Increased Expression of TGF-β and IFN-γ in Peripheral Blood Mononuclear Cells (PBMCs) Cultured in Conditioned Medium (CM) of K562 Cell Culture.

A. A. Mohamed Adil, Lavanya Vallinayagam, K. Chitra, Shazia Jamal, A. Pandurangan, Neesar Ahmed
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引用次数: 1

Abstract

In the present study, we investigated the effects of conditioned media (CM) collected from the cancer cell lines (K562, MCF-7, and HeLa) on peripheral blood mononuclear cells (PBMCs) isolated from the healthy human blood. The soluble factors in the CM are probably responsible for the differential mRNA expressions of Foxp3, Helios, Neuropilin- 1 (NRP-1), and glycoprotein A repetitions predominant (GARP), along with IFN-γ and TGF-β in PBMCs cultured with cancer cells CM. The PBMCs cultured with CM of K562 showed increased expression of Foxp3, Helios, NRP-1, GARP, IFN-γ, and TGF-β compared to PBMCs cultured with CM of MCF-7 and HeLa cells. In addition, the intracellular staining on PBMCs cultured with CM from cell lines were also evaluated for CD4, CD25, Foxp3, Helios, and NRP-1 by multicolor flow cytometry. The expression of CD4+CD25+Foxp3+, CD4+Helios+Foxp3+ and CD+NRP-1+Foxp3+ showed retarded cell population compared to control PBMCs. Our data suggest that soluble factors in CM of cancer cells may trigger the immune response in PBMCs resulting in a systematic response. Further research could lead to the identification of specific soluble factors that are involved in trafficking of cells into the immune cascades, which could be a safe and promising strategy for targeting human cancers.
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K562细胞培养条件培养基(CM)培养外周血单个核细胞(PBMCs)中TGF-β和IFN-γ的表达升高。
在本研究中,我们研究了从癌细胞系(K562、MCF-7和HeLa)中收集的条件培养基(CM)对从健康人血液中分离的外周血单个核细胞(PBMCs)的影响。CM中的可溶性因子可能是导致Foxp3、Helios、Neuropilin- 1 (NRP-1)、糖蛋白A重复显性(GARP)以及IFN-γ和TGF-β mRNA表达差异的原因。与MCF-7和HeLa细胞的CM培养的pbmc相比,K562 CM培养的pbmc中Foxp3、Helios、NRP-1、GARP、IFN-γ和TGF-β的表达增加。此外,采用多色流式细胞术检测细胞内CD4、CD25、Foxp3、Helios和NRP-1的表达。CD4+CD25+Foxp3+、CD4+Helios+Foxp3+和CD+NRP-1+Foxp3+的表达均表现为细胞群发育迟缓。我们的数据表明,癌细胞CM中的可溶性因子可能触发pbmc的免疫反应,导致系统性反应。进一步的研究可能会导致识别特定的可溶性因子,这些因子参与了细胞进入免疫级联的运输,这可能是针对人类癌症的一种安全和有希望的策略。
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