[Objective] This study aimed to investigate the immunological adjuvant function of aluminium phosphate and chicken IL-18 in NDV F gene vaccine. [Method] The vaccine (0.2 ml) containing aluminum phosphate adjuvant (...[Objective] This study aimed to investigate the immunological adjuvant function of aluminium phosphate and chicken IL-18 in NDV F gene vaccine. [Method] The vaccine (0.2 ml) containing aluminum phosphate adjuvant (90 μg), pcDNA/F (200μg), and pcDNA/chlL-18 (200 μg) was prepared. The 7 d old chick- ens to be tested were randomly divided into six groups (12 chickens in each group) and immunized through intramuscular injection with inactivated Newcastle disease vaccines, pcDNA/F+pcDNA/chlL-18+phosphate aluminum, pcDNA/F, pcDNA/F.+pcDNA/ chlL-18, pcDNA/F+aluminum phosphate, and physiological saline respectively; the secondary immunization was conducted with the same dose when the chickens were 21 d old. Their blood was sampled 0, 7, 14, 21, 28 d after first immunization. Anti- body titer was detected with ELISA and T cell transformation rate was measured with MIT. Experimental chicken will be challenged with 30 LD50 NDV virulence 28 d after first immunization. [Result] The survival rate of the chickens immunized with pcDNA/F+aluminium phosphate+pcDNA/chlL-18 achieved 8/12, higher than that of those immunized with pcDNA/F 4/12 and pcDNA/F+pcDNA/chlL-18 (6/12). The NDV antibody titer of the chickens immunized with pcDNA/F+ aluminum phosphate, pcD- NA/F+pcDNA/chlL-18 and pcDNA/F+pcDNA/chlL-18+aluminum phosphate is not differ- ent (P〉0.05), but significantly lower than that of the chickens immunized with tradi- tional vaccine (P〈0.05). The T cell transformation rate of the chickens immunized with pcDNA/F+pcDNA/chlL-18+aluminium phosphate was obviously higher than that of the chickens immunized with pcDNA/F (P〈0.05). The T cell transformation rates of chickens immunized with pcDNA/F and the traditional vaccine showed no signifi- cant difference (P〉0.05). [Conclusion] Combination of aluminium phosphate and pcD- NA/chlL-18 can significantly enhance the immune effect of NDV F gene vaccine.展开更多
Heat shock protein 65 (HSP65) is one of the most important protective immunogens against the tuberculosis infection. The signal sequence of antigen 85B and the whole HSP65 DNA sequence of human Mycobacterium tuberculo...Heat shock protein 65 (HSP65) is one of the most important protective immunogens against the tuberculosis infection. The signal sequence of antigen 85B and the whole HSP65 DNA sequence of human Mycobacterium tuberculosis (M. tuberculosis) were amplified from BCG genome and plasmid pCMV-MTHSP65 respectively by polymerase chain reactions (PCR). These two sequences were cloned into the plasmid pBCG-2100 under the control of the promoter of heat shock protein 70 (HSP70) from human M. tuberculosis, yielding the prokaryotic shuttle expression plasmid pBCG-SP-HSP65. Results of restriction endonuclease analysis, PCR detection and DNA sequencing analysis showed that the two cloned DNA sequences were consistent with those previously reported, and the direction of their inserting into the recombinant was correct and the reading frame had been maintained. The recombinants were electroporated into BCG to construct the recombinant BCG vaccine and induced by heating. The induced expression detected by SDS-PAGE showed that the content of 65 kD protein expressed in recombinant BCG was 35.69 % in total bacterial protein and 74.09 % in the cell lysate supernatants, suggesting that the recombinant HSP65 gene could express in BCG with high efficiency and the expressed proteins were mainly soluble. Western-blot showed that the secretive recombinant proteins could specifically combine with antibody against M. tuberculosis HSP65, indicating that the recombinant proteins possess the biological activity of HSP65.展开更多
Summary: The expression of foreign gene, Schistosoma Japonicum 26 ku antigen (Sj26GST), in Bacillus Calmette Guerin (BCG), Mycobacterium ( M. smegmatis ) and Escherichia coli ( E. coli ) were stud...Summary: The expression of foreign gene, Schistosoma Japonicum 26 ku antigen (Sj26GST), in Bacillus Calmette Guerin (BCG), Mycobacterium ( M. smegmatis ) and Escherichia coli ( E. coli ) were studied. The cDNA fragment encoding Sj26GST was amplified by PCR using plasmid pGEX, which could express Sj26GST in E. coli as template. The Sj26GST cDNA was cloned into the downstream of human M. tuberculosis heat shock protein (hsp) 70 promoter with correct reading frame, and then the DNA fragment containing hsp70 promoter and Sj26GST gene were subcloned together into E. coli Mycobacteria shuttle plasmid pBCG 2000 to construct the expression shuttle plasmid pBCG Sj26. The recombinant BCG and M. smegmatis mc 2 155, which were electroplated with pBCG Sj26, could express Sj26GST and the recombinant Schistosoma Japonicum vaccine BCG Sj26GST was made. The recombinant Sj26GST (rSj26GST) were soluble and could be observed on SDS PAGE at molecular weight of 26 ku. The content of rSj26GST accounted for 15 % and 10 % of total bacterial protein in BCG and M. smegmatis respectively. The results of Western blot showed the combination of rSj26GST with antibody of GST.展开更多
