scholarly journals Molecular Characterization of a Tetraspanin TSP11 Gene in Echinococcus granulosus and Evaluation Its Immunoprotection in Model Dogs

2021 ◽  
Vol 8 ◽  
Author(s):  
Jinwen Xian ◽  
Pengpeng Zhao ◽  
Ning Wang ◽  
Weiye Wang ◽  
Yanyan Zhang ◽  
...  
Keyword(s):  
Echinococcus Granulosus ◽  
Vaccine Development ◽  
Serum Levels ◽  
Disease Diagnosis ◽  
Vaccine Antigen ◽  
Interleukin 4 ◽  
Control Group ◽  
Potential Candidate ◽  
Intermediate Hosts ◽  
Wide Range

Cystic echinococcosis (CE) is a cosmopolitan zoonosis caused by the larval stage of Echinococcus granulosus, which affects humans and a wide range of mammalian intermediate hosts. Parasite tetraspanin proteins are crucial for host-parasite interactions, and therefore they may be useful for vaccine development or disease diagnosis. In the present study, the major antigen coding sequence of tetraspanin 11 (Eg-TSP11) from E. granulosus was determined. The results of immunolocalization showed that Eg-TSP11 was mainly located in the tegument of adult worms and protoscoleces. Western blotting analysis showed that the serum from dogs injected with recombinant Eg-TSP11 (rEg-TSP11) could recognize Eg-TSP11 among natural protoscolex proteins. Moreover, the serum from dogs with E. granulosus infection also recognized rEg-TSP11. Serum indirect enzyme-linked immunosorbent assays demonstrated that IgG levels gradually increased after the first immunization with rEg-TSP11 compared with those in the control group. Furthermore, the serum levels of interleukin 4, interleukin 5, and interferon gamma were significantly altered in the rEg-TSP11 group. Importantly, we found that vaccination with rEg-TSP11 significantly decreased worm burden and inhibited segment development in a dog model of E. granulosus infection. Based on these findings, we speculated that rEg-TSP11 might be a potential candidate vaccine antigen against E. granulosus infection in dogs.

scholarly journals Insights into Cross-species Evolution of Novel Human Coronavirus 2019-nCoV and Defining Immune Determinants for Vaccine Development

2020 ◽  
Author(s):  
Arunachalam Ramaiah ◽  
Vaithilingaraja Arumugaswami
Keyword(s):  
Binding Affinity ◽  
Vaccine Development ◽  
Asia Pacific ◽  
Potential Candidate ◽  
T Cell Epitopes ◽  
Live Animal ◽  
Synonymous Mutations ◽  
Wide Range ◽  
Novel Coronavirus ◽  
N Proteins

ABSTRACTNovel Coronavirus (nCoV) outbreak in the city of Wuhan, China during December 2019, has now spread to various countries across the globe triggering a heightened containment effort. This human pathogen is a member of betacoronavirus genus carrying 30 kilobase of single positive-sense RNA genome. Understanding the evolution, zoonotic transmission, and source of this novel virus would help accelerating containment and prevention efforts. The present study reported detailed analysis of 2019-nCoV genome evolution and potential candidate peptides for vaccine development. This nCoV genotype might have been evolved from a bat-CoV by accumulating non-synonymous mutations, indels, and recombination events. Structural proteins Spike (S), and Membrane (M) had extensive mutational changes, whereas Envelope (E) and Nucleocapsid (N) proteins were very conserved suggesting differential selection pressures exerted on 2019-nCoV during evolution. Interestingly, 2019-nCoV Spike protein contains a 39 nucleotide sequence insertion relative to SARS-like bat-SL-CoVZC45/2017. Furthermore, we identified eight high binding affinity (HBA) CD4 T-cell epitopes in the S, E, M and N proteins, which can be commonly recognized by HLA-DR alleles of Asia and Asia-Pacific Region population. These immunodominant epitopes can be incorporated in universal subunit CoV vaccine. Diverse HLA types and variations in the epitope binding affinity may contribute to the wide range of immunopathological outcomes of circulating virus in humans. Our findings emphasize the requirement for continuous surveillance of CoV strains in live animal markets to better understand the viral adaptation to human host and to develop practical solutions to prevent the emergence of novel pathogenic CoV strains.

