Tracking the Replication-Competent Zika Virus with Tetracysteine-Tagged Capsid Protein in Living Cells

Li, Shimin; Wang, Dianbing; Abbas, Ghulam; Li, Xia; Li, Min; Li, Qin; Ma, Yingxin; Wang, Lei; Wu, Hangshen; Cui, Zongqiang; Zhang, Xian-En

doi:10.1128/jvi.01846-21

“…Here, we address this issue by presenting a novel strategy for visualizing T cell-APC interactions via tetracysteine tagging of antigenic peptides. While tetracysteine tagging has been previously applied in cell biology and virology, these applications have been mostly limited to cell lines and in vitro studies in immunology-unrelated experimental settings ( Griffin et al, 1998 ; Mohl and Roy, 2017 ; Li et al, 2022 ; Ng et al, 2019 ). On the other hand, our technique labels antigenic peptides and tracks pMHCII complexes in primary APCs during their interactions with T cells both in vitro and in vivo.…”

Section: Discussionmentioning

confidence: 99%

Illuminating T cell-dendritic cell interactions in vivo by FlAsHing antigens

Akkaya,

Al Souz,

Williams

et al. 2024

eLife

0

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Delineating the complex network of interactions between antigen-specific T cells and antigen presenting cells (APCs) is crucial for effective precision therapies against cancer, chronic infections, and autoimmunity. However, the existing arsenal for examining antigen-specific T cell interactions is restricted to a select few antigen-T cell receptor pairs, with limited in situ utility. This lack of versatility is largely due to the disruptive effects of reagents on the immune synapse, which hinder real-time monitoring of antigen-specific interactions. To address this limitation, we have developed a novel and versatile immune monitoring strategy by adding a short cysteine-rich tag to antigenic peptides that emits fluorescence upon binding to thiol-reactive biarsenical hairpin compounds. Our findings demonstrate the specificity and durability of the novel antigen-targeting probes during dynamic immune monitoring in vitro and in vivo. This strategy opens new avenues for biological validation of T-cell receptors with newly identified epitopes by revealing the behavior of previously unrecognized antigen-receptor pairs, expanding our understanding of T cell responses.

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“…Here, we address this issue by presenting a novel strategy for visualizing T cell-APC interactions via tetracysteine tagging of antigenic peptides. While tetracysteine tagging has been previously applied in cell biology and virology, these applications have been mostly limited to cell lines and in vitro studies in immunology unrelated experimental settings [25][26][27][28]. On the other hand, our technique labels antigenic peptides and tracks pMHCII complexes in primary APCs during their interactions with T cells both in vitro and in vivo.…”

Section: Discussionmentioning

confidence: 99%

Illuminating T cell-dendritic cell interactions in vivo by FlAsHing antigens

Akkaya,

Souz,

Williams

et al. 2023

Preprint

0

View full text Add to dashboard Cite

Delineating the complex network of interactions between antigen-specific T cells and antigen presenting cells (APCs) is crucial for effective precision therapies against cancer, chronic infections, and autoimmunity. However, the existing arsenal for examining antigen-specific T cell interactions is restricted to a select few antigen-T cell receptor pairs, with limited in situ utility. This lack of versatility is largely due to the disruptive effects of reagents on the immune synapse, which hinder real-time monitoring of antigen-specific interactions. To address this limitation, we have developed a novel and versatile immune monitoring strategy by adding a short cysteine-rich tag to antigenic peptides that emits fluorescence upon binding to thiol-reactive biarsenical hairpin compounds. Our findings demonstrate the specificity and durability of the novel antigen-targeting probes during dynamic immune monitoring in vitro and in vivo. This strategy opens new avenues for biological validation of T-cell receptors with newly identified epitopes by revealing the behavior of previously unrecognized antigen-receptor pairs, expanding our understanding of T cell responses.

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“…The NS4AB of HCV and the PR of HIV were synthesized by Sangon Biotech (Beijing, China) with codon optimization. The fragment of NS2B-NS3 of ZIKV was amplified from the plasmid pACYC177-Natal-RGN ( 83 ). The above three proteases were all inserted into pCDNA3.1- (+) expression vectors with a C-terminal HA tag.…”

Section: Methodsmentioning

confidence: 99%