Fig 1.
The intestinal epithelial compartment of celiac disease contains subpopulations of NK cells and NKT cells characterized by increased levels of NKp30 and NKG2D and low levels of NKG2A.
Intraepithelial mononuclear cells isolated from duodenal biopsies of 7 normal controls (CTR), 10 inactive celiac disease (ICD) patients and 15 active celiac disease (ACD) patients were stained with CD45, CD103, CD3, CD56, NKp30, NKG2D and NKG2A antibodies. Each point in the graph indicates the percentage of positive cells in a single sample of a single patient. The horizontal bars represent the median values. A. Left graphs show the percentages of NK cells (CD45+,CD103+,CD56+) in CTR, ICD patients and ACD patients. Right graph shows the percentages of NKT cells (CD45+,CD103+,CD56+, CD3+) in CTR, ICD patients and ACD patients. B. Left graph shows the percentages of NKp30+ NK cells in CTR, ICD patients and ACD patients. Right graph shows the percentages of NKp30+ NKT cells in CTR, ICD patients and ACD patients. C. Left graph shows the percentages of NKG2D+ NK cells in CTR, ICD patients and ACD patients. Middle graph shows the percentages of NKG2D+ NKT cells in CTR, ICD patients and ACD patients. Right graph shows mean fluorescence intensity (MFI) values for NKG2D+ in NKT cells isolated from CTR, ICD patients and ACD patients. Data indicate mean and standard error. D. Left graph shows the percentages of NKG2A+ NK cells in CTR, ICD patients and ACD patients. Right graph shows the percentages of NKG2A+ NKT cells in CTR, ICD patients and ACD patients.
Fig 2.
NKp44/NKp46-double positive NK cells and NKT cells are decreased in active celiac disease (ACD).
A-C. Intraepithelial mononuclear cells isolated from duodenal biopsies of 13 normal controls (CTR), 6 inactive celiac disease (ICD) patients and 15 ACD patients were stained with CD45, CD103, CD56, CD3, NKp44 and NKp46 antibodies. Panel A shows the percentages of NKp46+ NK (CD45+,CD103+, CD56+; left) cells and NKT (CD45+,CD103+, CD56+, CD3+; right) cells in CTR, ICD patients and ACD patients. Panel B shows the percentages of NKp44+ NK (left) cells and NKT (right) cells in CTR, ICD patients and ACD patients. Panel C shows the percentages of NKp46/NKp44-double positive NK (left) cells and NKT (right) cells in CTR, ICD patients and ACD patients. Each point in the graph indicates the percentage of positive cells in a single sample of a single patient. The horizontal bars represent the median values. D. Representative contour-plots show NKp44 and NKp46 expression in NK cells and NKT cells of CTR, ICD patients and ACD patients.
Fig 3.
NKp44/NKp46-double positive NK cells and NKT cells are a source of granzyme B and IL-22.
Intraepithelial mononuclear cells isolated from duodenal biopsies of normal controls (CTR) were stained with CD45, CD103, CD56, CD3, NKp44, NKp46, granzyme B, IL-22 and TNF-α antibodies. A. Panel A shows representative histogram-plots of granzyme B, IL-22 and TNF-α expression in NKp44/NKp46-double positive NK cells and NKT cells. Data are representative of respectively 6, 5 and 3 separate experiments. B. Panel B shows the percentages of granzyme B, IL-22 and TNF-α expression in NKp46/NKp44-double positive cells. Each point in the graph indicates the percentage of positive cells in a single sample of a single patient.
Fig 4.
Stimulation of normal intra-epithelial cells with toll-like receptor ligands increases granzyme B expression in NKp44/NKp46-double positive NK cells and NKT cells.
Intraepithelial mononuclear cells, isolated from jejunal specimens of 3 controls, were either left unstimulated (Unst) or stimulated with LPS, poly I:C, PGN and CpG. Granzyme B expression was then evaluated in NKp44/NKp46-double positive NK (CD45+,CD103+, CD56+) cells or NKT (CD45+,CD103+, CD56+, CD3+) cells. A. Panel A shows representative histogram-plots of Granzyme B expression in intraepithelial NK and NKT cells either left unstimulated (Unst) or stimulated with LPS, poly I:C, PGN and CpG. B. Panel B shows the percentages of Granzyme B expression in intraepithelial NK and NKT cells either left unstimulated (Unst) or stimulated with LPS, poly I:C, PGN and CpG. Data are representative of 2 different experiments and show an increased expression of Granzyme B, that however do not reach statistical significance.