Department of Small Animal Medicine and Surgery, College of Veterinary Medicine, University of Georgia, Athens, GA, USA.
Department of Population Health, College of Veterinary Medicine, University of Georgia, Athens, GA, USA.
J Feline Med Surg. 2022 Jun;24(6):e43-e56. doi: 10.1177/1098612X221083074. Epub 2022 Mar 18.
Feline autologous mesenchymal stem cells (MSCs) show promise for immunomodulatory activity, but the functional impact of chronic kidney disease (CKD), concurrent immunosuppressive drug administration or infection is unknown. The study objectives compare endogenous cytokine gene expression (interleukin [IL]-6, IL-10, IL-12p40, IL-18 and transforming growth factor beta [TGF-β]) in adipose-derived MSCs (aMSCs) from cats with and without CKD, following in vitro exposure to microbial ligands and treatment with common immunosuppressive drugs.
Previously obtained aMSCs, phenotype CD44, CD90, CD105 and MHCII, from cats with (n = 6) and without (n = 6) CKD were compared via real-time PCR (RT-PCR) for immunomodulatory gene expression. aMSCs were exposed in vitro to lipopolysaccharide (LPS), peptidoglycan or polyinosinic:polycytidylic acid (Poly I:C), simulating bacterial or viral exposure, respectively. aMSCs were also exposed to ciclosporin, dexamethasone or methotrexate. Gene expression was measured using RT-PCR, and C was utilized after each run to calculate the delta cycle threshold.
aMSCs isolated from healthy and CKD cats showed no significant differences in gene expression in the five measured cytokines. No significant changes in measured gene expression after drug treatment or microbial ligand stimulation were observed between normal or CKD affected cats. Proinflammatory genes (IL-6, IL-12p40 and IL-18) showed altered expression in aMSCs from both groups when compared with the same cells in standard culture after exposure to methotrexate. Poly I:C altered IL-6 and TGF-β gene expression in aMSCs from both healthy and CKD cats when compared with the same cells in standard culture.
The five genes tested showed no statistical differences between aMSCs from healthy or CKD cats. There was altered cytokine gene expression between the control and treatment groups of both healthy and CKD cats suggesting feline aMSCs have altered function with immunosuppressive treatment or microbial ligand exposure. Although the current clinical relevance of this pilot study comparing brief exposure to select agents in vitro in aMSCs from a small number of cats is unknown, the study highlights a need for continued investigation into the effects of disease and concurrent therapies on use of cell-based therapies in feline patients.
猫自体间充质干细胞（MSCs）具有免疫调节活性的潜力，但慢性肾病（CKD）、同时使用免疫抑制药物或感染对其的功能影响尚不清楚。本研究的目的是比较有和无 CKD 的猫脂肪来源间充质干细胞（aMSCs）在体外暴露于微生物配体和常见免疫抑制药物治疗后的内源性细胞因子基因表达（白细胞介素[IL]-6、IL-10、IL-12p40、IL-18 和转化生长因子β[TGF-β]）。
通过实时 PCR（RT-PCR）比较有（n=6）和无（n=6）CKD 的猫的已获得的 aMSCs 的免疫调节基因表达，表型为 CD44、CD90、CD105 和 MHCII。aMSCs 分别暴露于脂多糖（LPS）、肽聚糖或聚肌苷酸：聚胞苷酸（Poly I:C），分别模拟细菌或病毒暴露。aMSCs 还暴露于环孢素、地塞米松或甲氨蝶呤。使用 RT-PCR 测量基因表达，每个运行后使用 C 来计算 delta 循环阈值。
从健康和 CKD 猫中分离的 aMSCs 在 5 种测量细胞因子的基因表达中没有显著差异。在正常或受 CKD 影响的猫中，药物治疗或微生物配体刺激后，未观察到测量的基因表达有显著变化。与暴露于甲氨蝶呤后的相同细胞相比，来自两组的 aMSCs 的促炎基因（IL-6、IL-12p40 和 IL-18）的表达发生改变。与标准培养中的相同细胞相比，Poly I:C 改变了健康和 CKD 猫的 aMSCs 中的 IL-6 和 TGF-β 基因表达。
在健康和 CKD 猫的 aMSCs 之间，测试的 5 个基因没有统计学差异。健康和 CKD 猫的对照组和治疗组之间的细胞因子基因表达发生改变，表明猫的 aMSCs 在接受免疫抑制治疗或微生物配体暴露时功能发生改变。虽然目前比较少数猫的 aMSCs 体外短暂暴露于选择剂的这项初步研究的临床相关性尚不清楚，但该研究强调需要继续研究疾病和同时进行的治疗对猫患者细胞治疗的应用的影响。
