In Vitro Differentiation of Naïve CD4⁺ T Cells: A Tool for Understanding the Development of Atherosclerosis

Methods Mol Biol. 2015:1339:177-89. doi: 10.1007/978-1-4939-2929-0_12.

Abstract

A complete knowledge of atherosclerosis requires a better understanding of how innate and adaptive immunity operate systemically and locally within the arterial wall. T-helper 1 (Th1) lymphocyte responses have proatherogenic effects in mice, contrasting with the responses of T regulatory cells (Tregs), which can suppress growth of atherosclerotic lesions. An imbalance in the differentiation of T-helper cells may therefore impact the development, size, and stability of atherosclerosis plaques. This chapter describes a method to isolate naïve CD4(+) T cells from atherosclerosis-prone mouse peripheral blood lymphocytes and to differentiate these CD4(+) T cells in vitro to various T helper cell lineages. These techniques allow the analysis of T lymphocytes in vitro, a necessary step in the study of molecular mechanisms involved in the inflammatory responses that trigger atherosclerosis.

Keywords: Atherosclerosis; Immunomagnetic isolation; T cell differentiation; Th1; Th17; Th2.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptive Immunity
  • Animals
  • Atherosclerosis / immunology*
  • Atherosclerosis / metabolism
  • Atherosclerosis / pathology
  • Biomarkers / metabolism
  • CD4-Positive T-Lymphocytes / immunology*
  • CD4-Positive T-Lymphocytes / metabolism
  • CD4-Positive T-Lymphocytes / pathology
  • Cell Culture Techniques
  • Cell Differentiation*
  • Cell Lineage*
  • Cells, Cultured
  • Immunity, Innate
  • Immunomagnetic Separation / methods*
  • Lymph Nodes / immunology*
  • Lymph Nodes / metabolism
  • Mice
  • Phenotype
  • Spleen / immunology*
  • Spleen / metabolism
  • T-Lymphocyte Subsets / immunology*
  • T-Lymphocyte Subsets / metabolism
  • T-Lymphocyte Subsets / pathology

Substances

  • Biomarkers