RESUMO
The alpha-fetoprotein derived growth inhibitory peptide (GIP) is a 34-amino acid peptide composed of three biologically active subfragments. GIP-34 and its three constituent segments have been synthesized, purified, and studied for biological activity. The GIP-34 and GIP-8 have been characterized as anticancer therapeutic peptides. An multicenter study was initiated to elucidate the means by which these peptide drugs could be targeted to tumor cells. The study first established which cancer types were specifically targeted by the GIP peptides in both in vitro and in vivo investigations. It was next demonstrated that radiolabeled peptide ((125)I GIP-34) is specifically localized to rodent breast tumors at 24 h post-injection. The radionuclide studies also provided evidence for a proposed cell surface receptor; this was confirmed in a further study using fluorescent-labeled GIP-nanobeads which localized at the plasma membrane of MCF-7 breast cancer cells. Finally, it was readily demonstrated that GIP conjugated to either fluorescein or doxorubicin (DOX) underwent tumor cell uptake; subsequently, DOX-GIP conjugates induced cytotoxic cell destruction indicating the utility of GIP segments as cancer therapeutic agents. Following a discussion of the preceding results, a candidate cell surface receptor family was proposed which correlated with previous published reports for a putative AFP/GIP receptor.
Assuntos
Antineoplásicos/administração & dosagem , Inibidores do Crescimento/administração & dosagem , Fragmentos de Peptídeos/administração & dosagem , alfa-Fetoproteínas/química , Antineoplásicos/química , Antineoplásicos/metabolismo , Linhagem Celular Tumoral , Sistemas de Liberação de Medicamentos , Inibidores do Crescimento/química , Inibidores do Crescimento/metabolismo , Humanos , Estudos Multicêntricos como Assunto , Fragmentos de Peptídeos/química , Fragmentos de Peptídeos/metabolismo , alfa-Fetoproteínas/administração & dosagem , alfa-Fetoproteínas/metabolismoRESUMO
PURPOSE: Classic Kaposi's sarcoma (KS) is rare in children. Although its etiology is not fully understood, human herpesvirus 8 (HHV-8) is present in the angiogenic lesions. We report an HIV-negative, 13-year-old patient of Sicilian descent with HHV-8-associated classic KS to facilitate the diagnosis and treatment of this entity in children. EXPERIMENTAL DESIGN: DNA was extracted from the skin specimen of the patient and analyzed via PCR assay and Southern blot hybridization for HHV-8 DNA. The amplified HHV-8 DNA was cloned, sequenced, and compared with the prototype HHV-8-KS330/BAM. RESULTS: The patient presented with purpuric lesions on the distal lower extremities and the tip of his nose, associated with thrombocytopenia and leukopenia, suggesting an immune-mediated cytopenia. While on prednisone, he developed marked vascular proliferation in the groins. Biopsy of the skin lesions showed KS, and HHV-8 was detected in the tissues by PCR. Sequence analysis of the amplified DNA was homologous to the prototype HHV-8-KS330/BAM. His HHV-8 strain was the A subgroup, the type associated with Mediterranean classic KS. Stopping prednisone and treatment with IFN-alpha and IgG resulted in regression of the groin lesions. CONCLUSIONS: This report emphasizes the importance of recognizing classic KS in children and avoiding immunosuppressive therapies in indolent classic KS. The diagnostic and therapeutic strategies were effective and well tolerated.
Assuntos
Herpesvirus Humano 8 , Sarcoma de Kaposi/patologia , Neoplasias Cutâneas/patologia , Adolescente , Antivirais/uso terapêutico , Sequência de Bases , DNA Viral/genética , Herpesvirus Humano 8/efeitos dos fármacos , Herpesvirus Humano 8/genética , Humanos , Imunoglobulinas Intravenosas/uso terapêutico , Interferon-alfa/uso terapêutico , Masculino , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , Sarcoma de Kaposi/tratamento farmacológico , Sarcoma de Kaposi/virologia , Homologia de Sequência do Ácido Nucleico , Pele/efeitos dos fármacos , Pele/patologia , Neoplasias Cutâneas/tratamento farmacológico , Neoplasias Cutâneas/virologiaRESUMO
In order to assess the prevalence rate of HTLV-1-associated T-cell lymphomas and human retrovirus infection in general, approximately 21,000 individuals representing various patient populations, retroviral risk groups, and blood donors were examined for HTLV-I, HTLV-II, HIV-1, or HIV-2 infection using serologic and PCR assays. The prevalence rates among volunteer blood donors were 0.02% and 0% for HTLV and HIV, respectively. Significantly increased HTLV prevalence rates were observed among paid blood donors, African American health care clinic patients, Amerindians, recipients of HTLV-positive cellular blood products, intravenous drug users, sexual contacts and family members of HTLV-positive people, and patients with primary thrombocytosis and other-than-low-grade non-Hodgkin's lymphoma (NHL). Among some of these groups there were significant differences in the prevalence of HTLV-I versus HTLV-II. The eight HTLV-positive NHL patients all had mature, high-grade, CD4+ T-cell lymphomas with clonally integrated HTLV-I, for a prevalence of 4% among other-than-low-grade NHL patients. Seven of the eight died from their disease within 2 years despite treatment. Interestingly, two groups at risk for HTLV infection, namely needle stick victims and recipients of HTLV-infected and/or pooled plasma products, showed no evidence for infection. Significantly increased HIV-1 prevalence was observed among paid blood donors, African Americans, homosexuals, female prostitutes, hemophiliacs, and other-than-low-grade NHL patients. Only one patient was infected with HIV-2. Of the nine HIV-positive, other-than-low-grade NHL patients, seven HIV-1 positives had B-cell lymphomas, one HIV-1 positive had an HTLV-I-positive CD4+ T-cell lymphoma, and one infected with HIV-2 had a CD4+ T-cell lymphoma that was HTLV negative. The data indicate that HTLV-I lymphoma, while uncommon, is not necessarily rare among other-than-low-grade NHL cases in the United States and, given its poor prognosis, should probably be studied separately in clinical trials.
