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1.
Anal Chim Acta ; 1302: 342514, 2024 May 08.
Artigo em Inglês | MEDLINE | ID: mdl-38580408

RESUMO

Monkeypox (mpox) is spreading around the world, and its rapid diagnosis is of great significance. In the present study, a rapid and sensitive fluorescent chromatography assisted with cloud system was developed for point-of-care diagnosis of mpox. To screen high affinity antibodies, nanoparticle antigen AaLS-A29 was generated by conjugating A29 onto scaffold AaLS. Immunization with AaLS-A29 induced significantly higher antibody titers and monoclonal antibodies were generated with the immunized mice. A pair of monoclonal antibodies, MXV 14 and MXV 15, were selected for fluorescence chromatography development. The Time-Resolved Fluorescence Immunoassay (TRFIA) was used to develop the chromatography assay. After optimization of the label and concentration of antibodies, a sensitive TRFIA assay with detection limit of 20 pg/mL and good repeatability was developed. The detection of the surrogate Vaccinia virus (VACA) strain Tian Tan showed that the TRFIA assay was more sensitive than the SYBR green I based quantitative PCR. In real samples, the detection result of this assay were highly consistent with the judgement of Quantitative Real-Time PCR (Concordance Rate = 90.48%) as well as the clinical diagnosis (Kappa Value = 0.844, P < 0.001). By combining the portable detection and online cloud system, the detection results could be uploaded and shared, making this detection system an ideal system for point-of-care diagnosis of mpox both in field laboratory and outbreak investigation.


Assuntos
Mpox , Animais , Camundongos , Sistemas Automatizados de Assistência Junto ao Leito , Fluorimunoensaio/métodos , Anticorpos Monoclonais
2.
Vaccine ; 42(8): 2072-2080, 2024 Mar 19.
Artigo em Inglês | MEDLINE | ID: mdl-38423815

RESUMO

Monkeypox (mpox) is a zoonotic disease caused by monkeypox virus (MPXV) of the orthopoxvirus genus. The emergence and global spread of mpox in 2022 was declared as a public health emergency by World Health Organization. This mpox pandemic alarmed us that mpox still threaten global public health. Live vaccines could be used for immunization for this disease with side effects. New alternative vaccines are urgently needed for this re-emerging disease. Specific antibody responses play key roles for protection against MPXV, therefore, vaccines that induce high humoral immunity will be ideal candidates. In the present study, we developed thermostable nanovaccine candidates for mpox by conjugating MPXV antigens with thermostable nanoscafolds. Three MPXV protective antigens, L1, A29, and A33, and the thermostable Aquafex aeolicus lumazine synthase (AaLS), were expressed in E. coli and purified by Ni-NTA methods. The nanovaccines were generated by conjugation of the antigens with AaLS. Thermal stability test results showed that the nanovaccines remained unchanged after one week storage under 37℃ and only partial degradation under 60℃, indicating high thermostability. Very interesting, one dose immunization with the nanovaccine could induce high potent antibody responses, and two dose induced 2-month high titers of antibodes. In vitro virus neutralization test showed that nanovaccine candidates induced significantly higher levels of neutralization antibodies than monomers. These results indicated that the AaLS conjugation nanovaccines of MPXV antigens are highly thermostable in terms of storage and antigenic, being good alternative vaccine candidates for this re-emerging disease.


Assuntos
Terapias Complementares , Mpox , Humanos , Nanovacinas , Escherichia coli , Adjuvantes Imunológicos , Anticorpos , Antígenos Virais , Monkeypox virus
3.
Vaccine ; 42(4): 732-737, 2024 Feb 06.
Artigo em Inglês | MEDLINE | ID: mdl-38220487

RESUMO

Foot-and-mouth disease (FMD) is an acute zoonosis causes significant economic losses. Vaccines able to stimulate efficient protective immune responses are urgently needed. In this study, Escherichia coli-derived recombinant VP1 of serotype A and O FMD virus (FMDV) was conjugated to thermostable scaffold lumazine synthase (LS) or Quasibacillus thermotolerans encapsulin (QtEnc) using a robust plug-and-display SpyTag/SpyCatcher system to generate multimeric nanovaccines. These nanovaccines induced highly potent antibody responses in vaccinated mice. On day 14 after the first immunisation, antibody titres were approximately 100 times higher than those of monomer antigens. Both vaccines induced high and long-term IgG antibody production. Moreover, the QtEnc-VP1 nanovaccine induced higher antibody titres than the LS-VP1 nanovaccine. The nanovaccines also induced Th1-biased immune responses and higher levels of neutralising antibodies. These data indicated that FMDV nanovaccines generated by conjugating VP1 with a thermostable scaffold are highly immunogenic and ideal candidates for FMDV control in low-resource areas.


Assuntos
Vírus da Febre Aftosa , Febre Aftosa , Vacinas Virais , Animais , Camundongos , Nanovacinas , Anticorpos Antivirais , Adjuvantes Imunológicos , Imunidade , Proteínas do Capsídeo
4.
Adv Sci (Weinh) ; 10(26): e2303049, 2023 09.
Artigo em Inglês | MEDLINE | ID: mdl-37395451