Background:The mobilization and redistribution of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)specific T-cells and neutralizing antibodies(nAbs)during exercise is purported to increase immune surveillan...Background:The mobilization and redistribution of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)specific T-cells and neutralizing antibodies(nAbs)during exercise is purported to increase immune surveillance and protect against severe coronavirus disease 2019(COVID-19).We sought to determine if COVID-19 vaccination would elicit exercise-responsive SARS-CoV-2 T-cells and transiently alter nAb titers.Methods:Eighteen healthy participants completed a 20-min bout of graded cycling exercise before and/or after receiving a COVID-19 vaccine.All major leukocyte subtypes were enumerated before,during,and after exercise by flow cytometry,and immune responses to SARS-CoV-2 were determined using whole blood peptide stimulation assays,T-cell receptor(TCR)-βsequencing,and SARS-CoV-2 nAb serology.Results:COVID-19 vaccination had no effect on the mobilization or egress of major leukocyte subsets in response to intensity-controlled graded exercise.However,non-infected participants had a significantly reduced mobilization of CD4+and CD8+naive T-cells,as well as CD4+central memory T-cells,after vaccination(synthetic immunity group);this was not seen after vaccination in those with prior SARS-CoV-2 infection(hybrid immunity group).Acute exercise after vaccination robustly mobilized SARS-CoV-2 specific T-cells to blood in an intensity-dependent manner.Both groups mobilized T-cells that reacted to spike protein;however,only the hybrid immunity group mobilized T-cells that reacted to membrane and nucleocapsid antigens.nAbs increased significantly during exercise only in the hybrid immunity group.Conclusion:These data indicate that acute exercise mobilizes SARS-CoV-2 specific T-cells that recognize spike protein and increases the redistribution of nAbs in individuals with hybrid immunity.展开更多
Despite the initial successes of the Bacillus Calmette-Guerin(BCG)vaccine in children,its efficacy against tuberculosis is highly variable.There is a lack of understanding about how mental conditions influence BCG vac...Despite the initial successes of the Bacillus Calmette-Guerin(BCG)vaccine in children,its efficacy against tuberculosis is highly variable.There is a lack of understanding about how mental conditions influence BCG vaccination.Here,we used the chronic social defeat stress(CSDS)model to explore the effects of depression on BCG vaccination efficacy.We observed higher lung and spleen bacterial loads and a lower organ index in depressed compared to BCG mice.Meanwhile,a relatively lower T cell protective efficacy was observed in both compared to control and BCG mice via a mycobacterium growth inhibition assay(MGIA).Cytokine expression of IL-12p40,IL-1β,IL-17,TNF-αand IFN-γwas reduced,whereas the expression of IL-10 and IL-5 was increased in the spleen of both compared to BCG mice.Moreover,the proportions of CD4^(+)IFN-γ^(+),CD8^(+)IFN-γ^(+)T lymphocytes and CD4^(+)effector/central memory T cells were reduced in the splenocytes of the depressed BCG mice.Depression promotes CD4^(+)regulatory T cells(Treg)and myeloid-derived suppressor cell(MDSC)generation in depressed mice,contributing to the reduced pro-inflammatory immune response upon BCG vaccination.This study provides insight into the decreased protective immunity by BCG vaccination attributable to depression in mice.展开更多
The BALB/c mice were immunized with rMS Sj26GST and rBCG Sj26GST vaccine in Schistosoma japonicum by subcutaneous injection After they were immunized for 8 weeks, the eyeballs were removed to get blood and macroph...The BALB/c mice were immunized with rMS Sj26GST and rBCG Sj26GST vaccine in Schistosoma japonicum by subcutaneous injection After they were immunized for 8 weeks, the eyeballs were removed to get blood and macrophages of abdominal cavity and spleen cells were harvested The lymphocytic stimulating index (SI) was used to measure the cellular proliferating ability and NO release was used to measure the phagocytic activity of the macrophages By using ELISA kit, the levels of interleukin 2 (IL 2) and interferon γ (IFN γ) in serum and the splenic lymphocytic cultured supernatant were detected The results showed that after the mice were immunized with 10 6 CFU of rMS Sj26GST and rBCG Sj26GST vaccine separately by subcutaneous injection, proliferating ability of splenic lymphocytes in the mice showed no difference ( P >0 05), but both were significantly increased as compared with that in the control group( P <0 05); The contents of NO in the intraperitoneal macrophages of rMS Sj26GST vaccine group were significantly lower than in the control group ( P <0 001) and rBCG Sj26GST vaccine group ( P <0 01); The levels of serum IL 2 in the rMS Sj26GST vaccine group were significantly increased as compared with that in the control group ( P <0 001), vector group ( P <0 01) and rBCG Sj26GST vaccine group ( P <0 05); The contents of serum IFN γ in the rMS Sj26GST vaccine group were significantly increased as compared with that in the control group ( P <0 01) and rBCG Sj26GST vaccine group ( P <0 05) The contents of IFN γ in the cultured supernatant were significantly lower than those of rBCG Sj26GST vaccine group ( P <0 001), but were significantly increased as compared with that in the control group ( P <0 01) It was indicated that both vaccines could enhance the immune response of the mice, but rMS Sj26GST vaccine had stronger immunogenicity than rBCG Sj26GST vaccine展开更多
Bacille Calmette Guerin (BCG), which has been used since 1921 as the only vaccine against tuber-culosis (TB), protects poorly, if at all, against pulmonary tuberculosis among adults in high incident developing countri...Bacille Calmette Guerin (BCG), which has been used since 1921 as the only vaccine against tuber-culosis (TB), protects poorly, if at all, against pulmonary tuberculosis among adults in high incident developing countries. This failure has been attributed to the possible down modulating action of T regulatory cells (Tregs), which can be stimulated by environmental mycobacteria and expanded by BCG vaccination. Tregs induced at the site of BCG vaccination may interfere with protection against tuberculosis. This communication describes the contribution of Tregs towards dampening the efficacy of BCG and plausible approaches to countering this down modulating effect of Tregs. Probably, antigen specific inhibition of the local recruitment of Tregs whilst avoiding generalised disturbance of immune homeostasis could prove to be worthwhile. Alternatively, drugs with short half life may achieve more acceptable transient inhibition of Tregs function than the prolonged action of monoclonal antibodies. Evolving novel safe strategies is a challenge for developing a better anti TB vaccine.展开更多