Assuntos
Leucemia-Linfoma de Células T do Adulto/epidemiologia , Infecções por Retroviridae/epidemiologia , Negro ou Afro-Americano , Agamaglobulinemia/epidemiologia , Doadores de Sangue , Comorbidade , DNA de Neoplasias/análise , DNA Viral/análise , Saúde da Família , Infecções por HIV/epidemiologia , Infecções por HIV/virologia , HIV-1/isolamento & purificação , HIV-2/isolamento & purificação , Infecções por HTLV-I/epidemiologia , Infecções por HTLV-II/epidemiologia , Hemofilia A/epidemiologia , Indígenas Norte-Americanos , Leucemia/epidemiologia , Leucemia-Linfoma de Células T do Adulto/etnologia , Linfoma/classificação , Linfoma/epidemiologia , Linfoma/etnologia , Linfoma/virologia , Linfoma Relacionado a AIDS/epidemiologia , Linfoma Relacionado a AIDS/etnologia , Linfoma Relacionado a AIDS/virologia , Ferimentos Penetrantes Produzidos por Agulha/complicações , Prevalência , Infecções por Retroviridae/etnologia , Infecções por Retroviridae/virologia , Doenças Reumáticas/epidemiologia , Fatores de Risco , Estudos Soroepidemiológicos , Comportamento Sexual , Abuso de Substâncias por Via Intravenosa , Trombocitose/epidemiologia , Reação Transfusional , Estados Unidos/epidemiologiaRESUMO
BACKGROUND: HTLV-I and HTLV-II are related exogenous pathogenic human retroviruses. Until recently, ELISAs based on HTLV-I antigens have been used to screen for the presence of HTLV-I or -II antibodies. The HTLV-I-based assays have not been as sensitive in detecting antibodies to HTLV-II as in detecting antibodies to HTLV-I. The Abbott HTLV-I/HTLV-II ELISA uses a combination of HTLV-I and HTLV-II antigens to detect antibodies to the whole HTLV group. The performance of this ELISA was compared to that of several HTLV-I-based serologic assays and an HTLV-II PCR assay in cohorts of South American Indians and New York City IV drug users (IVDUs) in whom HTLV-II is endemic. STUDY DESIGN AND METHODS: Sera from 429 South American Indians and New York City IVDUs were evaluated for HTLV antibodies by the use of three ELISAs (rgp21-enhanced HTLV-I/II, Cambridge Biotech; Vironostika HTLV-I/II, Organon Teknika; and HTLV-I/HTLV-II, Abbott), and a Western blot (WB) assay. Peripheral blood leukocyte DNA from each person was analyzed for HTLV-I and HTLV-II pol DNA via PCR. The HTLV-II PCR-positive samples were further subtyped via cloning and sequencing and/or oligomer restriction. RESULTS: Two hundred four samples (48%) were positive for HTLV-II by serologic and/or PCR assays. All of the positive samples from the Indians and approximately one-third of the positive samples from the IVDUs were of the HTLV-IIB subtype. Comparative analyses indicate that the sensitivity and specificity of the various assays were: PCR, 98 and 100 percent; Abbott HTLV-I/HTLV-II, 78 and 95 percent; Cambridge Biotech HTLV-I/II, 76 and 96 percent; Vironostika HTLV-I/II, 71 and 98 percent; and WB, 73 and 100 percent, respectively. CONCLUSION: There were no significant differences among the sensitivities and specificities of the HTLV-I/II ELISAs (p values, 0.056-0.438). The WB and PCR assays were much more specific than the other serologic assays (p=0.03). However, the PCR assay is significantly (p<0.001) more sensitive than any of the serologic assays in the detection of HTLV-II infection. Thus, optimal detection of HTLV-II infection would seem to require both serologic and DNA PCR assays.
