Islet amyloid polypeptide, islet amyloid, and diabetes mellitus

Islet amyloid polypeptide (IAPP, or amylin) is one of the major secretory products of β-cells of the pancreatic islets of Langerhans. It is a regulatory peptide with putative function both locally in the islets, where it inhibits insulin and glucagon secretion, and at distant targets. It has binding sites in the brain, possibly contributing also to satiety regulation and inhibits gastric emptying. Effects on several other organs have also been described. IAPP was discovered through its ability to aggregate into pancreatic islet amyloid deposits, which are seen particularly in association with type 2 diabetes in humans and with diabetes in a few other mammalian species, especially monkeys and cats. Aggregated IAPP has cytotoxic properties and is believed to be of critical importance for the loss of β-cells in type 2 diabetes and also in pancreatic islets transplanted into individuals with type 1 diabetes. This review deals both with physiological aspects of IAPP and with the pathophysiological role of aggregated forms of IAPP, including mechanisms whereby human IAPP forms toxic aggregates and amyloid fibrils.     

Islet amyloid, islet-amyloid polypeptide, and diabetes mellitus

Islet-amyloid deposits, which are a common feature of Type II diabetes mellitus, are derived from the polymerization of a putative hormone identified as IAPP. IAPP is synthesized by normal islet beta cells and probably is cosecreted with insulin. Although the physiologic function of IAPP and its role in the pathogenesis of Type II diabetes mellitus are just beginning to be unraveled, IAPP may play an important part in the development of this most common form of diabetes mellitus by opposing the action of insulin in peripheral tissues. The polymerization of IAPP to form extracellular islet-amyloid deposits may further contribute to the development of Type II diabetes mellitus by destroying islet cells and by disrupting the passage of glucose and hormones to and from them. Substantial evidence indicates that the propensity of IAPP to polymerize and form extracellular amyloid deposits in only certain species (e.g., humans, cats, and raccoons) is directly associated with an intrinsically amyloidogenic part of the molecule--i.e., positions 20 through 29 of IAPP. The inherent amyloidogenicity of IAPP in these species may be further facilitated by increased beta-cell production of IAPP, leading to a high local concentration that predisposes to polymerization. The latter possibility is supported by studies demonstrating that IAPP production by islet beta cells is increased in normoglycemic cats with impaired glucose tolerance. Although increased production of IAPP may initially cause insulin resistance, prolonged overproduction of IAPP may ultimately impair insulin secretion by leading to the progressive deposition of insoluble islet amyloid, a finding apparent in most subjects with overt diabetes. If, as these studies suggest, increased IAPP production is linked to the development of Type II diabetes mellitus, further studies must address the genetic and nongenetic factors that influence this important biologic change in humans and some animal species.     

Islet amyloid polypeptide: mechanisms of amyloidogenesis in the pancreatic islets and potential roles in diabetes mellitus.

Amyloid deposits characteristically associated with pancreatic islets of those species (e.g., humans, cats, and monkeys) that develop age-associated forms of diabetes have been shown to represent a concentrated and polymerized form of a previously unknown islet-derived protein identified either as IAPP or amylin. IAPP, a highly conserved and carboxy-terminally amidated 37 amino acid polypeptide with approximately 45% amino acid sequence identity to CGRP, is produced by islet beta cells and is cosecreted with insulin in response to glucose and other secretagogues. Prepro-IAPP is synthesized in beta cells as an 89 to 93 amino acid molecule, and mature IAPP appears to be formed by enzymatic processing similar to that involved in the formation of insulin. Glucose-stimulated IAPP secretion generally parallels that of insulin and, on a molar basis, IAPP represents about 1% of the amount of insulin secreted. A significant dissociation of IAPP and insulin secretion (associated with relatively greater upregulation of IAPP secretion) is observed in response to marked hyperglycemia, suggesting that IAPP and insulin expression are differentially regulated. The amyloidogenicity of IAPP in only a very limited number of species is importantly related to the amino acid residues inherently found in the 20-29 region of IAPP from those species. The 25-28 region of human and cat IAPP is identical in structure and appears to be the most important amyloidogenic sequence common to the human and cat. In vitro fibrillogenesis studies have shown that amino acid substitutions in this region especially affect the amyloidogenicity of IAPP. Studies in dogs and cats suggest that aberrations in beta cell synthesis (or processing) of IAPP may lead to an increased concentration of IAPP in the local milieu, thus providing a second prerequisite for the self aggregation of IAPP to form islet amyloid. IAPP has been implicated to have physiological roles in glucose regulation, hemodynamics, calcium homeostasis, and as an anorectic agent. The major current interest in IAPP concerns its potential relationships to glucose metabolism and the development of type 2 diabetes. Evidence has been provided which indicates that IAPP can inhibit glucose-stimulated insulin secretion by beta cells, and that IAPP can also potentially contribute to the pathogenesis of type 2 diabetes by increasing hepatic glucose output and by inducing peripheral insulin resistance.(ABSTRACT TRUNCATED AT 400 WORDS)     