Foot and mouth disease is the most contagious disease of mammals and has a great potential for causing severe economic loss in susceptible cloven-hoofed animals. Routine prophylactic vaccination in Egypt from 2012 til...Foot and mouth disease is the most contagious disease of mammals and has a great potential for causing severe economic loss in susceptible cloven-hoofed animals. Routine prophylactic vaccination in Egypt from 2012 till now has been conducted with Trivalent and/or hexavalent vaccine of MERIAL, against virus strains (O manisa + O-3039 + A Iran 05 + A Saudi 95 + Asia 1 + Sat2). This study aimed to evaluate the hemato-biochemical and immunological changes associated with the use of hexavalent vaccine alone or in combination with trivalent FMD vaccine. This study was carried out on 24 cattle divided into three groups eight cattle each. Group I served as control. Group II were selected from farms vaccinated with hexavalent FMD vaccine. Group III were selected from farms vaccinated with a combination of trivalent and hexavalent FMD vaccine. The results showed that both vaccinated groups showed a significant increase in total leukocytic count. Sera from hexavalent-trivalent vaccinated cattle demonstrated a significant increase in serum cortisol concentrations. Significant increase in serum activity of aspartate aminotransferase was recorded in animals vaccinated with a combination of hexavalent and trivalent vaccine. In addition, both regimes resulted in a significant increase in serum blood urea nitrogen compared to control. Both regimes induced a significant increase in serum levels of ceruloplasmin while phagocytic activity of neutrophils and phagocytic index was enhanced only by the hexavalent vaccine. Both vaccinated groups had significantly increased serum values of gamma globulins. These results suggested that hexavalent vaccine produced higher levels of safety and protective effects against FMD in cattle as compared to those produced by a combination of hexavalent and trivalent vaccines. It is also advisable to include the hexavalent vaccine within the program of obligatory and imperative vaccination against FMD.展开更多
Recalcitrant warts can accurately be defined as warts that persist after six months of conventional therapy. Up to one-third of non-genital warts, especially periungual and plantar warts, become recalcitrant. Traditio...Recalcitrant warts can accurately be defined as warts that persist after six months of conventional therapy. Up to one-third of non-genital warts, especially periungual and plantar warts, become recalcitrant. Traditional treatment options for warts include topical salicylic acid, cryotherapy, and electrocautery;however, patients with recalcitrant warts remain a major therapeutic challenge. There is evidence that immunotherapy can clear recalcitrant warts if traditional treatment fails. Given this, clinical studies published in PubMed and Google Scholar that used Bacillus Calmette-Guerin (BCG), Mycobacterium Indicus Pranii (Mw vaccine), and purified protein derivative (PPD) as immunotherapy for wart, were reviewed in this study. Neither of these treatments has been subjected to a randomized controlled trial, thus to date, there are no standardized protocols to use them. Our review highlights the scientific facts in the clinical applications of the previous options to treat recalcitrant warts and investigate the differences among them, concerning efficacy, adverse effects, dosage, and route of administration.展开更多
Objectives To explore the optimization method of vaccine clinical trial design based on immunological surrogate endpoint to improve the quality and efficiency of vaccine clinical research and development.Methods As to...Objectives To explore the optimization method of vaccine clinical trial design based on immunological surrogate endpoint to improve the quality and efficiency of vaccine clinical research and development.Methods As to the problems in the vaccine clinical research in China,the relevant guidelines and literatures of FDA and WHO were used to analyze and summarize the methods of optimizing the design of vaccine clinical trials.Results and Conclusion The adaptive design guidelines are established to guide clinical trial design,encourage the development and application of immunological surrogate endpoints,establish qualification process for drug development tools and information disclosure procedures to improve vaccine development efficiency.展开更多
Cytokines provided locally at tbe tumor site mayinitiate an effective anti-tumor immune responsewhich leads to rejection of a tumor which otherwisegrows progressively. Experimentillly, this can betested by gene transf...Cytokines provided locally at tbe tumor site mayinitiate an effective anti-tumor immune responsewhich leads to rejection of a tumor which otherwisegrows progressively. Experimentillly, this can betested by gene transfer into cultured tumor cellsfollowed by the analysis of the tumorigenicity of suchgenetically engincered cells. This approach allows展开更多
To study the kinetics in vivo of a Hantaan virus DNA vaccine, we constructed a fusion DNA vaccine, pEGFP/S, by cloning the S segment of Hantavirus into the vector, pEGFP-C1, which encodes Green fluorescent protein EGF...To study the kinetics in vivo of a Hantaan virus DNA vaccine, we constructed a fusion DNA vaccine, pEGFP/S, by cloning the S segment of Hantavirus into the vector, pEGFP-C1, which encodes Green fluorescent protein EGFP. In this report, we provide evidence that pEGFP/S was distributed and persistently expressed for more than 60 days in several organs after inoculation. Our findings suggest that the persistent immune responses induced by a Hantaan virus DNA vaccine are likely due to the plasmid pEGFP/S deposited in vivo, which acts as a booster immunization.展开更多
