Optimisation of the cultured ELISpot/Fluorospot technique for the selective investigation of SARS-CoV-2 reactive central memory T cells DOI Creative Commons

Jack R. Jerome,

Kirsty Wilson, Joshuah Fialho

и другие.

Frontiers in Immunology, Год журнала: 2025, Номер 16

Опубликована: Апрель 15, 2025

Introduction This study presents an optimised cultured ELISpot protocol for detecting central memory T-cell interferon gamma (IFNγ) responses against SARS-CoV-2 peptides following initial priming with either peptides, or whole spike protein. Methods Key variations include the culture length, timing of exogenous survival signals (IL-2), and endpoint analysis modality cell density to enhance assay sensitivity without compromising specificity IFNγ recall cognate antigen. Results We noted a duration 10 days, combined delayed IL-2 administration on day 5 while maintaining response towards antigen when compared shorter periods earlier signal provision. With regards lower-frequency interactions, as we observed our donor epitope responses, findings suggest Fluorospot be preferable chromogenic modality, immediate washing after collection better facilitate responses. enabled higher minimising generation visual artefacts, meanwhile was critical improving sensitivity. CCR7+ depletion used demonstrate selectively Lastly, provide evidence capacity delineate individual responding peptide pool priming, explore cross-reactivity between viral variant peptides. Conclusion work advances methodology investigating immunity, particularly in context SARS-CoV-2, emphasises balance enhancing specific limiting non-specific activation.

Язык: Английский

Clinical Characteristics During and After COVID‐19 Infection Among Healthcare Workers During the First Wave of Omicron in Chongqing, China DOI Creative Commons

Haoling Tang,

Zhiwei Chen,

Tianquan Huang

и другие.

Immunity Inflammation and Disease, Год журнала: 2025, Номер 13(1)

Опубликована: Янв. 1, 2025

Revealing the clinical manifestations and associations of COVID-19 before after negative transition remains an area significant uncertainty. The aim this study is to investigate characteristics observed during Omicron infection among a specific population, namely healthcare workers (HCWs). From November 4, 2022, January 15, 2023, HCWs in our hospital were enrolled document symptoms, prevention, treatment for using structured questionnaire. A total 1101 included, with SARS-CoV-2 detected 78.20% (861/1101) observation period. median duration nucleic acid conversion was 8 days. Forty-three symptoms identified (11 per individual). common fever, cough, headache, phlegm production, fatigue (60.67%-83.29%). These can be further categorized into five groups: fever type, upper respiratory tract influenza digestive system systemic type-all showing complex diverse patterns. Following infection, 19 recorded including four newly emerged ones: reduced lung capacity, memory loss, lethargy, inattention. Importantly, we association between gastrointestinal positive phase subsequent neuropsychiatric conversion. Interestingly, females experiencing menstruation or pregnancy exhibited higher risk while inhaled vaccines thymosin demonstrated protective effect against infection. displayed intricate patterns, shedding new light on interplay humans.

Язык: Английский

Процитировано

0

Optimisation of the cultured ELISpot/Fluorospot technique for the selective investigation of SARS-CoV-2 reactive central memory T cells DOI Creative Commons

Jack R. Jerome,

Kirsty Wilson, Joshuah Fialho

и другие.

Frontiers in Immunology, Год журнала: 2025, Номер 16

Опубликована: Апрель 15, 2025

Introduction This study presents an optimised cultured ELISpot protocol for detecting central memory T-cell interferon gamma (IFNγ) responses against SARS-CoV-2 peptides following initial priming with either peptides, or whole spike protein. Methods Key variations include the culture length, timing of exogenous survival signals (IL-2), and endpoint analysis modality cell density to enhance assay sensitivity without compromising specificity IFNγ recall cognate antigen. Results We noted a duration 10 days, combined delayed IL-2 administration on day 5 while maintaining response towards antigen when compared shorter periods earlier signal provision. With regards lower-frequency interactions, as we observed our donor epitope responses, findings suggest Fluorospot be preferable chromogenic modality, immediate washing after collection better facilitate responses. enabled higher minimising generation visual artefacts, meanwhile was critical improving sensitivity. CCR7+ depletion used demonstrate selectively Lastly, provide evidence capacity delineate individual responding peptide pool priming, explore cross-reactivity between viral variant peptides. Conclusion work advances methodology investigating immunity, particularly in context SARS-CoV-2, emphasises balance enhancing specific limiting non-specific activation.

Язык: Английский

Процитировано

0