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 Mycobacterium tuberculosis Culture Filtrate Proteins plus CpG Oligodeoxynucleotides Confer Protection to Mycobacterium bovis BCG-Primed Mice by Inhibiting Interleukin-4 Secretion

2009 ◽  
Vol 77 (12) ◽  
pp. 5311-5321 ◽  
Author(s):  
Denise Morais da Fonseca ◽  
Celio Lopes Silva ◽  
Pryscilla Fanini Wowk ◽  
Marina Oliveira e Paula ◽  
Simone Gusmão Ramos ◽  
...  
Keyword(s):  
Mycobacterium Tuberculosis ◽  
Mycobacterium Bovis ◽  
Culture Filtrate ◽  
Vaccine Development ◽  
Interleukin 4 ◽  
Interleukin 17 ◽  
Bcg Vaccination ◽  
Cpg Oligodeoxynucleotides ◽  
Culture Filtrate Proteins ◽  
Ifn Γ

ABSTRACT Culture filtrate proteins (CFP) are potential targets for tuberculosis vaccine development. We previously showed that despite the high level of gamma interferon (IFN-γ) production elicited by homologous immunization with CFP plus CpG oligodeoxynucleotides (CFP/CpG), we did not observe protection when these mice were challenged with Mycobacterium tuberculosis. In order to use the IFN-γ-inducing ability of CFP antigens, in this study we evaluated a prime-boost heterologous immunization based on CFP/CpG to boost Mycobacterium bovis BCG vaccination in order to find an immunization schedule that could induce protection. Heterologous BCG-CFP/CpG immunization provided significant protection against experimental tuberculosis, and this protection was sustained during the late phase of infection and was even better than that conferred by a single BCG immunization. The protection was associated with high levels of antigen-specific IFN-γ and interleukin-17 (IL-17) and low IL-4 production. The deleterious role of IL-4 was confirmed when IL-4 knockout mice vaccinated with CFP/CpG showed consistent protection similar to that elicited by BCG-CFP/CpG heterologous immunization. These findings show that a single dose of CFP/CpG can represent a new strategy to boost the protection conferred by BCG vaccination. Moreover, different immunological parameters, such as IFN-γ and IL-17 and tightly regulated IL-4 secretion, seem to contribute to the efficacy of this tuberculosis vaccine.

scholarly journals 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

2011 ◽  
Vol 2011 ◽  
pp. 1-10 ◽  
Author(s):  
Jia Lu ◽  
Chun Wang ◽  
Zhiguang Zhou ◽  
Ying Zhang ◽  
Tingting Cao ◽  
...  
Keyword(s):  
Fusion Protein ◽  
Dna Vaccine ◽  
Protective Efficacy ◽  
Bacterial Load ◽  
Control Group ◽  
Cell Mediated Immunity ◽  
Significant Protection ◽  
Booster Vaccine ◽  
Ifn Γ ◽  
Negative Population

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.

scholarly journals Virally Activated CD8 T Cells Home to Mycobacterium bovis BCG-Induced Granulomas but Enhance Antimycobacterial Protection Only in Immunodeficient Mice

2006 ◽  
Vol 75 (3) ◽  
pp. 1154-1166 ◽  
Author(s):  
Laura H. Hogan ◽  
Dominic O. Co ◽  
Jozsef Karman ◽  
Erika Heninger ◽  
M. Suresh ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Mycobacterium Bovis ◽  
Cd8 T Cells ◽  
Bacterial Load ◽  
Granulomatous Inflammation ◽  
Cd8 T Cell ◽  
Activated T Cells ◽  
Immunodeficient Mice ◽  
Ifn Γ

ABSTRACT The effect of secondary infections on CD4 T-cell-regulated chronic granulomatous inflammation is not well understood. Here, we have investigated the effect of an acute viral infection on the cellular composition and bacterial protection in Mycobacterium bovis strain bacille Calmette-Guérin (BCG)-induced granulomas using an immunocompetent and a partially immunodeficient murine model. Acute lymphocytic choriomeningitis virus (LCMV) coinfection of C57BL/6 mice led to substantial accumulation of gamma interferon (IFN-γ)-producing LCMV-specific T cells in liver granulomas and increased local IFN-γ. Despite traffic of activated T cells that resulted in a CD8 T-cell-dominated granuloma, the BCG liver organ load was unaltered from control levels. In OT-1 T-cell-receptor (TCR) transgenic mice, ovalbumin (OVA) immunization or LCMV coinfection of BCG-infected mice induced CD8 T-cell-dominated granulomas containing large numbers of non-BCG-specific activated T cells. The higher baseline BCG organ load in this CD8 TCR transgenic animal allowed us to demonstrate that OVA immunization and LCMV coinfection increased anti-BCG protection. The bacterial load remained substantially higher than in mice with a more complete TCR repertoire. Overall, the present study suggests that peripherally activated CD8 T cells can be recruited to chronic inflammatory sites, but their contribution to protective immunity is limited to conditions of underlying immunodeficiency.