scholarly journals Molecular characterization and immune protection of the 3-hydroxyacyl-CoA dehydrogenase gene in Echinococcus granulosus

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Jinwen Xian ◽  
Ning Wang ◽  
Pengpeng Zhao ◽  
Yanyan Zhang ◽  
Jimeng Meng ◽  
...  
Keyword(s):  
Echinococcus Granulosus ◽  
Interleukin 4 ◽  
Effective Vaccine ◽  
Control Group ◽  
Protective Effects ◽  
Immune Protection ◽  
Reading Frame ◽  
Reverse Transcription Pcr ◽  
Frame Sequence ◽  
Hydroxyacyl Coa Dehydrogenase

Abstract Background Cystic echinococcosis (CE) is a serious parasitic zoonosis caused by the larvae of the tapeworm Echinococcus granulosus. The development of an effective vaccine is one of the most promising strategies for controlling CE. Methods The E. granulosus 3-hydroxyacyl-CoA dehydrogenase (EgHCDH) gene was cloned and expressed in Escherichia coli. The distribution of EgHCDH in protoscoleces (PSCs) and adult worms was analyzed using immunofluorescence. The transcript levels of EgHCDH in PSCs and adult worms were analyzed using quantitative real-time reverse transcription PCR (RT-qPCR). The immune protective effects of the rEgHCDH were evaluated. Results The 924-bp open reading frame sequence of EgHCDH, which encodes a protein of approximately 34 kDa, was obtained. RT-qPCR analysis revealed that EgHCDH was expressed in both the PSCs and adult worms of E. granulosus. Immunofluorescence analysis showed that EgHCDH was mainly localized in the tegument of PSCs and adult worms. Western blot analysis showed that the recombinant protein was recognized by E. granulosus-infected dog sera. Animal challenge experiments demonstrated that dogs immunized with recombinant (r)EgHCDH had significantly higher serum IgG, interferon gamma and interleukin-4 concentrations than the phosphate-buffered saline (PBS) control group. The rEgHCDH vaccine was able to significantly reduce the number of E. granulosus and inhibit the segmental development of E. granulosus compared to the PBS control group. Conclusions The results suggest that rEgHCDH can induce partial immune protection against infection with E. granulosus and could be an effective candidate for the development of new vaccines. Graphical abstract

scholarly journals Probiotics Effect on Interleukin-4 (IL-4) and Imunoglobulin E (IgE) Levels on Asthmatic Patients

Sains Medika ◽  
2016 ◽  
Vol 6 (2) ◽  
pp. 61 ◽  
Author(s):  
Pujiati Pujiati
Keyword(s):  
Serum Level ◽  
Childhood Asthma ◽  
Immunoglobulin E ◽  
Controlled Trial ◽  
Intervention Group ◽  
Serum Levels ◽  
Interleukin 4 ◽  
Control Group ◽  
Asthmatic Patients ◽  
Therapeutic Benefits

Introduction: Probiotics may play a role in immune system maturation and may reduce the risk of allergies and asthma in childhood. However, the therapeutic benefits of probiotics in asthma depend on various factors such as strain of probiotics and dosing regimen. Objectives: The aims of this study was to evaluate the effect of probiotic (LactoB®) on Immunoglobulin E (IgE) and Interleukin (IL-4) serum level in childhood Asthma. Methods: Forty children aged 1-5 years with asthma were recruited into a randomized controlled trial. The children were assigned into a probiotic (Lactobacillus acidophilus, Bifidobacterium longun, Streptococcus; Lacto B®) or an equivalent volume of placebo, twice daily orally for 8 weeks. The IgE and IL-4 serum level were determined by ELISA. The differences between groups were analyzed by t-test dependent with confidence interval of 95%. Results: In intervention group, mean of IgE serum levels after the probiotics treatment was significantly lower compared to that of before the treatment (148.18 pg/mg; 283.20 pg/mg; respectively). Mean of IL-4 serum levels after the treatment was significantly lower compared with that of before the treatment (111.03 pg/mg; 142.08 pg/mg respectively). In control group, there were no significant differences between IgE serum levels mean before the administration of placebo and after the intervention (292.39 pg/ml; 286.94 pg/ml respectively). There were no significant differences between IL-4 serum levels mean before and after the treatment (136.76 pg/ml; 139.56 pg/ml).Conclusion: there was an effect of probiotics supplementation on IgE and IL-4 serum levels in childhood asthma.

