RESUMO
A serological survey was performed to determine the prevalence of antibodies against human bocavirus in an Apulian population. Anti-hBoV IgG antibodies were analysed in 1206 inhabitants (age range, 1month-84years) using a standardized ELISA test based on the use of recombinant hBoV VP2 virus-like particles. In total, 1075 (89.1%) of 1206 participants (mean age 32±24.8years) displayed anti-hBoV-IgG. The seroprevalence increased significantly (p<0.0001) in children from 2-4years (64.2%) to 5-9years (96.4%). A similar trend was observed in both male and female subjects. In conclusion, our results show that hBoV infection is common in this population, especially in children.
Assuntos
Anticorpos Antivirais/sangue , Bocavirus Humano/imunologia , Infecções por Parvoviridae/epidemiologia , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Antígenos Virais/imunologia , Proteínas do Capsídeo/imunologia , Criança , Pré-Escolar , Ensaio de Imunoadsorção Enzimática , Feminino , Bocavirus Humano/isolamento & purificação , Bocavirus Humano/patogenicidade , Humanos , Imunoglobulina G/sangue , Lactente , Itália/epidemiologia , Masculino , Pessoa de Meia-Idade , Infecções por Parvoviridae/imunologia , Infecções por Parvoviridae/virologia , Prevalência , Estudos Soroepidemiológicos , Adulto JovemRESUMO
BACKGROUND AND OBJECTIVES: Due to their high resistance to inactivation procedures, nonenveloped viruses such as parvovirus B19, human bocavirus (HBoV), human parvovirus 4 (PARV4), hepatitis A (HAV) and hepatitis E virus (HEV) pose a particular threat to blood products. Virus transmission to patients treated with blood products presents an additional burden to disease. We determined the frequency and the amount of nucleic acid specific for nonenveloped viruses in recently manufactured preparations of commercial coagulation factor concentrates. MATERIALS AND METHODS: At least three different batches of each of 13 different plasma-derived and recombinant coagulation factor products were tested for the presence and the amount of nucleic acid for parvovirus B19, HBoV, human parvovirus 4, hepatitis A virus and HEV by using quantitative polymerase chain reaction. RESULTS: Whereas none of the recombinant products tested positive for any of these viruses, parvovirus B19 DNA with amounts ranging between 2×10(1) and 1.3×10(3) genome equivalents/ml was detected in five plasma-derived products. In addition to parvovirus B19 genotype 1, genotypes 2 and 3 were observed in two batches of a factor VIII/von-Willebrand factor product. In two products (one factor VIII concentrate and one activated prothrombin complex concentrate), a combination of both genotypes 1 and 2 of parvovirus B19 was detected. CONCLUSION: The data show that nucleic acids from several relevant nonenveloped viruses are not found at detectable levels in coagulation factor concentrates. In some cases, parvovirus B19 DNA was detectable at low levels. Testing of the plasma pools for the full range of parvovirus genotypes is advocated for ensuring product safety.
Assuntos
Transfusão de Componentes Sanguíneos , DNA Viral/sangue , Vírus da Hepatite A Humana , Hepatite A/prevenção & controle , Vírus da Hepatite E , Hepatite E/prevenção & controle , Infecções por Parvoviridae/prevenção & controle , Parvovirus , Reação em Cadeia da Polimerase/métodos , RNA Viral/sangue , Hepatite A/sangue , Hepatite A/transmissão , Hepatite E/sangue , Hepatite E/transmissão , Humanos , Infecções por Parvoviridae/sangue , Infecções por Parvoviridae/transmissãoRESUMO
To determine the frequency and the impact of parvovirus B19 (B19V) infection and its influence on the course of haematological and/or oncological diseases in paediatric patients, consecutive serum and bone marrow samples from 110 were analyzed for markers of acute, past and persistent B19V-infection using qPCR, ELISA and WesternLine. Twenty-seven out of 110 (24.5%) children suffered from non-malignant diseases (anaemia, pancytopenia, autoimmune disorders); 68/110 (61.8%) patients had developed leukaemia, malignant lymphoma or solid malignant tumours; 15/110 patients (13.6%) presented with other symptoms. At admission, B19V-specific IgM and IgG indicating acute or previous B19V-infection were observed in 5 (4.5%) and 48 patients (43.6%), respectively. B19V-DNA (10(3) -10(9) geq/mL) was detectable in serum and/or bone marrow of 22 patients (20.0%). These suffered from leukaemia (5), non-Hodgkin lymphoma (2), solid tumours (6), autoimmune (4) and haematological (4) disease and fever (1). During clinical observation four further leukaemia patients developed viraemia and persistent B19V-infection was observed in 13/22 DNA-positive patients. Treatment of B19V-DNA-positive cancer patients was associated with more supportive therapy involving erythrocyte and thrombocyte transfusion and/or antibiotic therapy. Acute B19V-infection has been frequently observed in paediatric patients with haematological and/or oncological disease. In patients with non-malignant diseases anaemia or autoimmune disorders were diagnosed in association with B19V-infection. Furthermore, a significant number of cancer patients displayed markers for acute, recent or persistent B19V-infection. This association may be strengthened by frequent treatment with blood products combined with therapeutic immune suppression. In B19V-infected cancer patients supportive therapy was more complex.