Islet inflammation and hyperplasia induced by the pancreatic islet-specific overexpression of interleukin-6 in transgenic mice.

Interleukin-6 (IL-6) is thought to be involved in the pathogenesis of autoimmune insulin-dependent diabetes mellitus. To examine this possibility, we developed two lines of transgenic mice (termed RIP-IL6) which overexpressed IL-6 in the pancreatic islet beta cells. RIP-IL6 mice, while showing a modest reduction in body weight, remained normoglycemic throughout their lives. Furthermore, insulin gene expression and glucose tolerance were similar to non-transgenic littermates. Histopathological examination revealed significant changes in the pancreas but not other organs of RIP-IL6 animals, with marked alterations in the architecture of the islets, in the islet cells, and in surrounding tissues. In younger animals these changes included islet hyperplasia with increased mitotic figures, neo-ductular formation, fibrosis, and a scant mononuclear cell infiltration (insulitis). In addition, immunostaining for islet hormones revealed changes in both the topography and density of beta and alpha cells. In older RIP-IL6 mice, a more florid insulitis was observed which was composed predominantly of B220+ B lymphocytes and, to a lesser extent, Mac-1+ macrophages and CD4+ and CD8+ T lymphocytes. Immunostaining for mouse IgG revealed significant numbers of plasma cells in the peri-islet infiltrates, which suggested that IL-6 induced differentiation of the recruited B lymphocytes. Therefore, islet overexpression of IL-6 produces a complex, localized host response implicating this cytokine in not only inflammatory processes that occur in autoimmune diabetes but also cellular neogenesis, which may indicate a role in tissue repair.     

1型糖尿病小鼠胰岛胰岛淀粉样多肽和生长抑素阳性细胞表达的改变

目的探讨1型糖尿病(T1DM)小鼠胰岛表达胰岛淀粉样多肽(IAPP)、生长抑素(SS)的改变及可能的机制。方法取正常C57BL/6J小鼠,小剂量多次注射链脲菌素(STZ)建立1型糖尿病小鼠模型,分别于第3、7、10、14、21、28天取胰尾组织,通过免疫组织化学SABC法、激光扫描共焦显微镜免疫荧光检测法及图像分析、形态计量法进行研究。结果 1.IAPP阳性细胞面数密度(NA)自第3天开始减少(P0.05),平均吸光度自第7天开始增高,与正常及生理盐水对照组比较差异具有显著性(P0.05);2.1型糖尿病组小鼠胰岛SS阳性细胞NA自第3天开始增加(P0.05),平均吸光度自第7天开始增高(P0.05);3.免疫荧光双染法显示,小鼠胰岛IAPP和SS在部分细胞有共表达。结论 1型糖尿病小鼠胰岛IAPP阳性细胞数目减少,表达增强;SS阳性细胞数量及表达均上调;IAPP和SS在部分细胞有共表达。提示IAPP阳性细胞和SS阳性细胞均参与了1型糖尿病的病变过程。     

Defective induction of antigen-reactive proliferating T cells in B cell-deprived mice

Mice were injected from day of birth onward with rabbit anti-mouse IgM antiserum or purified rabbit anti-mouse IgM antibodies. These mice completely lacked Ig-positive cells or serum Ig, as analyzed by specific fluoresceinated antibodies on the fluorescence-activated cell sorter (FACS-II), by polyclonal B cell mitogens and by specific precipitation in agar. These animals were then primed in vivo by antigen emulsified in complete Freund's adjuvant, and, subsequently, their draining lymph nodes were tested for their T cell proliferative responses in vitro, to the relevant antigen and were found to be severely impaired. However, the antigen-presenting capacity of both spleen cells and thioglycollate-induced peritoneal cells was found to be intact.     

