Quantification of antigen specific CD8+ T cells using an ELISPOT assay

Miyahira, Yasushi; Murata, Kenichiro; Rodrı́guez, Dolores; Rodríguez, Juan; Esteban, Mariano; Rodrigues, Maurício M.; Zavala, Fidel

doi:10.1016/0022-1759(94)00327-s

Cited by 331 publications

(203 citation statements)

References 10 publications

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“…IFN-γ and IL-4 ELISPOT assays were performed using splenocytes from immunized mice as described [35][36][37][38]. Briefly, 96 wells multiscreen filter plates (Milipore Inc., Bedford, MA) were coated with monoclonal antibodies to mouse IFN-γ (R4-6A2, 10ug/ml) and IL-4 (11B11, 5 ug/ l) (BD Biosciences, San Diego, CA) overnight at room temp.…”

Section: Antigen Stimulated Elispot Assaysmentioning

confidence: 99%

Interleukin-6 has differential influence on the ability of adjuvant formulations to potentiate antibody reponses to a Plasmodium falciparum blood-stage vaccine

Hui

Hashimoto

2007

Vaccine

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The efficacy of vaccine adjuvants can be influenced by the immunological environment of the host, depending on the mechanism(s) by which they exert their immunopotentiating activities. Interleukin-6 is a pleiotropic cytokine that has a broad range of biological activities on immune and non-immune cells. We investigated the role of IL-6 on the ability of nine adjuvant formulations to induce antibody responses to the P. falciparum MSP1-19 malaria vaccine, using IL-6 -/-(KO) mice. Results showed that some adjuvants, ie. MPL-SE, CFA/IFA, ISA720/QS21/MPL, depended on IL-6 for their efficacy, while others exhibited increased potency in its absence. The efficacy of adjuvants in the IL-6 KO environment cannot be solely attributed to their ability to stimulate antigen-specific cellular responses, suggesting that other biological activities of IL-6 are also important. The results further suggest that two adjuvants utilized dissimilar pathways to potentiate the same type of immune response.

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Section: Antigen Stimulated Elispot Assaysmentioning

confidence: 99%

Interleukin-6 has differential influence on the ability of adjuvant formulations to potentiate antibody reponses to a Plasmodium falciparum blood-stage vaccine

Hui

Hashimoto

2007

Vaccine

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“…31,32 Spleen cells from mice that 14 days earlier had received their second or third immunization were used in these determinations. These measurements were made 36 h after being incubated at 371C and 5% CO 2 with target cells infected with recombinant poxvirus expressing P. falciparum proteins or incubated with peptides.…”

Section: Ifnc Elispotmentioning

confidence: 99%

Effect on antibody and T-cell responses of mixing five GMP-produced DNA plasmids and administration with plasmid expressing GM-CSF

et al. 2004

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One potential benefit of DNA vaccines is the capacity to elicit antibody and T-cell responses against multiple antigens at the same time by mixing plasmids expressing different proteins. A possible negative effect of such mixing is interference among plasmids regarding immunogenicity. In preparation for a clinical trial, we assessed the immunogenicity of GMP-produced plasmids encoding five Plasmodium falciparum proteins, PfCSP, PfSSP2, PfEXP1, PfLSA1, and PfLSA3, given as a mixture, or alone. The mixture induced higher levels of antibodies against whole parasites than did the individual plasmids, but was associated with a decrease in antibodies to individual P. falciparum proteins. T-cell responses were in general decreased by administration of the mixture. Immune responses to individual plasmids and mixtures were generally higher in inbred mice than in outbreds. In inbred BALB/c and C57BL/6 mice, coadministration of a plasmid expressing murine granulocyte-macrophage colony-stimulating factor (mGM-CSF), increased antibody and T-cell responses, but in outbred CD-1 mice, coadministration of mGM-CSF was associated with a decrease in antibody responses. Such variability in data from studies in different strains of mice underscores the importance of genetic background on immune response and carefully considering the goals of any preclinical studies of vaccine mixtures planned for human trials.

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“…33 Briefly, mouse spleen cells from each group were harvested at day 14 after vaccination. The splenocytes were cultured for 6 days in the presence of 1 g/ml E7-specific H-2D b CTL epitope (E7 49-57, RAHYNIVTF) 40 ELISPOT assay The ELISPOT assay described by Miyahira et al 41 and Murali-Krishna et al 13 was modified to detect HPV-16 E7-specific CD8 + T cells. The ELISPOT assay was used to determine the number of antigen-specific T cell precursors based on the secretion of IFN-␥ by individual activated T cells.…”

Section: Elisamentioning

confidence: 99%

Gene gun-mediated DNA vaccination induces antitumor immunity against human papillomavirus type 16 E7-expressing murine tumor metastases in the liver and lungs

Chen

Suh

et al. 1999

Gene Ther

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DNA vaccination has emerged as an attractive approach assess the E7-specific T cell-mediated and humoral for tumor immunotherapy. The aim of this study was to immunity. Mice vaccinated with Sig/E7/LAMP-1 DNA genevaluate the potency of DNA vaccines in preventing and erated the strongest E7-specific CTL activities, the highest treating the liver and lung metastases of a human papilnumbers of E7-specific CD8 + cell precursors and the highlomavirus-16 (HPV-16) E7-expressing murine tumor (TCest titers of E7-specific antibodies. While both E7 DNA and 1). We used the gene gun method to vaccinate C57BL/6Sig/E7/LAMP-1 DNA generated potent antitumor immunity mice intradermally with DNA vaccines containing the HPVin the liver and lung metastases models, the Sig/E7/LAMP-16 E7 gene, the E7 gene linked to the sorting signals of 1 DNA was more potent under stringent conditions. DNA the lysosome-associated membrane protein-1 (Sig/E7/ vaccination with E7-expressing plasmids was effective in LAMP-1), or the 'empty' plasmid vector. The in vivo anticontrolling liver and lung metastases of an E7-expressing tumor immunity was analyzed in both tumor prevention and murine tumor. Our data suggest that antigen-specific DNA tumor regression experiments. In addition, cytotoxic T lymvaccination can potentially be applied to control liver and phocyte (CTL) assays, enzyme-linked immunospot assay lung metastases of tumors with defined tumor-specific and enzyme-linked immunoabsorbent assay were used to antigens.

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Quantification of antigen specific CD8+ T cells using an ELISPOT assay

Cited by 331 publications

References 10 publications

Interleukin-6 has differential influence on the ability of adjuvant formulations to potentiate antibody reponses to a Plasmodium falciparum blood-stage vaccine

Interleukin-6 has differential influence on the ability of adjuvant formulations to potentiate antibody reponses to a Plasmodium falciparum blood-stage vaccine

Effect on antibody and T-cell responses of mixing five GMP-produced DNA plasmids and administration with plasmid expressing GM-CSF

Gene gun-mediated DNA vaccination induces antitumor immunity against human papillomavirus type 16 E7-expressing murine tumor metastases in the liver and lungs

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