Volume 115, Issue 4, Pages 978-987 (October 1998) T cell–mediated exocrine pancreatic damage in major histocompatibility complex class II–deficient mice  Bruce A. Vallance, Bryan R. Hewlett, Denis P. Snider, Stephen M. Collins  Gastroenterology  Volume 115, Issue 4, Pages 978-987 (October 1998) DOI: 10.1016/S0016-5085(98)70270-7 Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 1 Survival time course of C2D MHC II–deficient mice. Ten randomly chosen male and female C2D mice were followed up until they died of pancreatitis-associated complications or the animals became moribund and had to be killed. Lifespan was rounded to the nearest month. Gastroenterology 1998 115, 978-987DOI: (10.1016/S0016-5085(98)70270-7) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 2 Intestinal manifestations. (A) Macroscopic appearance of the gastrointestinal tracts removed from a sick female C2D mouse (8 months of age; left) compared with a healthy 4-month-old female C2D mouse (right). Note the distention of the gastrointestinal tract from the 8-month-old mouse, especially of the cecum, and the elongated appearance of the colon. Also note the pale color and large size of the fecal pellets in the colon of the 8-month-old C2D mouse compared with the normal appearance of those in the large bowel from the healthy mouse. (B) High-power view showing one of the cell clusters frequently found in the mesentery of affected C2D mice. The cells seem similar to neuroendocrine cells found in islets of Langerhans (original magnification 400×). Gastroenterology 1998 115, 978-987DOI: (10.1016/S0016-5085(98)70270-7) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 2 Intestinal manifestations. (A) Macroscopic appearance of the gastrointestinal tracts removed from a sick female C2D mouse (8 months of age; left) compared with a healthy 4-month-old female C2D mouse (right). Note the distention of the gastrointestinal tract from the 8-month-old mouse, especially of the cecum, and the elongated appearance of the colon. Also note the pale color and large size of the fecal pellets in the colon of the 8-month-old C2D mouse compared with the normal appearance of those in the large bowel from the healthy mouse. (B) High-power view showing one of the cell clusters frequently found in the mesentery of affected C2D mice. The cells seem similar to neuroendocrine cells found in islets of Langerhans (original magnification 400×). Gastroenterology 1998 115, 978-987DOI: (10.1016/S0016-5085(98)70270-7) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 3 Histopathology of C2D mice undergoing pancreatic disease. (A) Section of the pancreas of a 7-month-old C2D mouse showing that the injury progresses in a regional fashion. The upper right region shows only remnants of the exocrine pancreas, whereas the tissue in the upper left seems to be structurally normal (original magnification 40×). (B) High-power view of one of the lesions shows extensive immune and inflammatory cell infiltration into the body of the pancreas. Note the isolated acinar cells surrounded by infiltrate (original magnification 200×). (C) The pancreas of a male C2D mouse, 8 months old, showing extensive infiltrate within the body of the pancreas but little tissue destruction (original magnification 40×). (D) High-power view of the pancreas of a 7-month-old female C2D mouse showing both the endocrine and exocrine regions of the pancreas. An islet of Langerhans (center) is surrounded by infiltrating cells. Note the absence of acinar cells around the islet, the few isolated acinar cells within the infiltrated region, and the intact nature of the islet (original magnification 400×). (E) Histopathology of the pancreas from an 8-month-old female C2D mouse undergoing weight loss and passing pale pellets. Note the atrophied nature of the pancreas with only scattered islets of Langerhans remaining and the apparent replacement of the exocrine tissue with adipose tissue. A remnant of the exocrine pancreas is seen at the bottom left (original magnification 40×). (F) High-power view of the pancreas seen in E. The islets of Langerhans and ductal epithelial cells remain, whereas the acinar cells are gone, replaced by adipose tissue (original magnification 400×). Gastroenterology 1998 115, 978-987DOI: (10.1016/S0016-5085(98)70270-7) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 3 Histopathology of C2D mice undergoing pancreatic disease. (A) Section of the pancreas of a 7-month-old C2D mouse showing that the injury progresses in a regional fashion. The upper right region shows only remnants of the exocrine pancreas, whereas the tissue in the upper left seems to be structurally normal (original magnification 40×). (B) High-power view of one of the lesions shows extensive