scholarly journals Molecular Characterization and Expression Analysis of the Gene Encoding 3-Hydroxyacyl-CoA Dehydrogenase (EGR-03347) from Echinococcus Granulosus and the Evaluation of the Immune Protection of the Definitive Hosts (dogs)

2021 ◽  
Author(s):  
Jinwen Xian ◽  
Ning Wang ◽  
Pengpeng Zhao ◽  
Yanyan Zhang ◽  
Jimeng Meng ◽  
...  
Keyword(s):  
Echinococcus Granulosus ◽  
Post Infection ◽  
Tricarboxylic Acid Cycle ◽  
Interleukin 4 ◽  
Effective Vaccine ◽  
Control Group ◽  
Gene Encoding ◽  
Transcript Levels ◽  
Key Enzyme ◽  
Hydroxyacyl Coa Dehydrogenase

Abstract Background: Cystic echinococcosis, a serious parasitic zoonosis, is caused by tapeworm (Echinococcus granulosus) larvae. The development of an effective vaccine is a promising strategy to control echinococcosis. E. granulosus has a complete tricarboxylic acid cycle pathway, in which 3-hydroxyacyl-CoA dehydrogenase (EGR-03347) is a key enzyme.Methods: In the present study, the cDNA encoding EGR-03347 in Echinococcus granulosus (rEGR-03347) was successfully cloned and the molecular and biochemical characterizations carried out. The immunoreactivity of recombinant EGR-03347 (rEGR-03347) was investigated using western blotting. The immunolocalization of EGR-03347 in different life stages of E. granulosus was determined using specific polyclonal antibody, quantitative real-time reverse transcription polymerase chain reaction was used to analyze their transcript levels in PSCs and 28-day strobilated worms stages. In addition, recombinant protein rEGR-03347 was mixed with the adjuvant Quil A for vaccinating dogs, after three vaccine injections, all the dogs were orally challenge-infected with 100000 protoscoleces of E. granulosus. After 28 days of infection, all the dogs were euthanized and necropsied for collecting and counting E. granulosus worms, post-infection the antibody and cytokine were measured for the immunogenicity analysis of this protein.Results: rEGR-03347 is a highly conserved protein, consisting of 308 amino acids. Recombinant EGR-03347 could be identifed in the sera of patients with CE and in mouse anti-rEGR-03347 sera. Immunofluorescence analysis showed that EGR-03347 mainly localized in the tegument of protoscoleces and adults, and their transcript levels were high in the 28-day strobilated worms. Furthermore, enzyme-linked immunosorbent assays post‑infection showed that IgG gradually increased after the first immunization with rEGR-03347 compared with the control group, rEGR-03347 changed the interferon gamma and interleukin 4 levels. We observed an 87.2 % reduction in E. granulosus numbers and 66.7 % inhibition of the segmental development of E. granulosus in the rEGR‑03347‑vaccinated dogs compared with the nonvaccinated controls.Conclusions: This is the first report characterizing a 3-hydroxyacyl-CoA dehydrogenase from the tapeworm E. granulosus. We have characterized the sequence, structure and location of EGR‑03347 and investigated the immunoreactivity, immunogenicity and serodiagnostic potential of rEGR‑03347 . The results demonstrated rEGR-03347 as a potential vaccine against E. granulosus infection in dogs.

scholarly journals Protective Immunity Against Neospora Caninum Infection Induced by 14-3-3 Protein in Mice

2020 ◽  
Author(s):  
Shan Li ◽  
Nan Zhang ◽  
Shaoxiong Liu ◽  
Jianhua Li ◽  
Li Liu ◽  
...  
Keyword(s):  
Immune Responses ◽  
Vaccine Development ◽  
Neospora Caninum ◽  
Survival Rates ◽  
Parasite Burden ◽  
Mouse Serum ◽  
Intermediate Hosts ◽  
Caninum Infection ◽  
Wide Range ◽  
Ifn Γ