Assuntos
Neoplasias Hematológicas/virologia , Neoplasias/virologia , Infecções por Parvoviridae/sangue , Infecções por Parvoviridae/complicações , Parvovirus B19 Humano/isolamento & purificação , Adolescente , Anticorpos Antivirais/análise , Anticorpos Antivirais/sangue , Biomarcadores/análise , Biomarcadores/sangue , Medula Óssea/virologia , Distribuição de Qui-Quadrado , Criança , Pré-Escolar , Estudos de Coortes , DNA Viral , Feminino , Neoplasias Hematológicas/sangue , Humanos , Lactente , Masculino , Neoplasias/sangue , Transplante de Células-Tronco , Adulto JovemRESUMO
Human bocavirus (HBoV) was recently described as a new member of the Parvoviridae. In order to investigate the suggested association of HBoV with respiratory and gastric disease in infants and young children, sera of 357 paediatric patients hospitalized with infectious and non-infectious diseases were retrospectively analyzed for the presence of HBoV DNA and virus-specific antibodies using quantitative PCR and ELISA, respectively. HBoV seroprevalence was determined to range from 25% in infants younger than 1 year of age to 93% in children aged more than 3 years. Viral loads between 1 x 10(2) and 1.2 x 10(6) geq/mL were observed in 6.7% (20/297) of sera obtained preferentially from young children suffering from infectious diseases. HBoV genomes were furthermore detected in 5% (3/60) of sera collected from individuals with non-infectious illnesses. HBoV DNA was present most frequently in patients with respiratory disease (9.6%). Whereas only 5.2% of patients with upper respiratory tract disease were viraemic, HBoV DNA was found in 14.6% and 10.0% of patients with lower respiratory tract illness and pneumonia, respectively. Acute HBoV infections were also observed in 7.5% of patients with gastroenteritis and in one child with inflammatory bowel disease. None of 77 patients hospitalized for various other infectious diseases (e.g. rash, urinary tract infection, meningitis) displayed viraemia. In 60.9% and 47.8% of DNA-positive children, HBoV-specific IgM and IgG was observed, respectively. The present prospective study provides comprehensive data on the clinical association of acute HBoV infection with respiratory illness and on the seroprevalence of virus-specific antibodies in children.