Bacille Calmette-Gu6rin(BCG) vaccine is one of the most widely used vaccines in children.In Egypt,it is a part of the national compulsory childhood immunization program.The most controversial aspect of BCG is the vari...Bacille Calmette-Gu6rin(BCG) vaccine is one of the most widely used vaccines in children.In Egypt,it is a part of the national compulsory childhood immunization program.The most controversial aspect of BCG is the variable efficacy found in different studies.This study was to evaluate the efficacy status of the available BCG vaccine in Egypt within the last 10 years(BCG-Copenhagen).The pilot cross sectional study included 597 Egyptian children randomly selected.Their ages ranged from 6 months to 10 years old(mean_ 5 years,median: 3 years).All were assessed for history of BCG vaccine intake(primary at infancy and/or secondary at school age) and examined for the presence BCG scar.A group of the vaccinated children(62 children with BCG scar and 69 children without BCG scar) were further assessed with tuberculin skin test(TST).Prevalence of BCG vaccine intake in the studied children was 86.9%(519/597).Efficacy in term of BCG scar after vaccination was 66.6%(346/519).However,efficacy in term of post BCG vaccination tuberculin sensitization was only 3.8%(5/131).BCG vaccination program in Egypt seems to be widely prevalent;however, the immunological efficacy of the available strain is questionable.展开更多
With the increased prevalence of dengue infection in tropical countries, concerned members of the public are now pressing their local health ministries to act immediately and effectively in managing the rising numbers...With the increased prevalence of dengue infection in tropical countries, concerned members of the public are now pressing their local health ministries to act immediately and effectively in managing the rising numbers of reported cases. This includes reviews of the methodologies and the effectiveness of current combative systems to find other possible novel approaches that might yield better results. One of those novel approaches is the integration of a parasite into mosquito vector, manipulating the parasite-host interaction to reduce the transmission of dengue in endemic hotspots. Another alternative is by Sanofi-Pasteur’s dengue vaccine that showed over 60.8% success rate in reducing severe dengue infection in children aged 9 - 16 during its final clinical implementation phase. This report will compare and contrast these two novel ideas to determine which of the approaches are more likely to be effective in the long run. The aspects covered will include the application, effectiveness, functionality, and problems with these approaches. The results could then be utilised by governments or organizations to select precise and effective methods in reducing the prevalence of dengue infections in their countries.展开更多
A plasmid DNA vaccine is able to induce both humoral and cellular immune responses;however, the kinetic change of the Th1/Th2 response, antibody avidity, cytokine secretion, and neutralization activity after different...A plasmid DNA vaccine is able to induce both humoral and cellular immune responses;however, the kinetic change of the Th1/Th2 response, antibody avidity, cytokine secretion, and neutralization activity after different priming and boosting strategies have not been evaluated. A plasmid DNA, designated pCBD2 and previously shown to efficiently induce an immune response very similar to that by a wild type virus, was evaluated kinetically in this study. Our results suggest that a DNA vaccine delivered by the gene gun (gg) route produced higher and longer DENV-2-specific antibody titers than those induced through the intramuscular (im) route. Although the gg group induced a Th2 response and im delivery induced a Th1 response, priming by gg delivery, followed by a boosting by im delivery, did not shift the immune response from a Th2 to Th1 response. Furthermore, the antibody avidity (AI) measured by ELISA demonstrated a gradual increase of AI from low (AI range from 6.8% - 9.6%) on day 42 to high (AI value > 30) on day 119 in all but the gene-gun immunization group, in which an AI value of 23 was observed. Although there was lower avidity in the gg group, the mice sera from all three groups of mice demonstrated significant neutralization activity. This is the first report about the kinetics of immunogenicity of a DNA vaccine through different administration strategies, which suggests that gene gun delivery of a DNA vaccine can induce an immune response containing both neutralizing and nonneutralizing antibodies at high titers important for neutralization.展开更多
Introduction: Tuberculosis (TB) continues to be a global health challenge and currently only one licensed vaccine is available. For nearly 100 years, the Bacillus Calmette-Guérin (BCG) vaccine has been in use. Wh...Introduction: Tuberculosis (TB) continues to be a global health challenge and currently only one licensed vaccine is available. For nearly 100 years, the Bacillus Calmette-Guérin (BCG) vaccine has been in use. While it provides protection against disseminated TB in infants, its protection against adult and adolescent pulmonary tuberculosis (PTB) is variable. This literature review will provide an overview of the clinical status of candidate TB vaccines and discuss the challenges and future development trends of novel TB vaccine research, in combination with a general overview of the Tuberculosis (TB) disease and Mycobacterium tuberculosis itself. Methods: Bibliographic searches were carried out on medical journal databases, publishers, and aggregators. The most used databases were PubMed, NCBI and MDPI. Publications in English on these and other databases relating to novel TB vaccines were included in this review. Results: Currently, there are 12 main vaccine candidates