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1.
Vet Res ; 51(1): 110, 2020 Sep 03.
Artigo em Inglês | MEDLINE | ID: mdl-32883344

RESUMO

Canine parvovirus (CPV) can cause acute and highly contagious bloody enteritis in dog. To obtain antibodies against CPV, hens were immunized with virus-like particles (VLP) of CPV-VP2. The IgY single chain fragment variables (scFv) were generated by T7 phage display system and expressed in E. coli system. The titer of the primary scFv library reached to 1.5 × 106 pfu/mL, and 95% of the phages contained the target fragments. The CPV-VLP and CPV-VP2 protein showed similar reaction values to the purified scFv in the ELISA test, and the results of ELISA analysis using IgY-scFv toward CPV clinical samples were consistent with commercial immunochromatographic assay (ICA) and PCR detection, the scFv did not show cross reactivity with canine distemper virus (CDV) and canine coronavirus (CCV). IgY-scFv was successfully expressed in CRFK cells, and in the virus suppression assay, 55% of CPV infections were eliminated within 24 h. Docking results demonstrated that the number of amino acids of the binding sides between scFv and VP2 were AA37 and AA40, respectively. This study revealed the feasibility of a novel functional antibody fragment development strategy by generating diversified avian IgY-scFv libraries towards the pathogenic target of interest for both detection and therapeutic purposes in veterinary medicine.


Assuntos
Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , Proteínas do Capsídeo/imunologia , Doenças do Cão/virologia , Imunoglobulinas/imunologia , Infecções por Parvoviridae/veterinária , Parvovirus Canino/imunologia , Anticorpos de Cadeia Única/imunologia , Animais , Galinhas/imunologia , Doenças do Cão/diagnóstico , Doenças do Cão/imunologia , Cães , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Simulação de Acoplamento Molecular , Infecções por Parvoviridae/diagnóstico , Infecções por Parvoviridae/imunologia , Infecções por Parvoviridae/virologia , Anticorpos de Cadeia Única/genética
2.
Afr Health Sci ; 20(1): 219-226, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33402910

RESUMO

Background: Parvovirus B19 (B19) has tropism for cells of the erythroid lineage, which may lead to transient inhibition of erythropoiesis. Several studies and case reports suggested that B19 infection may contribute significantly to severe chronic anemia in HIV infected persons. Objective: To detect parvovirus B19 DNA in treatment-naïve HIV patients. Methods: This was a case control retrospective study. One hundred nineteen anemic and 81 non-anemic treatment-naïve HIV infected patients participated in the study at the Lagos University Teaching Hospital, Lagos, Nigeria. Polymerase chain reaction was used to detect B19 DNA. Results: Out of 200 patients analysed, 13(6.5%) had parvovirus B19 DNA. Eight HIV patients with anemia had B19 DNA while five non-anemic HIV patients had B19 DNA. This suggests that the presence of B19 DNA in the blood of HIV positive individuals may contribute to anemia because the majority (61.5%) who were positive for B19 DNA had anemia as compared to the non-anemic control group (38.5%). Conclusion: This study shows that the presence of B19 DNA in anemic HIV infected patients is not associated with chronic anaemia in HIV infection because no significant association exist.


Assuntos
Infecções Oportunistas Relacionadas com a AIDS/epidemiologia , Anemia/virologia , Infecções por HIV/complicações , Infecções por Parvoviridae/virologia , Parvovirus B19 Humano/isolamento & purificação , Infecções Oportunistas Relacionadas com a AIDS/sangue , Infecções Oportunistas Relacionadas com a AIDS/imunologia , Infecções Oportunistas Relacionadas com a AIDS/virologia , Adolescente , Adulto , Idoso , Anemia/epidemiologia , Anemia/imunologia , Anticorpos Antivirais/sangue , Estudos de Casos e Controles , Criança , DNA Viral/análise , Feminino , Infecções por HIV/epidemiologia , Humanos , Imunoglobulina G/sangue , Masculino , Pessoa de Meia-Idade , Nigéria/epidemiologia , Infecções por Parvoviridae/sangue , Infecções por Parvoviridae/epidemiologia , Infecções por Parvoviridae/imunologia , Parvovirus B19 Humano/genética , Parvovirus B19 Humano/imunologia , Reação em Cadeia da Polimerase , Estudos Retrospectivos , Adulto Jovem
3.
Medicina (B Aires) ; 79(6): 493-501, 2019.
Artigo em Espanhol | MEDLINE | ID: mdl-31829952

RESUMO

In recent years, gene therapy has been positioned as a real and safe option in the development of therapeutic alternatives for the cure and prevention of different diseases. It consists in the insertion of genetic material in a defective tissue or cell, through the use of a vector. There are several considerations for selecting the most appropriate vector, including the potential for binding and entry to the target cell, the ability of the genetic material to transfer to the nucleus, the ability to express the insert, and the absence of toxicity. In the current scenario, the most commonly used viral vectors are those derived from adeno-associated viruses (AAV). Characteristics such as biosafety, low toxicity and selective tropism have enabled its evaluation as a therapeutic option in many monogenic or complex diseases. Despite their advantages, AAV vectors have drawbacks, the most important being the patient's immune response to the vector, especially the response mediated by neutralizing antibodies (NAb). NAbs decrease the transduction of the vector and prevent the expression of the gene it transports, limiting its clinical application. Therefore, identifying and quantifying the presence and activity of NAbs is the first step in any gene therapy protocol with AAV vectors. The presence of NAbs depends mainly on exposure to the virus in nature and varies drastically according to age, geographic location and health status of the person evaluated.


