TcVac3 Induced Control of Trypanosoma cruzi Infection and Chronic Myocarditis in Mice

We characterized the immune responses elicited by a DNA-prime/MVA-boost vaccine (TcVac3) constituted of antigenic candidates (TcG2 and TcG4), shown to be recognized by B and T cell responses in Trypanosoma cruzi (Tc) infected multiple hosts. C57BL/6 mice immunized with TcVac3 elicited a strong antigen-specific, high-avidity, trypanolytic antibody response (IgG2b>IgG1); and a robust antigen- and Tc-specific CD8+T cell response with type-1 cytokine (IFN-γ+TNF-α>IL-4+IL-10) and cytolytic effector (CD8+CD107a+IFN-γ+Perforin+) phenotype. The vaccine-induced effector T cells significantly expanded upon challenge infection and provided >92% control of T. cruzi. Co-delivery of IL-12 and GMCSF cytokine adjuvants didn’t enhance the TcVac3-induced resistance to T. cruzi. In chronic phase, vaccinated/infected mice exhibited a significant decline (up to 70%) in IFN-γ+CD8+T cells, a predominance of immunoregulatory IL-10+/CD4+T and IL10+/CD8+T cells, and presented undetectable tissue parasitism, inflammatory infiltrate, and fibrosis in vaccinated/infected mice. In comparison, control mice responded to challenge infection by a low antibody response, mixed cytokine profile, and consistent activation of pro-inflammatory CD8+T cells associated with parasite persistence and pathologic damage in the heart. We conclude that TcVac3 elicited type-1 effector T cell immunity that effectively controlled T. cruzi infection, and subsequently, predominance of anti-inflammatory responses prevented chronic inflammation and myocarditis in chagasic mice.

[1]  S. Boscardin,et al.  A DNA‐priming protein‐boosting regimen significantly improves type 1 immune response but not protective immunity to Trypanosoma cruzi infection in a highly susceptible mouse strain , 2003, Immunology and cell biology.

[2]  Antonio Lanzavecchia,et al.  Maintenance of Serological Memory by Polyclonal Activation of Human Memory B Cells , 2002, Science.

[3]  C. Mady,et al.  TNF gene polymorphisms are associated with reduced survival in severe Chagas' disease cardiomyopathy patients. , 2006, Microbes and infection.

[4]  Y. Takashima,et al.  Immune Responses against a Single CD8+-T-Cell Epitope Induced by Virus Vector Vaccination Can Successfully Control Trypanosoma cruzi Infection , 2005, Infection and Immunity.

[5]  N. Garg,et al.  Prophylactic Efficacy of TcVac2 against Trypanosoma cruzi in Mice , 2010, PLoS neglected tropical diseases.

[6]  M. Lehtinen,et al.  Low avidity of human papillomavirus (HPV) type 16 antibodies is associated with increased risk of low-risk but not high-risk HPV type prevalence , 2011, BMC Research Notes.

[7]  A. Silber,et al.  Antibody Maturation in Trypanosoma cruzi-Infected Rats , 2001, Clinical Diagnostic Laboratory Immunology.

[8]  E. Malchiodi,et al.  Oral Vaccination with Salmonella enterica as a Cruzipain-DNA Delivery System Confers Protective Immunity against Trypanosoma cruzi , 2007, Infection and Immunity.

[9]  D. Engman,et al.  Chagas heart disease pathogenesis: one mechanism or many? , 2008, Current molecular medicine.

[10]  A. Reyes-Sandoval,et al.  Recombinant viruses as vaccines against viral diseases. , 2005, Brazilian journal of medical and biological research = Revista brasileira de pesquisas medicas e biologicas.

[11]  Melissa M. Eitzen,et al.  Vaccines for Biodefense and Emerging and Neglected Diseases , 2009 .

[12]  D. Moorhouse.  UNDP/World Bank/WHO: Special programme for research and training in tropical diseases 5th annual report , 1982 .

