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The Experimental Study on Inducing and Expanding CD56~+ Lymphocytes from Mononuclear Cells of Human Peripheral Blood and Bone Marrow with IL-2/anti-IL-2 Complex

Author: RenWeiFeng
Tutor: ZhaoChunTing；LiGuangLun
School: Qingdao University
Course: Internal Medicine
Keywords: IL-2 / IL-2 antibody was anti- CD56 ~ lymphocytes Cytotoxicity In vitro Amplification
CLC: R392
Type: Master's thesis
Year: 2009
Downloads: 35
Quote: 0
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Abstract


Objective To explore a from human peripheral blood mononuclear cells (PBMC) and bone marrow CD56 ~ efficient cytotoxic lymphocyte amplification method . Methods of stem cell growth medium (SCGM) based on the medium , the design IL-2 / anti- IL-2 antibody complexes (IL-2 and anti- IL-2 antibody was mixed at a mass ratio 1:20 ) experimental group , the corresponding concentration factor the IL-2, IL-2 antibody and anti- IL-2 IL-15 three control groups , respectively, from PBMC and bone marrow mononuclear cells induced amplification of CD56 ~ lymphocytes were detected by flow cytometry of peripheral blood -derived cells 7,10 7,14 days and bone marrow-derived cells in the first days of CD3/CD56 expression . With four methyl thiazolyl tetrazolium (MTT) assay and on tumor cell killing activity of K562 , respectively, compared with the control group activated killer cells for comparison. Results -based culture medium with SCGM , CD56 ~ lymphocytes by IL-2 / anti- IL-2 antibody for access to proliferate . In cultured for 7 and 10 days, peripheral blood CD56 ~ cells in the experimental group were amplified (24.67 ± 3.14) and (31.63 ± 5.01) times ; including CD3 ~ CD56 ~ and CD3 ~-CD56 ~ cells were amplified ( 110.93 ± 19.10), (157.60 ± 46.31) and (7.8 ± 1.17), (12.03 ± 1.64) times , compared with the control group , the difference was statistically significant (P <0.05 ) ; in the culture section 7 and 14 days, bone marrow cells in the experimental group were amplified CD56 ~ (11.08 ± 2.08) and (21.72 ± 2.19) times ; including CD3 ~ CD56 ~ and CD3CD56 ~ cells were amplified (30.30 ± 5.14), (60.18 ± 6.68) and (6.05 ± 1.05), (10.75 ± 1.51) times , compared with the control group , the difference was statistically significant (P < 0.05) ; cell efficiency / T ratio of 10:1 , the peripheral blood and bone marrow cells in the experimental group on K562 cell killing rates were (83.46 ± 1.56)% and (74.94 ± 2.28)%; compared with the same period in the control cells have a stronger lymphocyte cytotoxicity (P <0.05 ) . Conclusion IL-2 / IL-2 antibody anti (IL-2 / IL-2 antibody was anti- mass ratio of 1 / 20 ) under the action of found objects to SCGM based medium to obtain a large number of lymphocytes CD56 ~ cytotoxic immune effector cells for adoptive immunotherapy of the tumor offers a new CD56 ~ lymphocytes in vitro amplification methods .

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