Direct access to CD4+ T cells specific for defined antigens according to CD154 expression.
The direct assessment of T helper (T(H))-cell responses specific for antigens is essential to evaluate pathogenic and protective immunity. Presently, analysis and isolation of antigen-specific T(H) cells is restricted to cells that produce cytokines, or can be performed only with a rare selection of specific peptide major histocompatibility complex class II (MHC II) multimers. Here we report a new method that enables the assessment and isolation of T(H) cells specific for a defined antigen according to CD154 expression induced after stimulation in vitro. We show that antigen-induced CD154 expression is highly sensitive and specific for human and mouse antigen-specific T(H) cells. Moreover, the isolation of antigen-specific CD154(+) T(H) cells necessitates only surface staining with antibodies, thereby enabling the fast generation of antigen-specific T(H) cell lines. Our approach allows assessment of T(H) cells with a defined specificity for the combined quantitative and qualitative analysis of T(H)-cell immunity as well as for the isolation of specific T(H) cells for targeted cellular immunotherapies.
Frentsch
Marco
M
Clinical Immunology Group, Deutsches Rheuma-Forschungszentrum, Berlin, Germany.
Arbach
Olga
O
Kirchhoff
Dennis
D
Moewes
Beate
B
Worm
Margitta
M
Rothe
Martin
M
Scheffold
Alexander
A
Thiel
Andreas
A
D000818:Animals; D000941:Antigens / Q000276:immunology*; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology* / Q000378:metabolism*; D023201:CD40 Ligand / Q000032:analysis / Q000378:metabolism*; D002469:Cell Separation / Q000379:methods*; D005434:Flow Cytometry; D005786:Gene Expression Regulation; D006801:Humans; D051379:Mice; D008807:Mice, Inbred BALB C; D018345:Mice, Knockout; D013379:Substrate Specificity
D000941:Antigens; D023201:CD40 Ligand