Assuntos
Dependovirus/genética , Dependovirus/imunologia , Terapia Genética/métodos , Infecções por Parvoviridae/genética , Infecções por Parvoviridae/imunologia , Anticorpos Neutralizantes/análise , Anticorpos Antivirais/análise , Feminino , Vetores Genéticos , Humanos , Masculino , Infecções por Parvoviridae/virologia , Sorogrupo
4.
BMC Vet Res ; 15(1): 335, 2019 Sep 18.
Artigo em Inglês | MEDLINE | ID: mdl-31533724

RESUMO

BACKGROUND: Current guidelines recommend parvovirus revaccination of adult dogs no more frequently than every 3 years. The aim of this study was to determine the prevalence of dogs showing protective serum antibody titres against canine parvovirus 2 in breeding kennels in Northern Italy and to assess the effect of time from vaccination and the sex of the dog on antibody titres. The study was carried out on 370 animals of different breeds kept in 33 breeding kennels. Antibodies to canine parvovirus 2 in serum samples were measured with an indirect immunoenzymatic assay validated by the manufacturer in relation to the 'gold standard' haemagglutination inhibition test. The number of months that had elapsed since the last vaccination was calculated for each animal and categorized into the following classes: < 12 months; 13-24 months; 25-36 months; 37-48 months; and > 49 months. RESULTS: The prevalence of 'unprotected' dogs was 4.6%. A satisfactory solid herd immunity was present in the majority of breeding kennels, although some vaccination failures were detected. A significant negative correlation was found between antibody titre and months since last vaccination. Comparable antibody titres were found in the first 3 years after vaccination. Although the antibody titre over time was not affected by the sex of the dog, 'unprotected' females had been vaccinated more recently than males with analogous low titres. CONCLUSIONS: Parvovirus revaccination of adult dogs every 3 years, as currently recommended, is also the appropriate recommendation for breeding kennels. Serological tests could be a useful tool to assess the effectiveness of vaccination.


Assuntos
Anticorpos Antivirais/sangue , Doenças do Cão/imunologia , Infecções por Parvoviridae/veterinária , Animais , Doenças do Cão/prevenção & controle , Cães , Feminino , Itália , Masculino , Infecções por Parvoviridae/imunologia , Infecções por Parvoviridae/prevenção & controle , Parvovirus Canino/imunologia , Fatores de Tempo , Vacinação/estatística & dados numéricos , Vacinação/veterinária
5.
J Clin Virol ; 120: 17-19, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31521013

RESUMO

BACKGROUND: Diagnosis of human bocavirus 1 (HBoV1) has been based on qualitative PCRs detecting HBoV1 DNA or detection of HBoV1 mRNA. OBJECTIVE: This study aims to assess whether a rapid and automated HBoV1 antigen test is suitable for diagnosis of acute HBoV1 infection. STUDY DESIGN: HBoV1 antigen detection has been compared with quantitative HBoV1 DNA PCR and HBoV1 mRNA RT-PCR. RESULTS AND CONCLUSION: We conclude that HBoV1 antigen detection has higher clinical specificity and positive predictive value than HBoV1 DNA qualitative PCRs, yet a lower sensitivity than HBoV1 mRNA detection. Additionally, HBoV1 antigen detection is beneficial in its rapidity and availability as a point-of-care test.


Assuntos
Antígenos Virais/metabolismo , Bocavirus Humano/genética , Bocavirus Humano/imunologia , Infecções por Parvoviridae/diagnóstico , RNA Mensageiro/genética , Infecções Respiratórias/virologia , Automação , Criança , DNA Viral/genética , Feminino , Genótipo , Humanos , Masculino , Infecções por Parvoviridae/imunologia , Fenótipo , Sistemas Automatizados de Assistência Junto ao Leito , RNA Viral/genética , Infecções Respiratórias/imunologia , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sensibilidade e Especificidade , Carga Viral
6.
Sci Rep ; 9(1): 11266, 2019 08 02.
Artigo em Inglês | MEDLINE | ID: mdl-31375758

RESUMO

After its first identification in 1978, canine parvovirus (CPV) has been recognized all around the world as a major threat for canine population health. This ssDNA virus is characterized by a high substitution rate and several genetic and phenotypic variants emerged over time. Overall, the definition of 3 main antigenic variants was established based on specific amino acid markers located in a precise capsid position. However, the detection of several minor variants and incongruence observed between the antigenic classification and phylogeny have posed doubts on the reliability of this scheme. At the same time, CPV heterogeneity has favored the hypothesis of a differential virulence among variants, although no robust and consistent demonstration has been provided yet. The present study rejects the antigenic variant concept and attempts to evaluate the association between CPV strain phylogeny, reconstructed using the whole information contained in the VP2 coding gene, and several clinical and hemato-biochemical parameters, assessed from 34 CPV infected dogs at admission. By using different statistical approaches, the results of the present study show an association between viral phylogeny and host parameters ascribable to immune system, coagulation profile, acute phase response and, more generally, to the overall picture of the animal response. Particularly, a strong and significant phylogenetic signal was proven for neutrophil count and WBC. Therefore, despite the limited sample size, a relation between viral phylogeny and disease severity has been observed for the first time, suggesting that CPV virulence is an inherited trait. The likely existence of clades with different virulence highlights once more the relevance of intensive epidemiological monitoring and research on CPV evolution to better understand the virulence determinants, their epidemiology and develop adequate countermeasures.