Assuntos
Anticorpos Anti-HTLV-I/imunologia , Anticorpos Anti-HTLV-II/imunologia , Infecções por HTLV-II/genética , Ensaio de Imunoadsorção Enzimática , Humanos , Reação em Cadeia da Polimerase , Estatística como AssuntoRESUMO
The primate T-cell lymphoma/leukaemia viruses (PTLV) and bovine leukaemia virus (BLV) comprise a unique genus of retroviruses, infection with which induces seroreactivity in the host against conserved epitopes in their p24 gag and gp21 env cognate proteins. Herein, we have confirmed this serocrossreactivity. Patients with large granular lymphocyte (LGL) leukaemia have frequent seroreactivity to the p24 and gp21 env proteins of human T-cell lymphoma/leukaemia virus I (HTLV-I), one of the species in the genus. However, only a small minority of patients are actually infected with prototypic HTLV-I or HTLV-II, another species within the group. In an attempt to determine whether LGL leukaemia might be associated with other members of the PTLV/BLV genus, we examined the peripheral blood mononuclear cell DNA of 22 HTLV p24 and/or gp21 seropositive LGL leukaemia patients via PCR using degenerate and specific primer pair/probe systems capable of detecting all known members of the PTLV/BLV genus. None of the samples was positive. These data indicate that although HTLV-II may be associated with some cases of LGL leukaemia most patients are not infected with a PTLV or BLV virus.
Assuntos
DNA Viral/sangue , Deltaretrovirus/genética , Vírus da Leucemia Bovina/genética , Leucemia Mieloide/virologia , Autorradiografia , Reações Cruzadas , Primers do DNA , Sondas de DNA , Deltaretrovirus/imunologia , Produtos do Gene gag/imunologia , Humanos , Vírus da Leucemia Bovina/imunologia , Leucemia Mieloide/imunologia , Reação em Cadeia da Polimerase/métodos , Proteínas do Envelope Viral/imunologiaRESUMO
We describe a multiplex nucleic acid assay that identifies and determines the abundance of four different pathogenic retroviruses (HIV-1, HIV-2, and human T-lymphotrophic virus types I and II). Retroviral DNA sequences are amplified in a single, sealed tube by simultaneous PCR assays, and the resulting amplicons are detected in real time by the hybridization of four differently colored, amplicon-specific molecular beacons. The color of the fluorescence generated in the course of amplification identifies which retroviruses are present, and the number of thermal cycles required for the intensity of each color to rise significantly above background provides an accurate measure of the number of copies of each retroviral sequence that were present originally in the sample. Fewer than 10 retroviral genomes can be detected. Moreover, 10 copies of a rare retrovirus can be detected in the presence of 100, 000 copies of an abundant retrovirus. Ninety-six samples can be analyzed in 3 hr on a single plate, and the use of a closed-tube format eliminates crossover contamination. Utilizing previously well characterized clinical samples, we demonstrate that each of the pathogenic retroviruses can be identified correctly and no false positives occur. This assay enables the rapid and reliable screening of donated blood and transplantable tissues.
Assuntos
DNA Viral/sangue , Genes gag , HIV-1/isolamento & purificação , HIV-2/isolamento & purificação , Vírus Linfotrópico T Tipo 1 Humano/isolamento & purificação , Vírus Linfotrópico T Tipo 2 Humano/isolamento & purificação , Sequência de Bases , Sequência Conservada , Primers do DNA , DNA Viral/genética , HIV-1/genética , HIV-2/genética , Vírus Linfotrópico T Tipo 1 Humano/genética , Vírus Linfotrópico T Tipo 2 Humano/genética , Humanos , Reação em Cadeia da Polimerase/métodos , Reprodutibilidade dos Testes , Espectrometria de Fluorescência/métodos , Moldes GenéticosRESUMO
Lymphoma presenting as a soft tissue mass is rare and thus may be confused with the more common soft tissue sarcoma. No previous analysis of the clinical and radiologic features of lymphomas presenting as soft tissue masses is available because most of the cases reviewed are from the pathology literature. Four patients with diagnoses of extranodal lymphomas of the soft tissues were reviewed retrospectively with respect to their clinical features, primary tumor characteristics, stage, radiographic characteristics, treatment, and followup. Mean age was 72.5 years (range, 52-85 years). The soft tissue mass occurred in the thigh (three cases) and shoulder (one case). The median size of the soft tissue mass was 6.7 cm (range, 2-15 cm) in the largest dimension, as measured on magnetic resonance imaging. These patients each had evidence of lymphadenopathy at the time of diagnosis. Lactate dehydrogenase was increased significantly in two cases and increased slightly in two other cases. One case was Stage II(E) at presentation, one was Stage III(E), and two were Stage IV. All were B cell immunophenotype. All patients died between 2 and 24 months after diagnosis, despite the use of Cytoxan, vincristine, adriamycin, and prednisone chemotherapy in each case. Clinical and radiographic features that favor extranodal soft tissue lymphoma over sarcoma include pain and tenderness, lymphadenopathy (particularly when confluent radiologically), ipsilateral extremity swelling, and elevated lactate dehydrogenase.