Abstract BackgroundNeospora caninum causes infections in a wide range of intermediate hosts and remains a threatening disease worldwide because of the lack of effective drugs and vaccines. Our previous studies demonstrated that N. caninum 14-3-3 protein (Nc14-3-3), which is included in N. caninum extracellular vesicles (NEVs), can induce effective immune responses and stimulate cytokine expression in mouse peritoneal macrophages. However, whether Nc14-3-3 has a protective effect and its mechanisms are poorly understood.MethodsHere, we evaluated immune responses and protective effects of Nc14-3-3 against 2×107 Nc-1 tachyzoites. Antibody (IgG, IgGl and IgG2a) levels and Th1-type (IFN-γ and IL-12) and Th2-type (IL-4 and IL-10) cytokines in mouse serum; survival rates; survival time; and parasite burdens were detected.ResultsIn the present study, the immunostimulatory effect of Nc14-3-3 was confirmed, as it triggered Th1-type cytokine (IFN-γ and IL-12) production in mouse serum two weeks after the final immunization. Moreover, the immunization of C57BL/6 mice with Nc14-3-3 induced high IgG antibody levels and significant increases in CD8+ T lymphocytes in the spleens of mice, indicating that a significant cellular immune response was induced. Mouse survival rates and survival times were significantly prolonged after immunization survival rates were 40% for Nc14-3-3 immunization and 60% for NEV immunization, while mice that received GST, PBS, or blank control all died at 13, 9, and 8 days after intraperitoneal N. caninum challenge. In addition, qPCR analysis indicated that there was a lower parasite burden and milder pathological changes in the mice immunized with Nc14-3-3.ConclusionsOur data demonstrate the vaccination of mice with Nc14-3-3 elicits both cellular and humoural immune responses and provides partial protection against acute neosporosis. Thus, Nc14-3-3 could be an effective antigen candidate for vaccine development for neosporosis.

A novel CDA liquid biopsy technology for effective non-small cell lung cancer diagnosis.

2017 ◽  
Vol 35 (15_suppl) ◽  
pp. e23131-e23131
Author(s):  
Junjie Wu ◽  
Gengxi Jiang ◽  
Yutao Li ◽  
Xuedong Du ◽  
Hongmei Tao ◽  
...  
Keyword(s):  
Ct Scan ◽  
Liquid Biopsy ◽  
Critical Role ◽  
Area Under The Curve ◽  
Treatment Decision ◽  
Disease Diagnosis ◽  
Control Group ◽  
Threshold Values ◽  
Wide Range ◽  
Biopsy Technology

e23131 Background: Cancer Differentiation Analysis liquid biopsy (CDA) has been investigated as a viable clinical utility in NSCLC diagnosis, as well as a new candidate for part of an overall post-CT scan clinical diagnostic and treatment decision tool kit. It has been also evaluated in NSCLC diagnosis, combined with imaging technologies. Methods: This was a retrospective investigation in which 160 individuals were recruited at Changhai Hospital of Shanghai from July to Dec. in 2014. CDA and CT scan tests were performed on all samples before final confirmation by biopsy. For CDA tests, peripheral blood was drawn in EDTA tubes before operation. A performance predication model of CDA and CT scan test results was built using pROC package in R Language for the data of Area Under the Curve (AUC) and CDA threshold values. Further analysis was carried out based on the CDA threshold values. Results: Out of the 160 individuals, 40 were diagnosed as benign lung diseases selected as control group, 120 were confirmed as NSCLC by pathology. CDA and CT test data along with epidemiological information were collected and complied. Details of other statistical results were given in the Table below. Conclusions: CDA liquid biopsy is a novel, multi-level, multi-parameter based disease diagnosis technology with improvements in multiple areas including but not limited to higher sensitivity and specificity, ability to diagnose cancer early, no side effects, and covering a wide range of cancer sites. Results showed that AUC values of CDA were higher than those of CT scan for all NSCLC stage groups. CDA plus CT scan combination has the highest average AUC value for all stages. CDA test can provide more accurate and reliable diagnostic information and data for oncologists in making crucial clinical decisions. It is expected that CDA liquid biopsy technology will play an important and critical role in NSCLC diagnosis. [Table: see text]

scholarly journals Preliminary Study on Antifungal Mechanism of Aqueous Extract of Cnidium monnieri Against Trichophyton rubrum