Assuntos
Bocavirus Humano/isolamento & purificação , Infecções por Parvoviridae/epidemiologia , Infecções por Parvoviridae/patologia , Adolescente , Fatores Etários , Anticorpos Antivirais/sangue , Criança , Pré-Escolar , DNA Viral/sangue , Ensaio de Imunoadsorção Enzimática/métodos , Feminino , Gastroenteropatias/epidemiologia , Gastroenteropatias/patologia , Gastroenteropatias/virologia , Hospitalização , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Lactente , Masculino , Infecções por Parvoviridae/virologia , Reação em Cadeia da Polimerase/métodos , Prevalência , Infecções Respiratórias/epidemiologia , Infecções Respiratórias/patologia , Infecções Respiratórias/virologia , Estudos Retrospectivos , Soro/imunologia , Soro/virologia , Carga Viral , Virologia/métodosRESUMO
We, here, report the case of a parvovirus B19 infection in an immunocompetent male patient presenting with acute hepatitis and polyarthritis. To follow the course of infection, we used a previously established enzyme-linked immunosorbent spot assay (ELISPOT) technique to detect CD4+ T cells specific for viral proteins. Even though symptoms of arthritis and hepatitis resolved in the immunocompetent individual within a few weeks, viral DNA in serum and CD4+ T cells specific for the viral protein VP1 unique region were still detectable more than 6 month after the onset of symptoms, thus pointing to a persistent state of infection. On the basis of this observation, we hypothesize that the intensity of liver involvement correlates with the likelihood of developing persistent parvovirus B19 infection. The described ELISPOT technique to detect virus-specific CD4+ T cells provides an excellent tool to analyse the state of parvovirus B19 infection for future studies to test this hypothesis.
Assuntos
Artrite/virologia , Linfócitos T CD4-Positivos/imunologia , Hepatite Viral Humana/virologia , Infecções por Parvoviridae/diagnóstico , Parvovirus B19 Humano , Adulto , Artrite/imunologia , Biomarcadores/sangue , Western Blotting , Doença Crônica , DNA Viral/sangue , Ensaio de Imunoadsorção Enzimática/métodos , Hepatite Viral Humana/imunologia , Humanos , Imunidade Celular , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Masculino , Infecções por Parvoviridae/imunologia , Parvovirus B19 Humano/genéticaRESUMO
Acute parvovirus B19 infection is a risk for pregnant women. After vertical transmission the infected fetus may develop hydrops fetalis. Since B19 infection occurs mainly during childhood, children represent a main source for virus transmission. In order to determine whether certain groups in the German population show increased risks for B19 infection we analysed the seroprevalence using 6583 sera collected from adults in former Eastern and Western Germany during the German National Health Survey and 649 sera from healthy Thuringian children and adolescents. In adults the overall seroprevalence was 72.1%, rising from 20.4% in children (1-3 years) and 66.9% in adolescents (18-19 years) to 79.1% in the elderly (65-69 years). Significant differences were observed between females (73.3%) and males (70.9%) and between inhabitants of small (74.8%) and big cities (69.0%) but not between people of the former Eastern (72.8%) and Western states (72.0%) of Germany. For women during childbearing age (18-49 years) highest values were observed in those living together with two or more children (81.6%) and in women with occupational contact with children aged <6 years (88.9%). In contrast seroprevalence was significantly lower in age-matched female singles (64.8%) and in women with occupational contact with children aged >6 years and adolescents (63.8%).
Assuntos
Infecções por Parvoviridae/epidemiologia , Parvovirus B19 Humano/isolamento & purificação , Adolescente , Adulto , Fatores Etários , Idoso , Animais , Anticorpos Antivirais/sangue , Criança , Pré-Escolar , Feminino , Alemanha/epidemiologia , Humanos , Lactente , Recém-Nascido , Masculino , Pessoa de Meia-Idade , Estudos Soroepidemiológicos , Fatores Sexuais , População UrbanaAssuntos
Eritema Infeccioso/diagnóstico , Parvovirus B19 Humano , Leucemia-Linfoma Linfoblástico de Células Precursoras/terapia , Adolescente , Antineoplásicos/uso terapêutico , Terapia Combinada , Eritema Infeccioso/diagnóstico por imagem , Evolução Fatal , Transplante de Células-Tronco Hematopoéticas , Humanos , Masculino , Radiografia Torácica , Recidiva , Tiotepa/uso terapêutico , Tomografia Computadorizada por Raios X , Condicionamento Pré-Transplante/métodos , Transplante Homólogo , Vidarabina/análogos & derivados , Vidarabina/uso terapêutico , Irradiação Corporal TotalRESUMO
Parvovirus B19 infections may cause a widespread benign and self-limiting disease in children and adults known as erythema infectiosum (fifth disease). Several further manifestations are associated with B19 infections, such as arthralgias, arthritis, leucopenia and thrombocytopenia, anaemia and vasculitis and spontaneous abortion and hydrops fetalis in pregnant women. Persistent infections with continuous virus production may occur in immunocompetent as well as in immunosuppressed individuals. Parvovirus B19 infections have been frequently implicated as a cause or trigger of various forms of autoimmune diseases affecting joints, connective tissue and large and small vessels. Autoimmune neutropenia, thrombocytopenia and haemolytic anaemia are known as sequelae of B19 infections. The molecular basis of the autoimmune phenomena is unclear. Many patients with these long-lasting symptoms are not capable of eliminating the virus or controlling its propagation. Furthermore, latent viral genomes have been detected in cells of various organs and tissues by PCR. At present, it is not clear if these cells produce viral proteins and/or infectious B19 particles, if the virus genome can be reactivated to productive replication and if the presence of viral DNA indicates a causative role of parvovirus B19 with distinct diseases.
