In vivo assessment of specific cytotoxic T lymphocyte killing

Clemente, T.; Dominguez, Mariana Ribeiro; Vieira, N.J.; Rodrigues, Maurício M.; Amarante-Mendes, Gustavo P.

doi:10.1016/j.ymeth.2013.02.007

Cited by 26 publications

(23 citation statements)

References 10 publications

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“…Moreover, OVA re-stimulation strongly induced Th1 cytokine production in lymphocytes from DLNs of mice immunized with simvastatin-OVA ( Figure 6B), as well as enhancing lymphocyte cell proliferation ( Figure 6C). An in vivo cytotoxicity analysis (Clemente et al, 2013) conducted 28 days after OVA vaccination showed that simvastatin-treated (C) GGPPS deletion efficiency and specificity in immune cells isolated from GGPPS fl/fl /CD11c-cre and CD11c-cre mouse spleen and peritoneal cavity. (D) OVA-specific IgG on day 14 from GGPPS fl/fl /CD11c-Cre and CD11c-Cre mice immunized with OVA (100 mg) (n = 5).…”

Section: Mevalonate Pathway Inhibitors Are Suitable For Cancer Vaccinmentioning

confidence: 99%

The Mevalonate Pathway Is a Druggable Target for Vaccine Adjuvant Discovery

Xia

Xie

et al. 2018

Cell

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Section: Mevalonate Pathway Inhibitors Are Suitable For Cancer Vaccinmentioning

confidence: 99%

The Mevalonate Pathway Is a Druggable Target for Vaccine Adjuvant Discovery

Xia

Xie

et al. 2018

Cell

170

141

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“…We also attempted to detect IL-2 and TNF by IC expression assays. Finally, we also used an in vivo cytotoxicity assay to estimate the presence of cytotoxic T lymphocytes in vaccinated mice (24). The results were negative in most cases independently of the timing of the analyses.…”

Section: Specificity Of the Antibodies Elicited By The Homologous (Prmentioning

confidence: 99%

Immunogenicity of a Prime-Boost Vaccine Containing the Circumsporozoite Proteins of Plasmodium vivax in Rodents

et al. 2014

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g Plasmodium vivax is the most widespread and the second most prevalent malaria-causing species in the world. Current measures used to control the transmission of this disease would benefit from the development of an efficacious vaccine. In the case of the deadly parasite P. falciparum, the recombinant RTS,S vaccine containing the circumsporozoite antigen (CSP) consistently protects 30 to 50% of human volunteers against infection and is undergoing phase III clinical trials in Africa with similar efficacy. These findings encouraged us to develop a P. vivax vaccine containing the three circulating allelic forms of P. vivax CSP. Toward this goal, we generated three recombinant bacterial proteins representing the CSP alleles, as well as a hybrid polypeptide called PvCSP-All-CSP-epitopes. This hybrid contains the conserved N and C termini of P. vivax CSP and the three variant repeat domains in tandem. We also generated simian and human recombinant replication-defective adenovirus vectors expressing PvCSP-All-CSP-epitopes. Mice immunized with the mixture of recombinant proteins in a formulation containing the adjuvant poly(I·C) developed high and long-lasting serum IgG titers comparable to those elicited by proteins emulsified in complete Freund's adjuvant. Antibody titers were similar in mice immunized with homologous (protein-protein) and heterologous (adenovirus-protein) vaccine regimens. The antibodies recognized the three allelic forms of CSP, reacted to the repeated and nonrepeated regions of CSP, and recognized sporozoites expressing the alleles VK210 and VK247. The vaccine formulations described in this work should be useful for the further development of an anti-P. vivax vaccine.

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“…In vivo cytotoxicity was performed as previously described (34) with slight modifications and optimization. Briefly, after 7 days post-infection, spleens from WT donor mice were harvested and processed for single cell suspension by tweezing each spleen between the frosted ends of two sterile glass slides.…”

Section: In Vivo Cytotoxic Assaymentioning

confidence: 99%

RIPK3 and Caspase-1/11 Are Necessary for Optimal Antigen-Specific CD8 T Cell Response Elicited by Genetically Modified Listeria monocytogenes

et al. 2020

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Efficient induction of effector and long-term protective antigen-specific CD8 + T memory response by vaccination is essential to eliminate malignant and pathogen-infected cells. Intracellular infectious bacteria, including Listeria monocytogenes, have been considered potent vectors to carry multiple therapeutic proteins and generate antigenspecific CD8 + T cell responses. Although the role of molecules involved in inflammatory cell death pathways, such as necroptosis (RIPK3-mediated) and pyroptosis (Caspase-1/11-mediated), as effectors of immune response against intracellular bacteria are relatively well understood, their contribution to the adjuvant effect of recombinant bacterial vectors in the context of antigen-specific CD8 + T cell response remained obscure. Therefore, we evaluated the impact of RIPK3 and Caspase-1/11 (Casp-1/11) individual and combined deficiencies on the modulation of antigen-specific CD8 + T cell response during vaccination of mice with ovalbumin-expressing L. monocytogenes (LM-OVA). We observed that Casp-1/11 but not RIPK3 deficiency negatively impacts the capacity of mice to clear LM-OVA. Importantly, both RIPK3 and Casp-1/11 are necessary for optimal LM-OVA-mediated antigen-specific CD8 + T cell response, as measured by in vivo antigen-specific CD8 + T cell proliferation, target cell elimination, and cytokine production. Furthermore, Casp-1/11 and Casp-1/11/RIPK3 combined deficiencies restrict the early initiation of antigen-specific CD8 + T cell memory response. Taken together, our findings demonstrate that RIPK3 and Casp-1/11 influence the quality of CD8 + T cell responses induced by recombinant L. monocytogenes vectors.

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In vivo assessment of specific cytotoxic T lymphocyte killing

Cited by 26 publications

References 10 publications

The Mevalonate Pathway Is a Druggable Target for Vaccine Adjuvant Discovery

The Mevalonate Pathway Is a Druggable Target for Vaccine Adjuvant Discovery

Immunogenicity of a Prime-Boost Vaccine Containing the Circumsporozoite Proteins of Plasmodium vivax in Rodents

RIPK3 and Caspase-1/11 Are Necessary for Optimal Antigen-Specific CD8 T Cell Response Elicited by Genetically Modified Listeria monocytogenes

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