Assuntos
DNA de Cadeia Simples/genética , DNA Viral/genética , Interações Hospedeiro-Patógeno/genética , Infecções por Parvoviridae/veterinária , Parvovirus Canino/genética , Animais , Proteínas do Capsídeo/genética , DNA de Cadeia Simples/isolamento & purificação , DNA Viral/isolamento & purificação , Cães , Evolução Molecular , Feminino , Heterogeneidade Genética , Interações Hospedeiro-Patógeno/imunologia , Masculino , Infecções por Parvoviridae/sangue , Infecções por Parvoviridae/diagnóstico , Infecções por Parvoviridae/imunologia , Parvovirus Canino/isolamento & purificação , Parvovirus Canino/patogenicidade , Filogenia , Reação em Cadeia da Polimerase , Índice de Gravidade de Doença , Virulência/genética
7.
Gene Ther ; 26(9): 399-406, 2019 09.
Artigo em Inglês | MEDLINE | ID: mdl-31467408

RESUMO

Differences between mouse and human hearts pose a significant limitation to the value of small animal models when predicting vector behavior following recombinant adeno-associated viral (rAAV) vector-mediated cardiac gene therapy. Hence, sheep have been adopted as a preclinical animal, as they better model the anatomy and cardiac physiological processes of humans. There is, however, no comprehensive data on the shedding profile of rAAV in sheep following intracoronary delivery, so as to understand biosafety risks in future preclinical and clinical applications. In this study, sheep received intracoronary delivery of rAAV serotypes 2/6 (2 × 1012 vg), 2/8, and 2/9 (1 × 1013 vg) at doses previously administered in preclinical and clinical trials. This was followed by assessment over 96 h to examine vector shedding in urine, feces, nasal mucus, and saliva samples. Vector genomes were detected via real-time quantitative PCR in urine and feces up to 48 and 72 h post vector delivery, respectively. Of these results, functional vector particles were only detected via a highly sensitive infectious replication assay in feces samples up to 48 h following vector delivery. We conclude that rAAV-mediated gene transfer into sheep hearts results in low-grade shedding of non-functional vector particles for all excreta samples, except in the case of feces, where functional vector particles are present up to 48 h following vector delivery. These results may be used to inform containment and decontamination guidelines for large animal dealings, and to understand the biosafety risks associated with future preclinical and clinical uses of rAAV.


Assuntos
Dependovirus/genética , Vetores Genéticos , Eliminação de Partículas Virais , Animais , Anticorpos Neutralizantes/imunologia , Anticorpos Antivirais/imunologia , Cateterismo , Vasos Coronários , Dependovirus/imunologia , Dependovirus/fisiologia , Vetores Genéticos/administração & dosagem , Vetores Genéticos/imunologia , Células HeLa , Humanos , Injeções Intra-Arteriais , Masculino , Infecções por Parvoviridae/imunologia , Infecções por Parvoviridae/urina , Infecções por Parvoviridae/virologia , Reação em Cadeia da Polimerase em Tempo Real , Ovinos , Replicação Viral
8.
Vet Microbiol ; 235: 86-92, 2019 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-31282383

RESUMO

Although PCV2 infections generally cause mild disease in pigs, concurrent co-infections with other pathogens can damage the immune system and cause more severe diseases, collectively termed porcine circovirus associated diseases (PCVAD). Involvement of porcine parvovirus (PPV, a common cause of reproductive failure in naïve dams) in PCVAD caused by PCV2, has been reported. As this co-infection can be difficult to eliminate, there is a critical need to develop an effective vaccine to protect against PPV or synergistic effects of PCV2 and PPV under field conditions. In this study, we designed chimeric PCV2 virus-like particles (cVLPs) displaying a B-cell epitope derived from PPV1 structural protein around the surface of the 2-fold axes of PCV2 VLPs, based on 3D-structure analysis of the PCV2 capsid. The cVLPs were successfully prepared, verified by transmission electron microscopy and chromatography, with robust antibody titers against PCV2 and PPV1 produced in mice and guinea pigs. In addition, in guinea pigs challenged with 106 TCID50 PCV2, cVLPs conferred more effective immune protection (based on viral load) than a commercial PCV2 vaccine. Finally, antibody responses and immune protection against PPV were also evaluated. In guinea pigs vaccinated with cVLPs, although PPV antibodies detected by a hemagglutination inhibition (HI) assay appeared later after vaccination in the PCV2 cVLPs group than in the commercial PPV vaccine group, there were fewer PPV genomic DNA copies in the PCV2 cVLPs group than in a PBS group. In conclusion, guinea pigs vaccinated with cVLPs developed effective protective immunity against PCV2 challenge, with some protective immunity against PPV. This study provided valuable research data to pursue molecular design of chimeric epitopes PCV2 VLPs.


