Baibing Yang1, Joy M Davis1, Thomas H Gomez2, Mamoun Younes3,4, Xiurong Zhao5, Qiang Shen6, Run Wang1, Tien C Ko7, Yanna Cao8. 1. Department of Surgery, The University of Texas Health Science Center at Houston, Houston, TX, 77030, USA. 2. Center of Laboratory Animal Medicine and Care, The University of Texas Health Science Center at Houston, Houston, TX, 77030, USA. 3. Department of Pathology & Laboratory Medicine, The University of Texas Health Science Center at Houston, Houston, TX, 77030, USA. 4. Department of Pathology, George Washington University School of Medicine and Health Sciences, George Washington University Hospital, Washington, DC, 20037, USA. 5. Department of Neurology, The University of Texas Health Science Center at Houston, Houston, TX, 77030, USA. 6. Department of Genetics, Louisiana State University Health Sciences Center, New Orleans, LA, 70112, USA. 7. Department of Surgery, The University of Texas Health Science Center at Houston, Houston, TX, 77030, USA. Tien.C.Ko@uth.tmc.edu. 8. Department of Surgery, The University of Texas Health Science Center at Houston, Houston, TX, 77030, USA. Yanna.Cao@uth.tmc.edu.
Abstract
BACKGROUND: 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. METHODS: 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). RESULTS: 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. CONCLUSIONS: 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.
BACKGROUND: 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 APmouse models. METHODS: 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). RESULTS: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. CONCLUSIONS: 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.
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