Assuntos
Cardiomiopatia Dilatada , Coração/virologia , Infecções por Parvoviridae/complicações , Parvovirus B19 Humano , Animais , Doenças Autoimunes/etiologia , Doenças Autoimunes/virologia , Cardiomiopatia Dilatada/etiologia , Cardiomiopatia Dilatada/virologia , Feminino , Humanos , Parvovirus B19 Humano/classificação , Parvovirus B19 Humano/genética , Parvovirus B19 Humano/fisiologia , Gravidez , Complicações Infecciosas na GravidezRESUMO
To date cellular immune responses against parvovirus B19 (B19) have not been studied extensively. The aim of this study was to examine the T-cell response against the VP1-unique region as the immunodominant part of the viral structural protein VP1 in individuals with different courses of B19 infection. Therefore, a group of 13 parvovirus-positive probands was separated into subgroups characterized for recent or acute, past or persistent infection by means of the presence of specific immunoglobulin (Ig)M and IgG isotypes and of viral DNA in blood and tissue. Transiently transfected B-cells expressing VP1-unique region were used in ELISpot assays to investigate T-cell responses directed against the VP1-unique region in peripheral blood mononuclear cells (PBMC) of individual donors. Significant numbers of interferon-gamma (IFN-gamma) secreting lymphocytes were detectable in PBMC of all individuals with recent, acute or persistent B19 infection, but not in PBMC of donors with past B19 infection and seronegative individuals. A more detailed analysis of IFN-gamma producing cells by intracellular cytokine staining by flow cytometry revealed, that CD4(+) T cells but not CD8(+) cytotoxic lymphocytes (CTL) were the major subpopulation of IFN-gamma producing cells. These data strongly suggest the need of virus protein production for the maintenance of VP1-unique region-specific CD4(+) T-helper cell responses in B19-infected individuals.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Proteínas do Capsídeo/imunologia , Infecções por Parvoviridae/imunologia , Parvovirus B19 Humano/imunologia , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Western Blotting , Linfócitos T CD4-Positivos/metabolismo , Estudos de Casos e Controles , DNA Viral/sangue , Ensaio de Imunoadsorção Enzimática , Citometria de Fluxo , Humanos , Interferon gama/metabolismo , Ativação Linfocitária , Infecções por Parvoviridae/sangue , Infecções por Parvoviridae/virologia , Parvovirus B19 Humano/genética , Reação em Cadeia da PolimeraseRESUMO
The capsids of human parvovirus B19 consist of two structural proteins, the minor-capsid protein VP1 and the major-capsid protein VP2. The latter which constitutes for 95% of the capsid are able to form virus-like particles (VLPs) in yeast without the presence of VP1-proteins. VP2-proteins produced in Saccharomyces cerevisiae have the capacity to form VLPs in the absence of VP1-proteins. These yeast-derived VLPs resemble native virus or recombinant VP2-VLPs produced by baculovirus systems in respect of size, molecular weight and of antigenicity as shown by antigen-capture ELISA and T-cell proliferation tests. Regarding costs, yield and ease of handling particle production in yeast represents an alternative to the recombinant baculovirus expression system which is so far the source for VP2-VLPs of human parvovirus B19.
