Proliferation and Directional Differentiation of iNKT Cells Derived from  DBA/1 Mice Thymus

Dongzhi Chen; Zhao Li; Rui Liang; Huifang Liu; Yuanyuan Wang; Hongyun Shi; Ming Meng

doi:10.18502/ijaai.v20i5.7412

Dongzhi Chen Department of Immunology, School of Basic Medicine, Hebei University, Baoding, China
Zhao Li Department of Immunology, School of Basic Medicine, Hebei University, Baoding, China
Rui Liang Department of Immunology, School of Basic Medicine, Hebei University, Baoding, China
Huifang Liu Department of Immunology, School of Basic Medicine, Hebei University, Baoding, China
Yuanyuan Wang Department of Immunology, School of Basic Medicine, Hebei University, Baoding, China
Hongyun Shi Department of Radiotherapy, Affiliated Hospital of Hebei University, Baoding, China
Ming Meng Department of Immunology, School of Basic Medicine, Hebei University, Baoding, China

DOI: https://doi.org/10.18502/ijaai.v20i5.7412

Keywords: Alpha-galactosylceramide; Cell differentiation; Thymus gland

Abstract

The rates of invariant natural killer T (iNKT) cells in vivo are very low, and the amounts of cells obtained directly from the body are hard enough to fulfill their potential in clinical application.

To overcome this problem, we subcutaneously injected alpha-galactosylceramide (α-GalCer) into DBA/1 mice and thymic single cells were isolated and cultured in vitro. Fluorescence-activated cell sorting was used to detect the iNKT cells and their subsets in the thymus after the injection of α-GalCer by different methods. In addition, in vitro changes of single-cell suspensions and their cytokines in culture supernatants were assessed.

Compared with the α-GalCer multiple subcutaneous injection group, the rates of iNKT cells in the α-GalCer single subcutaneous injection group were markedly higher at each time point, while the highest levels of iNKT1 and iNKT2 cells were observed on day 4 and 8, respectively. In α-GalCer single subcutaneous injection for 8 days and thymic mononuclear cell cultured for 14 days group, the expansion rate of iNKT cells was significantly faster than the other groups, while it reached a peak for iNKT1 cells. Interferon-gamma was consistent with the development of iNKT1 cells, however no difference was found between the cultured iNKT cells in vitro and the natural iNKT cells in vivo in terms of cytokine production.

Herein, we introduced a method in which antigenic stimulation in vivo and directed induction in vitro yielded high levels of iNKT cells with specific functions.