Immunogenicity and Protective Efficacy against Murine Tuberculosis of a Prime-Boost Regimen with BCG and a DNA Vaccine Expressing ESAT-6 and Ag85A Fusion Protein

Heterologous prime-boost regimens utilizing BCG as a prime vaccine probably represent the best hope for the development of novel tuberculosis (TB) vaccines. In this study, we examined the immunogenicity and protective efficacy of DNA vaccine (pcD685A) expressing the fusion protein of Ag85A and ESAT-6 (r685A) and its booster effects in BCG-immunized mice. The recombinant r685A fusion protein stimulated higher level of antigen-specific IFN-γ release in tuberculin skin test- (TST-) positive healthy household contacts of active pulmonary TB patients than that in TST-negative population. Vaccination of C57BL/6 mice with pcD685A resulted in significant protection against challenge with virulentMycobacterium tuberculosisH37Rv when compared with the control group. Most importantly, pcD685A could act as a BCG booster and amplify Th1-type cell-mediated immunity in the lung of BCG-vaccinated mice as shown the increased expression of IFN-γ. The most significant reduction in bacterial load of both spleen and lung was obtained in mice vaccinated with BCG prime and pcD685A DNA booster when compared with BCG or pcD685A alone. Thus, our study indicates that pcD685A may be an efficient booster vaccine against TB with a strong ability to enhance prior BCG immunity.

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Fusion Cytokines IL-7-Linker-IL-15 Promote Mycobacterium Tuberculosis Subunit Vaccine to Induce Central Memory like T Cell-Mediated Immunity

Vaccines ◽

10.3390/vaccines8040715 ◽

2020 ◽

Vol 8 (4) ◽

pp. 715

Author(s):

Chunxiang Bai ◽

Lijun Zhou ◽

Junxia Tang ◽

Juanjuan He ◽

Jiangyuan Han ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Immune Responses ◽

Protective Efficacy ◽

Central Memory ◽

Control Group ◽

Cell Mediated Immunity ◽

Protein Subunit ◽

Subunit Vaccines ◽

Ifn Γ

Tuberculosis (TB), caused by Mycobacterium tuberculosis (M. tuberculosis), is among the most serious infectious diseases worldwide. Adjuvanted protein subunit vaccines have been demonstrated as a kind of promising novel vaccine. This study proposed to investigate whether cytokines interliukine-7 (IL-7) and interliukine-15 (IL-15) help TB subunit vaccines induce long-term cell-mediated immune responses, which are required for vaccination against TB. In this study, mice were immunized with the M. tuberculosis protein subunit vaccines combined with adnovirus-mediated cytokines IL-7, IL-15, IL-7-IL-15, and IL-7-Linker-IL-15 at 0, 2, and 4 weeks, respectively. Twenty weeks after the last immunization, the long-term immune responses, especially the central memory-like T cells (TCM like cell)-mediated immune responses, were determined with the methods of cultured IFN-γ-ELISPOT, expanded secondary immune responses, cell proliferation, and protective efficacy against Mycobacterium bovis Bacilli Calmette-Guerin (BCG) challenge, etc. The results showed that the group of vaccine + rAd-IL-7-Linker-IL-15 induced a stronger long-term antigen-specific TCM like cells-mediated immune responses and had higher protective efficacy against BCG challenge than the vaccine + rAd-vector control group, the vaccine + rAd-IL-7 and the vaccine + rAd-IL-15 groups. This study indicated that rAd-IL-7-Linker-IL-15 improved the TB subunit vaccine’s efficacy by augmenting TCM like cells and provided long-term protective efficacy against Mycobacteria.

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

Clinical and Vaccine Immunology ◽

10.1128/cvi.00151-06 ◽

2006 ◽

Vol 13 (8) ◽

pp. 930-935 ◽

Cited By ~ 15

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.

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Protection of Mice with a Divalent Tuberculosis DNA Vaccine Encoding Antigens Ag85B and MPT64

Acta Biochimica et Biophysica Sinica ◽

10.1093/abbs/36.4.269 ◽

2004 ◽

Vol 36 (4) ◽

pp. 269-276 ◽

Cited By ~ 13

Author(s):

Xia Tian ◽

Hong Cai ◽

Yu-Xian Zhu

Keyword(s):

