Making the effect visible – OX40 targeting nanobodies for in vivo imaging of activated T cells DOI Creative Commons
Desiree I. Frecot,

Simone Blaess,

Teresa R. Wagner

и другие.

Frontiers in Immunology, Год журнала: 2024, Номер 15

Опубликована: Окт. 15, 2024

Purpose Human OX40 (hOX40/CD134), a member of the TNF receptor superfamily, is mainly expressed on activated T lymphocytes. Triggered by its ligand OX40L (CD252), it provides costimulatory signals that support differentiation, proliferation and long-term survival cells. Besides being relevant therapeutic target, hOX40 also an important biomarker for monitoring presence or infiltration cells within tumor microenvironment (TME), inflammatory (IME) in immune-mediated diseases (IMIDs) lymphatic organs. Here, we developed novel single domain antibodies (nanobodies, Nbs) targeting to monitor activation status vivo molecular imaging. Methods Nbs against (hOX40-Nbs) were selected from immunized Nb-library phage display. The identified hOX40-Nbs characterized vitro , including determination their specificity, affinity, stability, epitope recognition impact signaling cell function. A lead candidate was site-specifically conjugated with fluorophore via sortagging applied noninvasive optical imaging (OI) hOX40-expressing xenograft mouse model. Results Our selection campaign revealed four unique exhibit strong binding affinities high stabilities under physiological conditions. Epitope binning mapping indicated at least two different epitopes hOX40. When analyzing signaling, agonistic effect excluded all validated Nbs. Incubation did not affect viability patterns, whereas differences cytokine release observed. In OI fluorophore-conjugated experimental mice xenografts demonstrated specificity functionality as probe. Conclusion Considering need advanced probes dynamics, propose, our have great potential longitudinal diagnostics. Quantification + TME IME will provide crucial insights into state infiltrating cells, offering valuable assessing immune responses, predicting treatment efficacy, guiding personalized immunotherapy strategies patients cancer IMIDs.

Язык: Английский

RNA aggregates harness the danger response for potent cancer immunotherapy DOI

Héctor R. Méndez‐Gómez,

Anna DeVries,

Paul Castillo

и другие.

Cell, Год журнала: 2024, Номер 187(10), С. 2521 - 2535.e21

Опубликована: Май 1, 2024

Язык: Английский

Процитировано

49
Altered receptor binding, antibody evasion and retention of T cell recognition by the SARS-CoV-2 XBB.1.5 spike protein DOI Creative Commons
Dhiraj Mannar, James W. Saville,

Chad Poloni

и другие.

Nature Communications, Год журнала: 2024, Номер 15(1)

Опубликована: Фев. 29, 2024

Abstract The XBB.1.5 variant of SARS-CoV-2 has rapidly achieved global dominance and exhibits a high growth advantage over previous variants. Preliminary reports suggest that the success stems from mutations within its spike glycoprotein, causing immune evasion enhanced receptor binding. We present binding studies demonstrate retention contacts with human ACE2 striking decrease in to mouse due revertant R493Q mutation. Despite extensive antibody binding, we highlight region on protein domain (RBD) is recognized by serum antibodies donor hybrid immunity, collected prior emergence variant. T cell assays reveal frequencies spike-specific CD4 + CD8 cells amongst donors skewed towards Th1 phenotype having attenuated effector cytokine secretion as compared ancestral protein-specific cells. Thus, while retained efficient gained antigenic alterations, it remains susceptible recognition induced via vaccination infection.

Язык: Английский

Процитировано

10
CD74 is a functional MIF receptor on activated CD4+ T cells DOI Creative Commons
Lin Zhang,

Iris Woltering,

Mathias Holzner

и другие.