2021 ◽  
Vol 12 ◽  
Author(s):  
Cao Yanyun ◽  
Tang Ying ◽  
Kong Wei ◽  
Fang Hua ◽  
Zhu Haijun ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Aqueous Extract ◽  
Trichophyton Rubrum ◽  
Economic Value ◽  
Differentially Expressed ◽  
Control Group ◽  
Potential Candidate ◽  
Aqueous Extracts ◽  
Antifungal Mechanism ◽  
Wide Range

Trichoderma rubrum (T. rubrum) is one of the important pathogens because it is the cause of most dermatomycosis. The treatment of Trichophyton rubrum infection is time-consuming and very expensive; it is easy for the infections to reoccur, leading to therapeutic failures, persistence, and chronic infection. These issues have inspired researchers to study natural alternative therapies instead. Cnidium monnieri (L.), as a kind of traditional Chinese medicine, has a variety of pharmacological activities and a wide range of applications, so it has a high potential for researching and economic value. We detected the effect of aqueous extract of C. monnieri (L.) on the activity of T. rubrum by Cell Count Kit-8 assay (CCK-8), and we found that 128 and 256 μg/ml of aqueous extracts of C. monnieri (L.) co-cultured with T. rubrum for 24 h showed the inhibitory effect on T. rubrum. The results of scanning electron microscopy (SEM) and transmission electron microscopy (TEM) confirmed that aqueous extract of C. monnieri (L.) damaged the T. rubrum. At the same time, mass spectrometry screening with T. rubrum before and after the treatment of 256 μg/ml of aqueous extracts of C. monnieri (L.) showed that 966 differentially expressed proteins were detected, including 524 upregulated differentially expressed genes (DEGs) and 442 downregulated DEGs. The most significantly downregulated protein was chitin synthase (CHS); and the results of qRT-PCR and Western blotting demonstrated that the expression level of CHS was downregulated in the 256 μg/ml group compared with the control group. The study showed that the aqueous extract of C. monnieri (L.) could destroy the morphology of mycelia and the internal structure of T. rubrum, and it could inhibit the growth of T. rubrum. The antifungal effect of aqueous extract of C. monnieri (L.) may be related to the downregulation of the expression of CHS in T. rubrum, and CHS may be one of the potential targets of its antifungal mechanism. We concluded that aqueous extract from C. monnieri (L.) may be a potential candidate for antifungal agents.

scholarly journals DNA Vaccine Using Mycobacterium bovis Ag85B Antigen Induces Partial Protection against Experimental Infection in BALB/c Mice

2006 ◽  
Vol 13 (8) ◽  
pp. 930-935 ◽  
Author(s):  
Francisco M. Teixeira ◽  
Henrique C. Teixeira ◽  
Ana Paula Ferreira ◽  
Michele F. Rodrigues ◽  
Vasco Azevedo ◽  
...  
Keyword(s):  
Dna Vaccine ◽  
Mycobacterium Bovis ◽  
Vaccine Development ◽  
Virulent Strain ◽  
Intracellular Cytokine Staining ◽  
Bacterial Load ◽  
Interleukin 4 ◽  
Control Group ◽  
Partial Protection ◽  
Ifn Γ

ABSTRACT Bovine tuberculosis is a major cause of economic loss in countries where it is endemic, and in some countries, it may be a significant zoonotic disease problem. Therefore, new strategies for vaccine development are required, and among them, genetic immunization has potential value. The main goal of this study was to test the Mycobacterium bovis Ag85B gene as a DNA vaccine following challenge with an M. bovis virulent strain (ATCC 19274). Groups of BALB/c mice (n = 10) were immunized four times intramuscularly with the pCI-Ag85B construct or the pCI vector alone as the control. High titers of total immunoglobulin G (IgG), IgG1, and IgG2a anti-Ag85B were measured in pCI-Ag85B immunized mice when compared to the pCI control group. Regarding cellular immunity, significant levels of gamma interferon (IFN-γ) (1,100 ± 157 pg/ml) and tumor necrosis factor alpha (650 ± 42 pg/ml) but not interleukin-4 were detected in splenocyte culture supernatants of pCI-Ag85B-vaccinated mice following stimulation with recombinant Ag85B. Further, the main source of IFN-γ is CD8+ T cells, as demonstrated by intracellular cytokine staining. As far as protection, a significant reduction in bacterial load in spleens (P < 0.05) was detected in pCI-Ag85B-immunized mice compared to the pCI vector control group. The results obtained here suggest that use of the Ag85B DNA vaccine is a promising strategy to control M. bovis infection due to its ability to induce a Th1 type of immune response. However, protective efficacy needs to be improved, since partial protection was achieved in spleens but not in lungs of vaccinated mice.