Dna Vaccine ◽

Protective Efficacy ◽

Vaccine Group ◽

Control Group ◽

Lung Weight ◽

Promising Tool ◽

Ifn Γ ◽

Immunodominant Antigens ◽

High Doses ◽

Tuberculosis Disease

Abstract DNA vaccine may be a promising tool for controlling tuberculosis development. However, vaccines encoding single antigens of mycobacterium did not produce protective effect as BCG did. In the present study, we evaluated the immunogenicity and protective efficacy of a divalent DNA vaccine encoding two immunodominant antigens Ag85B and MPT64 of Mycobacterium tuberculosis. We found that both humoral and Th1-type (high IFN-γ, low IL-4) cellular responses obtained from the divalent DNA vaccine group were significantly higher than that conferred by BCG. RT-PCR results showed that antigens were expressed differentially in various organs in divalent DNA vaccine group. The survival rate for mice treated with the divalent DNA vaccine after challenging with high doses of virulent M. tuberculosis H37Rv was significantly higher than that of the BCG group or any of the single DNA vaccine group. Significant differences were also found between the single and divalent DNA vaccinated mice in terms of body, spleen and lung weight. Bacterial loading decreased about 2000-fold in lungs and about 100-fold in spleens of divalent DNA vaccinated mice when compared with that of the control group. We conclude that our divalent DNA vaccine may be a better choice for controlling tuberculosis disease in animals.

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Improved immunogenicity and protective efficacy of a divalent DNA vaccine encoding Brucella L7/L12-truncated Omp31 fusion protein by a DNA priming and protein boosting regimen

Molecular Immunology ◽

10.1016/j.molimm.2015.04.015 ◽

2015 ◽

Vol 66 (2) ◽

pp. 384-391 ◽

Cited By ~ 24

Author(s):

Maryam Golshani ◽

Sima Rafati ◽

Seyed Davar Siadat ◽

Mehdi Nejati-Moheimani ◽

Fereshteh Shahcheraghi ◽

...

Keyword(s):

Fusion Protein ◽

Dna Vaccine ◽

Protective Efficacy

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Feline Leukemia Virus DNA Vaccine Efficacy Is Enhanced by Coadministration with Interleukin-12 (IL-12) and IL-18 Expression Vectors

Journal of Virology ◽

10.1128/jvi.75.18.8424-8433.2001 ◽

2001 ◽

Vol 75 (18) ◽

pp. 8424-8433 ◽

Cited By ~ 46

Author(s):

Linda Hanlon ◽

David Argyle ◽

Derek Bain ◽

Lesley Nicolson ◽

Stephen Dunham ◽

...

Keyword(s):

Dna Vaccine ◽

Leukemia Virus ◽

The Other ◽

Feline Leukemia Virus ◽

Expression Vectors ◽

Interleukin 12 ◽

Cell Mediated Immunity ◽

Empty Plasmid ◽

Ifn Γ ◽

Intraperitoneal Inoculation

ABSTRACT The expectation that cell-mediated immunity is important in the control of feline leukemia virus (FeLV) infection led us to test a DNA vaccine administered alone or with cytokines that favored the development of a Th1 immune response. The vaccine consisted of two plasmids, one expressing the gag/pol genes and the other expressing the env gene of FeLV-A/Glasgow-1. The genetic adjuvants were plasmids encoding the feline cytokines interleukin-12 (IL-12), IL-18, or gamma interferon (IFN-γ). Kittens were immunized by three intramuscular inoculations of the FeLV DNA vaccine alone or in combination with plasmids expressing IFN-γ, IL-12, or both IL-12 and IL-18. Control kittens were inoculated with empty plasmid. Following immunization, anti-FeLV antibodies were not detected in any kitten. Three weeks after the final immunization, the kittens were challenged by the intraperitoneal inoculation of FeLV-A/Glasgow-1 and were then monitored for a further 15 weeks for the presence of virus in plasma and, at the end of the trial, for latent virus in bone marrow. The vaccine consisting of FeLV DNA with the IL-12 and IL-18 genes conferred significant immunity, protecting completely against transient and persistent viremia, and in five of six kittens protecting against latent infection. None of the other vaccines provided significant protection.

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Vaccination with a DNA Vaccine Coding for Perforin-Like Protein 1 and MIC6 Induces Significant Protective Immunity against Toxoplasma gondii

Clinical and Vaccine Immunology ◽

10.1128/cvi.05578-11 ◽

2012 ◽

Vol 19 (5) ◽

pp. 684-689 ◽

Cited By ~ 26

Author(s):

Hai-Kuo Yan ◽

Zi-Guo Yuan ◽

Hui-Qun Song ◽

Eskild Petersen ◽

Yang Zhou ◽

...

