求助 Current protocols in immunology 一篇

woosheng

Curr Protoc Immunol. 2010 Aug;Chapter 14:Unit 14.11.3 v. M& L) ~7 s5 ~
Isolation and functional use of human NKT cells.
1 G' h3 ~4 w1 jExley MA, Wilson B, Balk SP.
1 i, G) y- Z- X: |& G5 C  m
4 x" _- G% k8 D  _0 b. |Cancer Biology Program, Hematology-Oncology Division, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA.
! c# Z# i- k  q5 ?/ a8 E9 d( EAbstract& S9 T1 K9 Q' j+ G
This unit details methods for the isolation, in vitro expansion, and functional characterization of human iNKT cells. The term iNKT derives from the fact that a large fraction of murine NKT cells recognize the MHC class I-like CD1d protein, are CD4+ or CD4-CD8- (double negative), and use an identical "invariant" TCRalpha chain, which is generated by precise Valpha14 and Jalpha281 (now renamed Jalpha18) rearrangements with either no N-region diversity or subsequent trimming to nearly identical amino-acid sequence (hence, 'iNKT'). Basic Protocol 1 and Alternate Protocol 1 use multi-color FACS analysis to identify and quantitate rare iNKT cells from human samples. Basic Protocol 2 describes iNKT cell purification. Alternate Protocol 2 describes a method for high-speed FACS sorting of iNKT cells. Alternate Protocol 3 employs a cell sorting approach to isolate iNKT cell clones. A Support Protocol for secondary stimulation and rapid expansion of iNKT cells is also included. Basic Protocol 3 explains functional analysis of iNKT.
' `* |1 @1 E9 A/ G: @- q- e* `# o4 Q. j2 b; s* j
PMID: 20814940

laolifeidao999

Isolation and functional use of human NKT cells.