Assuntos
Infecções por Circoviridae/veterinária , Coinfecção/veterinária , Epitopos de Linfócito B/imunologia , Imunidade Humoral , Infecções por Parvoviridae/veterinária , Vacinas Virais/imunologia , Animais , Anticorpos Neutralizantes/sangue , Anticorpos Antivirais/sangue , Linfócitos B/imunologia , Infecções por Circoviridae/imunologia , Infecções por Circoviridae/prevenção & controle , Circovirus/imunologia , Coinfecção/virologia , Feminino , Cobaias , Camundongos , Infecções por Parvoviridae/imunologia , Infecções por Parvoviridae/prevenção & controle , Parvovirus Suíno/imunologia , Suínos , Doenças dos Suínos/prevenção & controle , Doenças dos Suínos/virologia , Vacinas Atenuadas/imunologia , Vacinas de Partículas Semelhantes a Vírus/imunologia
9.
Transplant Proc ; 51(8): 2693-2696, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31351772

RESUMO

Parvovirus B19 (PVB19) has tropism to red blood cell progenitors and can be reactivated after organ transplantation. The aim of study was to describe clinical manifestations, laboratory findings, treatments used, and effectiveness in kidney recipients at Viet Duc hospital. A retrospective descriptive study was performed on 663 kidney recipients who were on regular follow-up from 2000 to 2018. PVB19 was detected by polymerase chain reaction PVB19-DNA. Effectiveness of therapy was assessed by Hemoglobin level. Nine out of 663 kidney recipients (1.4%) were diagnosed with PVB19-associated anemia. Eight of these 9 (89%) were diagnosed within the first 3 months following transplantation. All patients had normoscopic anemia; the average reticulocyte proportion and count were 0.15 ± 0.04% and 0.0039 ± 0.0011T/L, respectively. Graft dysfunction was observed in 4/9 (45%) patients. Treatment included reduction of immunosuppression, intravenous immunoglobulin (IVIG), and blood transfusion. All patients responded well to treatment except 1 (11%), who experienced relapse after using low dose of IVIG. PVB19-associated anemia usually occurred early after transplantation and was associated with very low reticulocyte proportion and count. Actual treatment was effective, but the risk of relapse was present.


Assuntos
Anemia/virologia , Transplante de Rim , Infecções por Parvoviridae , Adulto , Feminino , Humanos , Hospedeiro Imunocomprometido/imunologia , Transplante de Rim/efeitos adversos , Masculino , Pessoa de Meia-Idade , Infecções por Parvoviridae/complicações , Infecções por Parvoviridae/epidemiologia , Infecções por Parvoviridae/imunologia , Parvovirus B19 Humano , Estudos Retrospectivos
10.
PLoS One ; 14(5): e0216799, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31086415

RESUMO

Evidence has indicated that viral infection increases the risk of developing asthma. Although the association of human parvovirus B19 (B19V) or human bocavirus (HBoV) with respiratory diseases has been reported, little is known about the influence of the B19V-VP1u and HBoV-VP1u proteins on the symptoms of asthma. Herein, we investigated the systemic influence of subcutaneously injected B19V-VP1u and HBoV-VP1u recombinant proteins in an OVA-sensitized asthmatic mouse model. A significantly higher Penh ratio and IgE level were detected in the serum, bronchoalveolar lavage fluid (BALF) and the supernatant of a lymphocyte culture from mice treated with HBoV-VP1u or B19V-VP1u than in a lymphocyte culture from OVA-sensitized mice. Significantly higher levels of serum and BALF IgE, total IgG, IgG1, OVA-specific IgE and OVA-specific IgG1 were detected in mice treated with HBoV-VP1u or B19V-VP1u than in OVA-sensitized mice. Conversely, a significantly lower IgG2a level was detected in mice from the HBoV-VP1u or B19V-VP1u groups than in mice from the OVA group. The mice treated with HBoV-VP1u or B19V-VP1u exhibited more significant lung inflammatory indices, including elevated serum and BALF IL-4, IL-5, IL-10 and IL-13 levels; BALF lymphocyte, neutrophil and eosinophil counts, MMP-9 and MMP-2 activity; and the amount of lymphocyte infiltration, relative to those in the control mice or in those sensitized with OVA. These findings demonstrate that the subcutaneous injection of HBoV-VP1u or B19V-VP1u proteins in OVA-sensitized mice result in elevated asthmatic indices and suggest that human parvoviruses may increase the risk of developing airway inflammation in a mouse model of asthma.


Assuntos
Asma/virologia , Proteínas do Capsídeo/imunologia , Bocavirus Humano/imunologia , Infecções por Parvoviridae/complicações , Parvovirus B19 Humano/imunologia , Animais , Asma/etiologia , Asma/imunologia , Proteínas do Capsídeo/química , Modelos Animais de Doenças , Feminino , Humanos , Camundongos , Camundongos Endogâmicos BALB C , Infecções por Parvoviridae/imunologia
11.
Vet Microbiol ; 231: 177-182, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30955806