Keyword(s):

Toxoplasma Gondii ◽

Dna Vaccine ◽

Interleukin 2 ◽

Parasite Load ◽

Control Group ◽

Igg Antibody ◽

Content Type ◽

Humoral Immune ◽

Humoral Immune Responses ◽

Ifn Γ

ABSTRACTHost cell invasion byToxoplasma gondiiis tightly related to microneme protein 6 (MIC6) andT. gondiiperforin-like protein 1 (TgPLP1). In this study, we constructed a DNA vaccine expressing a TgPLP1/MIC6 fusion protein using the pIRESneo vector, and we evaluated the immune response induced by this vaccine in Kunming mice. Levels of IgG antibody, gamma interferon (IFN-γ), interleukin 2 (IL-2), IL-12, IL-4, and IL-10 were examined. Five mice were chosen randomly from every group (vaccinated groups or the nonvaccinated control group) and were challenged intragastrically with 80 cysts ofT. gondiistrain PRU (genotype II) in order to observe mortality daily. To analyze protection against a less-virulent challenge, eight mice of each group were orally infected with 20 cysts of strain PRU at the 14th day after the last immunization. The brain parasite load was evaluated 6 weeks after infection. The results demonstrated that immunization with pIRESneo/MIC6/PLP1 resulted in the lowest brain cyst count and prolonged the survival time of immunized mice. The levels ofToxoplasma-specific IgG, IFN-γ, IL-2, and IL-12 increased significantly, and the numbers of cysts in brains decreased more obviously, in the group immunized with plasmid pIRESneo/MIC6/PLP1 than in the other groups (P< 0.05). Compared with pIRESneo/MIC6/PLP1, coimmunization with pIRESneo/MIC6/PLP1 and adjuvant murine IL-18 promoted cellular and humoral immune responses but did not contribute significantly to cyst reduction (65.43% versus 61.60%) or the survival of immunized mice (45.0 ± 2.9 days versus 42.8 ± 2.9 days) (P> 0.05). Furthermore, the study also showed that the immune efficacy induced by pIRESneo/MIC6/PLP1 was better than that induced by pVAX/PLP1 or pVAX/MIC6 alone.

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Enhancement of the immunogenicity of a Mycobacterium tuberculosis fusion protein using ISCOMATRIX and PLUSCOM nano-adjuvants after nasal administration in mice

10.1101/2021.08.27.458002 ◽

2021 ◽

Author(s):

Arshid Yousefi Avarvand ◽

Zahra Meshkat ◽

Farzad Khademi ◽

Ehsan Aryan ◽

Mojtaba Sankian ◽

...

Keyword(s):

Mycobacterium Tuberculosis ◽

Immune Responses ◽

Fusion Protein ◽

Intranasal Administration ◽

Bacterial Infections ◽

Protective Efficacy ◽

Nasal Administration ◽

Mice Model ◽

Ifn Γ ◽

Nasal Immunization

Background: Tuberculosis (TB), a contagious disease caused by Mycobacterium tuberculosis (M. tuberculosis), remains a health problem worldwide and this infection has the highest mortality rate among bacterial infections. Current studies suggest that intranasal administration of new tuberculosis vaccines could enhance the immunogenicity of M. tuberculosis antigens. Hence, we aim to evaluate the protective efficacy and immunogenicity of HspX/EsxS fusion protein of M. tuberculosis along with ISCOMATRIX and PLUSCOM nano-adjuvants and MPLA through the intranasal administration in mice model. Methods:In present study, the recombinant fusion protein was expressed in Escherichia coli and purified and used to prepare different nanoparticle formulations in combination with ISCOMATRIX and PLUSCOM nano-adjuvants and MPLA. Mice were intranasally vaccinated with each formulation three times at an interval of 2 weeks. Finally, IFN-γ, IL-4. IL-17 and TGF-β concentration in supernatant of cultured splenocytes of vaccinated mice as well as serum titers of IgG1 and IgG2a and sIgA titers in nasal lavage were determined. Results:According to obtained results, intranasally vaccinated mice with formulations containing ISCOMATRIX and PLUSCOM nano-adjuvants and MPLA could effectively induced IFN-γ and sIgA responses. Moreover, both HspX/EsxS/ISCOMATRIX/MPLA and HspX/EsxS/PLUSCOM/MPLA and their BCG booster formulation could strongly stimulate the immune system and enhance the immunogenicity of M. tuberculosis antigens. Conclusion: The results demonstrate the potential of HspX/EsxS-fused protein in combination with ISCOMATRIX, PLUSCOM and MPLA after nasal administration in enhancing immune response against of M. tuberculosis antigens. So, nasal immunization with these formulations, could induce immune responses and considered as new TB vaccine or as BCG booster.

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Immunogenicity and protective efficacy of a tuberculosis DNA vaccine expressing a fusion protein of Ag85B-Esat6-HspX in mice

Vaccine ◽

10.1016/j.vaccine.2011.06.029 ◽

2012 ◽

Vol 30 (14) ◽

pp. 2490-2497 ◽

Cited By ~ 19

Author(s):

Wei Yuan ◽

Na Dong ◽

Lifang Zhang ◽

Jiangning Liu ◽

Shuzhu Lin ◽

...