Assuntos
Linfoma/diagnóstico , Neoplasias de Tecidos Moles/diagnóstico , Idoso , Idoso de 80 Anos ou mais , Evolução Fatal , Feminino , Humanos , L-Lactato Desidrogenase/análise , Linfoma/diagnóstico por imagem , Linfoma/enzimologia , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Radiografia , Estudos Retrospectivos , Neoplasias de Tecidos Moles/diagnóstico por imagem , Neoplasias de Tecidos Moles/enzimologiaRESUMO
PURPOSE: The aim of this study was to investigate the prognostic importance of codon 12 K-ras mutations in patients with early-stage non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: We identified 260 patients with surgically resected stage I (n = 193) and stage II (n = 67) NSCLC with at least a 5-year follow-up. We performed polymerase chain reaction analysis of DNA obtained from paraffin-embedded NSCLC tissue, using mutation-specific probes for codon 12 K-ras. RESULTS: K-ras mutations were detected in 35 of 213 assessable specimens (16.4%). K-ras mutations were detected in 27 of 93 adenocarcinomas (29.0%), one of 61 squamous cell carcinomas (1.6%), five of 39 large-cell carcinomas (12.8%), and two of 20 adenosquamous carcinomas (10%) (P = .001). G to T transversions accounted for 71% of the mutations. There was no statistically significant difference in overall survival for all patients with K-ras mutations (median survival, 39 months) compared with patients without K-ras mutations (median survival, 53 months; P = .33). There was no statistically significant difference in overall or disease-free survival for subgroups with stage I disease, adenocarcinoma, or non-squamous cell carcinoma or for specific amino acid substitutions. The median survival time for stage II patients with K-ras mutations was 13 months, compared with 38 months for patients without K-ras mutations (P = .03). CONCLUSION: Codon 12 K-ras mutations were more common in adenocarcinomas than in squamous cell carcinomas. For the subgroup with stage II NSCLC, there was a statistically significant adverse effect on survival for the presence of K-ras mutations. However, when the entire group was considered, the presence of K-ras mutations was not of prognostic significance in this cohort of patients with resected early-stage NSCLC.
Assuntos
Carcinoma Pulmonar de Células não Pequenas/genética , Carcinoma Pulmonar de Células não Pequenas/patologia , Códon , Genes ras , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , Mutação , Adulto , Idoso , Idoso de 80 Anos ou mais , Southern Blotting , Carcinoma Pulmonar de Células não Pequenas/cirurgia , DNA de Neoplasias/análise , DNA de Neoplasias/genética , Intervalo Livre de Doença , Feminino , Humanos , Neoplasias Pulmonares/cirurgia , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Reação em Cadeia da Polimerase , PrognósticoRESUMO
A polyclonal CD3(+), CD8(+) T-cell line, G2, was derived from the peripheral blood of a seropositive, PCR-positive, HTLV-IIB infected Guahibo Indian from Venezuela. The cell line is productively infected with HTLV-IIB. The entire HTLV-II G2 proviral DNA was sequenced via PCR using overlapping HTLV-II primer pairs. Phylogenetic analyses indicate that HTLV-II G2 is the most divergent HTLV-IIB strain identified to date. Characterization of its deduced proteins and its relationship to other members of the PTLV/BLV genus of retroviruses are discussed.
Assuntos
Genoma Viral , Vírus Linfotrópico T Tipo 2 Humano/genética , Indígenas Sul-Americanos , Sequência de Aminoácidos , Células Cultivadas , DNA Viral/análise , Vírus Linfotrópico T Tipo 2 Humano/classificação , Vírus Linfotrópico T Tipo 2 Humano/isolamento & purificação , Humanos , Imunofenotipagem , Dados de Sequência Molecular , Homologia de Sequência de Aminoácidos , VenezuelaRESUMO
Human herpesvirus 8 (HHV-8) has been proposed as a sexually transmitted etiologic agent of Kaposi's sarcoma (KS). In this study, by use of a sensitive polymerase chain reaction assay, HHV-8 DNA was detected in the skin lesions (92%), normal skin (23%), peripheral blood mononuclear cells (PBMC) (46%), plasma (7%), saliva (37%), and semen (12%) but not stool samples from KS patients. The average number of HHV-8 copies per microgram of positive target DNA was 64, 000, 9000, 40, 33,000, and 300 for skin, PBMC, plasma, saliva, and semen samples, respectively. Only 1 non-KS donor sample, of saliva, was positive for HHV-8. Sequencing showed 5% divergence among HHV-8 strains. The data suggest that saliva may be more important than semen or stool in the sexual transmission of HHV-8. The relatively high prevalence of HHV-8 in PBMC raises the question as to why there is no evidence for bloodborne virus transmission.
