Zhao Zhi-gang, Sun Li, Wang Xiao-fang, Zhang Yi-zhuo, Yu Yong, Yang Hong-liang, Zou Ping
Department of Hematology, Tianjin Medical University Cancer Institute and Hospital, Tianjin 300020, China.
Zhonghua Zhong Liu Za Zhi. 2011 Feb;33(2):105-9.
To study the immunomodulatory effects and mechanisms of mesenchymal stem cells (MSC) derived from the bone marrow in acute leukemia patients in vitro.
Bone marrow mononuclear cells from acute myeloid leukemia (AML) and acute lymphoblastic leukemia (ALL) were obtained and cultured in low serum medium. The immunophenotypes were assessed by FACS and immunol histochemistry. The levels of cytokines were evaluated by enzyme linked immunosorbant assay (ELISA). T-cell suppression ability was evaluated by Transwell chamber assay. Moreover, the immunoregulatory ability of AML- and ALL-derived MSC was detected by mixed lymphocyte culture assay.
ALL-derived MSC showed a typical fibroblast-like morphology. They were positive for CD29, CD44 and CD105, the positive rate were 98.81%, 99.25% and 90.52%, respectively, while negative for CD31, CD45 and CD34. Moreover, ALL- and AML-derived MSC didn't express HLA-DR and co-stirnulatory molecules (CD40, CD80 and CD86). ALL and AML derived MSC could secret several cytokines, such as TGF-β1 (567.58 ± 52.64 and 357.15 ± 33.52), HGF (647.27 ± 102.54 and 219.67 ± 62.37), IL-6 (59.67 ± 15.69 and 54.35 ± 12.31) and IL-11 (102.58 ± 23.54 and 78.21 ± 9.67), the level of secretion of TGF-β1 and HGF were higher in ALL bone marrow derived MSC than that of in AML bone marrow derived MSC. ALL and AML derived MSC significantly suppressed T lymphocyte proliferation in a dose-dependent manner, the counts per minute (CPM) were (3.58 ± 0.54) × 10(4), (2.87 ± 0.33) × 10(4), (1.78 ± 0.51) × 10(4) and (1.15 ± 0.15) × 10(4) for AML derived MSC, and CPM were (1.96 ± 0.31) × 10(4), (1.57 ± 0.28) × 10(4), (0.91 ± 0.41) × 10(4) and (0.22 ± 0.11) × 10(4) for ALL derived MSC when MSC were 0.5 × 10(4), 1 × 10(4), 2 × 10(4) and 5 × 10(4). In addition, the CPM was (4.01 ± 0.72) × 10(4) in control group. The immunosuppressive ability was different between MSCs derived from AML and ALL. The immunosuppressive effect of ALL derived MSC could be reversed by anti-TGF-β1 and anti-HGF antibody.
ALL-derived MSC show immunoregulatory effect in vitro and this effect is achieved through cytokines. But MSCs derived from AML display abnormal changes in T-cell suppression ability.
研究急性白血病患者骨髓间充质干细胞（MSC）的体外免疫调节作用及机制。
获取急性髓系白血病（AML）和急性淋巴细胞白血病（ALL）患者的骨髓单个核细胞，在低血清培养基中培养。通过流式细胞术（FACS）和免疫组织化学评估免疫表型。采用酶联免疫吸附测定（ELISA）评估细胞因子水平。通过Transwell小室测定评估T细胞抑制能力。此外，通过混合淋巴细胞培养测定检测AML和ALL来源的MSC的免疫调节能力。
ALL来源的MSC呈现典型的成纤维细胞样形态。它们CD29、CD44和CD105呈阳性，阳性率分别为98.81%、99.25%和90.52%，而CD31、CD45和CD34呈阴性。此外，ALL和AML来源的MSC不表达HLA-DR和共刺激分子（CD40、CD80和CD86）。ALL和AML来源的MSC可分泌多种细胞因子，如TGF-β1（567.58±52.64和357.15±33.52）、HGF（647.27±102.54和219.67±62.37）、IL-6（59.67±15.69和54.35±12.31）和IL-11（102.58±23.54和78.21±9.67），ALL骨髓来源的MSC中TGF-β1和HGF的分泌水平高于AML骨髓来源的MSC。ALL和AML来源的MSC以剂量依赖性方式显著抑制T淋巴细胞增殖，当MSC分别为0.5×10⁴、1×10⁴、2×10⁴和5×10⁴时，AML来源的MSC的每分钟计数（CPM）分别为（3.58±0.54）×10⁴、（2.87±0.33）×10⁴、（1.78±0.51）×10⁴和（1.15±0.15）×10⁴，ALL来源的MSC的CPM分别为（1.96±0.31）×10⁴、（1.57±0.28）×10⁴、（0.91±0.41）×10⁴和（0.22±0.11）×10⁴。此外，对照组的CPM为（4.01±0.72）×10⁴。AML和ALL来源的MSC的免疫抑制能力不同。ALL来源的MSC的免疫抑制作用可被抗TGF-β1和抗HGF抗体逆转。
ALL来源的MSC在体外具有免疫调节作用，且这种作用是通过细胞因子实现的。但AML来源的MSC在T细胞抑制能力方面表现出异常变化。
