GENERATION AND CHARACTERIZATION OF T CELL LINES AND T CELL CLONES FROM LAMINA PROPRIA OF CHILDREN WITH COW´S MILK ALLERGY

GUZMAN LUCIANA; DOCENA GUILLERMO; RUIZ MARIA CAROLINA; GARCIA MARCELA; CANZIANI, KARINA EVA; BERNEDO VIVIANA; MUGLIA CECILIA

Tucuman

Congreso; LVII REUNIÓN ANUAL SAI 2019; 2019

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 obtainedfrom peripheral lymphocytes. In this work we generated and characterized milk-specific T cell linesand T cell clones from lamina propia of colonic polyps of allergic patients.Lamina propia mononuclear cells (LPMC) obtained from colonic polyps were stimulated with milkproteins in medium with IL-2, IL-7 and IL-15 (T cell lines-TCL). T cell clones (TCC) were furthergenerated by limiting dilution. Antigen presenting cells were obtained by transforming PBMC withEpstein-Barr virus. Proliferation (CFSE), IL-13 and IFN-γ (ELISA) and TCRαβ and TCRγδ (flowcytometry) were assessed in recalled cells .We obtained 9 TCL and we assessed cell proliferation with milk proteins (25%±5 for CMP vs 5%±3for medium CFSElow cells, N=5) and cytokine secretion. We found a significant higher secretion ofIL-13 with CMP compared to medium (P˂0.01 N=9), whereas IFN-γ secretion remained unchangedupon stimulation. We obtained 19 TCC from one patient. CMP induced clone proliferation (54±4 vs71±5 MFI for CMP and medium, respectively). Based on IL-13/IFN-γ secretion ratio, 9 clones couldbe 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 frompatients with CMA. To the best of our knowledge this is the first report characterizing intestinalTCL and T clones from patients with food allergy.