Assuntos
Baculoviridae/metabolismo , Proteínas do Capsídeo/biossíntese , Parvovirus B19 Humano/fisiologia , Saccharomyces cerevisiae/metabolismo , Vírion/metabolismo , Baculoviridae/genética , Proteínas do Capsídeo/genética , Vetores Genéticos , Humanos , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/genética , Saccharomyces cerevisiae/genética , Vírion/genéticaRESUMO
Human parvovirus B19 is a small non-enveloped DNA virus with an icosahedral capsid consisting of proteins of only two species, the major protein VP2 and the minor protein VP1. VP2 is contained within VP1, which has an additional unique portion (VP1u) of 227 amino acids. We determined the ability of eukaryotically expressed parvovirus B19 virus-like particles consisting of VP1 and VP2 in the ratio recommended for vaccine use, or of VP2 alone, to stimulate, in an HLA class II restricted manner, peripheral blood mononuclear cells (PBMC) to proliferate and to secrete interferon gamma (IFN-gamma) and interleukin (IL)-10 cytokines among recently and remotely B19 infected subjects. PBMC reactivity with VP1u was determined specifically with a prokaryotically expressed VP1u antigen. In general, B19-specific IFN-gamma responses were stronger than IL-10 responses in both recent and remote infection; however, IL-10 responses were readily detectable among both groups, with the exception of patients with relapsed or persisting symptoms who showed strikingly low IL-10 responses. Whereas VP1u-specific IFN-gamma responses were very strong among the recently infected subjects, the VP1u-specific IFN-gamma and IL-10 responses were virtually absent among the remotely infected subjects. The disappearance of VP1u-specific IFN-gamma expression is surprising, as B-cell immunity against VP1u is well maintained.
Assuntos
Proteínas do Capsídeo/imunologia , Interferon gama/imunologia , Infecções por Parvoviridae/imunologia , Parvovirus B19 Humano/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Adulto , Divisão Celular/imunologia , Células Cultivadas , Endotoxinas/imunologia , Epitopos/imunologia , Feminino , Antígenos de Histocompatibilidade Classe II/imunologia , Humanos , Imunidade Celular/imunologia , Interleucina-10/imunologia , Leucócitos Mononucleares/imunologia , Masculino , Pessoa de Meia-Idade , Gravidez , Complicações Infecciosas na Gravidez/imunologiaRESUMO
This work presents conclusive evidence that connecting Pt and Co nanoparticles stabilized by an aluminum-organic shell with molecular spacers interacting with this shell can induce notable changes in the electronic structure of the metal. X-ray absorption spectroscopy measurements at the Al K-, the Pt L(III)-, and the Co K-edge provide consistent evidence for this effect. The changes induced by cross-linking with an acidic spacer are discussed in detail as an example to elucidate the mechanism of this effect. It turns out that a reconfiguration of the protection shell that occurs upon networking is responsible for the observed changes.
RESUMO
FKBP52, a multidomain peptidyl prolyl cis/trans-isomerase (PPIase), is found in complex with the chaperone Hsp90 and the co-chaperone p23. It displays both PPIase and chaperone activity in vitro. To localize these two activities to specific regions of the protein, we created and analyzed a set of fragments of FKBP52. The PPIase activity toward both peptides and proteins is confined entirely to domain 1 (amino acids 1-148). The chaperone activity, however, resides in the C-terminal part of FKBP52, mainly in the region between amino acids 264 and 400 (domain 3). Interestingly, this domain also contains the tetratricopeptide repeats, which are responsible for the binding to C-terminal amino acids of Hsp90. Competition assays with a C-terminal Hsp90 peptide suggest that the non-native protein and Hsp90 are bound by different regions within this domain.