RESUMO

Canine parvovirus (CPV) is one of the most important cause of mortality in young dogs and no specific treatment exists. Since prolonged leukopenia greatly increases the risk of death in infected pups, strategies to counteract this decline were investigated. The outcomes of CPV naturally infected pups treated with the recombinant canine granulocyte-colony stimulating factor (rcG-CSF), in combination with the routine therapy, were compared with similarly-managed infected pups not treated with rcG-CSF. A non-randomized prospective clinical trial was performed on 62 CPV infected pups with WBC counts <3000 cells/µL and two different groups were selected based on a non-randomized approach. Group A dogs (31/62) received 5 µg/Kg of rcG-CSF daily from the hospitalization day until WBC reached the reference range (3-5 days) and group B (31/62) received 1 ml of placebo injection. All dogs in group A recovered, while five dogs in group B died. The rcG-CSF treatment demonstrated a statistically significant effect on WBC counts (p < 0.0001) and, surprisingly, also on lymphocytes and monocytes counts (p < 0.0001). There was no significant effect of treatment on neutrophil count (p = 0.5502). Although lymphocytes and monocytes are not a specific target for rcG-CSF, our study highlights that rcG-CSF is able to improve haematological parameters compared to untreated dogs and a clear increase in their number was detected, as previously described for humans treated with the homologous molecule.


Assuntos
Doenças do Cão/prevenção & controle , Fator Estimulador de Colônias de Granulócitos/imunologia , Fator Estimulador de Colônias de Granulócitos/uso terapêutico , Contagem de Leucócitos/veterinária , Infecções por Parvoviridae/veterinária , Animais , Doenças do Cão/imunologia , Doenças do Cão/virologia , Cães , Feminino , Masculino , Infecções por Parvoviridae/imunologia , Infecções por Parvoviridae/prevenção & controle , Parvovirus Canino , Estudos Prospectivos , Proteínas Recombinantes/imunologia , Proteínas Recombinantes/uso terapêutico
12.
J Med Virol ; 91(7): 1351-1354, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30817853

RESUMO

Between September 2014 and December 2015, 298 sera from rash and fever patients from all over Cuba were investigated for specific IgM antibodies against measles, rubella, dengue, human parvovirus B19 (B19V) and human herpesvirus 6 (HHV6) using a commercial enzyme-linked immunosorbent assay kits. B19V IgM positive and equivocal samples were investigated by a polymerase chain reaction and genotyping. No measles, rubella or dengue cases were detected. HHV6-IgM antibodies were confirmed in 5.7% and B19V-IgM antibodies in 10.7% of the patients. A total of 31.3% of the B19V cases were between 5 and 9 years old and 34.4% were 20 years and older. The only B19V sequence obtained belonged to genotype 1a. Diagnosis was established for only 16% of the rash and fever patients, suggesting that other diseases such as Zika or Chikungunya may play a role.


Assuntos
Anticorpos Antivirais/sangue , Sarampo/virologia , Infecções por Parvoviridae/imunologia , Parvovirus B19 Humano/isolamento & purificação , Rubéola (Sarampo Alemão)/virologia , Adolescente , Adulto , Criança , Pré-Escolar , Coinfecção/virologia , Cuba , Ensaio de Imunoadsorção Enzimática , Exantema/virologia , Feminino , Febre/virologia , Humanos , Imunoglobulina M/sangue , Lactente , Masculino , Adulto Jovem
13.
Pak J Pharm Sci ; 32(1(Special)): 377-382, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30852473

RESUMO

Laboratory-prepared inactivated porcine parvovirus (PPV) vaccines and VP2 virus-like particles (VLPs) were utilized to immunize gilts. PPV BQ strain and SP2/0 cells were used. Hemagglutination-inhibiting (HI) antibody titers were measured in the immunized gilts and the differences in cytokine production of interferon gamma (IFN-γ, IL-2 and IL-4) were compared. CD4+ and CD8+ T cells proliferation were compared by flow cytometry. The variation between the immune response level induced by inactivated PPV vaccine and VP2 VLPs were determined. The results showed that all vaccinated gilts had HI antibody titers reaching 1:256 for at least one month post immunization and the peak level of antibody could be sustained for one month; further, PPV antibodies could be detected in the second week post immunization with VP2 VLPs. We also found that the level of cytokines (IFN-γ, IL-2 and IL-4) were all increased post immunization and continued to rise after the booster immunization; the level of increase in IFN-γ and IL-2 were significantly higher than IL-4. The flow cytometry results showed that the numbers of the CD4+ and CD8+ T cells subsets were significantly higher in the groups immunized with inactivated PPV vaccine or VP2 VLPs than those of negative control group (p<0.01); additionally, the number of CD4+ cells in the gilts that received VP2 VLP immunization was significantly higher than the inactivated vaccine group (p<0.01). In summary, the inactivated PPV vaccine and PPV VP2 VLPs were both able to induce humoral and cellular immunity, but the VP2 VLPs lead to better cellular immune responses in gilts compared to those of the inactivated vaccine..