Islet amyloid in type 2 human diabetes mellitus and adult diabetic cats contains a novel putative polypeptide hormone.

Amyloid deposition is a very typical alteration in the islets of Langerhans in human Type 2 (non-insulin-dependent) diabetes mellitus and in feline diabetes mellitus. Amyloid infiltration is also commonly found in insulin-producing pancreatic tumors. It was shown recently that amyloid purified from an insulinoma was composed mainly of a novel polypeptide (insulinoma amyloid polypeptide, IAPP), which had partial identity with the neuropeptide calcitonin gene-related peptide (CGRP). Cat islet amyloid contained a similar polypeptide. This finding is verified in the present study, and it is shown that the cat IAPP differs from the human peptide only in two of the 16 elucidated amino acid residues. The authors now also show by N-terminal amino acid sequence analysis that human islet amyloid is of IAPP origin. Although the significance of IAPP is unknown, its occurrence in pancreatic endocrine tissue and partial identity with a known neuropeptide suggests an endocrine regulatory function.     

Impaired glucose tolerance is associated with increased islet amyloid polypeptide (IAPP) immunoreactivity in pancreatic beta cells.

Adult cats determined by clinical laboratory evaluations to be normal, impaired glucose tolerant, or overtly diabetic were used to explore prospectively the relationships among pancreatic beta cell islet amyloid polypeptide (IAPP) immunoreactivity, islet amyloid (IA) deposition, and diabetogenesis. IAPP-derived IA was found in 11 of 14 (79%) diabetic cats, in four of nine (44%) impaired glucose tolerant cats, and in two of eight (25%) normal adult cats. The presence of IA even in very small amounts, therefore, predicts a very high probability (88%) that an animal has either impaired glucose tolerance or overt DM. Although all overtly diabetic cats had a marked decrease or absence of beta cell IAPP immunoreactivity, six of six cats with impaired glucose tolerance retained IAPP immunoreactivity with 1:15,000 dilutions of antisynthetic IAPP 7-17, whereas only one of seven normal cats had IAPP immunoreactivity beyond 1:10,000 dilutions. These findings suggest that increased IAPP production preceding the development of overt DM is linked to the progressive formation of insoluble IA deposits that are apparent in most overtly diabetic individuals. Of most importance, in that IAPP has been reported to inhibit both basal and insulin-stimulated rates of glycogen synthesis, is the possibility that increased production and release of IAPP by pancreatic beta cells plays a key role in the development of the insulin resistance and impaired glucose tolerance, both of which occur in Type 2 DM.     

Islet amyloid polypeptide in insulinoma and in the islets of the pancreas of non-diabetic and diabetic subjects

Summary Amyloid deposition is a common pathological feature in insulinoma and in the islets of the pancreas in type-2 diabetic patients. The present immunohisto-chemical study revealed that normal B-cells, insulinoma, and amyloid deposits in insulinoma and diabetic pancreatic islets were commonly immunoreactive with antiserum to C-terminal synthetic tetradecapeptide of human islet amyloid polypeptide (IAPP) (24–37). Amyloid fibrils in insulinoma were also positive to IAPP by immunoelectron microscopy. A high level of IAPP was detected in the plasma and tissue of a insulinoma patient by radioimmunoassay suggesting that amyloid deposition in insulinoma is due to overproduction of IAPP. Amyloid deposits immunoreactive to IAPP were also seen in all diabetic pancreatic islets, but in no non-diabetic islets. There was much amyloid deposition in the islets of severe diabetics, whose B-cells demonstrated decreased immunoreactivities for IAPP and insulin. The IAPP content of the pancreas was 649.0 and 847.7 pg/mg wet weight in each of two diabetic patients, and 1034.6 and 1447.7 pg/mg wet weight in two non-diabetic patients. The present study revealed that IAPP is a bioactive peptide secreted from islet B-cells and are amyloidogenic peptide concerned in diabetogenensis and/or the progression of type-2 diabetes mellitus.     

