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A universal MHCII technology platform to characterize antigen-specific CD4⁺ T cells.

ABSTRACT: CD4⁺ T cells are critical to the immune system and perform multiple functions; therefore, their identification and characterization are crucial to better understanding the immune system in both health and disease states. However, current methods rarely preserve their ex vivo phenotype, thus limiting our understanding of their in vivo functions. Here we introduce a flexible, rapid, and robust platform for ex vivo CD4⁺ T cell identification. By combining MHCII allele purification, allele-independent peptide loading, and multiplexed flow cytometry technologies, we can enable high-throughput personalized CD4⁺ T cell identification, immunophenotyping, and sorting. Using this platform in combination with single-cell sorting and multimodal analyses, we identified and characterized antigen-specific CD4⁺ T cells relevant to COVID-19 and cancer neoantigen immunotherapy. Overall, our platform can be used to detect and characterize CD4⁺ T cells across multiple diseases, with potential to guide CD4⁺ T cell epitope design for any disease-specific immunization strategy.

SUBMITTER: Vyasamneni R

PROVIDER: S-EPMC9939426 | biostudies-literature | 2023 Jan

REPOSITORIES: biostudies-literature

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A universal MHCII technology platform to characterize antigen-specific CD4+ T cells.

Vyasamneni Rohit R Kohler Victoria V Karki Binisha B Mahimkar Gauri G Esaulova Ekaterina E McGee Jonathan J Kallin Daniel D Sheen Joong Hyuk JH Harjanto Dewi D Kirsch Miles M Poran Asaf A Dong Jesse J Srinivasan Lakshmi L Gaynor Richard B RB Bushway Meghan E ME Srouji John R JR

Cell reports methods 20230113 1

CD4+ T cells are critical to the immune system and perform multiple functions; therefore, their identification and characterization are crucial to better understanding the immune system in both health and disease states. However, current methods rarely preserve their ex vivo phenotype, thus limiting our understanding of their in vivo functions. Here we introduce a flexible, rapid, and robust platform for ex vivo CD4+ T cell identification. By combining ...[more]

PMID: 36814840

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Project description:Background/objectivesAlthough obesity is associated with low-grade inflammation and metabolic disorders, clinical studies suggested some obese people were metabolically healthy with smaller adipocyte size compared with metabolically abnormal obese (MAO). This indicated adipocyte size may be an important predictor underlay the distinction between MAO and metabolically healthy obese. As recent study has shown that adipocytes expressed class II major histocompatibility complex (MHCII), which functioned as APCs during obesity. However, the relationship between adipocyte hypertrophy and MHCII expression was not involved. Here we hypothesize that hypertrophic adipocytes could be associated with upregulating MHCII to influence adipose tissue metabolism.MethodsAdipocytes were sorted by fluorescence-activated cell sorting (FACS) according to the cell size from MAO mice. The activation of MHCII, T cells and related signaling molecules were examined by FACS, ELISA and western blotting. 3T3-L1 cell line and primary adipocytes were used to examine the effect of free fatty acids (FFA) on adipocytes enlargement and MHCII expression.ResultsMAO mice had a significant increase in adipocytes size and FFA concentration. The large adipocytes from both obese and non-obese mice expressed higher levels of MHCII than small adipocytes. Importantly, large adipocytes from obese mice stimulated CD4(+) T cells to secrete more interferon (IFN)-γ. Furthermore, the activation of the JNK-STAT1 pathway was involved in upregulation of MHCII in large adipocytes. In vitro FFA treatment promoted adipocyte hypertrophy and expression of MHCII-associated genes.ConclusionsThis study demonstrates that large adipocytes highly express MHCII and function as APC to stimulate IFN-γ-expressing CD4(+) T cells, in which FFA may have important roles before IFN-γ elevated. These findings suggest that adipocyte hypertrophy, rather than overall obesity, is the major contributor to adipose tissue inflammation and insulin resistance.

Dataset Information

A universal MHCII technology platform to characterize antigen-specific CD4⁺ T cells.

Publications

A universal MHCII technology platform to characterize antigen-specific CD4<sup>+</sup> T cells.

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