Skip to main content
. 2006 Dec 20;1(1):e50. doi: 10.1371/journal.pone.0000050

Figure 1. Identification of NKT cell subsets in neonatal and adult blood PBMC.

Figure 1

(A) NKT cells in either neonatal or adult PBMC were identified by staining for the invariant TCR using antibodies against Vα24 (FITC) and Vβ11 (unlabelled followed by goat anti-mouse APC).

Double positive cells were identified as NKT cells and gated on for further analysis to determine CD4 (PE) and CD8 (biotin followed by strepavidin-PerCP-Cy5.5) expression.

(B) NKT cell subsets were identified by staining for Vα24, Vβ11, and CD4 as in 1A and then gated on for further analysis. Expression of the indicated cell surface marker on these subsets was then determined.

Combinations of the following antibodies were used, depending on the particular stain: Vα24 (FITC or unlabelled), Vβ11 (unlabelled or PE), CD4 (PE, biotin, or unlabelled), CCR7 (unlabelled), CD62L (PE), CD94 (unlabelled), CD161 (FITC), CD28 (biotin), goat anti-mouse (APC), and strepavidin (PerCP-Cy5.5). One representative donor is shown.

(C) Summary of NKT cell phenotype data for all neonatal and adult donors. P values are indicated.

For comparison of nCD4+ to either aCD4+ or aCD4− subsets, P values were generated using the Wilcoxon rank sum test.

For comparison of aCD4+ to aCD4− subsets, P values were generated using the Wilcoxon signed rank test.

(D) For CD4+ NKT cells of adult donors represented in 1C, the ratio of CD161 (% expression) to CD62L (% expression) was generated and compared to CCR7 expression using linear regression with a resulting equation of y = −19.277X+86.676 and p = 0.008.