in various phases of clinical trials, they include four protein or adjuvant vaccines, three viral-vectored vaccines, three mycobacterial whole cells or extract vaccines, and one each of the recombinant life and the attenuated Mycobacterium tuberculosis vaccine. Currently, the most likely candidate vaccines are the M72 + AS01E and Vaccae vaccines. M72 + AS01E is a recombinant fusion protein vaccine candidate, clinical trials showed that administering two doses of M72/AS01E was successful in reducing the development of active TB disease with 50% efficacy. Studies have also proven the efficacy of Vaccae (which is currently in phase III clinical trials) as an adjunctive therapy, with it being curative in conjunction with current therapy. Conclusion: Given the morbidity and mortality suffered globally by M. tuberculosis, it is time to realize the seriousness of the situation and accelerate our commitment and investment to the eradication of this infectious disease. With the number of vaccine candidates currently in clinical trials having promising results, it is imperative to continue these studies and accelerate towards phase III licensure trials if we are to achieve the milestone of “End TB Strategy” by 2035. Today, we are witnessing immense progress in both preclinical and clinical TB vaccine research despite disappointing results from some of the clinical efficacy trials like that of MVA85A. We can revisit the design of vaccines and learn from them. It is important not only to recognize and give credit to those that have tested well in human trials, such as M72 + AS01E, but to expedite and improve its efficacy through funding of its research.展开更多
Dental caries, the disease that causes tooth decay, is infectious, and the mutans streptococci bacteria have long been identified as the primary disease-causing agents. Caries vaccines showed promising results in expe...Dental caries, the disease that causes tooth decay, is infectious, and the mutans streptococci bacteria have long been identified as the primary disease-causing agents. Caries vaccines showed promising results in experimental studies;however, it remains far the effective use in humans due to political-economic and ethical issues. Progress towards practical vaccine development requires evaluation of candidate vaccines in clinical trials. Promising strategies of passive immunization also require further clinical evaluation. The purpose of this chapter is to review the literature on the main research projects aimed at developing caries vaccines.展开更多
文摘[Objective] This study aimed to investigate the immunological adjuvant function of aluminium phosphate and chicken IL-18 in NDV F gene vaccine. [Method] The vaccine (0.2 ml) containing aluminum phosphate adjuvant (90 μg), pcDNA/F (200μg), and pcDNA/chlL-18 (200 μg) was prepared. The 7 d old chick- ens to be tested were randomly divided into six groups (12 chickens in each group) and immunized through intramuscular injection with inactivated Newcastle disease vaccines, pcDNA/F+pcDNA/chlL-18+phosphate aluminum, pcDNA/F, pcDNA/F.+pcDNA/ chlL-18, pcDNA/F+aluminum phosphate, and physiological saline respectively; the secondary immunization was conducted with the same dose when the chickens were 21 d old. Their blood was sampled 0, 7, 14, 21, 28 d after first immunization. Anti- body titer was detected with ELISA and T cell transformation rate was measured with MIT. Experimental chicken will be challenged with 30 LD50 NDV virulence 28 d after first immunization. [Result] The survival rate of the chickens immunized with pcDNA/F+aluminium phosphate+pcDNA/chlL-18 achieved 8/12, higher than that of those immunized with pcDNA/F 4/12 and pcDNA/F+pcDNA/chlL-18 (6/12). The NDV antibody titer of the chickens immunized with pcDNA/F+ aluminum phosphate, pcD- NA/F+pcDNA/chlL-18 and pcDNA/F+pcDNA/chlL-18+aluminum phosphate is not differ- ent (P〉0.05), but significantly lower than that of the chickens immunized with tradi- tional vaccine (P〈0.05). The T cell transformation rate of the chickens immunized with pcDNA/F+pcDNA/chlL-18+aluminium phosphate was obviously higher than that of the chickens immunized with pcDNA/F (P〈0.05). The T cell transformation rates of chickens immunized with pcDNA/F and the traditional vaccine showed no signifi- cant difference (P〉0.05). [Conclusion] Combination of aluminium phosphate and pcD- NA/chlL-18 can significantly enhance the immune effect of NDV F gene vaccine.
文摘Heat shock protein 65 (HSP65) is one of the most important protective immunogens against the tuberculosis infection. The signal sequence of antigen 85B and the whole HSP65 DNA sequence of human Mycobacterium tuberculosis (M. tuberculosis) were amplified from BCG genome and plasmid pCMV-MTHSP65 respectively by polymerase chain reactions (PCR). These two sequences were cloned into the plasmid pBCG-2100 under the control of the promoter of heat shock protein 70 (HSP70) from human M. tuberculosis, yielding the prokaryotic shuttle expression plasmid pBCG-SP-HSP65. Results of restriction endonuclease analysis, PCR detection and DNA sequencing analysis showed that the two cloned DNA sequences were consistent with those previously reported, and the direction of their inserting into the recombinant was correct and the reading frame had been maintained. The recombinants were electroporated into BCG to construct the recombinant BCG vaccine and induced by heating. The induced expression detected by SDS-PAGE showed that the content of 65 kD protein expressed in recombinant BCG was 35.69 % in total bacterial protein and 74.09 % in the cell lysate supernatants, suggesting that the recombinant HSP65 gene could express in BCG with high efficiency and the expressed proteins were mainly soluble. Western-blot showed that the secretive recombinant proteins could specifically combine with antibody against M. tuberculosis HSP65, indicating that the recombinant proteins possess the biological activity of HSP65.
