Generation and characterization of T cell lines and T cell clones from lamina propria of children with cow?s milk allergy.

GUZMAN, LUCIANA; MUGLIA, CECILIA; RUÍZ, MARÍA CAROLINA; DOCENA, GUILLERMO ; CANZIANI, KARINA EVA; GARCÍA, MARCELA

San Miguel de Tucumán, Tucumán

Congreso; LXVII Reunión Anual de la Sociedad Argentina de Inmunología 2019; 2019

SAI

Cow´s milk allergy (CMA) is a frequent condition among children and it is mediated by T cells. Most of the information of food-specific T cells from patients with food allergy has been obtained from peripheral lymphocytes. In this work we generated and characterized milk-specific T cell lines and T cell clones from lamina propia of polyps of allergic patients .Lamina propia mononuclear cells (LPMC) obtained from colonic polyps were stimulated with milk proteins in medium with IL-2, IL-7 and IL-15 (T cell lines-TCL). T cell clones (TCC) were further generated by limiting dilution. Antigen presenting cells were obtained by transforming PBMC with Epstein-Barr virus. Proliferation (CFSE), IL-13 and IFN-γ (ELISA) and TCR TCRαβ and TCRγδ (flow cytometry) were assessed in recalled cells . We obtained 9 TCLand we assessed cell proliferation with milk proteins (25%±5 for CMP vs 5%±3 for medium CFSElow cells, N=5) and cytokine secretion. We found a significant higher secretion of IL-13 with CMP compared to medium (P˂0.01 N=9), whereas IFN-γ secretion remained unchanged upon stimulation. We obtained 1 TCC. 19 CMP specific LPTC were obtained. CMP induced clone proliferation (54±4 vs 71±5 MFI for CMP and medium, respectively). Based on IL-13/IFN-γ secretion ratio, 9 clones could be classified as TH2, while 5 clones were TH1. All clones were TCRαβ+. In conclusion, we obtained and characterized milk- specific lamina propria TCL and clones from patients with CMA. To the best of our knowledge this is the first report characterizing intestinal TCL and T clones from patients with food allergy.