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Generation and ex vivo expansion of HTLV‐1 specific CD8 + cytotoxic T‐lymphocytes for adoptive immunotherapy
Author(s) -
Peshwa Madhusudan V.,
Page Laura A.,
Qian Lichuan,
Yang Demao,
van Schooten Wim C. A.
Publication year - 2000
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/(sici)1097-0290(19960605)50:5<529::aid-bit7>3.0.co;2-h
Subject(s) - cytotoxic t cell , biology , cd8 , antigen , adoptive cell transfer , immunology , streptamer , immune system , antigen presenting cell , immunotherapy , microbiology and biotechnology , t cell , interleukin 21 , in vitro , virology , biochemistry
CD8 + cytotoxic T‐lymphocytes (CTLs) have been proven, in multiple animal models, to be the most powerful antiviral and antitumor components of the immune system. We have developed a protocol to activate and expand tumor and virus peptide‐specific CD8 + T‐lymphocytes from the peripheral blood of healthy, human trophic leukemia virus‐1 (HTLV‐1) seronegative human leucocyte antigen (HLA)‐A*0201 individuals. A combination of density‐based separation and culture conditions was employed to isolate dendritic cells (DCs), which are the most potent antigen‐presenting cells (APCs), and T‐lymphocytes. The DCs were pulsed with HLA‐A*0201 binding peptides and cultured with autologous T‐lymphocytes to generate peptide‐specific CTLs. The CTLs were generated against a nine‐amino‐acid peptide from the Tax protein of HTLV‐1. The CTLs were expanded according to a restimulation schedule employing peptide‐pulsed autologous monocytes and low‐dose interleukin‐2 (IL‐2) to numbers in excess of 100 × 10 6 cells following 5 weeks of culture. Expanded cells contained primarily CD3 + T‐cells, of which CD8 + T‐lymphocytes constituted greater than two‐thirds of the cell population. Obtained CTLs exhibited potent antigen‐specific lysis of peptide‐pulsed target cells in a dose‐dependent fashion in in vitro 51 Cr release cytotoxicity assay. This antigen‐specific killing was shown to be HLA class I restricted and mediated by CD8 + T‐lymphocytes. Since the T‐lymphocytes were obtained from HTLV‐1 seronegative donors, the generation of peptide‐specific CTLs represents reliable and reproducible elicitation of a primary immune response in vitro against naive antigens and subsequent expansion of generated CTLs for adoptive immunotherapy. © 1996 John Wiley & Sons, Inc.