scholarly journals CpG Oligodeoxynucleotides and Interleukin-12 Improve the Efficacy of Mycobacterium bovis BCG Vaccination in Mice Challenged with M. tuberculosis

2000 ◽  
Vol 68 (5) ◽  
pp. 2948-2953 ◽  
Author(s):  
Brenda L. Freidag ◽  
Genevieve B. Melton ◽  
Frank Collins ◽  
Dennis M. Klinman ◽  
Allen Cheever ◽  
...  
Keyword(s):  
Mycobacterium Bovis ◽  
Bacterial Load ◽  
Serial Passage ◽  
Interleukin 12 ◽  
Cpg Odn ◽  
Bcg Vaccination ◽  
Cpg Oligodeoxynucleotides ◽  
Enhanced Production ◽  
Ifn Γ

ABSTRACT Mycobacterium bovis bacillus Calmette-Guérin (BCG) is the only vaccine approved for prevention of tuberculosis. It has been postulated that serial passage of BCG over the years may have resulted in attenuation of its effectiveness. Because interleukin-12 (IL-12) and oligodeoxynucleotides (ODN) containing cytidine phosphate guanosine (CpG) motifs have been shown to enhance Th1 responses in vivo, they were chosen as adjuvants to increase the effectiveness of BCG vaccination. In this report, mice were vaccinated with BCG with or without IL-12 or CpG ODN and then challenged 6 weeks later via the aerosol route with the Erdman strain of M. tuberculosis. Mice vaccinated with BCG alone showed a 1- to 2-log reduction in bacterial load compared with control mice that did not receive any vaccination prior to M. tuberculosis challenge. Moreover, the bacterial loads of mice vaccinated with BCG plus IL-12 or CpG ODN were a further two- to fivefold lower than those of mice vaccinated with BCG alone. As an immune correlate, the antigen-specific production IFN-γ and mRNA expression in spleen cells prior to challenge were evaluated. Mice vaccinated with BCG plus IL-12 or CpG ODN showed enhanced production of IFN-γ compared with mice vaccinated with BCG alone. Finally, granulomas in BCG-vaccinated mice were smaller and more lymphocyte rich than those in unvaccinated mice; however, the addition of IL-12 or CpG ODN to BCG vaccination did not alter granuloma formation or result in added pulmonary damage. These observations support a role for immune adjuvants given with BCG vaccination to enhance its biologic efficacy.

scholarly journals A Single Dose of a DNA Vaccine Encoding Apa Coencapsulated with 6,6′-Trehalose Dimycolate in Microspheres Confers Long-Term Protection against Tuberculosis in Mycobacterium bovis BCG-Primed Mice

2013 ◽  
Vol 20 (8) ◽  
pp. 1162-1169 ◽  
Author(s):  
Dyego Carlétti ◽  
Denise Morais da Fonseca ◽  
Ana Flávia Gembre ◽  
Ana Paula Masson ◽  
Lívia Weijenborg Campos ◽  
...  
Keyword(s):  
Single Dose ◽  
Dna Vaccine ◽  
Mycobacterium Bovis ◽  
Bacterial Load ◽  
Lung Parenchyma ◽  
Content Type ◽  
Boost Regimen ◽  
Trehalose Dimycolate ◽  
Tb Infection Control

ABSTRACTMycobacterium bovisBCG prime DNA (Mycobacterium tuberculosisgenes)-booster vaccinations have been shown to induce greater protection against tuberculosis (TB) than BCG alone. This heterologous prime-boost strategy is perhaps the most realistic vaccination for the future of TB infection control, especially in countries where TB is endemic. Moreover, a prime-boost regimen using biodegradable microspheres seems to be a promising immunization to stimulate a long-lasting immune response. The alanine proline antigen (Apa) is a highly immunogenic glycoprotein secreted byM. tuberculosis. This study investigated the immune protection of Apa DNA vaccine against intratrachealM. tuberculosischallenge in mice on the basis of a heterologous prime-boost regimen. BALB/c mice were subcutaneously primed with BCG and intramuscularly boosted with a single dose of plasmid carryingapaand 6,6′-trehalose dimycolate (TDM) adjuvant, coencapsulated in microspheres (BCG-APA), and were evaluated 30 and 70 days after challenge. This prime-boost strategy (BCG-APA) resulted in a significant reduction in the bacterial load in the lungs, thus leading to better preservation of the lung parenchyma, 70 days postinfection compared to BCG vaccinated mice. The profound effect of this heterologous prime-boost regimen in the experimental model supports its development as a feasible strategy for prevention of TB.

