scholarly journals A novel whole-blood stimulation assay to detect and quantify memory T-cells in COVID-19 patients

Author(s):  
William Mouton ◽  
Christelle Compagnon ◽  
Kahina Saker ◽  
Soizic Daniel ◽  
Xavier Lacoux ◽  
...  

AbstractSARS-CoV-2 specific T-cells responses are essential for virus clearance. We present a novel and simple whole-blood assay allowing the detection of interferon-gamma-producing antiviral T-cells following peptide stimulation. We show that unlike neutralizing antibodies, antiviral memory T-cells persist at least 6 months in convalescent Covid-19 individuals.

2017 ◽  
Vol 152 (5) ◽  
pp. S615
Author(s):  
Laura Cook ◽  
May Q. Wong ◽  
Rosa Garcia ◽  
Megan E. Himmel ◽  
Brian Bressler ◽  
...  

Vaccines ◽  
2020 ◽  
Vol 8 (2) ◽  
pp. 225
Author(s):  
Eleonora E. Lambert ◽  
Véronique Corbière ◽  
Jacqueline A. M. van Gaans-van den Brink ◽  
Maxime Duijst ◽  
Prashanna Balaji Venkatasubramanian ◽  
...  

To advance research and development of improved pertussis vaccines, new immunoassays are needed to qualify the outcome of Bordetella pertussis (Bp) specific CD4+ T-cell differentiation. Here, we applied a recently developed whole blood assay to evaluate Bp specific CD4+ T-cell responses. The assay is based on intracellular cytokine detection after overnight in vitro Bp antigen stimulation of diluted whole blood. We show for the first time that CD4+ T-cell memory of Th1, Th2, and Th17 lineages can be identified simultaneously in whole blood. Participants ranging from 7 to 70 years of age with different priming backgrounds of whole-cell pertussis (wP) and acellular pertussis (aP) vaccination were analyzed around an acellular booster vaccination. The assay allowed detection of low frequent antigen-specific CD4+ T-cells and revealed significantly elevated numbers of activated and cytokine-producing CD4+ T-cells, with a significant tendency to segregate recall responses based on primary vaccination background. A stronger Th2 response hallmarked an aP primed cohort compared to a wP primed cohort. In conclusion, analysis of Bp specific CD4+ T-cell responses in whole blood showed separation based on vaccination background and provides a promising tool to assess the quantity and quality of CD4+ T-cell responses induced by vaccine candidates.


2020 ◽  
Author(s):  
Catherine Riou ◽  
Georgia Schäfer ◽  
Elsa du Bruyn ◽  
Rene T. Goliath ◽  
Cari Stek ◽  
...  

ABSTRACTRapid tests to evaluate SARS-CoV-2-specific T cell responses are urgently needed to decipher protective immunity and aid monitoring vaccine-induced immunity. Using a rapid whole blood assay requiring minimal amount of blood, we measured qualitatively and quantitatively SARS-CoV-2-specific CD4 T cell responses in 31 healthcare workers, using flow cytometry. 100% of COVID-19 convalescent participants displayed a detectable SARS-CoV-2-specific CD4 T cell response. SARS-CoV-2-responding cells were also detected in 40.9% of participants with no COVID-19-associated symptoms or who tested PCR negative. Phenotypic assessment indicated that, in COVID-19 convalescent participants, SARS-CoV-2 CD4 responses displayed an early differentiated memory phenotype with limited capacity to produce IFNγ. Conversely, in participants with no reported symptoms, SARS-CoV-2 CD4 responses were enriched in late differentiated cells, co-expressing IFNγ and TNFα and also Granzyme B. This proof of concept study presents a scalable alternative to PBMC-based assays to enumerate and phenotype SARS-CoV-2-responding T cells, thus representing a practical tool to monitor adaptive immunity in vaccine trials.SummaryIn this proof of concept study, we show that SARS-CoV-2 T cell responses are easily detectable using a rapid whole blood assay requiring minimal blood volume. Such assay could represent a suitable tool to monitor adaptive immunity in vaccine trials.


1983 ◽  
Vol 50 (04) ◽  
pp. 814-820 ◽  
Author(s):  
J A Bergeron ◽  
J M DiNovo ◽  
A F Razzano ◽  
W J Dodds

SummaryThe previously described native whole blood assay for materials in solution or suspension has been adapted to materials in a bead column configuration. These experiments showed that the glass itself accounts for little or none of the high blood-reactivity observed with conventional glass bead columns. Columns composed solely of soft glass that was “cleaned” by heat treatment (500-595° C 18 hr, electric oven) were benign toward flowing native whole blood for all variables measured (platelet count and platelet-free plasma [C14]-serotonin content, platelet factor 3 and factor XII activities, and recalcification time) with the standard contact protocol. In addition, the effluent successfully maintained perfusion of the isolated kidney, a measure of the ability of platelets to support vascular integrity. Prolonged (30 min) normothermic contact with titrated whole blood increased the subsequent reactivity of initially clean glass toward whole blood albeit to a level much less than that of conventional glass bead columns.


2021 ◽  
pp. 67-87
Author(s):  
Klaus Görlinger ◽  
James Iqbal ◽  
Daniel Dirkmann ◽  
Kenichi A. Tanaka

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