Induction of B- and T-cell responses to cruzipain in the murine model of Trypanosoma cruzi infection

Schnapp, Anita R.; Eickhoff, Christopher S.; Scharfstein, Júlio; Hoft, Daniel F.

doi:10.1016/s1286-4579(02)01600-3

Cited by 22 publications

(14 citation statements)

References 26 publications

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“…CTL assays were performed using a modified version of a previously reported method (35). Effector cell populations were prepared by culturing 3 ϫ 10 7 spleen cells with 1 ϫ 10 6 irradiated (6,000 rad) A20J-TS cells in upright T25 flasks in a total volume of 10 ml.…”

Section: In Vitro Ctl Assaysmentioning

confidence: 99%

Trans-Sialidase Recombinant Protein Mixed with CpG Motif-Containing Oligodeoxynucleotide Induces Protective Mucosal and Systemic Trypanosoma cruzi Immunity Involving CD8+ CTL and B Cell-Mediated Cross-Priming

Hoft

Eickhoff

Giddings

et al. 2007

The Journal of Immunology

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The Trypanosoma cruzi trans-sialidase (TS) is a unique enzyme with neuraminidase and sialic acid transfer activities important for parasite infectivity. The T. cruzi genome contains a large family of TS homologous genes, and it has been suggested that TS homologues provide a mechanism of immune escape important for chronic infection. We have investigated whether the consensus TS enzymatic domain could induce immunity protective against acute and chronic, as well as mucosal and systemic, T. cruzi infection. We have shown that: 1) TS-specific immunity can protect against acute T. cruzi infection; 2) effective TS-specific immunity is maintained during chronic T. cruzi infection despite the expression of numerous related TS superfamily genes encoding altered peptide ligands that in theory could promote immune tolerization; and 3) the practical intranasal delivery of recombinant TS protein combined with a ssDNA oligodeoxynucleotide (ODN) adjuvant containing unmethylated CpG motifs can induce both mucosal and systemic protective immunity. We have further demonstrated that the intranasal delivery of soluble TS recombinant Ag combined with CpG ODN induces both TS-specific CD4+ and CD8+ T cells associated with vaccine-induced protective immunity. In addition, optimal protection induced by intranasal TS Ag combined with CpG ODN requires B cells, which, after treatment with CpG ODN, have the ability to induce TS-specific CD8+ T cell cross-priming. Our results support the development of TS vaccines for human use, suggest surrogate markers for use in future human vaccine trials, and mechanistically identify B cells as important APC targets for vaccines designed to induce CD8+ CTL responses.

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Section: In Vitro Ctl Assaysmentioning

confidence: 99%

Trans-Sialidase Recombinant Protein Mixed with CpG Motif-Containing Oligodeoxynucleotide Induces Protective Mucosal and Systemic Trypanosoma cruzi Immunity Involving CD8+ CTL and B Cell-Mediated Cross-Priming

Hoft

Eickhoff

Giddings

et al. 2007

The Journal of Immunology

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“…Previous studies have documented the usefulness of Cz-based vaccines in eliciting protective immunity to T. cruzi (17,44,45). On the other hand, Salmonella is an ideal delivery system because is easy to handle and to manipulate genetically.…”

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confidence: 99%

Oral Vaccination withSalmonella entericaas a Cruzipain-DNA Delivery System Confers Protective Immunity againstTrypanosoma cruzi