Assuntos
Chaperonas Moleculares/metabolismo , Proteínas de Ligação a Tacrolimo/metabolismo , Animais , Ligação Competitiva , Citrato (si)-Sintase/metabolismo , Proteínas de Choque Térmico HSP90/metabolismo , Fragmentos de Peptídeos/metabolismo , Peptidilprolil Isomerase/metabolismo , Conformação Proteica , Estrutura Terciária de Proteína , Coelhos , Especificidade por Substrato , Proteínas de Ligação a Tacrolimo/químicaRESUMO
OBJECTIVE: We report an 11-year-old boy presenting with splenomegaly, chronic thrombocytopenia and concordant neutropenia. RESULTS: In contrast to autoantibodies against platelets, there were no detectable neutrophil-specific autoantibodies present in this patient. Extensive serologic investigations revealed increased IgM- and IgG-antibody titers against parvovirus B19. A nested polymerase chain reaction (PCR) showed parvovirus B19-specific sequences in the patient's bone-marrow cells but not in the serum. Specific antibodies against the structural proteins VP1 and VP2 in addition to those against non-structural protein NS1 of parvovirus B19 were detected by Western blot analysis. Thrombocytopenia and neutropenia responded to immunosuppressive therapy and subsequent splenectomy, the latter being necessary due to severe side-effects of steroid medication. CONCLUSION: Autoimmune thrombocytopenia/neutropenia may have been triggered and/or sustained by a chronic parvovirus B19 infection. Patients with this very rare disorder should be screened for this virus.
Assuntos
Doenças Autoimunes/virologia , Neutropenia/virologia , Infecções por Parvoviridae/virologia , Parvovirus B19 Humano/patogenicidade , Trombocitopenia/virologia , Anticorpos Antivirais/sangue , Doenças Autoimunes/terapia , Western Blotting , Criança , Doença Crônica , Humanos , Imunossupressores/farmacologia , Imunossupressores/uso terapêutico , Masculino , Neutropenia/terapia , Infecções por Parvoviridae/tratamento farmacológico , Parvovirus B19 Humano/efeitos dos fármacos , Reação em Cadeia da Polimerase , Testes Sorológicos , Trombocitopenia/terapiaRESUMO
Parvovirus B19 is the causative agent of erythema infectiosum in children, but the virus is associated with an increasing range of different diseases. These include acute and chronic arthritis, hydrops fetalis in pregnant women, aplastic anemia, and thrombocytopenia. The host's immune response is directed against the viral structural proteins VP1 and VP2. This study investigated the presence of IgG against the viral nonstructural protein NS1 using Western blot. Serum panels from healthy individuals, B19-infected pregnant women, and various disease groups were tested. The disease groups included patients with symptoms that may be linked to parvovirus B19 infection. The results showed that IgG against the NS1 protein was present in 22% of healthy individuals with past B19 infection. In cases of persistent or prolonged B19 infections, the prevalence of NS1-specific antibodies was as high as 80%. It is concluded that NS1-specific IgG may be used as an indicator of chronic or more severe courses of parvovirus B19 infections.
Assuntos
Anticorpos Antivirais/sangue , Proteínas do Capsídeo , Infecções por Parvoviridae/imunologia , Parvovirus B19 Humano/imunologia , Complicações Infecciosas na Gravidez/imunologia , Proteínas não Estruturais Virais/imunologia , Adolescente , Adulto , Idoso , Western Blotting , Capsídeo/imunologia , Feminino , Doenças Hematológicas/virologia , Humanos , Imunoglobulina G/sangue , Artropatias/virologia , Pessoa de Meia-Idade , Infecções por Parvoviridae/complicações , Infecções por Parvoviridae/virologia , Gravidez , Complicações Infecciosas na Gravidez/virologiaRESUMO
BACKGROUND: Since its discovery parvovirus B19-infections could be linked to a growing variety of diseases. Besides the harmless exanthema erythema infectiosum perferentially observed with B19-infections in childhood a panel of rather serious and also chronic courses that may be associated with anemia, thrombocytopenia, arthritis and others have been described. OBJECTIVE: In a 26-year-old female patient an acute parvovirus B19-infection was followed by a serious episode of systemic lupus erythematosus (SLE). Here we demonstrate the clinical and serological parameters which were observed in the patient during that episode in addition to the nucleotide sequence of the virus isolate. RESULTS AND CONCLUSION: In this patient parvovirus B19 was not the initial causative agent for SLE. However the B19 infection was followed by a severe flare of SLE and therefore may be considered as an enhancer of the autoimmune disease. The amount of nucleotide variability observed in the viral genome was in the range known from other B19 isolates. An elevated degree of mutations in antigenic domains was not detectable. Therefore, we would like to emphasize the possible role of parvovirus B19 in the aetiology or the enhancement of autoimmune diseases like SLE and the necessity of an according differential diagnosis.