scholarly journals Adoptive Transfer of CD4+ T Cells Specific for Subunit A of Helicobacter pylori Urease ReducesH. pylori Stomach Colonization in Mice in the Absence of Interleukin-4 (IL-4)/IL-13 Receptor Signaling

2001 ◽  
Vol 69 (3) ◽  
pp. 1714-1721 ◽  
Author(s):  
Bernadette Lucas ◽  
Dirk Bumann ◽  
Anna Walduck ◽  
Jan Koesling ◽  
Leyla Develioglu ◽  
...  
Keyword(s):  
T Cells ◽  
Helicobacter Pylori ◽  
Adoptive Transfer ◽  
Bacterial Load ◽  
Interleukin 4 ◽  
T Cell Epitopes ◽  
Necrosis Factor Alpha ◽  
Subunit A ◽  
Ifn Γ ◽  
H Pylori

ABSTRACT Protection in the murine model of Helicobacter pyloriinfection may be mediated by CD4+ T cells, but the mechanism remains unclear. To better understand how protection occurs in this model, we generated and characterized H. pyloriurease-specific CD4+ T cells from BALB/c mice immunized with Salmonella enterica serovar Typhimurium expressingH. pylori urease (subunits A and B). The CD4+ T cells were found to be specific for subunit A (UreA). Upon antigen-specific stimulation, expression of interleukin 4 (IL-4), IL-10, gamma interferon (IFN-γ), and tumor necrosis factor alpha was induced. Immunocytochemical analysis showed that the majority of cells produced IFN-γ and IL-10. Adoptive transfer of the UreA-specific CD4+ T cells into naive syngeneic recipients led to a threefold reduction in the number of bacteria in the recipient group when compared to that in the nonrecipient group. Stomach colonization was also reduced significantly after transfer of these cells into patently infected mice. Adoptive transfer of UreA-specific CD4+ T cells into IL-4 receptor α chain-deficient BALB/c mice indicated that IL-4 and IL-13 were not critical in the control of bacterial load. In addition, synthetic peptides were used to identify three functional T-cell epitopes present in subunit A which were recognized by the UreA-specific T cells. Analysis of H. pylori-specific cellular immune responses in recipient challenged and nonrecipient infected mice indicated a strong local restriction of the response in infected animals. The implications of these findings for the mechanism of protection and the development of peptide-based vaccination are discussed.

Expressions of IFN-γ and IL-4 before and after Treatment of Lupus Nephritis with Traditional Chinese Medicine Combined with Cyclophosphamide and Their Values for Efficacy Prediction and Evaluation

2020 ◽  
Author(s):  
Yan JIANG ◽  
Qichang ZHANG ◽  
Haoyu WANG ◽  
Dabei TANG ◽  
Yan ZHANG ◽  
...  
Keyword(s):  
Chinese Medicine ◽  
Traditional Chinese Medicine ◽  
Lupus Nephritis ◽  
Clinical Efficacy ◽  
Activity Index ◽  
Interleukin 4 ◽  
Control Group ◽  
Before And After ◽  
Ifn Γ ◽  
Efficacy Prediction

Background: To explore IFN-γ (interferon-γ) and IL-4 (interleukin-4) expressions before and after the treatment of LN (lupus nephritis) and their values for efficacy prediction and evaluation. Methods: Altogether 107 patients with LN treated in the First Hospital of Qiqihaer City, Qiqihar, China from March 2017 to September 2018 were enrolled. Sixty-two patients were treated with cyclophosphamide and prednisolone (control group), while another 45 patients were treated with Qing Shen Fang based on the control group (observation group). Their clinical efficacy and changes in immune indices after treatment were observed. Results: Compared with those in the control group, clinical efficacy, IFN-γ, IL-4, hemoglobin, complements C3 and C4, ESR (erythrocyte sedimentation rate), serum IgG, SLEDAI (Systemic Lupus Erythematosus Disease Activity Index) score, and TCMSSS (Traditional Chinese Medicine Syndrome Score Scale) score were significantly improved after treatment in the study group. Based on the observation, IFN-γ and IL-4 could be used as potential indicators for evaluating clinical efficacy. Conclusion: The combination of cyclophosphamide, prednisolone, and Qing Shen Fang improves conditions of patients with LN and significantly reduces their IFN-γ and IL-4 levels in serum. IFN-γ and IL-4 can be used as potential indicators for the efficacy prediction and evaluation of the disease.

scholarly journals Signaling via Interleukin-4 Receptor α Chain Is Required for Successful Vaccination against Schistosomiasis in BALB/c Mice