et al. 2008

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To stimulate both local and systemic immune responses against Trypanosoma cruzi, Salmonella enterica serovar Typhimurium aroA was exploited as a DNA delivery system for cruzipain (SCz). In a murine model we compared SCz alone (GI) or coadministered with Salmonella carrying a plasmid encoding granulocyte-macrophage colony-stimulating factor (GII), as well as protocols in which SCz priming was followed by boosting with recombinant cruzipain (rCz) admixed with either CpG-ODN (GIII) or MALP-2, a synthetic derivative of a macrophage-activating lipopeptide of 2 kDa from Mycoplasma fermentans (GIV). The results showed that protocols that included four oral doses of SCz (GI) elicited mainly a mucosal response characterized by immunoglobulin A (IgA) secretion and proliferation of gut-associated lymphoid tissue cells, with weak systemic responses. In contrast, the protocol that included a boost with rCz plus CpG (GIII) triggered stronger systemic responses in terms of Cz-specific serum IgG titers, splenocyte proliferation, gamma interferon (IFN-␥) secretion, and delayed-type hypersensitivity response. Trypomastigote challenge of vaccinated mice resulted in significantly lower levels of parasitemia compared to controls. Protection was abolished by depletion of either CD4 ؉ or CD8 ؉ T cells. Parasite control was also evident from the reduction of tissue damage, as revealed by histopathologic studies and serum levels of enzymes that are markers of muscle injury in chronic Chagas' disease (i.e., creatine kinase, aspartate aminotransferase, and lactate dehydrogenase). Enhanced release of IFN-␥ and interleukin-2 was observed in GI and GII upon restimulation of splenocytes in the nonparasitic phase of infection. Our results indicate that Salmonella-mediated delivery of Cz-DNA by itself promotes the elicitation of an immune response that controls T. cruzi infection, thereby reducing parasite loads and subsequent damage to muscle tissues.

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“…Antibody-mediated protection has been reported with P. falciparum SERA/SERP H antigens in Aotus monkeys and mice (11,45), with Leishmania major cysteine protease in mice (49), with Entamoeba histolytica cysteine protease antigens in vitro (23), and also with Schistosoma mansoni calpains (12). Possibilities for anti-cysteine protease vaccines against parasitic organisms has been reported for Fasciola hepatica and Haemonchus contorius infections (6,42,43), as well as for Trypanosoma cruzi (34,35). Protection of mice against gastric colonization by Helicobacter pylori by immunization with urease subunits A and B has also demonstrated vaccine potential of proteases (5).…”

Section: Discussionmentioning

confidence: 99%

Falcipain-1, aPlasmodium falciparumCysteine Protease with Vaccine Potential

Kumar

Korde

et al. 2007

Infect Immun

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Cysteine proteases (falcipains) of Plasmodium falciparum are potential targets for antimalarial chemotherapy, since they have been shown to be involved in important cellular functions such as hemoglobin degradation and invasion/rupture of red blood cells during parasite life cycle. The role of falcipain-1 at the asexual blood stages of the parasite still remains uncertain. This is mainly due to a lack of methods to prepare this protein in an active form. In order to obtain biologically active falcipain-1, a number of falcipain-1 constructs were designed and a systematic assessment of the refolding conditions was done. We describe here the expression, purification, and characterization of a falcipain-1 construct encoding mature falcipain-1 and 35 amino acids from the C-terminal of the pro domain. Recombinant falcipain-1 was overexpressed in the form of inclusion bodies, solubilized, and purified by Ni 2؉ -nitrilotriacetic acid affinity chromatography under denaturing conditions. A systemic approach was then followed to optimize refolding parameters. An optimum refolding condition was obtained, and the yield of the purified refolded falcipain-1 was ϳ1 mg/liter. Activity of the protein was analyzed by fluorometric and gelatin degradation assays. Immunolocalization studies using anti-falcipain-1 sera revealed a distinct staining at the apical end of the P. falciparum merozoites. Previous studies using falcipain-1-specific inhibitors have suggested a role of falcipain-1 in merozoite invasion. Based on its localization and its role in invasion, we analyzed the immunogenicity of falcipain-1 in mice, followed by heterologous challenge with Plasmodium yoelii sporozoites. Our results suggest a possible role of falcipain-1 in merozoite invasion.

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Induction of B- and T-cell responses to cruzipain in the murine model of Trypanosoma cruzi infection

Cited by 22 publications

References 26 publications

Trans-Sialidase Recombinant Protein Mixed with CpG Motif-Containing Oligodeoxynucleotide Induces Protective Mucosal and Systemic Trypanosoma cruzi Immunity Involving CD8+ CTL and B Cell-Mediated Cross-Priming

Trans-Sialidase Recombinant Protein Mixed with CpG Motif-Containing Oligodeoxynucleotide Induces Protective Mucosal and Systemic Trypanosoma cruzi Immunity Involving CD8+ CTL and B Cell-Mediated Cross-Priming

Oral Vaccination withSalmonella entericaas a Cruzipain-DNA Delivery System Confers Protective Immunity againstTrypanosoma cruzi

Falcipain-1, aPlasmodium falciparumCysteine Protease with Vaccine Potential

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