T cell stimulation and expansion using anti-CD3/CD28 beads.
Following appropriate stimulation, T lymphocytes will proliferate extensively in vitro. Traditionally, mitogenic lectins such as phytohemagglutinin (PHA) and concanavalin A (Con A) have been used for polyclonal T cell stimulation. A more physiologically relevant approach uses beads coated with anti-CD3 and anti-CD28 to stimulate T cells in a manner that partially mimics stimulation by antigen-presenting cells. This protocol describes the steps involved in T cell stimulation and their subsequent in vitro expansion using anti-CD3/CD28 beads.
Trickett
Annette
A
Clinical Haematology and Haematology-SEALS, St. George Hospital, Sydney, NSW, Australia. a.trickett@unsw.edu.au
Kwan
Yiu Lam
YL
D000911:Antibodies, Monoclonal / Q000008:administration & dosage*; D018106:CD28 Antigens; D017252:CD3 Complex; D006801:Humans; D007158:Immunologic Techniques; D066298:In Vitro Techniques; D008213:Lymphocyte Activation / Q000187:drug effects; D008863:Microspheres; D010835:Phytohemagglutinins / Q000494:pharmacology; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology*
D000911:Antibodies, Monoclonal; D018106:CD28 Antigens; D017252:CD3 Complex; D010835:Phytohemagglutinins