immune and inflammatory cell infiltration into the body of the pancreas. Note the isolated acinar cells surrounded by infiltrate (original magnification 200×). (C) The pancreas of a male C2D mouse, 8 months old, showing extensive infiltrate within the body of the pancreas but little tissue destruction (original magnification 40×). (D) High-power view of the pancreas of a 7-month-old female C2D mouse showing both the endocrine and exocrine regions of the pancreas. An islet of Langerhans (center) is surrounded by infiltrating cells. Note the absence of acinar cells around the islet, the few isolated acinar cells within the infiltrated region, and the intact nature of the islet (original magnification 400×). (E) Histopathology of the pancreas from an 8-month-old female C2D mouse undergoing weight loss and passing pale pellets. Note the atrophied nature of the pancreas with only scattered islets of Langerhans remaining and the apparent replacement of the exocrine tissue with adipose tissue. A remnant of the exocrine pancreas is seen at the bottom left (original magnification 40×). (F) High-power view of the pancreas seen in E. The islets of Langerhans and ductal epithelial cells remain, whereas the acinar cells are gone, replaced by adipose tissue (original magnification 400×). Gastroenterology 1998 115, 978-987DOI: (10.1016/S0016-5085(98)70270-7) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 3 Histopathology of C2D mice undergoing pancreatic disease. (A) Section of the pancreas of a 7-month-old C2D mouse showing that the injury progresses in a regional fashion. The upper right region shows only remnants of the exocrine pancreas, whereas the tissue in the upper left seems to be structurally normal (original magnification 40×). (B) High-power view of one of the lesions shows extensive immune and inflammatory cell infiltration into the body of the pancreas. Note the isolated acinar cells surrounded by infiltrate (original magnification 200×). (C) The pancreas of a male C2D mouse, 8 months old, showing extensive infiltrate within the body of the pancreas but little tissue destruction (original magnification 40×). (D) High-power view of the pancreas of a 7-month-old female C2D mouse showing both the endocrine and exocrine regions of the pancreas. An islet of Langerhans (center) is surrounded by infiltrating cells. Note the absence of acinar cells around the islet, the few isolated acinar cells within the infiltrated region, and the intact nature of the islet (original magnification 400×). (E) Histopathology of the pancreas from an 8-month-old female C2D mouse undergoing weight loss and passing pale pellets. Note the atrophied nature of the pancreas with only scattered islets of Langerhans remaining and the apparent replacement of the exocrine tissue with adipose tissue. A remnant of the exocrine pancreas is seen at the bottom left (original magnification 40×). (F) High-power view of the pancreas seen in E. The islets of Langerhans and ductal epithelial cells remain, whereas the acinar cells are gone, replaced by adipose tissue (original magnification 400×). Gastroenterology 1998 115, 978-987DOI: (10.1016/S0016-5085(98)70270-7) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 3 Histopathology of C2D mice undergoing pancreatic disease. (A) Section of the pancreas of a 7-month-old C2D mouse showing that the injury progresses in a regional fashion. The upper right region shows only remnants of the exocrine pancreas, whereas the tissue in the upper left seems to be structurally normal (original magnification 40×). (B) High-power view of one of the lesions shows extensive immune and inflammatory cell infiltration into the body of the pancreas. Note the isolated acinar cells surrounded by infiltrate (original magnification 200×). (C) The pancreas of a male C2D mouse, 8 months old, showing extensive infiltrate within the body of the pancreas but little tissue destruction (original magnification 40×). (D) High-power view of the pancreas of a 7-month-old female C2D mouse showing both the endocrine and exocrine regions of the pancreas. An islet of Langerhans (center) is surrounded by infiltrating cells. Note the absence of acinar cells around the islet, the few isolated acinar cells within the infiltrated region, and the intact nature of the islet (original magnification 400×). (E) Histopathology of the pancreas from an 8-month-old female C2D mouse undergoing weight loss and passing pale pellets. Note the atrophied nature of the pancreas with only scattered islets of Langerhans remaining and the apparent replacement of the exocrine tissue with adipose tissue. A remnant of the exocrine pancreas is seen at the bottom left (original magnification 40×). (F) High-power view of the pancreas seen in