iNKT Cel Transfer: The Use of Cell Sorting Combined with Flow Cytometry Validation Approach

Methods Mol Biol. 2021:2388:113-122. doi: 10.1007/978-1-0716-1775-5_10.

Abstract

Natural killer T (NKT) cells are an innate-like T cell subset that recognize lipid antigens presented by CD1d-expressing antigen presenting cells (APCs), such as dendritic cells, macrophages, and B cells. They can be subdivided into two different subsets according to the variation in αβ TCR chains: type I and type II NKT cells. Type I, also called invariant NKT cells (iNKT), express restricted TCRs with an invariant α-chain (Vα24-Jα18 in humans and Vα14-Jα18 in mice) and limited β-chains. Here we have established a protocol in which iNKT cells are isolated from a donor wild-type mouse and transferred into iNKT KO (Jα18-/-) mouse. Below we will explore the methods for cell sorting of splenic iNKTs, iNKT cells transfer, and detection of transferred cells into the liver using flow cytometry technique.

Keywords: Cell sorting; Cell transfer; Flow cytometry; Isolation of splenocytes; Jα18−/− mice; Liver lymphocytes isolation; Retro-orbital injection; iNKT cell.

MeSH terms

  • Animals
  • Antigen-Presenting Cells
  • Antigens, CD1d
  • Flow Cytometry
  • Mice
  • Natural Killer T-Cells*
  • Receptors, Antigen, T-Cell
  • T-Lymphocyte Subsets

Substances

  • Antigens, CD1d
  • Receptors, Antigen, T-Cell