Characteristic pancreatic and splenic immune cell infiltration patterns in mouse acute pancreatitis.

A systemic evaluation of immune cell infiltration patterns in experimental acute pancreatitis (AP) is lacking. Using multi-dimensional flow cytometry, this study profiled infiltrating immune cell types in multiple AP mouse models. Three AP models were generated in C57BL/6 mice via cerulein (CAE) injection, alcohol and palmitoleic acid (EtOH + POA) injection, and alcohol diet feeding and cerulein (EtOH + CAE) injection. Primary pancreatic cells and splenocytes were prepared, and multi-dimensional flow cytometry was performed and analyzed by manual gating and computerized PhenoGraph, followed by visualization with t-distributed stochastic neighbor embedding (t-SNE). CAE treatment induced a time-dependent increase of major innate immune cells and a decrease of follicular B cells, and TCD4+ cells and the subtypes in the pancreas, whereas elicited a reversed pattern in the spleen. EtOH + POA treatment resulted in weaker effects than CAE treatment. EtOH feeding enhanced CAE-induced amylase secretion, but unexpectedly attenuated CAE-induced immune cell regulation. In comparison with manual gating analysis, computerized analysis demonstrated a remarkable time efficiency and reproducibility on the innate immune cells and B cells. The reverse pattern of increased innate and decreased adaptive immune cells was consistent in the pancreas in CAE and EtOH + POA treatments. Alcohol feeding opposed the CAE effect on immune cell regulation. Together, the immune profiling approach utilized in this study provides a better understanding of overall immune responses in AP, which may facilitate the identification of intervention windows and new therapeutic strategies. Computerized analysis is superior to manual gating by dramatically reducing analysis time.

目前缺乏对实验性急性胰腺炎（AP）中免疫细胞浸润模式的系统性评估。本研究利用多维流式细胞术，对多种AP小鼠模型中的浸润免疫细胞类型进行了分析。 通过雨蛙肽（CAE）注射、酒精和棕榈油酸（EtOH + POA）注射以及酒精饮食喂养和雨蛙肽（EtOH + CAE）注射，在C57BL/6小鼠中构建了三种AP模型。制备原代胰腺细胞和脾细胞，进行多维流式细胞术检测，并通过手动设门和计算机化的PhenoGraph进行分析，随后利用t分布随机邻域嵌入（t - SNE）进行可视化。 CAE处理导致胰腺中主要的固有免疫细胞随时间增加，滤泡B细胞、TCD4 +细胞及其亚型减少，而在脾脏中则呈现相反的模式。EtOH + POA处理的效果比CAE处理弱。EtOH喂养增强了CAE诱导的淀粉酶分泌，但意外地减弱了CAE诱导的免疫细胞调节。与手动设门分析相比，计算机化分析在固有免疫细胞和B细胞方面显示出显著的时间效率和可重复性。 在CAE和EtOH + POA处理中，胰腺中固有免疫细胞增加和适应性免疫细胞减少的相反模式是一致的。酒精喂养抵消了CAE对免疫细胞调节的作用。总之，本研究中使用的免疫分析方法有助于更好地理解AP中的整体免疫反应，这可能有助于确定干预窗口和新的治疗策略。计算机化分析通过大幅缩短分析时间优于手动设门。