1998 Fiscal Year Final Research Report Summary
Development of a novel method for the selective expansion of human NK cells using immobilized lgG
| Project/Area Number |
09671215
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General surgery
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| Research Institution | The University of Tokyo |
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Principal Investigator |
MIKAMO Shinsuke The Institute of Medical Science, The University of Tokyo, Research Associate, 医科学研究所, 助手 (10107435)
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| Project Period (FY) |
1997 – 1998
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| Keywords | NK cells / lgG / CD16 / CD25 / Adhesion / cell death / expansion / lL12 |
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| Research Abstract |
Vujanovic and colleagues, described a procedure to expand antitumor effector cells from rat, and the technique was applied to human cancer therapy. The A-NK cells represented a small subset of PBMC, but expanded up to 1000-fold. Recently they tried to combine PBSCT and A-NK cell transfer, with better clinical outcome. We developed a new method to expand similar NK cells from resting NK cells from human PBMC, using immobilized IgG(ilgG).
Although NK cells were activated with ilgG, cell death was frequently observed. Overcontaminated Mo killed NK cells, A few % contamination showed dual effect, supporting NK cell growth and killing them, at the same time. .There might be exist activation induced cell death also.
The procedure consists of three steps. 1) Initial cell preparation and stimulation. 2) Activated NK cells became selectively adherent to the plastic and non-adherent T cells could be separated. 3) Adherent NK cells spontaneously detached from the plastic during the several hours of stimulation period and could be separated from contaminated Mo. The 3 steps offered differential treatments of the cultures.
Culture conditions, including co-existence of Mo and T cells, addition of IL2 and/or IL12, were tested. Then, a novel method was developed for activation without cell death.
NK cells from a healthy donor were activated by the method and a NK culture of high purity was obtained expanding 1000 fold. This method might bring forward a new strategy for adoptive immunotherapy.
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