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

CANZIANI, K.; BERNEDO, V; MUGLIA C.I.; GUZMAN, L; DOCENA, G.; RUIZ, MC; GARCIA, M

Tucumán

Congreso; LXVII Congreso Argentino de Inmunología; 2019

Sociedad Argentina de Inmunología

Cow´s milk protein allergy (CMPA) is a frequent condition among children. Most information regarding T cells from individuals with food allergy comes from studying peripheral blood lymphocytes. In this work we have generated and characterized CMP specific T cell lines and T cell clones, from the lamina propia of polyps of individuals with CMPA.Lamina propia mononuclear cells were obtained from polyps. Cells were incubated in milk supplemented medium with IL-2, IL-7 and IL-15 (T cell lines). T cell clones were generated by limiting dilution. Antigen presenting cells were obtained by transforming PBMC with Epstein Bahr virus. Proliferation (CFSE), IL-13 and IFN-γ (ELISA) and TCR TCRαβ and TCRγδ (flow cytometry) were assessed after incubation with CMP or medium. We obtained 9 lamina propria milk specific T cell lines (LPTCL), from patients with CMPA. Milk induced proliferation of LPTCL compared to medium(25%±5 for CMP vs 5%±3 for medium CFSElow cells, N=5). IL-13 secretion was significantly higher for CMP stimulated compared to medium in all LPTCL (P˂0.01 N=9 for LPTC, T-Student Paired test). No significant induction of IFN-γ secretion was found. Cloning was performed with LPTC cells from one patient. 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, 9 clones could be classified as TH2, while 5 clones were TH1. All clones were TCRαβ+. In conclusion, we have obtained and preliminary characterized CMP specific LPTCL and clones form patients with CMPA. To our knowledge this is the first report for LPTCL and clones specific for a food allergen.