Leung Gabriella, Petri Björn, Reyes José Luis, Wang Arthur, Iannuzzi Jordan, McKay Derek M
Gastrointestinal Research Group, Department of Physiology and Pharmacology, Snyder Institute for Chronic Diseases, University of Calgary, Calgary, Alberta, Canada.
Mouse Phenomics Resource Laboratory, Department of Microbiology, Immunology and Infectious Diseases, Snyder Institute for Chronic Diseases, Cumming School of Medicine, University of Calgary, Calgary, Alberta, Canada.
Mol Med. 2016 May;21(1):924-936. doi: 10.2119/molmed.2015.00193. Epub 2015 Dec 9.
The adoptive transfer of alternatively activated macrophages (AAMs) has proven to attenuate inflammation in multiple mouse models of colitis; however, the effect of cryopreservation on AAMs, the ability of previously frozen AAMs to block dinitrobenzene sulfonic acid (DNBS) (Th1) and oxazolone (Th2) colitis and their migration postinjection remains unknown. Here we have found that while cryopreservation reduced mRNA expression of canonical markers of interleukin (IL)-4-treated macrophages [M(IL-4)], this step did not translate to reduced protein or activity, and the cells retained their capacity to drive the suppression of colitis. The anticolitic effect of M(IL-4) adoptive transfer required neither T or B cell nor peritoneal macrophages in the recipient. After injection into the peritoneal cavity, M(IL-4)s migrated to the spleen, mesenteric lymph nodes and colon of DNBS-treated mice. The chemokines CCL2, CCL4 and CX3CL1 were expressed in the colon during the course of DNBS-induced colitis. The expression of integrin β7 on transferred M(IL-4)s was required for their anticolitic effect, whereas the presence of the chemokine receptors CCR2 and CX3CR1 were dispensable in this model. Collectively, the data show that M(IL-4)s can be cryopreserved M(IL-4)s and subsequently used to suppress colitis in an integrin β7-dependent manner, and we suggest that these proof-of-concept studies may lead to new cellular therapies for human inflammatory bowel disease.
在多种结肠炎小鼠模型中，过继转移替代性活化巨噬细胞（AAM）已被证明可减轻炎症；然而，冷冻保存对AAM的影响、先前冷冻的AAM阻断二硝基苯磺酸（DNBS）（Th1型）和恶唑酮（Th2型）结肠炎的能力及其注射后的迁移情况仍不清楚。在此，我们发现，虽然冷冻保存降低了白细胞介素（IL）-4处理的巨噬细胞[M(IL-4)]的典型标志物的mRNA表达，但这一步骤并未转化为蛋白质减少或活性降低，且这些细胞保留了驱动结肠炎抑制的能力。M(IL-4)过继转移的抗结肠炎作用在受体中既不需要T细胞、B细胞，也不需要腹腔巨噬细胞。注射到腹腔后，M(IL-4)迁移至DNBS处理小鼠的脾脏、肠系膜淋巴结和结肠。在DNBS诱导的结肠炎病程中，趋化因子CCL2、CCL4和CX3CL1在结肠中表达。转移的M(IL-4)上整合素β7的表达是其抗结肠炎作用所必需的，而趋化因子受体CCR2和CX3CR1的存在在该模型中是可有可无的。总体而言，数据表明M(IL-4)可被冷冻保存，随后用于以整合素β7依赖的方式抑制结肠炎，我们认为这些概念验证研究可能会带来针对人类炎症性肠病的新细胞疗法。