2001 ◽  
Vol 69 (1) ◽  
pp. 228-236 ◽  
Author(s):  
Adrian P. Mountford ◽  
Karen G. Hogg ◽  
Patricia S. Coulson ◽  
Frank Brombacher
Keyword(s):  
Immune Responses ◽  
Protective Immunity ◽  
Interleukin 4 ◽  
Lymphoid Tissues ◽  
Partial Protection ◽  
Exposure Site ◽  
Ifn Γ ◽  
Α Chain ◽  
Interleukin 4 Receptor ◽  
Effector Response

ABSTRACT Although protective immunity in C57BL/6 mice induced by a single dose of the radiation-attenuated schistosome vaccine is believed to be mediated by Th1-type immune responses, we here report that in BALB/c mice protection can also depend upon signaling via the interleukin-4 (IL-4) receptor which conventionally governs the development of Th2-type immune responses. We show that in BALB/c mice deficient for the IL-4 receptor α chain (IL-4Rα−/−), which are unresponsive to IL-4 and IL-13, vaccine-induced protection is abrogated compared with that in wild-type (WT) mice. In vaccinated IL-4Rα−/− mice, IL-12p40 production by cells from the skin exposure site was elevated, although gamma interferon (IFN-γ) production in draining lymphoid tissues was similar in WT and IL-4Rα−/− mice. Nevertheless, the effector response in IL-4Rα−/− mice was Th1 biased with elevated IFN-γ in the lungs and higher immunoglobulin G2a (IgG2a) and IgG2b titers but negligible quantities of Th2-associated IgG1 and IgE. Interestingly, levels of IL-4 were equivalent in WT and IL-4Rα−/−mice, indicating that Th2 responses were not dependent upon signaling by IL-4 or IL-13. No differences in the phenotype and composition of the pulmonary effector mechanism that might explain the failure to induce protection in IL-4Rα−/− mice were detected. However, passive transfer of partial protection to naive IL-4Rα−/− mice, using serum from vaccinated WT mice, indicates that Th2-associated antibodies such as IgG1 have a role in parasite elimination in BALB/c strain mice and that signaling via IL-4R can be an important factor in the generation of protection.

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

2012 ◽  
Vol 19 (5) ◽  
pp. 684-689 ◽  
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.

scholarly journals Predominance of CD4 Th1 and CD8 Tc1 Cells Revealed by Characterization of the Cellular Immune Response Generated by Immunization with a DNA Vaccine Containing a Trypanosoma cruzi Gene

1999 ◽  
Vol 67 (8) ◽  
pp. 3855-3863 ◽  
Author(s):  
Mauricio M. Rodrigues ◽  
Marcelo Ribeirão ◽  
Vera Pereira-Chioccola ◽  
Laurent Renia ◽  
Fabio Costa
Keyword(s):  
T Cells ◽  
Trypanosoma Cruzi ◽  
Dna Vaccine ◽  
Cd8 T Cells ◽  
Catalytic Domain ◽  
Therapeutic Vaccine ◽  
Cell Types ◽  
Interleukin 4 ◽  
Ifn Γ

ABSTRACT Immunization with a plasmid DNA containing the gene encoding the catalytic domain of trans-sialidase (TS) elicits protective immune responses against experimental Trypanosoma cruziinfection. As several studies provided strong evidence that during infection CD4 Th1 and CD8 T cytotoxic type 1 (Tc1) cells are important factors in host resistance, the present study was designed to evaluate which T-cell types were activated in DNA-vaccinated BALB/c mice. We found that bulk cells from DNA-immunized mice had CD4 and CD8 T cells that produced gamma interferon (IFN-γ) but not interleukin-4 (IL-4) or IL-10. To characterize the TS-specific T cells at the clonal level, we generated CD4 and CD8 clones. We obtained cytotoxic CD4 clones of the Th1 type that secreted large amounts of IFN-γ but not IL-4 or IL-10. Unexpectedly, we obtained other CD4 clones with a Th2 phenotype, secreting IL-4 and IL-10 but not IFN-γ. All CD8 clones were cytotoxic and produced IFN-γ. IL-4 and IL-10 were not secreted by these cells. Using synthetic peptides, we determined a CD8 epitope recognized by several clones as being represented by amino acids IYNVGQVSI. The antiparasitic activity of a CD4 Th1 and a CD8 Tc1 clone was assessed in vitro. CD4 or CD8 T cells significantly inhibited T. cruzidevelopment in infected macrophages or fibroblasts, respectively. We concluded that DNA vaccine efficiently generates potentially protective CD4 Th1 and CD8 Tc1 cells specific for a T. cruzi antigen, therefore reinforcing the possibility of using this strategy for developing a preventive or therapeutic vaccine against Chagas’ disease.

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