[13]  C. Staib,et al.  Modified vaccinia virus Ankara as antigen delivery system: how can we best use its potential? , 2004, Current Opinion in Biotechnology.

[14]  R. Corrêa-Oliveira,et al.  Anti-Trypanosoma cruzi Immunoglobulin G1 Can Be a Useful Tool for Diagnosis and Prognosis of Human Chagas' Disease , 2001, Clinical Diagnostic Laboratory Immunology.

[15]  K. Marsh,et al.  Immunogenicity of the candidate malaria vaccines FP9 and modified vaccinia virus Ankara encoding the pre-erythrocytic antigen ME-TRAP in 1-6 year old children in a malaria endemic area. , 2006, Vaccine.

[16]  R. Corrêa-Oliveira,et al.  T‐Cell Repertoire Analysis in Acute and Chronic Human Chagas'Disease: Differentail Frequencies of Vb5 Expressing T Cells , 2000, Scandinavian journal of immunology.

[17]  R. Gazzinelli,et al.  Protective immunity against trypanosoma cruzi infection in a highly susceptible mouse strain after vaccination with genes encoding the amastigote surface protein-2 and trans-sialidase. , 2004, Human gene therapy.

[18]  Adeeba Kamarulzaman,et al.  AIDS Res Hum Retroviruses , 2006 .

[19]  T. Greenhalgh,et al.  Construction, characterization, and immunogenicity of a multigene modified vaccinia Ankara (MVA) vaccine based on HIV type 1 subtype C. , 2008, AIDS research and human retroviruses.

[20]  P. Earl,et al.  Broad immunogenicity of a multigene, multiclade HIV-1 DNA vaccine boosted with heterologous HIV-1 recombinant modified vaccinia virus Ankara. , 2008, The Journal of infectious diseases.

[21]  B. Caetano,et al.  Recombinant viruses as tools to induce protective cellular immunity against infectious diseases. , 2004, International microbiology : the official journal of the Spanish Society for Microbiology.

[22]  N. Garg,et al.  NADPH oxidase inhibition ameliorates Trypanosoma cruzi‐induced myocarditis during Chagas disease , 2011, The Journal of pathology.

[23]  D. Webster,et al.  Synergistic DNA-MVA prime-boost vaccination regimes for malaria and tuberculosis. , 2006, Vaccine.

[24]  Eva Acosta-Rodriguez,et al.  Polyclonal B cell activation in infections: infectious agents’ devilry or defense mechanism of the host? , 2007, Journal of Leukocyte Biology.

[25]  M. Merchlinsky,et al.  Prime-boost immunization with DNA and modified vaccinia virus ankara vectors expressing herpes simplex virus-2 glycoprotein D elicits greater specific antibody and cytokine responses than DNA vaccine alone. , 2002, The Journal of infectious diseases.

[26]  N. Garg,et al.  Delivery of antigenic candidates by a DNA/MVA heterologous approach elicits effector CD8(+)T cell mediated immunity against Trypanosoma cruzi. , 2012, Vaccine.

[27]  C. Alonso,et al.  DNA Immunization with Trypanosoma cruzi HSP70 Fused to the KMP11 Protein Elicits a Cytotoxic and Humoral Immune Response against the Antigen and Leads to Protection , 2001, Infection and Immunity.

[28]  N. Garg,et al.  Chapter 69 – American Trypanosomiasis , 2009 .

[29]  S. Williams-Blangero,et al.  Potential role of CD4+CD25HIGH regulatory T cells in morbidity in Chagas disease. , 2007, Frontiers in bioscience : a journal and virtual library.

[30]  D. Webster,et al.  Safety of recombinant fowlpox strain FP9 and modified vaccinia virus Ankara vaccines against liver-stage P. falciparum malaria in non-immune volunteers. , 2006, Vaccine.

[31]  S. Akira,et al.  Survival of lethal poxvirus infection in mice depends on TLR9, and therapeutic vaccination provides protection. , 2008, The Journal of clinical investigation.

[32]  B. Maciel,et al.  Pathogenesis of Chronic Chagas Heart Disease , 2007, Circulation.

