Shenouda Mira M, Gillgrass Amy, Nham Tina, Hogg Richard, Lee Amanda J, Chew Marianne V, Shafaei Mahsa, Aarts Craig, Lee Dean A, Hassell John, Bane Anita, Dhesy-Thind Sukhbinder, Ashkar Ali A
Department of Pathology and Molecular Medicine, McMaster Immunology Research Centre, McMaster University, 1280 Main Street West, MDCL 4015, Hamilton, ON, Canada.
Cellular Therapy and Cancer Immunology Program, Department of Hematology/Oncology and BMT, Nationwide Children's Hospital, The Ohio State University Comprehensive Cancer Center, Ohio, USA.
Breast Cancer Res. 2017 Jul 1;19(1):76. doi: 10.1186/s13058-017-0867-9.
Natural killer (NK) cells play a critical role in cancer immunosurveillance. Recent developments in NK cell ex-vivo expansion makes it possible to generate millions of activated NK cells from a small volume of peripheral blood. We tested the functionality of ex vivo expanded NK cells in vitro against breast cancer cell lines and in vivo using a xenograft mouse model. The study aim was to assess functionality and phenotype of expanded NK cells from breast cancer patients against breast cancer cell lines and autologous primary tumours.
We used a well-established NK cell co-culture system to expand NK cells ex vivo from healthy donors and breast cancer patients and examined their surface marker expression. Moreover, we tested the ability of expanded NK cells to lyse the triple negative breast cancer and HER2-positive breast cancer cell lines MDA-MB-231 and MDA-MB-453, respectively. We also tested their ability to prevent tumour growth in vivo using a xenograft mouse model. Finally, we tested the cytotoxicity of expanded NK cells against autologous and allogeneic primary breast cancer tumours in vitro.
After 3 weeks of culture we observed over 1000-fold expansion of NK cells isolated from either breast cancer patients or healthy donors. We also showed that the phenotype of expanded NK cells is comparable between those from healthy donors and cancer patients. Moreover, our results confirm the ability of ex vivo expanded NK cells to lyse tumour cell lines in vitro. While the cell lines examined had differential sensitivity to NK cell killing we found this was correlated with level of major histocompatibility complex (MHC) class I expression. In our in vivo model, NK cells prevented tumour establishment and growth in immunocompromised mice. Finally, we showed that NK cells expanded from the peripheral blood of breast cancer patients show high cytotoxicity against allogeneic and autologous patient-derived tumour cells in vitro.
NK cells from breast cancer patients can be expanded similarly to those from healthy donors, have a high cytotoxic ability against breast cancer cell lines and patient-derived tumour cells, and can be compatible with current cancer treatments to restore NK cell function in cancer patients.
自然杀伤（NK）细胞在癌症免疫监视中发挥关键作用。NK细胞体外扩增技术的最新进展使得从小体积外周血中产生数百万活化的NK细胞成为可能。我们在体外针对乳腺癌细胞系以及在体内使用异种移植小鼠模型测试了体外扩增的NK细胞的功能。本研究的目的是评估乳腺癌患者来源的扩增NK细胞针对乳腺癌细胞系和自体原发性肿瘤的功能及表型。
我们使用成熟的NK细胞共培养系统从健康供体和乳腺癌患者体外扩增NK细胞，并检测其表面标志物表达。此外，我们分别测试了扩增的NK细胞裂解三阴性乳腺癌细胞系MDA-MB-231和HER2阳性乳腺癌细胞系MDA-MB-453的能力。我们还使用异种移植小鼠模型测试了它们在体内预防肿瘤生长的能力。最后，我们在体外测试了扩增的NK细胞对自体和异体原发性乳腺癌肿瘤的细胞毒性。
培养3周后，我们观察到从乳腺癌患者或健康供体分离的NK细胞扩增了1000倍以上。我们还表明，健康供体来源的扩增NK细胞与癌症患者来源的扩增NK细胞在表型上具有可比性。此外，我们的结果证实了体外扩增的NK细胞在体外裂解肿瘤细胞系的能力。虽然所检测的细胞系对NK细胞杀伤的敏感性不同，但我们发现这与主要组织相容性复合体（MHC）I类分子的表达水平相关。在我们的体内模型中，NK细胞可防止免疫缺陷小鼠体内肿瘤的形成和生长。最后，我们表明从乳腺癌患者外周血扩增的NK细胞在体外对异体和自体患者来源的肿瘤细胞具有高细胞毒性。
乳腺癌患者的NK细胞与健康供体的NK细胞一样能够扩增，对乳腺癌细胞系和患者来源的肿瘤细胞具有高细胞毒性，并且可以与当前的癌症治疗方法兼容，以恢复癌症患者的NK细胞功能。
