ELISPOT Assay for Monitoring Cytotoxic T Lymphocytes (CTL) Activity in Cancer Vaccine Clinical Trials

Malyguine, Anatoli; Strobl, Susan; Dunham, Kimberly; Shurin, Michael R.; Sayers, Thomas J.

doi:10.3390/cells1020111

Cited by 56 publications

(33 citation statements)

References 57 publications

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“…CTLs can be key effector cells in the immunomediated destruction of virally infected cells. Therefore, we measured both the frequency and function of antigen‐specific CTLs with an IFN‐ γ ELISPOT assay by stimulating PBMCs with 1 mg ml −1 293 cell lysate prepared from cells transduced with the AdhAQP1 vector (Malyguine et al , ). There were no statistically significant differences in the mean number of activated PBMCs between grouped responders and non‐responders in any of the time points measured (Figures and ).…”

Section: Resultsmentioning

confidence: 99%

Immune reactivity after adenoviral‐mediated aquaporin‐1 cDNA transfer to human parotid glands

et al. 2017

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OBJECTIVES The purpose of this study was to examine the humoral and cellular immune reactivity to adenoviral vector (AdhAQP1) administration in the human parotid gland over the first 42 days of a clinical gene therapy trial. METHODS Of eleven treated subjects, five were considered as positive responders (Baum et al, 2012). Herein, we measured serum neutralizing antibody titers, circulating cytotoxic lymphocytes, and lymphocyte proliferation in peripheral blood mononuclear cells. Additionally, after adenoviral vector stimulation of lymphocyte proliferation, we quantified secreted cytokine levels. RESULTS Responders showed little to modest immune reactivity during the first 42 days following gene transfer. Additionally, baseline serum neutralizing antibody titers to serotype 5-adenovirus generally were not predictive of a subject’s response to parotid gland administration of AdhAQP1. Cytokine profiling from activated peripheral blood mononuclear cells could not distinguish responders and non-responders. CONCLUSIONS The data are the first to describe immune responses after adenoviral vector administration in a human parotid gland. Importantly, we found that modest (2–3 fold) changes in systemic cell-mediated immune reactivity did not preclude positive subject responses to gene transfer. However, changes beyond that level likely impeded the efficacy of gene transfer.

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Section: Resultsmentioning

confidence: 99%

Immune reactivity after adenoviral‐mediated aquaporin‐1 cDNA transfer to human parotid glands

et al. 2017

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“…Enzyme-linked immunospot (ELISpot) assay, relying on a colorimetric enzymatic reaction of antibodies, is one of the most extensively used methods for measuring the cellular secretory activities in vitro 4 . In particular, secretory activities of individual cells may correlate with pathological developments and may provide important information for disease diagnosis.…”

Section: Introductionmentioning

confidence: 99%

Microfluidic compartments with sensing microbeads for dynamic monitoring of cytokine and exosome release from single cells

Son

Rahimian

Shin

et al. 2016

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Monitoring activity of single cells has high significance for basic science and diagnostic applications. Here we describe a reconfigurable microfluidic device for confining single cells along with antibody-modified sensing beads inside 20 picoliter (pL) microcompartments for monitoring cellular secretory activity. An array of ∼7000 microchambers fabricated in the roof of the reconfigurable microfluidic device could be raised or lowered by applying negative pressure. The floor of the device was micropatterned to contain cell attachment sites in registration with the microcompartments. Using this set-up, we demonstrated the detection of inflammatory cytokine IFN-γ and exosomes from single immune cells and cancer cells respectively. The detection scheme was similar in both cases: cells were first captured on the surface inside the microfluidic device, then sensing microbeads were introduced into the device so that, once the microcompartments were lowered, single cells and microbeads became confined together. The liquid bathing the beads and the cells inside the compartments also contained fluorescently-labeled secondary antibodies (Abs). The capture of cell-secreted molecules onto microbeads was followed by binding of secondary antibodies - this caused microbeads to become fluorescent. The fluorescence intensity of the microbeads changed over time, providing dynamics of single cell secretory activity. The microdevice described here may be particularly useful in the cases where panning upstream of sensing is required or to analyze secretory activity of anchorage-dependent cells.

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“…For example, although ELISPOT is commonly used to evaluate cytotoxicity in vaccine trials [33], it simply measures cytokine production, and usually only IFN-γ, being an indirect marker of CTL activity at best. Future DC trials may benefit from the use of multiparameter flow cytometry to obtain a more accurate panel of CTL- specific response [34].…”

Section: Discussionmentioning

confidence: 99%

Dendritic Cell-Based Immunotherapies to Fight HIV: How Far from a Success Story? A Systematic Review and Meta-Analysis

Coelho

Moura

Kamada

et al. 2016

IJMS

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The scientific community still faces the challenge of developing strategies to cure HIV-1. One of these pursued strategies is the development of immunotherapeutic vaccines based on dendritic cells (DCs), pulsed with the virus, that aim to boost HIV-1 specific immune response. We aimed to review DCs-based therapeutic vaccines reports and critically assess evidence to gain insights for the improvement of these strategies. We performed a systematic review, followed by meta-analysis and meta-regression, of clinical trial reports. Twelve studies were selected for meta-analysis. The experimental vaccines had low efficiency, with an overall success rate around 38% (95% confidence interval = 26.7%–51.3%). Protocols differed according to antigen choice, DC culture method, and doses, although multivariate analysis did not show an influence of any of them on overall success rate. The DC-based vaccines elicited at least some immunogenicity, that was sometimes associated with plasmatic viral load transient control. The protocols included both naïve and antiretroviral therapy (ART)-experienced individuals, and used different criteria for assessing vaccine efficacy. Although the vaccines did not work as expected, they are proof of concept that immune responses can be boosted against HIV-1. Protocol standardization and use of auxiliary approaches, such as latent HIV-1 reservoir activation and patient genomics are paramount for fine-tuning future HIV-1 cure strategies.

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ELISPOT Assay for Monitoring Cytotoxic T Lymphocytes (CTL) Activity in Cancer Vaccine Clinical Trials

Cited by 56 publications

References 57 publications

Immune reactivity after adenoviral‐mediated aquaporin‐1 cDNA transfer to human parotid glands

Immune reactivity after adenoviral‐mediated aquaporin‐1 cDNA transfer to human parotid glands

Microfluidic compartments with sensing microbeads for dynamic monitoring of cytokine and exosome release from single cells

Dendritic Cell-Based Immunotherapies to Fight HIV: How Far from a Success Story? A Systematic Review and Meta-Analysis

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