[33]  H. Kikuta,et al.  Analysis of Mumps Vaccine Failure by Means of Avidity Testing for Mumps Virus-Specific Immunoglobulin G , 1998, Clinical Diagnostic Laboratory Immunology.

[34]  M. O. Rocha,et al.  Trypanosoma cruzi Infection Induces Differential Modulation of Costimulatory Molecules and Cytokines by Monocytes and T Cells from Patients with Indeterminate and Cardiac Chagas' Disease , 2007, Infection and Immunity.

[35]  R. Mccabe,et al.  Gamma interferon suppresses acute and chronic Trypanosoma cruzi infection in cyclosporin-treated mice , 1991, Infection and immunity.

[36]  A. McMichael,et al.  Safety and tolerability of recombinant modified vaccinia virus Ankara expressing an HIV-1 gag/multiepitope immunogen (MVA.HIVA) in HIV-1-infected persons receiving combination antiretroviral therapy. , 2007, Vaccine.

[37]  A. Barbabosa‐Pliego,et al.  Testing the Efficacy of a Multi-Component DNA-Prime/DNA-Boost Vaccine against Trypanosoma cruzi Infection in Dogs , 2011, PLoS neglected tropical diseases.

[38]  R. Gazzinelli,et al.  Perforin and Gamma Interferon Expression Are Required for CD4+ and CD8+ T-Cell-Dependent Protective Immunity against a Human Parasite, Trypanosoma cruzi, Elicited by Heterologous Plasmid DNA Prime-Recombinant Adenovirus 5 Boost Vaccination , 2009, Infection and Immunity.

[39]  S. Reed.  In vivo administration of recombinant IFN-gamma induces macrophage activation, and prevents acute disease, immune suppression, and death in experimental Trypanosoma cruzi infections. , 1988, Journal of immunology.

[40]  T. Mosmann.  Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. , 1983, Journal of immunological methods.

[41]  P. Allen,et al.  Cardiac gene expression profiling provides evidence for cytokinopathy as a molecular mechanism in Chagas' disease cardiomyopathy. , 2005, The American journal of pathology.

[42]  J. C. Vazquez-Chagoyán,et al.  Vaccine development against Trypanosoma cruzi and Chagas disease. , 2011, Advances in parasitology.

[43]  Lin Chen,et al.  Neutrophil depletion exacerbates experimental Chagas' disease in BALB / c, but protects C57BL / 6 mice through modulating the Th1 / Th2 dichotomy in different directions , 2001, European journal of immunology.

[44]  M. O. Rocha,et al.  Monocytes from Patients with Indeterminate and Cardiac Forms of Chagas' Disease Display Distinct Phenotypic and Functional Characteristics Associated with Morbidity , 2004, Infection and Immunity.

[45]  Z. Cohn,et al.  Trypanosoma cruzi: in vivo and in vitro correlation between T-cell activation and susceptibility in inbred strains of mice. , 1981, Experimental parasitology.

[46]  M. Sinha,et al.  Utility of the Trypanosoma cruzi Sequence Database for Identification of Potential Vaccine Candidates by In Silico and In Vitro Screening , 2004, Infection and Immunity.

[47]  P. Earl,et al.  Recombinant modified vaccinia virus Ankara provides durable protection against disease caused by an immunodeficiency virus as well as long-term immunity to an orthopoxvirus in a non-human primate. , 2007, Virology.

[48]  N. Garg,et al.  Previously Unrecognized Vaccine Candidates Control Trypanosoma cruzi Infection and Immunopathology in Mice , 2008, Clinical and Vaccine Immunology.

[49]  N. Garg,et al.  Gene expression analysis in mitochondria from chagasic mice: alterations in specific metabolic pathways. , 2004, The Biochemical journal.

[50]  Z. Brener,et al.  Lytic antibody titre as a means of assessing cure after treatment of Chagas disease: a 10 years follow-up study. , 1993, Transactions of the Royal Society of Tropical Medicine and Hygiene.