In Vitro Propagation and Dynamics of T cells from Skin Biopsies by Methods Using Interleukins-2 and -4 or Anti-CD3/CD28 Antibody-coated Microbeads
Hideo Hashizume, Anker Hansen, Lars K. Poulsen, Allan Randrup Thomsen, Masahiro Takigawa, Kristian Thestrup-Pedersen
In order to explore the mechanisms of inflammatory skin disorders, we established two methods of expanding skin-derived lymphocytes, one using high levels of interleukin (IL)-2 and IL-4 (method A) and the other using low levels of cytokines and anti-CD3/CD28 microbeads (method B). Both methods provide advantages for functional studies. With either of these two, we could obtain more than 107 cells/ from a 3 mm skin biopsy in 21 days from 23 out of 26 biopsies of various skin diseases. The relevance of these cells was confirmed by shifted T-cell receptor β chain variable region (TCR-Vβ) repertoire and antigen-dependent proliferation in antigen-driven skin disorders. The propagation of skin-resident lymphocytes, seen especially in method A, seems to be mediated by a functional defect of regulatory T cells residing in skin sequentially expanding under the conditions of our methods.