Cellular and Molecular Life Sciences, Год журнала: 2024, Номер 81(1)

Опубликована: Июль 11, 2024

Abstract Next to its classical role in MHC II-mediated antigen presentation, CD74 was identified as a high-affinity receptor for macrophage migration inhibitory factor (MIF), pleiotropic cytokine and major determinant of various acute chronic inflammatory conditions, cardiovascular diseases cancer. Recent evidence suggests that is expressed T cells, but the functional relevance this observation poorly understood. Here, we characterized regulation expression MIF chemokine receptors during activation human CD4 + cells studied links MIF-induced T-cell migration, function, COVID-19 disease stage. profiling resting primary via flow cytometry revealed high surface CXCR4, while CD74, CXCR2 ACKR3/CXCR7 were not measurably expressed. However, constitutively intracellularly, which upon significantly upregulated, post-translationally modified by chondroitin sulfate could be detected on cell surface, determined cytometry, Western blot, immunohistochemistry, re-analysis available RNA-sequencing proteomic data sets. Applying 3D-matrix-based live cell-imaging pathway-specific inhibitors, causal involvement CXCR4 migration. Mechanistically, proximity ligation assay visualized CD74/CXCR4 heterocomplexes activated diminished after treatment, pointing towards MIF-mediated internalization process. Lastly, cohort 30 patients, found upregulated CD8 patients with severe compared only mild course. Together, our study characterizes network course reveals novel II-independent marker cells.

Язык: Английский

Процитировано

10
Chimeric antigen receptor with novel intracellular modules improves antitumor performance of T cells DOI Creative Commons
Pengju Wang, Yiyi Wang,

Xiaojuan Zhao

и другие.

Signal Transduction and Targeted Therapy, Год журнала: 2025, Номер 10(1)

Опубликована: Янв. 14, 2025

Abstract The excessive cytokine release and limited persistence represent major challenges for chimeric antigen receptor T (CAR-T) cell therapy in diverse tumors. Conventional CARs employ an intracellular domain (ICD) from the ζ subunit of CD3 as a signaling module, it is largely unknown how alternative chains potentially contribute to CAR design. Here, we obtained series CAR-T cells against HER2 mesothelin using comprising single immunoreceptor tyrosine-based activation motif different subunits ICD CARs. While these reconstituted conferred sufficient antigen-specific cytolytic activity on equipped cells, they elicited low tonic signal, ameliorated exhaustion promoted memory differentiation cells. Intriguingly, CD3ε-derived outperformed others generation that produced minimized cytokines. Mechanistically, CD3ε-based displayed restrained cytomembrane expression engineered which was ascribed endoplasmic reticulum retention mediated by carboxyl terminal basic residues. present study demonstrated applicability reconstitution modules subunits, depicted novel pattern reduces release, thus paving way preparation displaying improved safety tumor antigens.

Язык: Английский

Процитировано

2
A quest for universal anti-SARS-CoV-2 T cell assay: systematic review, meta-analysis, and experimental validation DOI Creative Commons
Akshay Binayke,

Aymaan Zaheer,

Siddhesh Vishwakarma

и другие.

npj Vaccines, Год журнала: 2024, Номер 9(1)

Опубликована: Янв. 2, 2024

Abstract Measuring SARS-CoV-2-specific T cell responses is crucial to understanding an individual’s immunity COVID-19. However, high inter- and intra-assay variability make it difficult define cells as a correlate of protection against To address this, we performed systematic review meta-analysis 495 datasets from 94 original articles evaluating using three assays – Activation Induced Marker (AIM), Intracellular Cytokine Staining (ICS), Enzyme-Linked Immunospot (ELISPOT), defined each assay’s quantitative range. We validated these ranges samples 193 SARS-CoV-2-exposed individuals. Although IFNγ ELISPOT was the preferred assay, our experimental validation suggested that under-represented repertoire. Our data indicate combination AIM ICS or FluoroSpot assay would better represent frequency, polyfunctionality, compartmentalization antigen-specific responses. Taken together, results contribute defining propose choice can be employed understand cellular immune response viral diseases.

Язык: Английский

Процитировано

9
Harnessing the biology of regulatory T cells to treat disease DOI
Christine M. Wardell, Dominic A. Boardman, Megan K. Levings

и другие.

Nature Reviews Drug Discovery, Год журнала: 2024, Номер unknown

Опубликована: Дек. 16, 2024

Язык: Английский

Процитировано

7
Deconvoluting TCR-dependent and -independent activation is vital for reliable Ag-specific CD4 + T cell characterization by AIM assay DOI Creative Commons
Ming Z. M. Zheng, Lauren Burmas, Hyon‐Xhi Tan

и другие.