E. The islets of Langerhans and ductal epithelial cells remain, whereas the acinar cells are gone, replaced by adipose tissue (original magnification 400×). Gastroenterology 1998 115, 978-987DOI: (10.1016/S0016-5085(98)70270-7) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 3 Histopathology of C2D mice undergoing pancreatic disease. (A) Section of the pancreas of a 7-month-old C2D mouse showing that the injury progresses in a regional fashion. The upper right region shows only remnants of the exocrine pancreas, whereas the tissue in the upper left seems to be structurally normal (original magnification 40×). (B) High-power view of one of the lesions shows extensive immune and inflammatory cell infiltration into the body of the pancreas. Note the isolated acinar cells surrounded by infiltrate (original magnification 200×). (C) The pancreas of a male C2D mouse, 8 months old, showing extensive infiltrate within the body of the pancreas but little tissue destruction (original magnification 40×). (D) High-power view of the pancreas of a 7-month-old female C2D mouse showing both the endocrine and exocrine regions of the pancreas. An islet of Langerhans (center) is surrounded by infiltrating cells. Note the absence of acinar cells around the islet, the few isolated acinar cells within the infiltrated region, and the intact nature of the islet (original magnification 400×). (E) Histopathology of the pancreas from an 8-month-old female C2D mouse undergoing weight loss and passing pale pellets. Note the atrophied nature of the pancreas with only scattered islets of Langerhans remaining and the apparent replacement of the exocrine tissue with adipose tissue. A remnant of the exocrine pancreas is seen at the bottom left (original magnification 40×). (F) High-power view of the pancreas seen in E. The islets of Langerhans and ductal epithelial cells remain, whereas the acinar cells are gone, replaced by adipose tissue (original magnification 400×). Gastroenterology 1998 115, 978-987DOI: (10.1016/S0016-5085(98)70270-7) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 3 Histopathology of C2D mice undergoing pancreatic disease. (A) Section of the pancreas of a 7-month-old C2D mouse showing that the injury progresses in a regional fashion. The upper right region shows only remnants of the exocrine pancreas, whereas the tissue in the upper left seems to be structurally normal (original magnification 40×). (B) High-power view of one of the lesions shows extensive immune and inflammatory cell infiltration into the body of the pancreas. Note the isolated acinar cells surrounded by infiltrate (original magnification 200×). (C) The pancreas of a male C2D mouse, 8 months old, showing extensive infiltrate within the body of the pancreas but little tissue destruction (original magnification 40×). (D) High-power view of the pancreas of a 7-month-old female C2D mouse showing both the endocrine and exocrine regions of the pancreas. An islet of Langerhans (center) is surrounded by infiltrating cells. Note the absence of acinar cells around the islet, the few isolated acinar cells within the infiltrated region, and the intact nature of the islet (original magnification 400×). (E) Histopathology of the pancreas from an 8-month-old female C2D mouse undergoing weight loss and passing pale pellets. Note the atrophied nature of the pancreas with only scattered islets of Langerhans remaining and the apparent replacement of the exocrine tissue with adipose tissue. A remnant of the exocrine pancreas is seen at the bottom left (original magnification 40×). (F) High-power view of the pancreas seen in E. The islets of Langerhans and ductal epithelial cells remain, whereas the acinar cells are gone, replaced by adipose tissue (original magnification 400×). Gastroenterology 1998 115, 978-987DOI: (10.1016/S0016-5085(98)70270-7) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 4 Immunohistochemical staining for T-cell markers. Immunohistochemical staining of the pancreas of a 7-month-old female C2D mice before weight loss or other signs of illness. (A) CD3 staining of pancreas and adjacent duodenum showing numerous CD3+ cells in an area of heavy infiltrate and interspersed between acinar cells (original magnification 200×). (B) CD8 staining showing numerous CD8+ cells in an area of heavy infiltrate within the pancreas, as well as surrounding scattered acinar cells (original magnification 200×). (C) CD4 staining of pancreas showing several positive cells on the periphery of an islet of Langerhans (original magnification 200×). Gastroenterology 1998 115, 978-987DOI: (10.1016/S0016-5085(98)70270-7) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 4 Immunohistochemical staining for T-cell markers. Immunohistochemical