Abstract
We recently developed a new culture system based on dialysis perfusion (designated JCC-device) for the generation and expansion of human lymphokine-activated killer (LAK) cells (Murata et al., 1990). More recently we have scaled up the volume of the culture vessel of the JCC-device from 100 ml to 400 ml for clinical use. In the present study, using this new 400 ml JCC-device, we cultured human lymph node lymphocytes (LNL) obtained from 8 surgical patients with primary lung cancer, and investigated the cellular characteristics in comparison with a conventional batchwise culture system using tissue culture dishes. With the JCC-device, the cell density reached a maximum 2.7×107 cells/ml with greater than 90% viability by the appropriate exchange of perfusion medium and by making additions at the appropriate intervals for recombinant interleukin-2 (rIL-2). The expansion fold of LNL with the JCC-device, ranging 6.6- to 19.2-fold (mean 13.8-fold), was not significantly different from that in dish cultures. There was no marked difference in cell surface phenotypes between the two culture systems in 7 out of 8 cases. As for LAK activity of LNL, the JCC culture was either superior or equal in 4 out of 8 cases, but inferior in the other 4 cases to the conventional dish cultures. In the latter cases, the usage of serum for the JCC culture was limited, which might have resulted in the low LAK activity. The JCC-device was able to reduce the consumption of basal medium, rIL-2 and serum by 20%, 84% and 96%, respectively compared to the conventional tissue culture systems. The JCC-device improved the routine performance of adoptive immunotherapy with LAK cells and rIL-2.
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Abbreviations
- LAK:
-
lymphokine-activated killer
- rIL-2:
-
recombinant interleukin-2
- LNL:
-
lymph node lymphocytes
- BM:
-
basal medium
- CM:
-
complete medium
- HBSS:
-
Hanks balanced salt solution
- JRU:
-
Japan reference unit
References
EpsteinMA, AchongBG, BarrYM, ZajacB, HenleG and HenleW (1966) Morphological and virological investigations on cultured Burkitt tumor lymphoblasts (strain Raji). J. Natl. Cancer Inst. 37: 549–559.
HoyerM, MeinekeT, LewisW, ZwillingB and ReinhartJ (1986) Characterization and modulation of human lymphokine (interleukin 2) activated killer cells induction. Cancer. Res. 46: 2834–2838.
KatoK, YamadaT, KawaharaK, OndaH, AsanoT, SuginoH and KakinumaA (1985) Purification and characterization of recombinant human interleukin-2 produced Esherichia coli. Biochem. Biophys. Res. Commun. 130: 692–699.
LozzioCB and LozzioBB (1973) Cytotoxicity of a factor isolated from human spleen. J. Natl. Cancer Inst. 50: 535–545.
MurataM, YanoT, TogamiM, YasumotoK, SugimachiK, KimuraG and NomotoK (1990) Development of a new culture system for human lymphokine activated killer cells. J. Immunol. Methods 129: 89–95.
OchoaAC, GromoG, AlterBJ, SondelPM and BachFH (1987) Long-term growth of lymphokine-activated killer (LAK) cells: role of anti-CD3, β-IL-1, interferon-γ, and β. J. Immunol. 138: 2728–2733.
RosenbergSA, LotzeMT, MuulLM, ChangAE, AvisFP, LeitmanS, LinehanWM, RobertsonCN, RobertaEL, RubinJT, SeippCA, SimpsonCG and WhiteDE (1987) A progress report on the treatment of 157 patients with advanced cancer using lymphokine-activated killer cells and interleukin-2 or high dose interleukin-2 alone. N. Engl. J. Med. 316: 889–897.
WestWH, TauerKW, YannelliJR, MarshallGD, OrrDW, ThurmanGB and OldhamRK (1987) Constant infusion recombinant interleukin-2 in adoptive immunotherapy of advanced cancer. N. Engl. J. Med. 316: 898–905.
YanoT, YasumotoK and NomotoK (1989) Generation and expansion of lymphokine-activated killer cells from lymph node lymphocytes in human lung cancer. Eur. J. Cancer Clin. Oncol. 25: 201–208.
YanoT, MurataM, IshidaT, MitsudomiT, KimuraG, SugimachiK and NomotoK (1989) Phenotypic characterization of lymphokine-activated killer cells from human lymph node lymphocytes. Cell. Immunol. 122: 281–294.
YanoT, IshidaT, YoshinoI, MurataM, YasumotoK, KimuraG, NomotoK and SugimachiK (1991) A regimen of surgical adjuvant immunotherapy for cancer with interleukin-2 and lymphokine-activated killer cells. Biotherapy. 3: 245–251.
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Murata, M., Yano, T., Yoshino, I. et al. Development of a new culture system for human lymphokine-activated killer cells: Comparison with a conventional static culture method. Cytotechnology 7, 75–83 (1991). https://doi.org/10.1007/BF00350913
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DOI: https://doi.org/10.1007/BF00350913