文摘Summary: The expression of foreign gene, Schistosoma Japonicum 26 ku antigen (Sj26GST), in Bacillus Calmette Guerin (BCG), Mycobacterium ( M. smegmatis ) and Escherichia coli ( E. coli ) were studied. The cDNA fragment encoding Sj26GST was amplified by PCR using plasmid pGEX, which could express Sj26GST in E. coli as template. The Sj26GST cDNA was cloned into the downstream of human M. tuberculosis heat shock protein (hsp) 70 promoter with correct reading frame, and then the DNA fragment containing hsp70 promoter and Sj26GST gene were subcloned together into E. coli Mycobacteria shuttle plasmid pBCG 2000 to construct the expression shuttle plasmid pBCG Sj26. The recombinant BCG and M. smegmatis mc 2 155, which were electroplated with pBCG Sj26, could express Sj26GST and the recombinant Schistosoma Japonicum vaccine BCG Sj26GST was made. The recombinant Sj26GST (rSj26GST) were soluble and could be observed on SDS PAGE at molecular weight of 26 ku. The content of rSj26GST accounted for 15 % and 10 % of total bacterial protein in BCG and M. smegmatis respectively. The results of Western blot showed the combination of rSj26GST with antibody of GST.
文摘Background:The mobilization and redistribution of severe acute respiratory syndrome coronavirus 2(SARS-CoV-2)specific T-cells and neutralizing antibodies(nAbs)during exercise is purported to increase immune surveillance and protect against severe coronavirus disease 2019(COVID-19).We sought to determine if COVID-19 vaccination would elicit exercise-responsive SARS-CoV-2 T-cells and transiently alter nAb titers.Methods:Eighteen healthy participants completed a 20-min bout of graded cycling exercise before and/or after receiving a COVID-19 vaccine.All major leukocyte subtypes were enumerated before,during,and after exercise by flow cytometry,and immune responses to SARS-CoV-2 were determined using whole blood peptide stimulation assays,T-cell receptor(TCR)-βsequencing,and SARS-CoV-2 nAb serology.Results:COVID-19 vaccination had no effect on the mobilization or egress of major leukocyte subsets in response to intensity-controlled graded exercise.However,non-infected participants had a significantly reduced mobilization of CD4+and CD8+naive T-cells,as well as CD4+central memory T-cells,after vaccination(synthetic immunity group);this was not seen after vaccination in those with prior SARS-CoV-2 infection(hybrid immunity group).Acute exercise after vaccination robustly mobilized SARS-CoV-2 specific T-cells to blood in an intensity-dependent manner.Both groups mobilized T-cells that reacted to spike protein;however,only the hybrid immunity group mobilized T-cells that reacted to membrane and nucleocapsid antigens.nAbs increased significantly during exercise only in the hybrid immunity group.Conclusion:These data indicate that acute exercise mobilizes SARS-CoV-2 specific T-cells that recognize spike protein and increases the redistribution of nAbs in individuals with hybrid immunity.
基金funded by the National Natural Science Foundation of China(Grant No.U21A20259,31602061,31872470)the National Key Research and Development Program of China(Grant No.2021YFD1800401).
文摘Despite the initial successes of the Bacillus Calmette-Guerin(BCG)vaccine in children,its efficacy against tuberculosis is highly variable.There is a lack of understanding about how mental conditions influence BCG vaccination.Here,we used the chronic social defeat stress(CSDS)model to explore the effects of depression on BCG vaccination efficacy.We observed higher lung and spleen bacterial loads and a lower organ index in depressed compared to BCG mice.Meanwhile,a relatively lower T cell protective efficacy was observed in both compared to control and BCG mice via a mycobacterium growth inhibition assay(MGIA).Cytokine expression of IL-12p40,IL-1β,IL-17,TNF-αand IFN-γwas reduced,whereas the expression of IL-10 and IL-5 was increased in the spleen of both compared to BCG mice.Moreover,the proportions of CD4^(+)IFN-γ^(+),CD8^(+)IFN-γ^(+)T lymphocytes and CD4^(+)effector/central memory T cells were reduced in the splenocytes of the depressed BCG mice.Depression promotes CD4^(+)regulatory T cells(Treg)and myeloid-derived suppressor cell(MDSC)generation in depressed mice,contributing to the reduced pro-inflammatory immune response upon BCG vaccination.This study provides insight into the decreased protective immunity by BCG vaccination attributable to depression in mice.
基金ThisprojectwassupportedbyagrantfromNationalNaturalSciencesFoundationofChina (No 39870 6 6 3)
文摘The BALB/c mice were immunized with rMS Sj26GST and rBCG Sj26GST vaccine in Schistosoma japonicum by subcutaneous injection After they were immunized for 8 weeks, the eyeballs were removed to get blood and macrophages of abdominal cavity and spleen cells were harvested The lymphocytic stimulating index (SI) was used to measure the cellular proliferating ability and NO release was used to measure the phagocytic activity of the macrophages By using ELISA kit, the levels of interleukin 2 (IL 2) and interferon γ (IFN γ) in serum and the splenic lymphocytic cultured supernatant were detected The results showed that after the mice were immunized with 10 6 CFU of rMS Sj26GST and rBCG Sj26GST vaccine separately by subcutaneous injection, proliferating ability of splenic lymphocytes in the mice showed no difference ( P >0 05), but both were significantly increased as compared with that in the control group( P <0 05); The contents of NO in the intraperitoneal macrophages of rMS Sj26GST vaccine group were significantly lower than in the control group ( P <0 001) and rBCG Sj26GST vaccine group ( P <0 01); The levels of serum IL 2 in the rMS Sj26GST vaccine group were significantly increased as compared with that in the control group ( P <0 001), vector group ( P <0 01) and rBCG Sj26GST vaccine group ( P <0 05); The contents of serum IFN γ in the rMS Sj26GST vaccine group were significantly increased as compared with that in the control group ( P <0 01) and rBCG Sj26GST vaccine group ( P <0 05) The contents of IFN γ in the cultured supernatant were significantly lower than those of rBCG Sj26GST vaccine group ( P <0 001), but were significantly increased as compared with that in the control group ( P <0 01) It was indicated that both vaccines could enhance the immune response of the mice, but rMS Sj26GST vaccine had stronger immunogenicity than rBCG Sj26GST vaccine
文摘Bacille Calmette Guerin (BCG), which has been used since 1921 as the only vaccine against tuber-culosis (TB), protects poorly, if at all, against pulmonary tuberculosis among adults in high incident developing countries. This failure has been attributed to the possible down modulating action of T regulatory cells (Tregs), which can be stimulated by environmental mycobacteria and expanded by BCG vaccination. Tregs induced at the site of BCG vaccination may interfere with protection against tuberculosis. This communication describes the contribution of Tregs towards dampening the efficacy of BCG and plausible approaches to countering this down modulating effect of Tregs. Probably, antigen specific inhibition of the local recruitment of Tregs whilst avoiding generalised disturbance of immune homeostasis could prove to be worthwhile. Alternatively, drugs with short half life may achieve more acceptable transient inhibition of Tregs function than the prolonged action of monoclonal antibodies. Evolving novel safe strategies is a challenge for developing a better anti TB vaccine.