Assuntos
Antígenos Virais/imunologia , Proteínas do Capsídeo/imunologia , Infecções por Parvoviridae/prevenção & controle , Parvovirus Suíno/imunologia , Doenças dos Suínos/prevenção & controle , Vacinas de Partículas Semelhantes a Vírus/imunologia , Animais , Anticorpos Antivirais/sangue , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD8-Positivos/imunologia , Proliferação de Células/efeitos dos fármacos , Citocinas/sangue , Feminino , Imunidade Celular/efeitos dos fármacos , Imunidade Humoral/efeitos dos fármacos , Infecções por Parvoviridae/sangue , Infecções por Parvoviridae/imunologia , Infecções por Parvoviridae/veterinária , Suínos , Doenças dos Suínos/sangue , Doenças dos Suínos/imunologia , Vacinas de Produtos Inativados/imunologia
14.
J Med Virol ; 91(7): 1224-1231, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30851123

RESUMO

Usually transmitted via respiratory droplets, parvovirus B19 (B19V) can also be acquired by blood transfusion especially because of viral persistence, resistance to blood treatment procedures, and high viral load during the early infection phase. This is particularly problematic in immunocompromised or anemic patients where the infection can have a severe outcome. As B19V DNA was detected in blood donations from South Brazil during a viral metagenomic survey performed by Next-Generation Sequencing, the objective of this retrospective study was to evaluate the seroprevalence, B19V DNA presence and circulating genotypes in a Hospital Blood Transfusion Service in Santa Maria city in South Brazil (Rio Grande do Sul state). Among 480 volunteer blood donors, 53.9% (n = 258 of 479) were anti-B19V IgG-positive, and 9 (1.9%) plasma samples presented B19V DNA. In almost all cases (n = 7 of 9, 77.8%), B19V DNA load was accompanied by the presence of anti-B19V IgG suggesting a persistent infection. The sequencing of the strains demonstrated that all belong to genotype 1 which is the most prevalent worldwide. The analysis of the recipient information of the positive for B19V DNA units revealed no related posttransfusion adverse effects. Our results demonstrate for the first time, B19V seroprevalence, viral load, and genotypes among blood donors from South Brazil and give a light for the circulation and impact of this B19V in this part of the country.


Assuntos
Anticorpos Antivirais/sangue , Doadores de Sangue , Infecções por Parvoviridae/epidemiologia , Parvovirus B19 Humano/isolamento & purificação , Carga Viral , Adolescente , Adulto , Idoso , Brasil/epidemiologia , DNA Viral/sangue , Feminino , Genótipo , Humanos , Imunoglobulina G/sangue , Imunoglobulina M/sangue , Masculino , Pessoa de Meia-Idade , Infecções por Parvoviridae/imunologia , Parvovirus B19 Humano/genética , Parvovirus B19 Humano/imunologia , Prevalência , Estudos Retrospectivos , Estudos Soroepidemiológicos , Adulto Jovem
15.
Exp Clin Transplant ; 17(Suppl 1): 195-197, 2019 01.
Artigo em Inglês | MEDLINE | ID: mdl-30777553

RESUMO

Parvovirus B19 is a single-stranded DNA virus that typically has an affinity for erythroid progenitor cells in bone marrow and leads to pure red cell aplasia. This is a common pathogen in humans, and the expression of the infection depends on the host's hematologic and immunologic status. Here, we report a female patient who developed severe and persistent anemia after kidney transplant while being on immunosuppressive therapy. The parvovirus B19 immunoglobulin M test was positive, and the virus was detected by polymerase chain reaction as parvovirus B19 (23.5 million copies/mL) in the blood sample. Bone marrow examination revealed giant pronormoblasts. She responded well to intravenous immunoglobulin without adverse event. Hemoglobin levels gradually increased, and normal levels were achieved at 3 months posttreatment. Although her renal function did not deteriorate, severe anemia (with hemoglobin level 5 g/dL) recurred 3 times during 12 months posttransplant.


Assuntos
Imunoglobulinas Intravenosas/administração & dosagem , Fatores Imunológicos/administração & dosagem , Transplante de Rim/efeitos adversos , Infecções Oportunistas/tratamento farmacológico , Infecções por Parvoviridae/tratamento farmacológico , Parvovirus B19 Humano/efeitos dos fármacos , Aplasia Pura de Série Vermelha/tratamento farmacológico , Adulto , Esquema de Medicação , Feminino , Humanos , Hospedeiro Imunocomprometido , Imunossupressores/administração & dosagem , Imunossupressores/efeitos adversos , Infecções Oportunistas/diagnóstico , Infecções Oportunistas/imunologia , Infecções Oportunistas/virologia , Infecções por Parvoviridae/diagnóstico , Infecções por Parvoviridae/imunologia , Infecções por Parvoviridae/virologia , Parvovirus B19 Humano/imunologia , Parvovirus B19 Humano/patogenicidade , Recidiva , Aplasia Pura de Série Vermelha/diagnóstico , Aplasia Pura de Série Vermelha/imunologia , Aplasia Pura de Série Vermelha/virologia , Resultado do Tratamento , Ativação Viral
16.
Methods ; 158: 44-53, 2019 04 01.
Artigo em Inglês | MEDLINE | ID: mdl-30703462

