Comparison of two flow cytometric methods for detection of human invariant natural killer T cells (iNKT).
BACKGROUND
Invariant natural killer cells (iNKT) are an important immunoregulatory T cell subset. Currently several flow cytometry-based approaches exist for the identification of iNKT cells, which rely on using the 6B11 monoclonal antibody or a combination of anti-Valpha24 and anti-Vbeta11 antibodies.


OBJECTIVE
The aim of this study was to compare the ability of two flow cytometry-based methods for detecting the frequency of circulating iNKT cells.


METHODS
The frequency of iNKT cells was detected in the peripheral blood of 37 healthy adult donors by flow cytometry using the 6B11 antibody or a combination of anti-Valpha24 and anti-Vbeta11 antibodies.


RESULTS
The frequency of iNKT cells detected by 6B11 antibody or by combination of anti-Valpha24 and anti-Vbeta11 antibodies was significantly different (0.54% vs. 0.31%, respectively, p<0.001) but the values were highly correlated (Spearman r = 0.742, p<0.0001).


CONCLUSIONS
The results of this study indicate that different combinations of mAbs detect different frequencies of peripheral blood iNKT cells and a consensus in the field needs to be established to allow better assessment of iNKT-related studies and suggest using different methods for accurate identification of iNKT cells.
Fereidouni
Mohammad
M
Department of Allergy and Immunology, Ghaem Hospital, Mashhad University of Medical Sciences, Mashhad, Iran.
Jabbari Azad
Farahzad
F
Mahmoudi
Mahmoud
M
Varasteh
Abdolreza
A
Farid Hosseini
Reza
R
D000328:Adult; D000911:Antibodies, Monoclonal / Q000737:chemistry* / Q000276:immunology; D005260:Female; D005434:Flow Cytometry / Q000379:methods*; D006801:Humans; D018655:Lymphocyte Count / Q000379:methods; D008297:Male; D055611:Natural Killer T-Cells / Q000166:cytology* / Q000276:immunology; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000276:immunology; D012680:Sensitivity and Specificity
D000911:Antibodies, Monoclonal; D016693:Receptors, Antigen, T-Cell, alpha-beta