staining of the pancreas of a 7-month-old female C2D mice before weight loss or other signs of illness. (A) CD3 staining of pancreas and adjacent duodenum showing numerous CD3+ cells in an area of heavy infiltrate and interspersed between acinar cells (original magnification 200×). (B) CD8 staining showing numerous CD8+ cells in an area of heavy infiltrate within the pancreas, as well as surrounding scattered acinar cells (original magnification 200×). (C) CD4 staining of pancreas showing several positive cells on the periphery of an islet of Langerhans (original magnification 200×). Gastroenterology 1998 115, 978-987DOI: (10.1016/S0016-5085(98)70270-7) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 4 Immunohistochemical staining for T-cell markers. Immunohistochemical staining of the pancreas of a 7-month-old female C2D mice before weight loss or other signs of illness. (A) CD3 staining of pancreas and adjacent duodenum showing numerous CD3+ cells in an area of heavy infiltrate and interspersed between acinar cells (original magnification 200×). (B) CD8 staining showing numerous CD8+ cells in an area of heavy infiltrate within the pancreas, as well as surrounding scattered acinar cells (original magnification 200×). (C) CD4 staining of pancreas showing several positive cells on the periphery of an islet of Langerhans (original magnification 200×). Gastroenterology 1998 115, 978-987DOI: (10.1016/S0016-5085(98)70270-7) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 5 Adoptive transfer of pancreatitis. Histological appearance of the pancreas removed from an athymic mouse 3 months after adoptive transfer of C2D splenic and lymph node–derived mononuclear cells. Note the loss of exocrine tissue and the numerous mononuclear cells infiltrating and surrounding the remaining acinar cells (original magnification 400×). Gastroenterology 1998 115, 978-987DOI: (10.1016/S0016-5085(98)70270-7) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 6 Exocrine and endocrine function over the course of pancreatitis. Serum levels of amylase, lipase, and glucose from female C2D mice at 4 months of age (no signs of disease), at 6-7 months of age (no signs of disease), and at 7-8 months of age (sick, defecation of pale pellets) as well as 7-8-month-old female C57BL/6 mice (no signs of disease). (A) Serum amylase levels in international units per liter as assessed by an enzymatic rate assay. (B) Serum lipase levels in international units per liter as assessed by an enzymatic rate assay. (C) Serum glucose levels in millimoles per liter as assessed by a colorimetric assay. The results are from 4 to 6 mice per group and are expressed as the mean ± 1SEM. Significance (*) was found to be P < 0.01 for amylase and P < 0.001 for lipase. Gastroenterology 1998 115, 978-987DOI: (10.1016/S0016-5085(98)70270-7) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 6 Exocrine and endocrine function over the course of pancreatitis. Serum levels of amylase, lipase, and glucose from female C2D mice at 4 months of age (no signs of disease), at 6-7 months of age (no signs of disease), and at 7-8 months of age (sick, defecation of pale pellets) as well as 7-8-month-old female C57BL/6 mice (no signs of disease). (A) Serum amylase levels in international units per liter as assessed by an enzymatic rate assay. (B) Serum lipase levels in international units per liter as assessed by an enzymatic rate assay. (C) Serum glucose levels in millimoles per liter as assessed by a colorimetric assay. The results are from 4 to 6 mice per group and are expressed as the mean ± 1SEM. Significance (*) was found to be P < 0.01 for amylase and P < 0.001 for lipase. Gastroenterology 1998 115, 978-987DOI: (10.1016/S0016-5085(98)70270-7) Copyright © 1998 American Gastroenterological Association Terms and Conditions

Fig. 6 Exocrine and endocrine function over the course of pancreatitis. Serum levels of amylase, lipase, and glucose from female C2D mice at 4 months of age (no signs of disease), at 6-7 months of age (no signs of disease), and at 7-8 months of age (sick, defecation of pale pellets) as well as 7-8-month-old female C57BL/6 mice (no signs of disease). (A) Serum amylase levels in international units per liter as assessed by an enzymatic rate assay. (B) Serum lipase levels in international units per liter as assessed by an enzymatic rate assay. (C) Serum glucose levels in millimoles per liter as assessed by a colorimetric assay. The results are from 4 to 6 mice per group and are expressed as the mean ± 1SEM. Significance (*) was found to be P < 0.01 for amylase and P < 0.001 for lipase. Gastroenterology 1998 115, 978-987DOI: (10.1016/S0016-5085(98)70270-7) Copyright © 1998 American Gastroenterological Association Terms and Conditions