[Biological characteristics and antitumor activity of CIK cells activated by recombinant human fibronectin for human lung cancer cell lines in vitro].
OBJECTIVE
The CIK cell is one of the most important means of the adoptive cellular immunotherapy, and it is a hotspot of which to simplify its culture procedure and to promote its inhibition rate. The aim of this study is to observe the biological function of the CIK cells cultivated by the recombinant human fibronectin (RN) and to establish an effective and simple way of cells expansion.


METHODS
We separated the mononuclear cells (PBMCs) in 50 mL peripheral blood from 10 healthy persons with density gradient centrifugation in the lymphocyte-separating medium, and the PBMCs were divided into two groups, of which were cultivated by RN-introduced and conventional method separately. Then we estimated the proliferation ability, and analyzed the immunologic type, IFN-gamma, IL-4, perforin and granzyme B of them with flow cytometry. Besides that, we tested the inhibition rate of CIKs cells to four kinds of human lung cancer cell lines in vitro by MTT assay.


RESULTS
The RN-induced group had a higher proliferation rate that was 2.0-3.5 times of the conventional group, and there was an obvious statistical difference between the two (P < 0.05). The proliferation rates of CD3+CD16+CD56+T cells in each group were 3 778 and 2 068 times of the initial number, respectively. There was also a higher percentage of CD3+CD8+ T cells in RN-induced group (P < 0.05), while the percentage of CD3+CD4+T cells had no significant statistical difference (P > 0.05). We found a similar inhibition rate of the CIK cells to all this human lung cancer cell lines (P > 0.05). The cells which secreted IFN-gamma increased, while the cells which secreted IL-4 did not. The cells which secreted granzyme B and perforin were positive.


CONCLUSIONS
It is an effective and simple way to cultivate the CIK cells with RN, which should be adopted.
Wang
Shiyong
S
Department of Biotherapy, Fourth Affiliated Hospital of China Medical University, Shenyang 110032, China. sywang66@yahoo.com
Du
Weili
W
Zhang
Hui
H
Wulan
Tuya
T
Zhang
Yuan
Y
He
Ying
Y
Yang
Yunfeng
Y
Liu
Sa
S
Zhang
Zhe
Z
Wang
Jialing
J
D017252:CD3 Complex / Q000378:metabolism; D015496:CD4-Positive T-Lymphocytes / Q000276:immunology; D019002:CD56 Antigen / Q000378:metabolism; D045744:Cell Line, Tumor; D002478:Cells, Cultured; D055612:Cytokine-Induced Killer Cells / Q000187:drug effects* / Q000378:metabolism*; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D005434:Flow Cytometry; D006801:Humans; D016219:Immunotherapy, Adoptive / Q000379:methods*; D015847:Interleukin-4 / Q000378:metabolism; D008175:Lung Neoplasms / Q000628:therapy*; D017452:Receptors, IgG / Q000378:metabolism; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D013601:T-Lymphocytes / Q000276:immunology
D017252:CD3 Complex; D019002:CD56 Antigen; D005353:Fibronectins; D017452:Receptors, IgG; D011994:Recombinant Proteins; D015847:Interleukin-4