Granzyme B ELISPOT assay for ex vivo measurements of T cell immunity

Rininsland, Frauke; Helms, Thomas; Asaad, Robert; Boehm, Bernhard O.; Tary‐Lehmann, Magdalena

doi:10.1016/s0022-1759(00)00191-5

Cited by 98 publications

(66 citation statements)

References 21 publications

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“…In the present study we took advantage of this observation to determine the frequency of CTL effectors using a novel technique that measures the number of granzyme B secreting cells by Elispot [27]. We observed mean increases of 145 ± 56 SFC/10 6 PBMC (range: 83-256) in subjects immunized CVD 909.…”

Section: Discussionmentioning

confidence: 87%

“…Because the granule-dependent pathway is largely responsible for the killing of S. Typhiinfected targets by specific CTLs, we determine the magnitude of the specific cytotoxic response induced by CVD 909 by measuring the ex vivo frequency of CTL effectors by using granzyme-B Elispot assay [27]. Following infection with S. Typhi, autologous blasts were cultured with ex vivo PBMC and the frequency of S. Typhi specific T cells measured by their release of Granzyme-B using a commercially available Elispot assay.…”

Section: Granzyme-b Productionmentioning

confidence: 99%

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Cell-mediated immune responses in humans after immunization with one or two doses of oral live attenuated typhoid vaccine CVD 909

Wahid

Salerno-Gonçalves

Tacket

et al. 2007

Vaccine

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CVD 909 is a novel live attenuated S. Typhi oral vaccine candidate derived from strain CVD 908-htrA which constitutively expresses Vi. Herein we investigated whether the genetic manipulations involved in modifying CVD 908-htrA altered its ability to induce potent T-cell immune responses (CMI) after a single dose (5 subjects) and, in a separate trial, whether a second dose (8 subjects) further enhanced its immunogenicity. In these clinical trials we observed that CVD 909 immunization elicits a wide array of CMI, including cytotoxic T cells (CTL), IFN-γ, TNF-α and IL-10 (but not IL-2, IL-4 or IL-5) production, and proliferation to S. Typhi antigens. However, the administration of a second dose did not result in increases in CMI. These results suggest that the genetic manipulations to constitutively express Vi did not adversely affect the ability of CVD 909 to elicit a wide array of CMI responses. These observations add impetus for the continuing evaluation of CVD 909 as a typhoid vaccine candidate.

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Section: Discussionmentioning

confidence: 87%

Section: Granzyme-b Productionmentioning

confidence: 99%

Cell-mediated immune responses in humans after immunization with one or two doses of oral live attenuated typhoid vaccine CVD 909

Wahid

Salerno-Gonçalves

Tacket

et al. 2007

Vaccine

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show abstract

“…In this degranulation process only a fraction of the GzB stored per cell is secreted (enabling the CD8 + cells to be "serial killers") [22] which makes it challenging to detect the loss of GzB through degranulation by flow cytometry. In ELISPOT assays, on the contrary, the actually released GzB can be measured [25].…”

Section: Introductionmentioning

confidence: 99%

Granzyme B production distinguishes recently activated CD8+ memory cells from resting memory cells

Nowacki

Küerten

Zhang³

et al. 2007

Cellular Immunology

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For immune diagnostic purposes it would be critical to be able to distinguish between ongoing immune processes, such as active infections, and long-term immune memory, for example imprinted by infections that have been cleared a long time ago or by vaccinations. We tested the hypothesis that the secretion of Granzyme B, as detected in ex vivo ELISPOT assays, permits this distinction. We studied EBV-, flu-and CMV-specific CD8 + cells in healthy individuals, Vaccinia virus-reactive CD8 + cells in the course of vaccination, and HIV-specific CD8 + cells in HIV-infected individuals. Antigen-specific ex vivo GzB production was detected only transiently after Vaccinia immunization, and in HIV-infected individuals. Our data suggest that ex vivo ELISPOT measurements of granzyme B permit the identification of actively ongoing CD8 + cell responses -a notion that is pertinent to the immune diagnostic of infections, transplantation, allergies, autoimmune diseases, tumors, and vaccine development.

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“…14 With the introduction of a test for granzyme B-production by mononuclear blood cells it is now possible to study virus-specific cytotoxicity in response to vaccinations. [15][16][17] It appears that cellular immunity against influenza virus is less strain-specific than are serum antibodies, and that the ensuing cross-reactivity may then render vaccines active against different microbial strains. 18,19 In this study, we intended to compare the immunogenic effects of a simple non-living vaccine formulation consisting of beta-propiolactone-inactivated influenza virus in saline, with the effects of formulations of virus also containing B. pertussis, or a thixotropic substance, after being given intranasally.…”

Section: Introductionmentioning

confidence: 99%

A Non-Living Nasal Influenza Vaccine Can Induce Major Humoral and Cellular Immune Responses in Humans without the Need for Adjuvants

Samdal¹,

Bakke²,

Oftung³

et al. 2005

Human Vaccines

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Twenty-eight healthy adult volunteers were immunized intranasally with an inactivated whole-virus influenza vaccine based on the strain A/New Caledonia/20/99 (H1N1), either in saline or mixed with formaldehyde-inactivated Bordetella pertussis as a mucosal adjuvant, or in a thixotropic vehicle with mucoadhesive properties. After four doses, all groups of vaccinees developed significant IgG-and IgA-antibody responses, measured by ELISA, in respectively serum and nasal secretions. None of the volunteers had demonstrable hemagglutination inhibition (HAI) antibodies in serum before being immunized, whereas more than 80% of them reached HAI titers > 40, considered protective, after immunizations. In addition, cellular immune responses, measured as significant increases in CD4 + T-cell proliferation and granzyme B-producing cytotoxic T-cells, were detected against the vaccine strain as well as against heterologous virus strains (H3N2). However, no additive effect on these responses could be demonstrated with use of B. pertussis or the thixotropic substance in the present vaccines. It appeared, actually, that the mucoadhesive vehicle containing the thixotropic substance was less efficient than were the two other formulations. An influenza vaccine made as a simple particulate formulation of inactivated virus, and given repeatedly onto the nasal mucosa, may thus be an attractive alternative to currently available vaccines.

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Granzyme B ELISPOT assay for ex vivo measurements of T cell immunity

Cited by 98 publications

References 21 publications

Cell-mediated immune responses in humans after immunization with one or two doses of oral live attenuated typhoid vaccine CVD 909

Cell-mediated immune responses in humans after immunization with one or two doses of oral live attenuated typhoid vaccine CVD 909

Granzyme B production distinguishes recently activated CD8+ memory cells from resting memory cells

A Non-Living Nasal Influenza Vaccine Can Induce Major Humoral and Cellular Immune Responses in Humans without the Need for Adjuvants

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