文摘Foot and mouth disease is the most contagious disease of mammals and has a great potential for causing severe economic loss in susceptible cloven-hoofed animals. Routine prophylactic vaccination in Egypt from 2012 till now has been conducted with Trivalent and/or hexavalent vaccine of MERIAL, against virus strains (O manisa + O-3039 + A Iran 05 + A Saudi 95 + Asia 1 + Sat2). This study aimed to evaluate the hemato-biochemical and immunological changes associated with the use of hexavalent vaccine alone or in combination with trivalent FMD vaccine. This study was carried out on 24 cattle divided into three groups eight cattle each. Group I served as control. Group II were selected from farms vaccinated with hexavalent FMD vaccine. Group III were selected from farms vaccinated with a combination of trivalent and hexavalent FMD vaccine. The results showed that both vaccinated groups showed a significant increase in total leukocytic count. Sera from hexavalent-trivalent vaccinated cattle demonstrated a significant increase in serum cortisol concentrations. Significant increase in serum activity of aspartate aminotransferase was recorded in animals vaccinated with a combination of hexavalent and trivalent vaccine. In addition, both regimes resulted in a significant increase in serum blood urea nitrogen compared to control. Both regimes induced a significant increase in serum levels of ceruloplasmin while phagocytic activity of neutrophils and phagocytic index was enhanced only by the hexavalent vaccine. Both vaccinated groups had significantly increased serum values of gamma globulins. These results suggested that hexavalent vaccine produced higher levels of safety and protective effects against FMD in cattle as compared to those produced by a combination of hexavalent and trivalent vaccines. It is also advisable to include the hexavalent vaccine within the program of obligatory and imperative vaccination against FMD.
文摘Recalcitrant warts can accurately be defined as warts that persist after six months of conventional therapy. Up to one-third of non-genital warts, especially periungual and plantar warts, become recalcitrant. Traditional treatment options for warts include topical salicylic acid, cryotherapy, and electrocautery;however, patients with recalcitrant warts remain a major therapeutic challenge. There is evidence that immunotherapy can clear recalcitrant warts if traditional treatment fails. Given this, clinical studies published in PubMed and Google Scholar that used Bacillus Calmette-Guerin (BCG), Mycobacterium Indicus Pranii (Mw vaccine), and purified protein derivative (PPD) as immunotherapy for wart, were reviewed in this study. Neither of these treatments has been subjected to a randomized controlled trial, thus to date, there are no standardized protocols to use them. Our review highlights the scientific facts in the clinical applications of the previous options to treat recalcitrant warts and investigate the differences among them, concerning efficacy, adverse effects, dosage, and route of administration.
文摘Objectives To explore the optimization method of vaccine clinical trial design based on immunological surrogate endpoint to improve the quality and efficiency of vaccine clinical research and development.Methods As to the problems in the vaccine clinical research in China,the relevant guidelines and literatures of FDA and WHO were used to analyze and summarize the methods of optimizing the design of vaccine clinical trials.Results and Conclusion The adaptive design guidelines are established to guide clinical trial design,encourage the development and application of immunological surrogate endpoints,establish qualification process for drug development tools and information disclosure procedures to improve vaccine development efficiency.
文摘Cytokines provided locally at tbe tumor site mayinitiate an effective anti-tumor immune responsewhich leads to rejection of a tumor which otherwisegrows progressively. Experimentillly, this can betested by gene transfer into cultured tumor cellsfollowed by the analysis of the tumorigenicity of suchgenetically engincered cells. This approach allows
文摘To study the kinetics in vivo of a Hantaan virus DNA vaccine, we constructed a fusion DNA vaccine, pEGFP/S, by cloning the S segment of Hantavirus into the vector, pEGFP-C1, which encodes Green fluorescent protein EGFP. In this report, we provide evidence that pEGFP/S was distributed and persistently expressed for more than 60 days in several organs after inoculation. Our findings suggest that the persistent immune responses induced by a Hantaan virus DNA vaccine are likely due to the plasmid pEGFP/S deposited in vivo, which acts as a booster immunization.