Science Advances, Год журнала: 2025, Номер 11(17)

Опубликована: Апрель 25, 2025

Activation-induced marker (AIM) assays identify antigen (Ag)–specific T cells, but recent studies revealed AIM + helper cell 17 (T H 17)–like (CCR6 ) and circulating follicular cells (cTfh) were not associated with peptide/HLA tetramer staining. We show that CD39 regulatory reg )–like CD26 hi 22–like undergo receptor (TCR)–independent activation by cytokines during Ag stimulation, leading to nonspecific up-regulation of readouts. Transcriptional analysis enabled discrimination bona fide Ag-specific from cytokine-activated 22 cells. CXCR4 down-regulation emerged as a hallmark clonotypic expansion TCR-dependent in memory CD4 cTfh. By tracking tetramer-binding upon restimulation, we demonstrated CXCR4−CD137 provided more accurate measure Ag-specificity than standard This modified assay excluded the predominantly CCR6 contributed an average 12-fold overestimation population. Our findings provide approach characterize genuine

Язык: Английский

Процитировано

1
The MegaPool Approach to Characterize Adaptive CD4+ and CD8+ T Cell Responses DOI Creative Commons
Ricardo da Silva Antunes, Daniela Weiskopf, John Sidney

и другие.

Current Protocols, Год журнала: 2023, Номер 3(11)

Опубликована: Ноя. 1, 2023

Abstract Epitopes recognized by T cells are a collection of short peptide fragments derived from specific antigens or proteins. Immunological research to study cell responses is hindered the extreme degree heterogeneity epitope targets, which usually multiple antigens; within given antigen, hundreds different epitopes can be recognized, differing one individual next because recognition restricted epitopes’ ability bind MHC molecules, extremely polymorphic in individuals. Testing large pools encompassing peptides technically challenging logistical considerations regarding solvent‐induced toxicity. To address this issue, we developed MegaPool (MP) approach based on sequential lyophilization numbers that used variety assays measure responses, including ELISPOT, intracellular cytokine staining, and activation‐induced marker assays, has been validated infectious diseases, allergies, autoimmunity. Here, describe procedures for generating testing MPs, starting with synthesis lyophilization, as well step‐by‐step guide recommendations their handling experimental usage. Overall, MP powerful strategy studying understanding immune system's role health disease. © 2023 The Authors. Current Protocols published Wiley Periodicals LLC. Basic Protocol 1 : Generation (“MegaPools”) 2 quantitation antigen‐specific

Язык: Английский

Процитировано

16
Respiratory pathogenic microbial infections: a narrative review DOI Creative Commons

Yiyin Long,

Yan Zheng, Changlin Li

и другие.

International Journal of Medical Sciences, Год журнала: 2024, Номер 21(5), С. 826 - 836

Опубликована: Янв. 1, 2024

Respiratory infectious diseases have long been recognised as a substantial global healthcare burden and are one of the leading causes death worldwide, particularly in vulnerable individuals. In post COVID-19 era, there has surge prevalence influenza virus A other multiple known viruses causing cold compared with during same period previous three years, which coincided countries easing restrictions worldwide. This article aims to review community-acquired respiratory illnesses covering broad spectrum viruses, bacteria, atypical microorganisms focuses on cluster pathogens China, thereby providing effective prevention control measures.

Язык: Английский

Процитировано

5
Increased frequencies of highly activated regulatory T cells skewed to a T helper 1-like phenotype with reduced suppressive capacity in dengue patients DOI Creative Commons

Sotheary Sann,

Borita Heng,

Hoa Thi My Vo

и другие.

mBio, Год журнала: 2024, Номер 15(6)

Опубликована: Май 16, 2024

The pathogenesis of dengue involves a complex interplay between the viral factor and host immune response. A mismatch infecting serotype adaptive memory response is hypothesized to lead exacerbated responses resulting in severe dengue. Here, we aim define detail phenotype function different regulatory T cell (Treg) subsets their association with disease severity cohort acute virus (DENV)-infected Cambodian children. Treg frequencies proliferation Tregs are increased patients compared age-matched controls. from skewed Th1-type phenotype. Interestingly, produce more interleukin-10 after

Язык: Английский

Процитировано

4