RESUMO

Antigen delivery based on non-virus-like particle self-associating protein nanoscffolds, such as Aquifex aeolicus lumazine synthase (AaLS), is limited due to the immunotoxicity and/or premature clearance of antigen-scaffold complex resulted from triggering unregulated innate immune responses. Here, using rational immunoinformatics prediction and computational modeling, we screen the T epitope peptides from thermophilic nanoproteins with the same spatial structure as hyperthermophilic icosahedral AaLS, and reassemble them into a novel thermostable self-assembling nanoscaffold RPT that can specifically activate T cell-mediated immunity. Tumor model antigen ovalbumin T epitopes and the severe acute respiratory syndrome coronavirus 2 receptor-binding domain are loaded onto the scaffold surface through the SpyCather/SpyTag system to construct nanovaccines. Compared to AaLS, RPT -constructed nanovaccines elicit more potent cytotoxic T cell and CD4+ T helper 1 (Th1)-biased immune responses, and generate less anti-scaffold antibody. Moreover, RPT significantly upregulate the expression of transcription factors and cytokines related to the differentiation of type-1 conventional dendritic cells, promoting the cross-presentation of antigens to CD8+ T cells and Th1 polarization of CD4+ T cells. RPT confers antigens with increased stability against heating, freeze-thawing, and lyophilization with almost no antigenicity loss. This novel nanoscaffold offers a simple, safe, and robust strategy for boosting T-cell immunity-dependent vaccine development.


Assuntos
Linfócitos T CD8-Positivos , COVID-19 , Humanos , Imunidade Celular , Linfócitos T Citotóxicos , Antígenos de Neoplasias
5.
Diagn Microbiol Infect Dis ; 105(4): 115890, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-36739792

RESUMO

Golgi protein 73 (GP73) has been recognized as a biomarker for evaluating liver diseases, although the serum profile of patients with HIV remains unclear. This study was designed to investigate the diagnostic values of serum GP73 in patients with HIV. A total of 92 patients with HIV and 60 healthy participants were selected, and serum samples were collected; 51 of 92 patients were followed up and all indicators were re-tested after 1 year. Patients with HIV had significantly lower GP73 concentration, lower viral load, and higher CD4+ T cell counts after antiretroviral treatment. A significant correlation between the changes of GP73 level and CD4+ T cell count was observed. The CD4+ T cell count was significantly correlated with the glycosylated GP73 level. The area under the ROC curve (AUC) of GP73 to predict negative viral load-negative conversion alone was 0.705. When the cut-off value was set at 146.7 ng/mL, the sensitivity and specificity were 73% and 70% respectively. These results indicate that serum GP73 may have predictive ability for negative viral load-negative conversion.


Assuntos
Carcinoma Hepatocelular , Infecções por HIV , Neoplasias Hepáticas , Humanos , Carcinoma Hepatocelular/diagnóstico , Neoplasias Hepáticas/diagnóstico , Seguimentos , Proteínas de Membrana , Infecções por HIV/diagnóstico , Infecções por HIV/tratamento farmacológico
6.
One Health ; 15: 100449, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36532675

RESUMO

Brucellosis is a typical zoonosis driven by various risk factors, including environmental ones. The present study aimed to explore the driving effect of environmental factors on human brucellosis in a high incidence rate area, which provides understanding and implications in mitigating disease transmission risk in a multi-system between the human-animal-environment interface for preventing and controlling brucellosis based on the One Health concept. Based on the monthly time series data of human brucellosis and environmental variables, a Seasonal Autoregressive Integrated Moving Average Model with explanatory variables (SARIMAX) was applied to assess the association between environmental indicators and human brucellosis incidence (IHB). The results indicated distinct seasonal fluctuation during the study duration, tending to climb from April to August. Atmospheric pressure, precipitation, relative humidity, mean temperature, sunshine duration, and normalized difference vegetation index significantly drive IHB. Moreover, the well-fitting and predicting capability were performed and assessed in the optimal model was the SARIMAX (0,1,1) (0,1,1)12 model with the normalized difference vegetation index (ß = 0.349, P = 0.036) and mean temperature (ß = 0.133, P = 0.046) lagged in 6 months, and the precipitation lagged in 1 month (ß = -0.090, P = 0.004). Our study suggests the association between environmental risk factors and human brucellosis infection, which can be contributed to mitigating the transmission risk in the environmental drivers in a multi-system interface through comprehensive prevention and intervention strategies based on the One Health concept.

7.
Front Public Health ; 10: 856103, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-35784246

RESUMO

Objective: This article aims to study the influencing factors of pgRNA and its change magnitude based on the real world. Methods: A total of 421 patients who were tested for pgRNA were selected. According to the baseline data, the subjects were divided into negative and positive groups. The Chi-square test and logistic regression were used to analyze the influencing factors of pgRNA status. Based on the follow-up data, the rank-sum test and linear regression were used to analyze the influencing factors of pgRNA change magnitude. Results: A total of 153 (36.3%) of the 421 subjects were pgRNA-negative and 268 (63.7%) were pgRNA-positive. Logistic regression analysis showed that positive HBV DNA (OR: 40.51), positive HBeAg (OR: 66.24), tenofovir treatment (OR: 23.47), and entecavir treatment (OR: 14.90) were the independent risk factors for positive pgRNA. Univariate linear regression showed that the pgRNA change magnitude of patients treated with entecavir was higher than that of patients treated with tenofovir. Multivariate linear regression showed that age was an independent factor influencing pgRNA change magnitude. Conclusions: The pgRNA of patients who were young, female, HBV DNA-positive, high-HBsAg, HBeAg-positive is higher than the detection line. HBV DNA and HBeAg are the independent risk factors of positive pgRNA. Different antiviral regimens and disease stages have significantly different effects on pgRNA status. There was a significant correlation between pgRNA and FIB-4, suggesting that pgRNA is related to liver fibrosis. The decrease in pgRNA was greater in young patients than in non-young patients. The decrease in pgRNA was greater in patients treated with tenofovir than in patients treated with entecavir.


Assuntos
Vírus da Hepatite B , Hepatite B , DNA Viral/análise , DNA Viral/genética , Feminino , Antígenos E da Hepatite B , Vírus da Hepatite B/genética , Humanos , RNA Viral/análise , Tenofovir/uso terapêutico
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