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.
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 10(7) 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 beta chain variable region (TCR-Vbeta) 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.
Hashizume
Hideo
H
Department of Dermatology, Aarhus University Hospital, Aarhus, Denmark. hihashiz@hama-med.ac.jp
Hansen
Anker
A
Poulsen
Lars K
LK
Thomsen
Allan Randrup
AR
Takigawa
Masahiro
M
Thestrup-Pedersen
Kristian
K
D000328:Adult; D000368:Aged; D000906:Antibodies; D001706:Biopsy; D018106:CD28 Antigens / Q000276:immunology*; D017252:CD3 Complex / Q000276:immunology*; D018929:Cell Culture Techniques; D002460:Cell Line; D049109:Cell Proliferation; D002469:Cell Separation; D003602:Cytotoxicity, Immunologic; D003718:Denmark; D005260:Female; D005434:Flow Cytometry; D006801:Humans; D007376:Interleukin-2 / Q000378:metabolism*; D015847:Interleukin-4 / Q000378:metabolism*; D007564:Japan; D008297:Male; D008863:Microspheres; D008875:Middle Aged; D010641:Phenotype; D016693:Receptors, Antigen, T-Cell, alpha-beta / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D012867:Skin / Q000276:immunology* / Q000473:pathology; D013601:T-Lymphocytes / Q000276:immunology* / Q000473:pathology; D013997:Time Factors
D000906:Antibodies; D018106:CD28 Antigens; D017252:CD3 Complex; C508594:IL2 protein, human; C508598:IL4 protein, human; D007376:Interleukin-2; D016693:Receptors, Antigen, T-Cell, alpha-beta; D011994:Recombinant Proteins; D015847:Interleukin-4