RESUMO

Serological assays detecting antibodies in serum or plasma samples are useful and versatile instruments to investigate an individual's infection and vaccination history, e.g. for clinical diagnosis, personal risk evaluation, and seroepidemiological studies. Multiplex Serology is a suspension bead array-based high-throughput methodology for simultaneous measurement of antibodies against multiple pathogens in a single reaction vessel, thus economizing sample volume, measurement time, and costs. We developed and validated bead-based pathogen-specific Monoplex Serology assays, i.e. assays including only antigens for the respective pathogen, to detect antibodies against Corynebacterium diphtheriae and Clostridium tetani toxins, rubella virus and parvovirus B19. The developed assays expand the portfolio of existing pathogen-specific bead-based serology assays and can be efficiently incorporated into larger Multiplex Serology panels. The newly developed Monoplex Serology assays consist of only one antigen per infectious agent, expressed as Glutathione S-transferase-fusion proteins in E. coli. Specificity, sensitivity and Cohen's kappa statistics in comparison with routine clinical diagnostic assays were calculated for serum dilutions 1:100 and 1:1000. All pathogen-specific assays were successfully validated at both serum dilutions with the exception of rubella Monoplex Serology which showed impaired sensitivity (57.6%) at dilution 1:1000. Specificities of successfully validated Monoplex Serology assays ranged from 85.6% to 100.0% (median: 91.7%), and sensitivities from 81.3% to 95.8% (median: 90.9%); agreement with the reference assays ranged from substantial to almost perfect (kappa: 0.66-0.86, median: 0.78). Statistical performance and slim assay design enable efficient incorporation of the developed assays into Multiplex Serology.


Assuntos
Anticorpos Antibacterianos/isolamento & purificação , Anticorpos Antivirais/isolamento & purificação , Ensaios de Triagem em Larga Escala/métodos , Testes Sorológicos/métodos , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/imunologia , Anticorpos Antivirais/sangue , Anticorpos Antivirais/imunologia , Antígenos de Bactérias/genética , Antígenos de Bactérias/imunologia , Antígenos Virais/genética , Antígenos Virais/imunologia , Clostridium tetani/imunologia , Corynebacterium diphtheriae/imunologia , Difteria/sangue , Difteria/diagnóstico , Difteria/imunologia , Difteria/microbiologia , Ensaio de Imunoadsorção Enzimática/instrumentação , Ensaio de Imunoadsorção Enzimática/métodos , Ensaios de Triagem em Larga Escala/instrumentação , Humanos , Imunoglobulina G/sangue , Imunoglobulina G/imunologia , Imunoglobulina G/isolamento & purificação , Fenômenos Magnéticos , Microesferas , Modelos Animais , Infecções por Parvoviridae/sangue , Infecções por Parvoviridae/diagnóstico , Infecções por Parvoviridae/imunologia , Infecções por Parvoviridae/virologia , Parvovirus B19 Humano/imunologia , Proteínas Recombinantes/genética , Proteínas Recombinantes/imunologia , Rubéola (Sarampo Alemão)/sangue , Rubéola (Sarampo Alemão)/diagnóstico , Rubéola (Sarampo Alemão)/imunologia , Rubéola (Sarampo Alemão)/virologia , Vírus da Rubéola/imunologia , Sensibilidade e Especificidade , Testes Sorológicos/instrumentação , Tétano/sangue , Tétano/diagnóstico , Tétano/imunologia , Tétano/microbiologia , Toxina Tetânica/genética , Toxina Tetânica/imunologia
17.
Viruses ; 11(2)2019 01 25.
Artigo em Inglês | MEDLINE | ID: mdl-30691064

RESUMO

Decades ago, Friedmann and Roblin postulated several barriers to gene therapy, including tissue targeting, delivery across the blood⁻brain barrier (BBB), and host immune responses. These issues remain pertinent till today. Since then, several advances have been made in elucidating structures of adeno-associated virus (AAV) serotypes, antibody epitopes, and ways to modify antibody-binding sites. AAVs capsid has also been engineered to re-direct tissue tropism, reduce ubiquitination, and promote passage across the BBB. Furthermore, the use of high(er) dose recombinant AAV (rAAV) has been accompanied by a better understanding of immune responses in both experimental animals and early clinical trials, and novel work is being performed to modulate the immune response. While the immune responses to rAAV remains a major challenge in translating experimental drugs to approved medicine, and will likely require more than a single solution, we now better understand the hurdles to formulate and test experimental solutions to surmount them.


Assuntos
Dependovirus/imunologia , Terapia Genética , Vetores Genéticos/imunologia , Interações entre Hospedeiro e Microrganismos/imunologia , Imunidade Inata , Infecções por Parvoviridae/imunologia , Imunidade Adaptativa , Animais , Proteínas do Capsídeo/genética , Proteínas do Capsídeo/imunologia , Ensaios Clínicos como Assunto , Humanos , Camundongos
18.
J Pediatric Infect Dis Soc ; 8(1): 73-76, 2019 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-29415165

RESUMO

Single gene defects that impair lymphocyte cytotoxicity can predispose to severe viral infection that normally remains subclinical. The classic severe presentation is hemophagocytic lymphohistiocytosis. Here, we report the case of a neonate who presented with cytomegalovirus palatal ulceration and bocavirus pneumonitis secondary to impaired cytotoxicity caused by biallelic mutations in the UNC13D gene.


