Immortalized B Cells Transfected with mRNA of Antigen Fused to MITD (IBMAM): An Effective Tool for Antigen-Specific T-Cell Expansion and TCR Validation

Authors

Zhe Wang
Tiantian Zhang
Aaron Anderson
Vincent Lee
Szymon Szymura
Zhenyuan Dong
Benjamin Kuang
Elizabeth Oh
Jingwei Liu
Sattva S Neelapu
Larry Kwak
Soung-Chul Cha

Publication Date

3-6-2023

Journal

Biomedicines

Abstract

Peripheral mononuclear blood cells (PBMCs) are the most widely used study materials for immunomonitoring and antigen-specific T-cell identification. However, limited patient PBMCs and low-frequency antigen-specific T cells remain as significant technical challenges. To address these limitations, we established a novel platform comprised of optimized HLA-matched immortalized B cells transfected with mRNA of a prototype viral or tumor antigen conjugated to MHC class-I trafficking domain protein (MITD) to increase the efficiency of epitope expression in antigen-presenting cells (APCs) essential to expanding antigen-specific T cells. When applied to CMV as a model, the IBMAM platform could successfully expand CMV-specific T cells from low-frequency CMV PBMCs from seropositive donors. Additionally, this platform can be applied to the validation of antigen specific TCRs. Together, compared to using APCs with synthesized peptides, this platform is an unlimited, highly efficient, and cost-effective resource in detecting and expanding antigen-specific T cells and validating antigen-specific TCRs.

Keywords

B cell immortalization, CMV, MITD, TCR validation and discovery, antigen-specific T cells, immunomonitoring, mRNA, novel platform

DOI

10.3390/biomedicines11030796

PMID

36979775

PMCID

PMC10045729

PubMedCentral® Posted Date

3-6-2023

PubMedCentral® Full Text Version

Post-print

Published Open-Access

yes