Exploring the dynamics of T‐cell responses: a combined approach using EdU incorporation and proliferation dye dilution assay DOI Open Access

Hilde Raaphorst,

Sinéad M. Lougheed,

Latifa Saou

et al.

Immunology and Cell Biology, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 30, 2024

Understanding antigen-specific T-cell responses is crucial for advancing immunotherapies and vaccine development. This study proposes a novel approach combining two complementary assays: the 5-ethynyl-2'-deoxyuridine (EdU) incorporation assay (tracking proliferation over 0-48 h) VPD450 dye dilution 4-6 days). Integrating these techniques provides additional insights into kinetics. Both assays were independently optimized using anti-CD3 anti-CD28 polyclonal T cell stimulation. 1 μM suitable assessing proliferation. The EdU concentration should match stimulation strength, requiring higher concentrations to efficiently track DNA replication detection during increased cellular division. Day 5 was optimal read-out day assay. We then combined assays. As proof of principle, we stimulated PBMCs from healthy donors with tetanus toxoid assess responses. Additionally, demonstrated assay's application in drug research by evaluating mixed lymphocyte reaction abatacept, an agonistic anti-CTLA-4 antibody. offers qualitative kinetics, beneficial efficiency or designing new treatments targeting proliferation, such as autoimmune settings.

Language: Английский

Age and Latent Cytomegalovirus Infection Do Not Affect the Magnitude of De Novo SARS‐CoV‐2‐Specific CD8+ T Cell Responses DOI Creative Commons
Jet van den Dijssel,

Veronique A. L. Konijn,

Mariël C Duurland

et al.

European Journal of Immunology, Journal Year: 2025, Volume and Issue: 55(3)

Published: March 1, 2025

ABSTRACT Immunosenescence, age‐related immune dysregulation, reduces immunity upon vaccinations and infections. Cytomegalovirus (CMV) infection results in declining naïve (T ) increasing terminally differentiated emra T cell populations, further aggravating aging. Both immunosenescence CMV have been speculated to hamper the formation of protective T‐cell against novel or emerging pathogens. The SARS‐CoV‐2 pandemic presented a unique opportunity examine impact age and/or on generation de novo SARS‐CoV‐2‐specific CD8 + responses 40 younger (22–40 years) 37 older (50–66 convalescent individuals. Heterotetramer combinatorial coding combined with phenotypic markers were used study 35 epitope‐specific populations directly ex vivo. Neither nor affected frequencies, despite reduced total cells ‐ Robust central memory cm detected adults regardless status. Our data demonstrate that aging status did not response. However, individuals displayed lowest stem scm ), highest PD1 suggesting age, CMV, may long‐term immunity.

Language: Английский

Citations

0
Beyond Suppression: Peripheral T Cell Responses to Vaccination in Inflammatory Bowel Disease Patients Undergoing Anti-Tumor-Necrosis-Factor Therapy DOI Creative Commons
Martin Qui, Ennaliza Salazar

Vaccines, Journal Year: 2024, Volume and Issue: 12(11), P. 1280 - 1280

Published: Nov. 14, 2024

Alimentary tract inflammation in inflammatory bowel disease (IBD) is treated by systemically administered drugs that alter fundamental host immune responses. Biologics target tumor necrosis factor (TNF) are first-line biologics IBD, used widely for their effectiveness, steroid-sparing quality, and lower cost. While they enable a significant proportion of patients to achieve clinical remission, carry an increased risk infection poor serological responses vaccination. Conversely, our understanding adaptive T cell anti-TNF-treated IBD remains limited. The introduction COVID-19 vaccines has prompted research both challenges refines view on immunomodulatory therapy its potential implications immunity protection. Here, we review these emergent findings, evaluate how shape vaccine-induced the context anti-TNF provide perspective highlighting need holistic evaluation cellular humoral this population.

Language: Английский

Citations

0
Advances in Functional Organic-based Nanosystems for RNA Delivery, Targeting Different Organs DOI Creative Commons

Mahzad Motalleb,

Samira Malyen,

Geyou Ao

et al.

Materials chemistry horizons., Journal Year: 2024, Volume and Issue: 3(2), P. 1 - 14

Published: Nov. 1, 2024

Language: Английский

Citations

0
Exploring the dynamics of T‐cell responses: a combined approach using EdU incorporation and proliferation dye dilution assay DOI Open Access

Hilde Raaphorst,

Sinéad M. Lougheed,

Latifa Saou

et al.

Immunology and Cell Biology, Journal Year: 2024, Volume and Issue: unknown

Published: Dec. 30, 2024

Understanding antigen-specific T-cell responses is crucial for advancing immunotherapies and vaccine development. This study proposes a novel approach combining two complementary assays: the 5-ethynyl-2'-deoxyuridine (EdU) incorporation assay (tracking proliferation over 0-48 h) VPD450 dye dilution 4-6 days). Integrating these techniques provides additional insights into kinetics. Both assays were independently optimized using anti-CD3 anti-CD28 polyclonal T cell stimulation. 1 μM suitable assessing proliferation. The EdU concentration should match stimulation strength, requiring higher concentrations to efficiently track DNA replication detection during increased cellular division. Day 5 was optimal read-out day assay. We then combined assays. As proof of principle, we stimulated PBMCs from healthy donors with tetanus toxoid assess responses. Additionally, demonstrated assay's application in drug research by evaluating mixed lymphocyte reaction abatacept, an agonistic anti-CTLA-4 antibody. offers qualitative kinetics, beneficial efficiency or designing new treatments targeting proliferation, such as autoimmune settings.

Language: Английский

Citations

0