Assuntos
Infecções por Citomegalovirus/imunologia , Citotoxicidade Imunológica , Bocavirus Humano/isolamento & purificação , Linfócitos/imunologia , Proteínas de Membrana/genética , Palato Duro/imunologia , Infecções por Parvoviridae/imunologia , Pneumonia Viral/imunologia , Úlcera/imunologia , Infecções por Citomegalovirus/patologia , Humanos , Recém-Nascido , Masculino , Mutação , Palato Duro/patologia , Palato Duro/virologia , Infecções por Parvoviridae/genética , Infecções por Parvoviridae/patologia , Pneumonia Viral/genética , Pneumonia Viral/patologia , Úlcera/patologia , Úlcera/virologia
19.
J Appl Microbiol ; 126(1): 49-57, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30288879

RESUMO

AIMS: To evaluate the effect of a DNA priming and protein boosting immunization scheme in ducks. METHODS AND RESULTS: Pekin ducks were immunized with pTCY/VP2 DNA vaccine; on day 14 (D14) after primary immunization, the ducks were boosted with either the same vaccine (DNA + DNA) or the rVP2 vaccine (DNA + rVP2). CpG oligodeoxynucleotides containing three copies of GACGTT motifs were used as the adjuvant in the vaccines. Compared with unimmunized controls, both immunization schemes significantly increased the titre of antigen-specific antibodies, lymphocyte proliferation index, percentage of CD4+ and CD8+ cells in peripheral blood mononuclear cells (PBMCs) and mRNA expression of interferon (IFN)-α, IFN-γ, interleukin (IL)-6 and IL-12 in antigen-stimulated PBMCs. Furthermore, compared with the DNA + DNA homologous scheme, the DNA + rVP2 heterologous scheme significantly increased lymphocyte proliferation, percentage of CD4+ and CD8+ cells in PBMCs and upregulation of mRNA expression of cytokines 2 weeks after the boost (D28). CONCLUSIONS: The DNA + rVP2 immunization scheme enhanced immune responses, mainly Th1 type, against parvovirus in ducks. SIGNIFICANCE AND IMPACT OF THE STUDY: The DNA priming and protein boosting heterologous immunization strategy can be applied to develop vaccines against viral infections in ducks. It can potentially be used in breeding ducks because of long-term immunity may confer protection for ducklings.


Assuntos
Infecções por Parvoviridae/veterinária , Parvovirus/imunologia , Doenças das Aves Domésticas/prevenção & controle , Vacinas de DNA/administração & dosagem , Proteínas Virais/administração & dosagem , Adjuvantes Imunológicos/administração & dosagem , Animais , Citocinas/genética , Citocinas/imunologia , Patos , Imunização , Imunização Secundária , Leucócitos Mononucleares/imunologia , Infecções por Parvoviridae/imunologia , Infecções por Parvoviridae/prevenção & controle , Infecções por Parvoviridae/virologia , Parvovirus/genética , Doenças das Aves Domésticas/genética , Doenças das Aves Domésticas/imunologia , Doenças das Aves Domésticas/virologia , Células Th1/imunologia , Vacinas de DNA/genética , Vacinas de DNA/imunologia , Proteínas Virais/genética , Proteínas Virais/imunologia
20.
BMC Vet Res ; 14(1): 348, 2018 Nov 16.
Artigo em Inglês | MEDLINE | ID: mdl-30445957

RESUMO

BACKGROUND: Canine Distemper Virus (CDV) and Canine Parvovirus (CPV) lead to infections with high mortality rates in dogs. These viruses affect unvaccinated dogs or dogs with incomplete vaccination protocols. Vaccination plays an important role in reducing death rates, preventing clinical cases and controlling the spread of virus However, the efficacy of vaccination might be affected by different factors including vaccine scheduling and the neutralization of the vaccine targets by maternal antibodies. In face of these factors, the main goals of this study are (i) to investigate the antibody responses of puppies undergoing different primary vaccination protocols against CPV and CDV and (ii) to estimate the time until seroreversion in adult dogs unvaccinated for at least 3 years. RESULTS: Antibody protection against CDV and CPV was evaluated in a total of 20 dogs: 5 puppies that initiated immunization at 6 weeks after birth (group A), 8 animals that started vaccination between 8 and 12 weeks of age (group B), and 7 adult dogs that have not been vaccinated for at least 3 years (group C). Blood samples were collected from each animal, with 3 to 4 weeks apart. Antibody responses were measured using indirect ELISA. In the second immunization point, no significant differences were found between the seroconversion of groups A and B for each viral infection (p = 0.81 and 0.20 for CDV and CPV, respectively). In the third immunization, there was evidence for a shorter time to achieve a protective titer against CPV in group B when compared to group A (p = 0.015). Similar evidence was not found for CDV (p-value = 0.41). In Group C, the average time until seroveversion was estimated at 2.86 years and 7.63 years for CDV and CPV, respectively. CONCLUSION: Vaccine response to CDV and CPV is specific in each individual. Effective immune protection in primary vaccination depends mainly on the initial titer of maternal antibodies acquired by the neonate. Other factors such as environmental exposure, immunization schedules and immune system activity influence the duration of immunity in adult dogs. The variability found reinforces the need to determine individual humoral immunity levels in order to assess vaccine efficacy.


Assuntos
Vírus da Cinomose Canina/imunologia , Cinomose/imunologia , Doenças do Cão/imunologia , Imunidade Humoral , Infecções por Parvoviridae/veterinária , Parvovirus Canino/imunologia , Vacinas Virais/uso terapêutico , Animais , Anticorpos Antivirais/sangue , Cinomose/prevenção & controle , Doenças do Cão/prevenção & controle , Doenças do Cão/virologia , Cães , Ensaio de Imunoadsorção Enzimática/veterinária , Feminino , Imunidade Humoral/imunologia , Masculino , Infecções por Parvoviridae/imunologia , Infecções por Parvoviridae/prevenção & controle , Projetos Piloto
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