Assuntos
DNA Viral/análise , Herpesvirus Humano 8/isolamento & purificação , Sarcoma de Kaposi/virologia , DNA Viral/sangue , Soronegatividade para HIV , Soropositividade para HIV/sangue , Soropositividade para HIV/virologia , Infecções por Herpesviridae/complicações , Infecções por Herpesviridae/diagnóstico , Infecções por Herpesviridae/transmissão , Herpesvirus Humano 8/genética , Humanos , Masculino , Monócitos/virologia , Filogenia , Reação em Cadeia da Polimerase/métodos , Valores de Referência , Saliva/virologia , Sarcoma de Kaposi/etiologia , Sêmen/virologia , Sensibilidade e Especificidade , Comportamento Sexual , Infecções Sexualmente Transmissíveis/transmissão , Infecções Sexualmente Transmissíveis/virologia , Pele/virologiaRESUMO
Sera from approximately 50% of patients with large granular lymphocyte (LGL) leukaemia react with a recombinant human T-cell leukaemia/lymphoma virus (HTLV) transmembrane envelope protein, p21e. Two immunodominant epitopes within env p21e have been defined by reactivity against recombinant proteins GD21 and BA21. In this study sera from 41 patients with LGL leukaemia were examined for reactivity against these recombinant HTLV env proteins. Overall, 21/41 (51%) sera reacted to p21e. Only two sera reacted to GD21. The predominant immunoreactivity against p21e was directed against the BA21 epitope, with 19/41 (46%) sera being BA21 positive. Seroconversion to BA21 protein was also documented. PCR analyses confirmed the low incidence of protypical HTLV sequences (2/41, 5%). These data document an association between BA21 seroreactivity and LGL leukaemia. This finding raises the possibility that such BA21 seroreactivity could be due to cross-reactivity to a cellular or retroviral antigen sharing some amino acid homology with the transmembrane glycoprotein of HTLV.
Assuntos
Mapeamento de Epitopos , Epitopos/imunologia , Produtos do Gene env/imunologia , Vírus Linfotrópico T Tipo 1 Humano/imunologia , Leucemia Linfoide/imunologia , Proteínas Oncogênicas de Retroviridae/imunologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Western Blotting , Feminino , Humanos , Leucemia-Linfoma de Células T do Adulto/imunologia , Masculino , Pessoa de Meia-Idade , Produtos do Gene env do Vírus da Imunodeficiência HumanaRESUMO
Post-transplantation lymphoproliferative disorders (PTLD) are a clinicopathologically heterogeneous group of lymphoid proliferations. The majority are of B-cell origin and associated with Epstein-Barr virus (EBV) infection. In contrast, the development of T-cell PTLD is much less common and EBV does not appear to be involved in pathogenesis. In this report we describe three patients who developed large granular lymphocyte (LGL) leukaemia after renal transplantation. These patients had clonal expansion of CD3+, CD8+, CD57+, CD56- LGL. We were unable to detect CMV antigen or find evidence for EBV or human T-cell leukaemia/lymphoma virus genome in the LGL from these patients. These data show that LGL leukaemia should be included as one of the types of T-cell proliferations which can occur post transplant.
Assuntos
Transplante de Rim/efeitos adversos , Leucemia Linfoide/virologia , Adulto , Idoso , Antígenos CD/análise , Western Blotting , Citomegalovirus/isolamento & purificação , Infecções por Citomegalovirus/complicações , DNA Viral/análise , Feminino , Rearranjo Gênico da Cadeia beta dos Receptores de Antígenos dos Linfócitos T , Genoma Viral , Infecções por HTLV-I/complicações , Infecções por HTLV-II/complicações , Infecções por Herpesviridae/complicações , Herpesvirus Humano 4/isolamento & purificação , Vírus Linfotrópico T Tipo 1 Humano/isolamento & purificação , Vírus Linfotrópico T Tipo 2 Humano/isolamento & purificação , Humanos , Terapia de Imunossupressão/efeitos adversos , Masculino , Infecções Oportunistas/complicações , Receptores de Antígenos de Linfócitos T/análiseRESUMO
Peripheral blood mononuclear cells from asymptomatic HTLV-II-infected and uninfected Gran Chaco Amerindians were analyzed using polymerase chain reaction (PCR) for expansions of T-cell receptor (TCR) V-beta gene clonotypes. Analyses were performed using primer pairs designed to identify expanded T-cell familial clonotypes based on their unique TCR beta gene rearrangements. Of the 30 HTLV-IIB-positive samples tested, five showed evidence of V-beta clonotypic T-cell expansion. Of the five expansions, two were monoclonotypic and the remaining three were oligoclonotypic. In comparison, 30 HTLV-II-negative Amerindians showed no evidence of clonotypic T-cell expansion. Amplified DNA from one of the monoclonotypic samples was subsequently cloned and sequenced and was found to have uniform variable/ diversity/joining sequences confirming its unique monoclonal T-cell expansion. This method of detecting clonal TCR beta gene rearrangements has the advantage over traditional Southern blot techniques of being more sensitive and specific even with suboptimal specimens. The prognostic significance of clonotypic T-cell expansion in a group such as the HTLV-II-infected Gran Chaco Amerindians remains to be determined.
