Response definition criteria for ELISPOT assays revisited

Moodie, Zoe; Price, Leah; Gouttefangeas, Cécile; Mander, Ann; Janetzki, Sylvia; Löwer, Martin; Welters, Marij J.P.; Ottensmeier, Christian; Burg, Sjoerd H. van der; Britten, Cedrik M.

doi:10.1007/s00262-010-0875-4

Cited by 199 publications

(172 citation statements)

References 22 publications

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“…For multiple group comparisons, nonparametric Kruskal-Wallis and Dunn multiple-comparison tests were used. For ELISpot data, statistical significance was evaluated with the DFR(eq) script (20). Differences were considered statistically significant when P < 0.05.…”

Section: Discussionmentioning

confidence: 99%

Intravenous Injection of MVA Virus Targets CD8+ Lymphocytes to Tumors to Control Tumor Growth upon Combinatorial Treatment with a TLR9 Agonist

Fend

Gatard-Scheikl

Kintz

et al. 2014

Cancer Immunology Research

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Effector T-cell access to tumor tissue is a limiting step for clinical efficacy of antigen-specific T cell-based immunotherapies. Ectopic mouse tumor models, in which a subcutaneously (s.c.) implanted tumor is treated with s.c. or intramuscular therapeutic immunization, may not be optimal for targeting effector T cells to an organ-borne tumor. We used an orthotopic renal carcinoma model to evaluate the impact of injection routes on therapeutic efficacy of a Modified Vaccinia virus Ankara viral vector expressing the human mucin 1 tumor-associated xeno-antigen (MVA-MUC1). We show that intravenous (i.v.) administration of MVA-MUC1 displayed enhanced efficacy when compared with s.c. injection. Therapeutic efficacy of MVA-MUC1 was further enhanced by i.v. injection of a TLR9 agonist. In all cases, infiltration of tumor-bearing kidney by CD8

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Section: Discussionmentioning

confidence: 99%

Intravenous Injection of MVA Virus Targets CD8+ Lymphocytes to Tumors to Control Tumor Growth upon Combinatorial Treatment with a TLR9 Agonist

Fend

Gatard-Scheikl

Kintz

et al. 2014

Cancer Immunology Research

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“…We defined responses using empirical response methods: positive wells were required to contain a minimum of 10 spots/2 Â 10 5 cells and have at least 3-fold more spots than negative control wells. These criteria have been shown to yield very low false positive rates (36).…”

Section: Ifn-g Elispotmentioning

confidence: 99%

Surveillance of the Tumor Mutanome by T Cells during Progression from Primary to Recurrent Ovarian Cancer

Wick

Webb

Nielsen

et al. 2014

Clinical Cancer Research

144

125

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Purpose: Cancers accumulate mutations over time, each of which brings the potential for recognition by the immune system. We evaluated T-cell recognition of the tumor mutanome in patients with ovarian cancer undergoing standard treatment.Experimental Design: Tumor-associated T cells from 3 patients with ovarian cancer were assessed by ELISPOT for recognition of nonsynonymous mutations identified by whole exome sequencing of autologous tumor. The relative levels of mutations and responding T cells were monitored in serial tumor samples collected at primary surgery and first and second recurrence.Results: The vast majority of mutations (78/79) were not recognized by tumor-associated T cells; however, a highly specific CD8 þ T-cell response to the mutation hydroxysteroid dehydrogenase-like protein 1 (HSDL1) L25V was detected in one patient. In the primary tumor, the HSDL1 L25V mutation had low prevalence and expression, and a corresponding T-cell response was undetectable. At first recurrence, there was a striking increase in the abundance of the mutation and corresponding MHC class I epitope, and this was accompanied by the emergence of the HSDL1 L25V -specific CD8 þ T-cell response. At second recurrence, the HSDL1 L25V mutation and epitope continued to be expressed; however, the corresponding T-cell response was no longer detectable. Conclusion:The immune system can respond to the evolving ovarian cancer genome. However, the T-cell response detected here was rare, was transient, and ultimately failed to prevent disease progression. These findings reveal the limitations of spontaneous tumor immunity in the setting of standard treatments and suggest a high degree of ignorance of tumor mutations that could potentially be reversed by immunotherapy.

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“…The spots were counted using an ELISpot 4.0 reader (AID Diagnostika GmbH, San Diego, CA). The positive responses to each antigen were measured by an empirical rule (ER) which defines a positive response as an at least 2-fold increase of spot number in the experimental wells over the background with a minimum threshold of 5 spots per 100,000 PBMCs in the experimental wells (24). An M. tuberculosis ESAT-6-specific positive response was defined as spots appearing in M. tuberculosis ESAT-6/CFP-10 and either M. tuberculosis ESAT-6 1-57 or M. tuberculosis ESAT-6 55-95 stimulation alongside the positive response to anti-CD3 antibody.…”

Section: Ifn-␥ Elispot Assaymentioning

confidence: 99%

Identification of Immunological Biomarkers Which May Differentiate Latent Tuberculosis from Exposure to Environmental Nontuberculous Mycobacteria in Children

Hur

Crampin

Chisambo

et al. 2014

Clin Vaccine Immunol

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Response definition criteria for ELISPOT assays revisited

Cited by 199 publications

References 22 publications

Intravenous Injection of MVA Virus Targets CD8+ Lymphocytes to Tumors to Control Tumor Growth upon Combinatorial Treatment with a TLR9 Agonist

Intravenous Injection of MVA Virus Targets CD8+ Lymphocytes to Tumors to Control Tumor Growth upon Combinatorial Treatment with a TLR9 Agonist

Surveillance of the Tumor Mutanome by T Cells during Progression from Primary to Recurrent Ovarian Cancer

Identification of Immunological Biomarkers Which May Differentiate Latent Tuberculosis from Exposure to Environmental Nontuberculous Mycobacteria in Children

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