Assuntos
Proteínas do Capsídeo , Lúpus Eritematoso Sistêmico/complicações , Infecções por Parvoviridae/complicações , Parvovirus/isolamento & purificação , Adulto , Capsídeo/sangue , DNA Viral/sangue , Feminino , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Lúpus Eritematoso Sistêmico/virologia , Dados de Sequência Molecular , Infecções por Parvoviridae/virologia , Parvovirus/genética , Parvovirus/imunologia , Reação em Cadeia da Polimerase , Testes SorológicosRESUMO
We investigated CD8(+) T cell frequencies of five different Epstein-Barr virus-specific cytotoxic T lymphocyte epitopes located within proteins of the replicative cycle and the latent state in healthy long-term virus carriers with IFN-gamma enzyme-linked immunospot assay. Frequencies of the HLA-A3-restricted epitope RVRAYTYSK (RVR) whose minimal length was mapped in this study to amino acid position 148-156 of the immediate-early protein BRLF1 were compared with those of a further known HLA-A3-restricted epitope within EBNA3A, RLRAEAQVK (RLR). Determination of frequencies of CD8(+) T lymphocytes directed against lytic antigen epitope RVR revealed that only one of eight donors recognized this epitope. Frequency was calculated to be 65 RVR-specific CD8(+) T lymphocytes per 10(6) PBMC. None of the HLA-A3-positive donors exhibited IFN-gamma release after antigenic stimulation with the EBNA3A-specific peptide epitope RLR. Furthermore, we chose three known HLA-B8-restricted epitopes, RAKFKQLL (RAK), FLRGRAYGL (FLR), and QAKWRLQTL (QAK), of the lytic protein BZLF1 and the latent protein EBNA3A. Examination of eight HLA-B8-positive virus carriers revealed that the BZLF1-specific epitope RAK was recognized by all donors with a median frequency of 233 RAK-specific CD8(+) T lymphocytes per 10(6) PBMC. Only 50% of these donors reacted against EBNA3A-specific epitope FLR and a minority (25%) reacted against EBNA3A-specific epitope QAK.
Assuntos
Linfócitos T CD8-Positivos/imunologia , Portador Sadio/imunologia , Infecções por Vírus Epstein-Barr/imunologia , Herpesvirus Humano 4/imunologia , Linfócitos T CD8-Positivos/citologia , Portador Sadio/sangue , Portador Sadio/virologia , Linhagem Celular , Epitopos de Linfócito T/imunologia , Infecções por Vírus Epstein-Barr/sangue , Infecções por Vírus Epstein-Barr/virologia , Antígenos Nucleares do Vírus Epstein-Barr/imunologia , Células-Tronco Hematopoéticas/imunologia , Humanos , Proteínas Imediatamente Precoces/imunologia , Linfócitos T Citotóxicos/citologia , Linfócitos T Citotóxicos/imunologia , Transativadores/imunologia , Proteínas ViraisRESUMO
BiP, a resident endoplasmic reticulum member of the HSP70 family of molecular chaperones, associates transiently with a wide variety of newly synthesized exocytotic proteins. In addition to immunoglobulin heavy and light chains, the first natural substrates identified for BiP, a number of viral polypeptides including the human immunodeficiency virus type 1 envelope glycoprotein gp160 interact with BiP during their passage through the endoplasmic reticulum. We have used a computer algorithm developed to predict BiP-binding sites within protein primary sequences to identify sites within gp160 that might mediate its association with BiP. Analysis of the ability of 22 synthetic heptapeptides corresponding to predicted binding sites to stimulate the ATPase activity of BiP or to compete with an unfolded polypeptide for binding to BiP indicated that about half of them are indeed recognized by the chaperone. All of the confirmed binding sites are localized within conserved regions of gp160, suggesting a conserved role for BiP in the folding of gp160. Information on the characteristics of confirmed BiP-binding peptides gained in this and previous studies has been utilized to improve the predictive power of the BiP Score algorithm and to investigate the differences in peptide binding specificities of HSP70 family members.