Assuntos
Rearranjo Gênico da Cadeia beta dos Receptores de Antígenos dos Linfócitos T/genética , Indígenas Sul-Americanos , Leucemia de Células T/etnologia , Leucemia de Células T/imunologia , Linfócitos T/imunologia , Argentina/epidemiologia , Sequência de Bases , Clonagem Molecular , DNA/análise , Primers do DNA/química , Genes Codificadores da Cadeia beta de Receptores de Linfócitos T/genética , Vírus Linfotrópico T Tipo 2 Humano , Humanos , Imunofenotipagem , Dados de Sequência Molecular , Paraguai/epidemiologia , Reação em Cadeia da Polimerase , Receptores de Antígenos de Linfócitos T alfa-beta/genéticaRESUMO
The peripheral blood of 41 Yaruro and Guahibo Indians from Venezuela was examined for HTLV antibodies and DNA. Twenty-five samples (61%) were found to be infected with HTLV-IIB. The sensitivities of the serologic and DNA polymerase chain reaction (PCR) analyses were 80% and 96%, respectively. Epidemiologic studies supported both sexual and perinatal transmission of the virus. Sequence analyses of the HTLV-IIB strains from these Indians indicate that they are unique relative to HTLV-II detected in other groups of humans. HTLV-IIB-G2 isolated from a Guahibo Indian is the most divergent HTLV-IIB strain relative to the prototype HTLV-II NRA.
Assuntos
Infecções por HTLV-II/etnologia , Vírus Linfotrópico T Tipo 2 Humano/genética , Indígenas Sul-Americanos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Animais , Sequência de Bases , Cricetinae , Transmissão de Doença Infecciosa , Infecções por HTLV-II/genética , Infecções por HTLV-II/transmissão , Vírus Linfotrópico T Tipo 2 Humano/classificação , Humanos , Transmissão Vertical de Doenças Infecciosas , Pessoa de Meia-Idade , Dados de Sequência Molecular , Linhagem , Filogenia , Reação em Cadeia da Polimerase , Prevalência , Homologia de Sequência do Ácido Nucleico , Estudos Soroepidemiológicos , Venezuela/epidemiologiaRESUMO
Natural killer (NK) cells are CD3- large granular lymphocytes (LGL) responsible for immunity against viral infections. A chronic lymphoproliferative disorder of NK cells has been described in which the expanded NK cells display a restricted phenotype and cytotoxic activity. These data raise the hypothesis that proliferating LGL in these patients result from discrete expansions of NK cells responding to an unknown, perhaps viral, antigen. Recently, it was found that mice transgenic for the tax gene of human T-cell leukemia/lymphoma virus (HTLV) develop NK leukemia. Therefore, we studied 15 patients with chronic NK lymphoproliferative disorder for evidence of HTLV infection. Sera were tested using an HTLV-I/II-enzyme linked immunosorbent assay and a modified Western blot assay containing recombinant env proteins. None of the sera met conventional criteria for HTLV seroreactivity. However, sera from 11 patients (73%) reacted with the recombinant HTLV env protein p21E. The anti-p21E reactivity of these sera was then mapped employing the recombinant proteins GD21 and BA21. No reactivity to the immunodominant HTLV epitope GD21 was observed, suggesting that prototypical HTLV infection is unlikely in these patients. This was confirmed by finding no evidence for HTLV nucleic acids by PCR analyses employing primers specific for conserved regions in the env, pol, and pX genes. In contrast, 10 of the 15 sera reacted with the epitope BA21, documenting for the first time an association between a unique seroreactivity and disease. The high incidence of BA21 seroreactivity in these patients suggests that exposure to a protein containing homology to BA21 may be important in the pathogenesis of this lymphoproliferative disorder.
