LMSSitesePortfolio註冊登入
P-17 Apoptotic neuroblastoma cells could induce mesenchymal stromal cells into immuno-stimulatory phenotype with cytotoxic potential
by 陳輔卿, 2015-10-25 07:48, 人氣(742)
P-17 
 

Apoptotic neuroblastoma cells could induce mesenchymal stromal cells into immuno-stimulatory phenotype with cytotoxic potential

 

Jiamin Cao, Godfrey Chi-Fung Chan

Department of Paediatrics & Adolescent medicine, The University of Hong Kong, HKSAR, China

Background

Tumors exist in an inflammatory microenvironment, and mesenchymal stromal cells (MSCs, or mesenchymal stem cells) is part of this complex. We previously reported that MSCs can engulf apoptotic bodies of lymphoid cells and turn into immuno-stimulatory cells. In patients undergo chemotherapy, large amounts of apoptotic cells will be released to the cancer microenvironment. Little is known about the impact of these apoptotic particles on MSCs.  Therefore, in this study, we aim to explore the role of apoptotic cells on MSCs as in the cancer microenvironment.

Method

Luciferase tranasduced neuroblastoma cells (SKNLP) were treated with cisplatin in vitro and yielded a collection of ACs as defined by flow cytometry (FC-PI). The ACs were then collected by ultracentrifugation and washed, then co-cultured with human derived MSCs. The co-culture supernatant was harvested 3 days after co-culture, and used as conditional medium (CM). SKNLP was then treated with CM and assayed using flow cytometry (AV-PI) to assess the degree of apoptosis. Caspase-3 colorimetric protease assay kit was utilized to investigate the activation of caspase dependent pathway. To study the immune-modulatory effect of AC stimulated MSCs on macrophages, the CM was added to macrophages and the cytokines production was measured by ELISA assay.

Result

We found that the CM could induce significant cell-death of neuroblastoma cells as shown by increase AV+/PI- and AV+/PI+ cells. It is mediated via the caspase dependent apoptotic pathway as shown by increase caspase-3 activation. In addition, CM yielded from the co-culture of MSCs and APs would stimulate TNF-a but not IL-10 secretion by macrophages.

Conclusion

MSCs stimulated by APs derived from post-chemotherapy treated neuroblastoma cells exhibited cytotoxicity and pro-inflammatory potential. Hence, APs stimulated MSCs may serve as immunomodulator for the host’s macrophages at the tumor microenvironment.