scholarly journals Effects of copper nanoparticles on the morphological and physiological changes in chicken

2019 ◽  
Vol 41 (2) ◽  
Author(s):  
Nguyen Thi Xuan ◽  
Phi Thi Thu Trang ◽  
Le Thi Thu Hien
Keyword(s):  
Copper Nanoparticles ◽  
Serum Levels ◽  
Feed Additive ◽  
Control Group ◽  
Cu Content ◽  
Wide Range ◽  
Body Weights ◽  
Liver And Kidney ◽  
Kidney Functions ◽  
Poultry Farming

Copper nanoparticles (nCu) have a wide range of applications in many different fields of life. In poultry farming, nCu is well-known as a feed additive to stimulate the development, a good antimicrobial agent and a suppressor of diarrhea. The aim of this study is to perform the analysis of biosafety of nCu. In this study, chickens were treated with drinking water containing 2 mg/L or 1000 mg/L nCu. After 5 weeks of treatment, the survival rate, dry body weight, Cu content in serum and liver tissues of chickens were examined. Liver and kidney functions were also determined by measuring serum levels of ALT, AST, and creatinine. As a result, chickens treated with drinking containing 2 mg/L nCu had no changes in liver and kidney functions as well as their body weights. However, treatment of chickens with 1000 mg/L nCu caused accumulationof Cu in the serum and liver and increase of the serum levels of ALT and AST compared with control group. Apparently they sick and did not develop properly. In conclusion, nCu concentration at 2 mg/L may be recommended for the agricultural, feed and medicine formulations to meet biological safety.ALT, AST, creatinine, nCu, chicken.

scholarly journals iTRAQ-based comparative proteomic analysis in different developmental stages of Echinococcus granulosus

Parasite ◽  
2021 ◽  
Vol 28 ◽  
pp. 15
Author(s):  
Xin Li ◽  
Song Jiang ◽  
Xuhai Wang ◽  
Wenqiao Hui ◽  
Bin Jia
Keyword(s):  
Echinococcus Granulosus ◽  
Metabolic Pathways ◽  
Developmental Stages ◽  
Vaccine Development ◽  
Absolute Quantification ◽  
Differentially Expressed ◽  
Specific Gene ◽  
Differentially Expressed Proteins ◽  
Intermediate Hosts ◽  
Differential Proteins

Cystic echinococcosis, caused by infection with the larval stage of the cestode Echinococcus granulosus, is a chronic zoonosis. The lifecycle of the E. granulosus parasite includes three consecutive stages that require specific gene regulation or protein expression to survive environmental shifts between definitive hosts and intermediate hosts. The aim of the present study is to screen and analyze the stage differential antigens to be considered for vaccine development against E. granulosus. By using the iTRAQ (isobaric tags for relative and absolute quantification) method, the differentially expressed proteins were selected from the three consecutive developmental stages of E. granulosus: oncosphere, adult tapeworms, and protoscolex. Through a bioinformatics analysis including Clusters of Orthologous Groups (COG), Gene Ontology (GO), and pathway metabolic annotation, we identified some proteins of interest from each stage. The results showed that a large number of differentially expressed proteins (375: oncosphere vs. adult, 346: oncosphere vs. protoscolex, and 391: adult vs. protoscolex) were identified from the three main lifecycle stages. Analysis of the differential protein pathways showed that these differential proteins are mainly enriched in metabolic pathways, Huntington’s diseases, Alzheimer’s diseases, and ribosome metabolic pathways. Interestingly, among these differential proteins, expression levels of paramyosin, HSP60, HSP70, HSP90, cathepsin L1, cathepsin D, casein kinase, and calmodulin were significantly higher in the oncosphere than in the adult or protoscolex (p < 0.05). We hope our findings will help to identify potential targets for diagnosis or for therapeutic and prophylactic intervention.

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