Infiltration of Foxp3- and Toll-like receptor-4-positive cells in the intestines of children with food allergy.
OBJECTIVE
Regulatory T (Treg) cells together with intestinal microflora play a central role in controlling allergic inflammation. We examined the markers related to Treg cells, and bacterial signaling, such as Toll-like receptors (TLR)-2 and -4, in the duodenal mucosa of patients with food allergy (FA).


METHODS
Small intestinal samples were collected from patients with FA on a normal or an elimination diet, from healthy controls and patients with untreated celiac disease. Single and double immunohistochemistry were used to enumerate the densities of Foxp3-positive cells and TLR2- and TLR4-positive cells in the mucosa and evaluate the colocalization of Foxp3 expression in CD4, CD25, and CTLA-4 cells. The mRNA expression of CD25, Foxp3, TLR2, and TLR4 was measured by reverse transcriptase-polymerase chain reaction.


RESULTS
The densities of Foxp3 and TLR4 cells were significantly increased in patients with untreated FA compared with healthy controls (P = 0.003, P = 0.033), and the Foxp3 cells were higher in untreated than in treated allergic patients (P < 0.001). The immense majority of Foxp3 cells were CD4 (median 100%), CTLA-4 (100%), or CD25 (81%). The ratio of Foxp3 mRNA to Foxp3 cells was decreased in patients with FA and in patients with celiac disease compared with controls (P = 0.036, P = 0.035).


CONCLUSIONS
Foxp3 cells are increased in the duodenum of patients with untreated FA, but these cells are not able to suppress the harmful immune response, indicated by the low expression of Foxp3 transcripts. The increase of TLR4 cells and their correlation with TCRgammadelta intraepithelial lymphocytes suggest a role for the innate immunity and intestinal microbiota in FA.
Westerholm-Ormio
Mia
M
Hospital for Children and Adolescents, University of Helsinki, Finland. mia.westerholm-ormio@hus.fi
Vaarala
Outi
O
Tiittanen
Minna
M
Savilahti
Erkki
E
D015703:Antigens, CD / Q000378:metabolism; D015415:Biomarkers; D002446:Celiac Disease / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D002648:Child; D002675:Child, Preschool; D004032:Diet; D004386:Duodenum / Q000276:immunology / Q000378:metabolism*; D005260:Female; D005512:Food Hypersensitivity / Q000235:genetics / Q000378:metabolism*; D051858:Forkhead Transcription Factors / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D006801:Humans; D007223:Infant; D007413:Intestinal Mucosa / Q000276:immunology / Q000378:metabolism*; D008297:Male; D012333:RNA, Messenger / Q000378:metabolism; D016692:Receptors, Antigen, T-Cell, gamma-delta / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050378:T-Lymphocytes, Regulatory / Q000378:metabolism*; D051195:Toll-Like Receptor 2 / Q000235:genetics / Q000378:metabolism; D051197:Toll-Like Receptor 4 / Q000235:genetics / Q000378:metabolism*
D015703:Antigens, CD; D015415:Biomarkers; C418974:FOXP3 protein, human; D051858:Forkhead Transcription Factors; D012333:RNA, Messenger; D016692:Receptors, Antigen, T-Cell, gamma-delta; D051195:Toll-Like Receptor 2; D051197:Toll-Like Receptor 4