Large-scale culture system of human CD4+ helper/killer T cells for the application to adoptive tumour immunotherapy.
A simple method for the rapid expansion of human CD4+ T cells with both helper and killer functions was established. CD4+ T cells separated from peripheral blood mononuclear cells using immunomagnetic beads were stimulated with immobilised OKT-3 monoclonal antibody (mAb) plus recombinant interleukin 2 (rIL-2) in 96 well culture plates. After 6 day-culture, the CD4+ T cells were restimulated by immobilised OKT-3 mAb for an additional 24 h, then inoculated into concentrated rotary-tissue culture bag and cultured for further 9 days. This procedure yielded a 3000-fold increase in cell number (about 3-5 x 10(9) per bag). Most of the cells (over 96%) continued to express CD4+ antigen and retained their capacity to produce IL-2. The activated CD4+ T cells showed marked cytotoxicity against Fc receptor positive tumour cells in the presence of OKT-3 mAb. Moreover, we succeeded in a specific targeting of the expanded CD4+ helper/killer T cells to c-erb B-2 positive tumour cells by means of anti-CD3 x anti-c-erb B-2 bispecific antibody. These results suggested that our established simple system will be available for the expansion of large number of CD4+ helper/killer T cells which may provide an efficient strategy for adoptive tumour immunotherapy.
Blood Transfusion Service Center, Tokai University School of Medicine, Isehara, Japan.
Y
Y
Nakamura
Y
Y
Tokuda
M
M
Iwasawa
H
H
Tsukamoto
M
M
Kidokoro
N
N
Kobayashi
S
S
Kato
T
T
Mitomi
S
S
Habu
T
T
Nishimura
D000911:Antibodies, Monoclonal; D000918:Antibody Specificity; D014408:Biomarkers, Tumor / Q000032:analysis; D015704:CD4 Antigens / Q000032:analysis / Q000276:immunology*; D002455:Cell Division; D002478:Cells, Cultured; D046508:Culture Techniques / Q000295:instrumentation* / Q000379:methods; D005434:Flow Cytometry; D006801:Humans; D016219:Immunotherapy, Adoptive; D007694:Killer Cells, Natural / Q000166:cytology* / Q000637:transplantation; D009369:Neoplasms / Q000276:immunology / Q000628:therapy*; D011518:Proto-Oncogene Proteins / Q000032:analysis / Q000235:genetics; D011519:Proto-Oncogenes; D018719:Erb-b2 Receptor Tyrosine Kinases; D016176:T-Lymphocyte Subsets / Q000166:cytology* / Q000637:transplantation; D006377:T-Lymphocytes, Helper-Inducer / Q000166:cytology* / Q000637:transplantation
D000911:Antibodies, Monoclonal; D014408:Biomarkers, Tumor; D015704:CD4 Antigens; D011518:Proto-Oncogene Proteins; D018719:Erb-b2 Receptor Tyrosine Kinases