Institute of Cellular Therapeutics, Allegheny Health Network, Pittsburgh, PA, United States.
Department of Biological Sciences, Carnegie Mellon University, Pittsburgh, PA, United States.
Front Immunol. 2018 May 3;9:894. doi: 10.3389/fimmu.2018.00894. eCollection 2018.
Dendritic cells (DC) are important in the onset and severity of inflammatory bowel disease (IBD). Tolerogenic DC induce T-cells to become therapeutic Foxp3+ regulatory T-cells (Tregs). We therefore asked if experimental IBD could be prevented by administration of bone marrow-derived DC generated under conventional GM-CSF/IL-4 conditions but in the presence of a mixture of antisense DNA oligonucleotides targeting the primary transcripts of CD40, CD80, and CD86. These cell products (which we call AS-ODN BM-DC) have demonstrated tolerogenic activity in preventing type 1 diabetes and preserving beta cell mass in new-onset type 1 diabetes in the NOD mouse strain, in earlier studies. In addition to measuring efficacy in prevention of experimental IBD, we also sought to identify possible mechanism(s) of action. Weight, behavior, stool frequency, and character were observed daily for 7-10 days in experimental colitis in mice exposed to dextran sodium sulfate (DSS) following injection of the AS-ODN BM-DC. After euthanasia, the colons were processed for histology while spleen and mesenteric lymph nodes (MLNs) were made into single cells to measure Foxp3+ Treg as well as IL-10+ regulatory B-cell (Breg) population frequency by flow cytometry. AS-ODN BM-DC prevented DSS-induced colitis development. Recipients of these cells exhibited significant increases in Foxp3+ Treg and IL-10+ Breg in MLN and spleen. Histological examination of colon sections of colitis-free mice remained largely architecturally physiologic and mostly free of leukocyte infiltration when compared with DSS-treated animals. Although DSS colitis is mainly an innate immunity-driven condition, our study adds to the growing body of evidence showing that Foxp3+ Treg and IL-10 Bregs can suppress a mainly innate-driven inflammation. The already-established safety of human DC generated from monocytic progenitors in the presence of the mixture of antisense DNA targeting the primary transcripts of CD40, CD80, and CD86 in humans offers the potential to adapt them for clinical IBD therapy.
树突状细胞(DC)在炎症性肠病(IBD)的发病和严重程度中起重要作用。耐受原性 DC 诱导 T 细胞成为治疗性 Foxp3+调节性 T 细胞(Treg)。因此,我们询问是否可以通过给予在常规 GM-CSF/IL-4 条件下生成但存在针对 CD40、CD80 和 CD86 主要转录本的混合反义 DNA 寡核苷酸的骨髓来源的 DC 来预防实验性 IBD。这些细胞产物(我们称之为 AS-ODN BM-DC)在早期研究中已证明在预防 1 型糖尿病和保留新发生的 1 型糖尿病 NOD 小鼠中的胰岛β细胞质量方面具有耐受原性活性。除了测量预防实验性 IBD 的功效外,我们还试图确定可能的作用机制。在注射 AS-ODN BM-DC 后,用葡聚糖硫酸钠(DSS)暴露的小鼠中观察到实验性结肠炎的 7-10 天内,每天观察体重、行为、粪便频率和特征。安乐死后,对结肠进行组织学处理,同时将脾和肠系膜淋巴结(MLN)制成单细胞,通过流式细胞术测量 Foxp3+Treg 以及 IL-10+Breg 的频率。AS-ODN BM-DC 可预防 DSS 诱导的结肠炎发展。这些细胞的接受者在 MLN 和脾中 Foxp3+Treg 和 IL-10+Breg 的频率显著增加。与 DSS 处理的动物相比,无结肠炎的小鼠结肠切片的组织学检查在结构上仍然保持生理功能,并且大部分没有白细胞浸润。尽管 DSS 结肠炎主要是一种固有免疫驱动的疾病,但我们的研究增加了越来越多的证据,表明 Foxp3+Treg 和 IL-10 Bregs 可以抑制主要的固有免疫驱动的炎症。已经在人类中建立了从单核细胞祖细胞生成的人类 DC 存在针对 CD40、CD80 和 CD86 的主要转录本的混合反义 DNA 的安全性,为将其适应用于临床 IBD 治疗提供了潜力。