma/microbiología , Papillomaviridae/clasificación , Condiloma Acuminado/microbiología , Neoplasias de la Conjuntiva/diagnóstico , Neoplasias de la Conjuntiva/patología , Femenino , Humanos , Lactante , Papiloma/diagnóstico , Papiloma/patología , Papiloma/transmisión , Embarazo , RecurrenciaRESUMEN
Genital tract papillomas in five children were examined for the presence of human papillomavirus (HPV) DNA by molecular hybridization. Papillomavirus DNA was detected in each sample and was identified as HPV-6 (three cases), HPV-6 or HPV-11 (one case), or HPV-16 (one case). These viruses are the same as are responsible for genital papillomas (condylomata) of adults. The transmission of adult genital tract viruses to children occurs primarily by a venereal route but may occur by a nonvenereal route.
Asunto(s)
Condiloma Acuminado/microbiología , Neoplasias de los Genitales Femeninos/microbiología , Neoplasias de los Genitales Masculinos/microbiología , Niño , Abuso Sexual Infantil , Preescolar , Condiloma Acuminado/transmisión , Femenino , Neoplasias de los Genitales Femeninos/transmisión , Neoplasias de los Genitales Masculinos/transmisión , Humanos , Masculino , Papillomaviridae/aislamiento & purificaciónRESUMEN
Warty lesions of the oral cavity were examined for etiologic association with genital tract papillomaviruses HPV-6, HPV-11, and HPV-16. DNAs extracted from ten oral biopsies were screened for HPV genomic sequences by Southern transfer hybridization with 32P-labeled viral DNA probes. Nonstringent hybridization with an HPV-6 probe revealed papillomavirus DNA sequences in four of seven tissues with histologic evidence of papillomatosis, in none of two tissues without histologic evidence of papillomatosis, and in one tissue that was not examined by histology. Stringent hybridization tests with HPV-6 and HPV-16 probes identified the genome in one tissue as being HPV-16, in a second tissue as being HPV-6 subtype a, and in a third tissue as HPV-6 (subtype unidentified); papillomavirus DNA sequences in two tissues are as yet not identified. An additional case of HPV-6 or HPV-11 related oral cavity lesion was diagnosed by in situ hybridization of paraffin sections with a 35S-labeled, mixed HPV-6 + HPV-11 probe. The hybridization in the positive section was extensive and confined to epithelial nuclei. The oral lesions associated with genital tract papillomaviruses were asymptomatic, multiple or single, and were located in different parts of the oral cavity, for example, on the gingivae, on the tongue, on the lip, on the tonsillar pillar, and on the floor of the mouth.
Asunto(s)
Genitales Femeninos/microbiología , Genitales Masculinos/microbiología , Enfermedades de la Boca/microbiología , Papillomaviridae/aislamiento & purificación , Verrugas/microbiología , Adolescente , Adulto , Antígenos Virales/análisis , Secuencia de Bases , Niño , ADN Viral/análisis , Femenino , Humanos , Masculino , Neoplasias de la Boca/microbiología , Hibridación de Ácido Nucleico , Papillomaviridae/genética , Papillomaviridae/inmunología , Infecciones Tumorales por Virus/microbiologíaRESUMEN
Cervical Papanicolaou smears and paraffin sections of biopsy specimens obtained from women attending dysplasia clinics were examined for viral DNA sequences by in situ hybridization technique using 35S-labeled cloned recombinant DNA probes of human papillomavirus (HPV) types 6, 11, and 16. These and one unrelated DNA probe complementary to measles virus RNA were labeled by nick translation using either one or two 35S-labeled nucleotides. The radiolabeled probes were reduced in size with DNase to 60-160 nucleotides. Paraffin sections and cervical smears were collected on pretreated slides, hybridized with the probes under stringent or nonstringent conditions for 50 h, and autoradiographed. Additional cervical specimens from the same women were examined for the presence of genus-specific papillomavirus capsid antigen by the immunoperoxidase technique. Preliminary results may be summarized as follows. The infecting virus could be identified in smears as well as in sections. Viral DNA sequences were detected only when there were condylomatous cells in the specimen and in only a proportion of the condylomatous cells. Even under stringent conditions, some specimens reacted with both HPV-6 and HPV-11. None of the specimens hybridized with HPV-16 or with the unrelated probe. In some instances, the cells did not hybridize with any of the three probes even when duplicate specimens contained frankly condylomatous, capsid antigen-positive cells. In situ hybridization of Papanicolaou smears or of tissue sections is a practical method for diagnosis and follow-up of specific papillomavirus infection using routinely collected material.
Asunto(s)
Cuello del Útero/microbiología , ADN , Hibridación de Ácido Nucleico , Prueba de Papanicolaou , Papillomaviridae/aislamiento & purificación , Radioisótopos de Azufre , Frotis Vaginal , Adulto , Femenino , Marcadores Genéticos , Humanos , Parafina , Enfermedades del Cuello del Útero/diagnósticoRESUMEN
Transfer into mouse and rat embryo fibroblasts in primary culture of cloned polyoma virus genes encoding only the large T protein led to the establishment of flat colonies in sparse subcultures at a frequency equal to that of transformation by wild-type virus. Cell lines could be derived from such colonies and maintained in culture for large numbers of generations without entering crisis. They exhibited a normal phenotype, by the criteria of growth on plastic to a low saturation density and of anchorage dependency. However, they required a lower serum concentration for growth than spontaneously established 3T3 cells. Similar results were obtained after transfer of recombinant DNA molecules encoding only the amino-terminal 40% of the large T protein, suggesting that this "immortalization" function corresponds to the activity of an amino-terminal domain of the protein. Immunoprecipitation analysis of T antigens in cell lines established after transfer of the full-size and of the truncated large T genes demonstrated the expression of the full-size large T protein and of a Mr 40,000 antigen expressed from the amino-terminal part of the gene, respectively. After transfer of a "large T only" plasmid that carries a tsa mutation, cell lines were established at 33 degrees C with the same efficiency as with the wild-type large T gene, but their growth was arrested after a shift to 40 degrees C, with a progressive loss in cell viability. This result indicates a continuous requirement for a large T function in the maintenance of "immortality."
Asunto(s)
Transformación Celular Neoplásica , Poliomavirus/genética , Proteínas Virales/genética , Animales , Antígenos Transformadores de Poliomavirus , Células Cultivadas , Enzimas de Restricción del ADN , ADN Recombinante/metabolismo , Embrión de Mamíferos , Fibroblastos/fisiología , Ratones , Plásmidos , RatasRESUMEN
A total of 284 isolates of gram-negative enteric bacilli associated with urinary tract infections or normal fecal flora were examined for resistance to antimicrobial agents and the presence of transmissible R factors. Klebsiella pneumoniae and Proteus isolates exhibited the highest frequencies of resistance to antimicrobial agents. The incidence of drug resistance among clinical isolates of Escherichia coli was considerably higher than its incidence among E. coli strains isolated from healthy individuals. Transmissible R factors mediating resistance to antimicrobial agents were demonstrated in 43% of resistant strains of E. coli isolated from urinary infections and 21 - 26% of those isolated from healthy individuals. Frequencies of R factor transfer among drug-resistant strains of K. pneumoniae and Proteus species were 26 and 28%, respectively.