Assuntos
Anticorpos Antivirais/sangue , Vírus Linfotrópico T Tipo 1 Humano/imunologia , Células Matadoras Naturais/imunologia , Transtornos Linfoproliferativos/imunologia , Proteínas do Envelope Viral/imunologia , Animais , Anticorpos Antivirais/imunologia , Antígenos Virais/imunologia , Complexo CD3 , Feminino , Granulócitos/imunologia , Granulócitos/patologia , Humanos , Células Matadoras Naturais/patologia , Transtornos Linfoproliferativos/sangue , Camundongos , Proteínas Recombinantes/imunologiaRESUMO
Degenerate and specific PCR assays were developed for bovine leukaemia virus (BLV) and/or primate T cell leukaemia/lymphoma viruses (PTLV). The degenerate assays detected all major variants of the BLV/PTLV genus at a sensitivity of 10-100 copies of input DNA; the specific systems detected 1-10 copies of input target. Sensitivity was 100% in specific DNA-PCR assays done on peripheral blood from seropositive BLV-infected cattle and HTLV-I- or HTLV-II-infected humans, and 62% in RNA/DNA-PCR assays on sera from BLV seropositive cattle. The pol fragments from 21 different BLV strains, isolated from cattle in North and Central America, were cloned and sequenced, and compared to other published BLV and PTLV pol sequences. BLV and PTLV sequences differed by 42%. Sequence divergence was up to 6% among the BLV strains, and up to 36% among the PTLV strains (with PTLV-I and PTLV-II differing among themselves by 15% and 8%, respectively). Some cows were infected with several BLV strains. Among retroviruses, BLV and PTLV sequences formed a distinct clade. The data support the interpretation that BLV and PTLV evolved from a common ancestor many millennia ago, and some considerable time before the PTLV-I and PTLV-II strains diverged from each other. The dissemination of the BLV strains studied probably resulted from the export of European cattle throughout the world over the last 500 years. The relatively similar mutation rates of BLV and PTLV, after their various points of divergence, suggest that there could be a much wider genetic range of BLV than has currently been defined.
Assuntos
Vírus da Leucemia Bovina/isolamento & purificação , Reação em Cadeia da Polimerase , Primatas/virologia , Retroviridae/isolamento & purificação , Sequência de Aminoácidos , Animais , Sequência de Bases , Bovinos , Linhagem Celular , Humanos , Vírus da Leucemia Bovina/classificação , Vírus da Leucemia Bovina/genética , Dados de Sequência Molecular , Filogenia , Retroviridae/classificação , Retroviridae/genéticaRESUMO
The etiology of large granular lymphocyte (LGL) leukemia is uncertain. Recently, a Kaposi's sarcoma-associated herpes virus, denoted as human herpes virus 8 (HHV-8), has been identified. Some data suggest that HHV-8 and Epstein-Barr virus (EBV) may interact to induce malignant transformation. Infection with EBV has been implicated in the pathogenesis of some cases of LGL leukemia. Therefore, we performed PCR analyses for HHV-8 detection in samples from nineteen patients with LGL leukemia; three of these samples contained the EBV genome. We could not detect HHV-8 sequences in any of these patients. Therefore, HHV-8 infection is not involved in the pathogenesis of T-LGL leukemia.
Assuntos
DNA Viral/genética , Herpesvirus Humano 4/isolamento & purificação , Herpesvirus Humano 8/isolamento & purificação , Leucemia Linfoide/virologia , Herpesvirus Humano 4/crescimento & desenvolvimento , Herpesvirus Humano 8/genética , HumanosRESUMO
BACKGROUND: Human T-cell lymphoma/leukemia viruses types I and II (HTLV-I and HTLV-II) are related exogenous human retroviruses. The former is definitely pathogenic while disease association with the latter is unclear. There are two subtypes of HTLV-II, A and B. Currently, enzyme-linked immunosorbent assays (ELISAs) based on HTLV-I antigens are used to screen for the presence of HTLV-I and -II antibodies. Confirmation and subtyping are accomplished by Western blot (WB) or ELISAs based on HTLV-I whole viral antigens and/or HTLV-I and HTLV-IIA peptides. The sensitivity and specificity of these serologic assays were compared to those of HTLV-I and-II-specific polymerase chain reaction (PCR) assays in tests on samples from Indians from South America in whom the HTLV-IIB subtype is endemic. STUDY DESIGN AND METHODS: Sera from 246 Gran Chaco Indians were evaluated for HTLV antibodies with the use of four ELISAs (Retrotek HTLV-I; Cambridge Biotech rgp21 enhanced HTLV-I/II; Vironostika HTLV-I/II; and Select HTLV-I/II), and a WB assay. Peripheral blood leukocyte DNA from each Indian was analyzed for HTLV-I or HTLV-II pol DNA via PCR. Fifteen of the PCR-positive samples were further subtyped via cloning and sequencing and/or oligomer restriction. RESULTS: Ninety-seven samples (39%) were positive for HTLV-II by serologic and/or PCR assays. All 15 positive DNA samples that were further analyzed were of the HTLV-IIB subtype and were clustered as a highly conserved phylogenetic group. Comparative analyses indicate that the sensitivity and specificity of the various assays were: PCR, 97 and 100 percent; Retrotek, 70 and 91 percent; Cambridge Biotech, 74 and 96 percent; Vironostika, 73 and 99 percent; Select 72 and 98 percent; and WB, 70 and 100 percent. CONCLUSION: The sensitivities of the tested HTLV serologic assays were comparable. However, the specificity of the Retrotek ELISA was significantly lower than that of the others. When positive, the subtyping assays were very specific. However, PCR assays would seem preferable or to be a necessary adjunct for the sensitive detection of HTLV-IIB infection.