Keyword(s):

Fusion Protein ◽

Dna Vaccine ◽

Protective Efficacy

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Co-Expression of Chicken IL-2 and IL-7 Enhances the Immunogenicity and Protective Efficacy of a VP2-Expressing DNA Vaccine against IBDV in Chickens

Viruses ◽

10.3390/v11050476 ◽

2019 ◽

Vol 11 (5) ◽

pp. 476 ◽

Cited By ~ 1

Author(s):

Shanshan Huo ◽

Jianlou Zhang ◽

Jinghui Fan ◽

Xing Wang ◽

Fengyang Wu ◽

...

Keyword(s):

Dna Vaccine ◽

Protective Efficacy ◽

T Cell Proliferation ◽

Gene Vector ◽

Safe Vaccine ◽

Bursal Disease ◽

Gene Vectors ◽

Antibody Titers ◽

Protection Efficacy ◽

Ifn Γ

Chicken infectious bursal disease (IBD) is still incompletely controlled worldwide. Although IBD virus (IBDV) VP2 DNA vaccine was considered a safe vaccine for IBD prevention, the immunogenicity by itself remains poor, resulting in the failure of effectively protecting chickens from infection. We and others demonstrated that chicken IL-2 (chIL-2) and chIL-7 have the capacity to enhance the immunogenicity of the VP2 DNA vaccine. However, whether chIL-2 and chIL-7 can mutually enhance the immunogenicity of VP2 DNA vaccine and thereby augment the latter’s protection efficacy remains unknown. By using chIL-2/chIL-7 bicistronic gene vector to co-immunize the chickens together with the VP2 DNA vaccine, we now show that chIL-2 and chIL-7 significantly increased IBDV VP2-specific antibody titers, T cell proliferation, and IFN-γ production, resulting in the ultimate enhancement of vaccine-induced protection efficacy relative to that of chIL-2 or chIL-7 gene vectors alone. These results suggest that chIL-2 and chIL-7 can mutually enhance VP2 DNA vaccine’s efficacy, thereby establishing a concrete foundation for future optimization of IBDV VP2 DNA vaccine to prevent/treat chicken IBD.

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Synthetic Long Peptide Derived from Mycobacterium tuberculosis Latency Antigen Rv1733c Protects against Tuberculosis

Clinical and Vaccine Immunology ◽

10.1128/cvi.00271-15 ◽

2015 ◽

Vol 22 (9) ◽

pp. 1060-1069 ◽

Cited By ~ 19

Author(s):

Mariateresa Coppola ◽

Susan J. F. van den Eeden ◽

Louis Wilson ◽

Kees L. M. C. Franken ◽

Tom H. M. Ottenhoff ◽

...

Keyword(s):

T Cells ◽

Mycobacterium Tuberculosis ◽

Protective Efficacy ◽

Bacterial Load ◽

Effective Vaccine ◽

Protective Capacity ◽

Content Type ◽

Ifn Γ ◽

Vaccine Approach

ABSTRACTResponsible for 9 million new cases of active disease and nearly 2 million deaths each year, tuberculosis (TB) remains a global health threat of overwhelming dimensions.Mycobacterium bovisBCG, the only licensed vaccine available, fails to confer lifelong protection and to prevent reactivation of latent infection. Although 15 new vaccine candidates are now in clinical trials, an effective vaccine against TB remains elusive, and new strategies for vaccination are vital. BCG vaccination fails to induce immunity againstMycobacterium tuberculosislatency antigens. Synthetic long peptides (SLPs) combined with adjuvants have been studied mostly for therapeutic cancer vaccines, yet not for TB, and proved to induce efficient antitumor immunity. This study investigated an SLP derived from Rv1733c, a majorM. tuberculosislatency antigen which is highly expressed by “dormant”M. tuberculosisand well recognized by T cells from latentlyM. tuberculosis-infected individuals. In order to assess itsin vivoimmunogenicity and protective capacity, Rv1733c SLP in CpG was administered to HLA-DR3 transgenic mice. Immunization with Rv1733c SLP elicited gamma interferon-positive/tumor necrosis factor-positive (IFN-γ+/TNF+) and IFN-γ+CD4+T cells and Rv1733c-specific antibodies and led to a significant reduction in the bacterial load in the lungs ofM. tuberculosis-challenged mice. This was observed both in a pre- and in a post-M. tuberculosischallenge setting. Moreover, Rv1733c SLP immunization significantly boosted the protective efficacy of BCG, demonstrating the potential ofM. tuberculosislatency antigens to improve BCG efficacy. These data suggest a promising role forM. tuberculosislatency antigen Rv1733c-derived SLPs as a novel TB vaccine approach, both in a prophylactic and in a postinfection setting.

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