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Isolation and Functional Use of Human NKT Cells
Mark A Exley1,2, S Brian Wilson3, Steven P Balk4
1Gastroenterology, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts.
Current Protocols in Immunology
|November 2, 2017
Summary
This study provides detailed methods for isolating, expanding, and functionally characterizing human invariant natural killer T (iNKT) cells. These protocols enable researchers to identify, purify, and analyze these rare immune cells for further study.
Area of Science:
- Immunology
- Cell Biology
Background:
- Invariant natural killer T (iNKT) cells are a unique T cell population recognizing the CD1d molecule.
- Human iNKT cells typically express an invariant TCRα chain (Vα24-Jα18) and are mostly CD4+ or double-negative (DN).
Purpose of the Study:
- To provide comprehensive protocols for the isolation, in vitro expansion, and functional characterization of human iNKT cells.
- To enable researchers to identify, quantify, and purify rare iNKT cells from human samples.
Main Methods:
- Multi-color Fluorescence-Activated Cell Sorting (FACS) analysis for identification and quantitation.
- Purification methods including high-speed FACS sorting.
- Functional assays and cell sorting for iNKT cell clone isolation.
Main Results:
- Detailed protocols for identifying and isolating iNKT cells using FACS.
- Methods for in vitro expansion and secondary stimulation of iNKT cells.
- Established procedures for functional characterization of iNKT cells and clones.
Conclusions:
- The presented protocols offer a robust framework for studying human iNKT cells.
- These methods facilitate the research of iNKT cell biology and potential therapeutic applications.