Assuntos
Infecções por HTLV-II/etnologia , Infecções por HTLV-II/epidemiologia , Indígenas Sul-Americanos , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Sequência de Aminoácidos , Western Blotting , Criança , Pré-Escolar , Ensaio de Imunoadsorção Enzimática , Reações Falso-Negativas , Soronegatividade para HIV , Soropositividade para HIV/diagnóstico , Infecções por HTLV-II/diagnóstico , Humanos , Pessoa de Meia-Idade , Dados de Sequência Molecular , Reação em Cadeia da Polimerase , PrevalênciaRESUMO
The DNA sequences of a novel human gamma-herpesvirus type 8 (HHV-8) have recently been detected in Kaposi's sarcoma (KS) lesions obtained from different populations in whom this neoplasm occurs, suggesting that this virus may be implicated in the etiology and/or pathogenesis of KS. To study the distribution and possible means of transmission of the putative viral agent, specimens of KS skin lesions, matched uninvolved skin, peripheral blood mononuclear cells (PB-MCs), and semen were collected from 12 HIV-positive homosexual men with acquired immune deficiency syndrome (AIDS)-related KS (AIDS-KS) and 2 human immunodeficiency virus (HIV)-negative homosexual men with KS. HHV-8 virus DNA was detected by polymerase chain reaction (PCR) studies in all 14 of these KS specimens and in 6 of 14 biopsies of normal-appearing skin distant from any KS lesions including 1 uninvolved skin specimen from an HIV-negative homosexual male with KS. In addition, 3 of 12 PBMC samples and 3 of 12 semen samples from the AIDS-KS patients were positive for HHV-8. The DNA sequences of HHV-8 were not detected in the matched semen and PBMC specimens obtained from 2 HIV-negative homosexual men with KS, 4 HIV-positive homosexual patients without KS, 2 HIV-seronegative healthy homosexual men, 5 HIV-positive heterosexual male intravenous drug users, or 5 healthy HIV-negative heterosexual donors. Using PCR in situ, positive signals for HHV-8 were demonstrated in the B lymphocyte subsets of PBMCs and/or in spermatozoa and mononuclear cells in the semen from some of the PCR-positive specimens from the AIDS-KS patients examined. These data show that HHV-8 is present in and could possibly be transmitted via semen and/or blood from some homosexual men with AIDS-KS.
Assuntos
Síndrome da Imunodeficiência Adquirida/sangue , Síndrome da Imunodeficiência Adquirida/virologia , DNA Viral/sangue , Herpesvirus Humano 8/genética , Sarcoma de Kaposi/sangue , Sarcoma de Kaposi/virologia , Sêmen/virologia , Síndrome da Imunodeficiência Adquirida/patologia , Herpesvirus Humano 8/isolamento & purificação , Humanos , Hibridização In Situ , Masculino , Reação em Cadeia da Polimerase , Sarcoma de Kaposi/patologia , Sêmen/química , Neoplasias Cutâneas/sangue , Neoplasias Cutâneas/patologia , Neoplasias Cutâneas/virologiaRESUMO
Human T cell leukemia/lymphoma virus (HTLV-II) type II infection was detected by polymerase chain reaction or serologic analyses (or both) in 22% of 697 Indians of six different ethnic back-grounds inhabiting the Argentinean and Paraguayan Gran Chaco. None was infected with HTLV-I. The prevalence of HTLV-II increased with age (14% in those < 13 years and 23% in those > or = 13 years). HTLV-II infection was found in all 20 Gran Chaco communities studied, but marked differences (44%-4%) in the rate of infection were observed even in communities separated by only a few miles. These variations correlated closely with ethnicity. In the high-incidence communities, infection clustered within families, with evidence for both sexual and perinatal transmission, primarily via breast-feeding. By contrast, only 2% of 94 Mapuche Indians from southern Argentina were positive for HTLV-II. Analyses of pol and long terminal repeat sequences from 15 Gran Chaco HTLV-II strains indicated that they constitute a highly conserved branch of the HTLV-IIB substrain.