Epithelium derived interleukin 15 regulates intraepithelial lymphocyte Th1 cytokine production, cytotoxicity, and survival in coeliac disease.
OBJECTIVE
Epithelium derived interleukin (IL)-15 signalling via IL-15Ralpha is critical for the development, activation, and survival of intraepithelial lymphocytes (IEL). We aimed to better understand the IL-15 driven effects on IEL underlying mucosal damage and lymphomagenesis in coeliac disease (CD).


METHODS
Enterocytes, IEL, and lamina propria mononuclear cells (LPMC) were isolated from 46 patients with uncomplicated CD (25 untreated and 21 treated) and 22 controls. IL-15 and IL-15Ralpha expression were determined by immunoblotting. Secretion of IL-15, interferon gamma (IFN-gamma), tumour necrosis factor alpha (TNF-alpha), and granzyme B into cell culture supernatants was assessed by ELISA. The ability of IL-15 to regulate IEL proliferation, perforin/granzyme dependent cytotoxicity, and apoptosis was tested by adding different combinations of IL-15, IL-15 blocking antibody, or chloroquine to IEL cultured alone or with Caco-2 cells as target. IL-15 mucosal levels were also determined by ELISA in five patients with complicated CD (two ulcerative jejunoileites, one refractory sprue, and two enteropathy associated T cell lymphomas) tested for T cell receptor gamma chain clonality.


RESULTS
IL-15 was overexpressed in untreated CD enterocytes and LPMC, and in the mucosa of complicated CD patients and uncomplicated untreated CD patients, where its levels correlated with the degree of mucosal damage. Enterocytes from untreated, but not treated, CD patients and controls secreted IL-15. Untreated CD IEL, characterised by higher IL-15Ralpha expression, showed increased proliferation, production of IFN-gamma and TNF-alpha, and perforin/granzyme dependent cytotoxicity, and a decreased propensity to apoptosis in response to IL-15.


CONCLUSIONS
Our findings suggest that IL-15 plays a crucial role in the generation of epithelial damage in active CD. Its promotion of IEL survival in CD may predispose to the emergence of T cell clonal proliferations. Blocking IL-15, by suppressing uncontrolled IEL activation and survival, has the potential to provide new therapeutic tools to prevent tissue damage and lymphomagenesis in CD.
Di Sabatino
A
A
First Department of Medicine, IRCCS Policlinico San Matteo, University of Pavia, Pavia, Italy.
Ciccocioppo
R
R
Cupelli
F
F
Cinque
B
B
Millimaggi
D
D
Clarkson
M M
MM
Paulli
M
M
Cifone
M G
MG
Corazza
G R
GR
D000328:Adult; D000368:Aged; D000906:Antibodies / Q000276:immunology; D017209:Apoptosis / Q000276:immunology; D018938:Caco-2 Cells; D002446:Celiac Disease / Q000150:complications / Q000276:immunology*; D002455:Cell Division / Q000276:immunology; D002478:Cells, Cultured; D002738:Chloroquine / Q000494:pharmacology; D016207:Cytokines / Q000276:immunology* / Q000378:metabolism; D003602:Cytotoxicity, Immunologic / Q000187:drug effects / Q000276:immunology; D020895:Enterocytes / Q000276:immunology / Q000378:metabolism; D004847:Epithelial Cells / Q000276:immunology; D005260:Female; D006801:Humans; D007371:Interferon-gamma / Q000276:immunology / Q000378:metabolism; D019409:Interleukin-15 / Q000276:immunology* / Q000378:metabolism; D007413:Intestinal Mucosa / Q000276:immunology / Q000378:metabolism; D007421:Intestine, Small / Q000276:immunology; D008214:Lymphocytes / Q000276:immunology* / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000276:immunology; D008875:Middle Aged; D054353:Perforin; D052899:Pore Forming Cytotoxic Proteins; D053647:Receptors, Interleukin-15; D015375:Receptors, Interleukin-2 / Q000276:immunology / Q000378:metabolism; D018417:Th1 Cells / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000276:immunology
D000906:Antibodies; D016207:Cytokines; C508024:IL15RA protein, human; D019409:Interleukin-15; D008562:Membrane Glycoproteins; D052899:Pore Forming Cytotoxic Proteins; D053647:Receptors, Interleukin-15; D015375:Receptors, Interleukin-2; D014409:Tumor Necrosis Factor-alpha; D054353:Perforin; D007371:Interferon-gamma; D002738:Chloroquine