文摘Bacille Calmette-Gu6rin(BCG) vaccine is one of the most widely used vaccines in children.In Egypt,it is a part of the national compulsory childhood immunization program.The most controversial aspect of BCG is the variable efficacy found in different studies.This study was to evaluate the efficacy status of the available BCG vaccine in Egypt within the last 10 years(BCG-Copenhagen).The pilot cross sectional study included 597 Egyptian children randomly selected.Their ages ranged from 6 months to 10 years old(mean_ 5 years,median: 3 years).All were assessed for history of BCG vaccine intake(primary at infancy and/or secondary at school age) and examined for the presence BCG scar.A group of the vaccinated children(62 children with BCG scar and 69 children without BCG scar) were further assessed with tuberculin skin test(TST).Prevalence of BCG vaccine intake in the studied children was 86.9%(519/597).Efficacy in term of BCG scar after vaccination was 66.6%(346/519).However,efficacy in term of post BCG vaccination tuberculin sensitization was only 3.8%(5/131).BCG vaccination program in Egypt seems to be widely prevalent;however, the immunological efficacy of the available strain is questionable.
文摘With the increased prevalence of dengue infection in tropical countries, concerned members of the public are now pressing their local health ministries to act immediately and effectively in managing the rising numbers of reported cases. This includes reviews of the methodologies and the effectiveness of current combative systems to find other possible novel approaches that might yield better results. One of those novel approaches is the integration of a parasite into mosquito vector, manipulating the parasite-host interaction to reduce the transmission of dengue in endemic hotspots. Another alternative is by Sanofi-Pasteur’s dengue vaccine that showed over 60.8% success rate in reducing severe dengue infection in children aged 9 - 16 during its final clinical implementation phase. This report will compare and contrast these two novel ideas to determine which of the approaches are more likely to be effective in the long run. The aspects covered will include the application, effectiveness, functionality, and problems with these approaches. The results could then be utilised by governments or organizations to select precise and effective methods in reducing the prevalence of dengue infections in their countries.
文摘A plasmid DNA vaccine is able to induce both humoral and cellular immune responses;however, the kinetic change of the Th1/Th2 response, antibody avidity, cytokine secretion, and neutralization activity after different priming and boosting strategies have not been evaluated. A plasmid DNA, designated pCBD2 and previously shown to efficiently induce an immune response very similar to that by a wild type virus, was evaluated kinetically in this study. Our results suggest that a DNA vaccine delivered by the gene gun (gg) route produced higher and longer DENV-2-specific antibody titers than those induced through the intramuscular (im) route. Although the gg group induced a Th2 response and im delivery induced a Th1 response, priming by gg delivery, followed by a boosting by im delivery, did not shift the immune response from a Th2 to Th1 response. Furthermore, the antibody avidity (AI) measured by ELISA demonstrated a gradual increase of AI from low (AI range from 6.8% - 9.6%) on day 42 to high (AI value > 30) on day 119 in all but the gene-gun immunization group, in which an AI value of 23 was observed. Although there was lower avidity in the gg group, the mice sera from all three groups of mice demonstrated significant neutralization activity. This is the first report about the kinetics of immunogenicity of a DNA vaccine through different administration strategies, which suggests that gene gun delivery of a DNA vaccine can induce an immune response containing both neutralizing and nonneutralizing antibodies at high titers important for neutralization.
文摘Introduction: Tuberculosis (TB) continues to be a global health challenge and currently only one licensed vaccine is available. For nearly 100 years, the Bacillus Calmette-Guérin (BCG) vaccine has been in use. While it provides protection against disseminated TB in infants, its protection against adult and adolescent pulmonary tuberculosis (PTB) is variable. This literature review will provide an overview of the clinical status of candidate TB vaccines and discuss the challenges and future development trends of novel TB vaccine research, in combination with a general overview of the Tuberculosis (TB) disease and Mycobacterium tuberculosis itself. Methods: Bibliographic searches were carried out on medical journal databases, publishers, and aggregators. The most used databases were PubMed, NCBI and MDPI. Publications in English on these and other databases relating to novel TB vaccines were included in this review. Results: Currently, there are 12 main vaccine candidates in various phases of clinical trials, they include four protein or adjuvant vaccines, three viral-vectored vaccines, three mycobacterial whole cells or extract vaccines, and one each of the recombinant life and the attenuated Mycobacterium tuberculosis vaccine. Currently, the most likely candidate vaccines are the M72 + AS01E and Vaccae vaccines. M72 + AS01E is a recombinant fusion protein vaccine candidate, clinical trials showed that administering two doses of M72/AS01E was successful in reducing the development of active TB disease with 50% efficacy. Studies have also proven the efficacy of Vaccae (which is currently in phase III clinical trials) as an adjunctive therapy, with it being curative in conjunction with current therapy. Conclusion: Given the morbidity and mortality suffered globally by M. tuberculosis, it is time to realize the seriousness of the situation and accelerate our commitment and investment to the eradication of this infectious disease. With the number of vaccine candidates currently in clinical trials having promising results, it is imperative to continue these studies and accelerate towards phase III licensure trials if we are to achieve the milestone of “End TB Strategy” by 2035. Today, we are witnessing immense progress in both preclinical and clinical TB vaccine research despite disappointing results from some of the clinical efficacy trials like that of MVA85A. We can revisit the design of vaccines and learn from them. It is important not only to recognize and give credit to those that have tested well in human trials, such as M72 + AS01E, but to expedite and improve its efficacy through funding of its research.
文摘Dental caries, the disease that causes tooth decay, is infectious, and the mutans streptococci bacteria have long been identified as the primary disease-causing agents. Caries vaccines showed promising results in experimental studies;however, it remains far the effective use in humans due to political-economic and ethical issues. Progress towards practical vaccine development requires evaluation of candidate vaccines in clinical trials. Promising strategies of passive immunization also require further clinical evaluation. The purpose of this chapter is to review the literature on the main research projects aimed at developing caries vaccines.