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ISSN 打印: 1040-8401
ISSN 在线: 2162-6472

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DOI: 10.1615/CritRevImmunol.2017018316
pages 359-378

Flow Cytometric Characterization of Antigen-Specific T Cells Based on RNA and Its Advantages in Detecting Infections and Immunological Disorders

Felix Radford
Department of Molecular, Cellular, and Developmental Biology, Yale University, New Haven, CT 06520
Sanjay Tyagi
Public Health Research Institute, New Jersey Medical School, Rutgers University, Newark, NJ 07103
Maria Laura Gennaro
Public Health Research Institute, New Jersey Medical School, Rutgers University, Newark, NJ 07103
Richard Pine
Public Health Research Institute, New Jersey Medical School, Rutgers University, Newark, NJ 07103
Yuri Bushkin
Public Health Research Institute, New Jersey Medical School, Rutgers University, Newark, NJ 07103

ABSTRACT

Fluorescence in situ hybridization coupled with flow cytometry (FISH-Flow) is a highly quantitative, high-throughput platform allowing precise quantification of total mRNA transcripts in single cells. In undiagnosed infections posing a significant health burden worldwide, such as latent tuberculosis or asymptomatic recurrent malaria, an important challenge is to develop accurate diagnostic tools. Antigen-specific T cells create a persistent memory to pathogens, making them useful for diagnosis of infection. Stimulation of memory response initiates T-cell transitions between functional states. Numerous studies have shown that changes in protein levels lag real-time T-cell transitions. However, analysis at the single-cell transcriptional level can determine the differences. FISH-Flow is a powerful tool with which to study the functional states of T-cell subsets and to identify the gene expression profiles of antigen-specific T cells during disease progression. Advances in instrumentation, fluorophores, and FISH methodologies will broaden and deepen the use of FISH-Flow, changing the immunological field by allowing determination of functional immune signatures at the mRNA level and the development of new diagnostic tools.


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