Kumar Amrendra, Bezbradica Jelena S, Stanic Aleksandar K, Joyce Sebastian
Veterans Administration, Tennessee Valley Healthcare System, Nashville, Tennessee.
Vanderbilt University Medical Center, Nashville, Tennessee.
Curr Protoc Immunol. 2017 Apr 3;117:14.13.1-14.13.55. doi: 10.1002/cpim.22.
Semi-invariant natural killer T (iNKT) cells are CD1d-restricted innate-like lymphocytes that recognize lipid agonists. Activated iNKT cells have immunoregulatory properties. Human and mouse iNKT cell functions elicited by different glycolipid agonists are highly conserved, making the mouse an excellent animal model for understanding iNKT cell biology in vivo. This unit describes basic methods for the characterization and quantification (see Basic Protocol 1) and functional analysis of mouse iNKT cells in vivo or in vitro. This unit also contains protocols that describe enrichment and purification of iNKT cells, generation of CD1d tetramer, and lipid antigen loading onto cell-bound and soluble CD1d for activation of NKT cell hybridomas. © 2017 by John Wiley & Sons, Inc.
半不变自然杀伤T(iNKT)细胞是受CD1d限制的固有样淋巴细胞,可识别脂质激动剂。活化的iNKT细胞具有免疫调节特性。不同糖脂激动剂引发的人和小鼠iNKT细胞功能高度保守,这使得小鼠成为在体内理解iNKT细胞生物学的优秀动物模型。本单元描述了小鼠iNKT细胞在体内或体外的表征与定量(见基本方案1)以及功能分析的基本方法。本单元还包含描述iNKT细胞富集和纯化、CD1d四聚体的生成以及将脂质抗原加载到细胞结合型和可溶性CD1d上以激活NKT细胞杂交瘤的方案。©2017约翰威立父子公司。