Cephalic phase pancreatic polypeptide responses to liquid and solid stimuli in humans

The hormone, pancreatic polypeptide (PP) is postulated to be involved in body weight regulation. PP release is dependent on vagal activation and is a marker of vagal efferent activity. Because vagal activity plays a role in glucose homeostasis, elucidating the conditions of activation has important implications for nutrient metabolism. In humans, modified sham-feeding is known to elicit vagally-mediated hormonal responses. We present results of 3 studies in which healthy human subjects tasted various stimuli including sweet and salty liquids, unflavored and flavored gum and mixed nutrient foods flavored with either sweet or salt and rendered palatable or unpalatable. We examined the effects of these stimuli on PP levels relative to fasting. We found that liquids flavored with either glucose or salt, did not elicit an increase in PP levels greater than fasting. Similarly, chewing gum, whether unflavored or flavored with a non-nutritive sweetener or the sweetener paired with a mint flavor, did not significantly increase PP levels. In contrast, when subjects tasted mixed nutrient foods, these reliably elicited increases in PP levels at 4 min post-stimulus (sweet palatable, p < 0.002; sweet unpalatable, p < 0.001; salty, palatable, p < 0.05, salty unpalatable, p < 0.05). The magnitude of release was influenced by the flavor, i.e. a sweet palatable stimulus (320.1 ± 93.7 pg/ml/30 min) elicited the greatest increase in PP compared with a salty palatable stimulus (142.4 ± 88.7 pg/ml/30 min; p < 0.05). These data suggest that liquids and chewing gum do not provide adequate stimulation for vagal efferent activation in humans and that mixed nutrient foods are the optimal stimuli.     

Immunofluorescent studies of the development of pre-B cells, B lymphocytes and immunoglobulin isotype diversity in humans.

Immunofluorescent techniques were used to examine several aspects of B cell ontogeny in humans. Large lymphoid cells containing intracytoplasmic IgM (pre-B cells) were present in fetal liver as early as 7 weeks of gestation, approximately 2 weeks prior to the appearance of surface IgM positive (sIgM⁺) B lymphocytes. Pre-B cells outnumbered sIgM⁺ B lymphocytes in fetal liver up until the 13th week of gestation. In fetuses older than 13 weeks, pre-B cells and sIgM⁺ B lymphocytes were present in approximately equal proportions in liver and bone marrow. Pre-B cells in fetal liver, and fetal and adult marrow, were large and small (indicating a heterogeneous population of cytoplasmic IgM⁺. SIg^- cells in these sites), while only the small pre-B cells were present in fetal spleen, blood and lymph node. Lymphocytes bearing sIgG were detected earlier than those bearing sIgD or sIgA, which were present by the 12th gestational week. Using double-staining techniques, we determined that during fetal life, (a) the proportion of B lymphocytes bearing only sIgM, as opposed to those bearing both sIgM and sIgD, was much higher in liver and bone marrow than in spleen, blood and lymph node, and (b) sIgG, sIgA and sIgD appear independently on lymphocytes bearing sIgM. Studies of the frequency of double-stained cells for each combination of the four sIg isotypes indicated that B lymphocytes from neonatal humans may simultaneously bear three or more sIg isotypes, whereas sIgG⁺ and sIgA⁺ B lymphocytes in adult blood usually express only the single isotype. Lower concentrations of anti-y antibodies were required for modulation of sIgM from B lymphocytes of fetal liver and adult bone marrow than for equivalent removal of sIgM from circulating B cells of mature individuals. In conjunction with data obtained in mice, our observations indicate that (a) the presence of large and small pre-B cells, (b) a high ratio of sIgM single to sIgM.sIgD double B lymphocytes, and (c) increased sensitivity to modulation of B cell sIgM by divalent anti-μ antibodies define the fetal liver and adult bone marrow as bursa-equivalent sites in humans. Our results are consistent with a model of isotype diversification in which immature sIgM⁺ cells give rise to B cell sublines devoted to synthesis of each of the Ig classes, and sIgD is secondarily expressed on unstimulated B cells of all of these sublines.     

Coexistence of glucagon and pancreatic polypeptide in human and rat pancreatic endocrine cells

Pancreatic islet cells containing both glucagon and pancreatic polypeptide simultaneously (glucagon/PP cells) were identified in the rat and human normal pancreas using immunocy-tochemical staining on consecutive serial sections and double-immunolabeling techniques on the same sections. Numerous glucagon/PP cells were found at the periphery of the islets in all regions of the pancreas, particularly in the rat. As a whole, these bipeptide-containing cells appeared in higher proportions than the cells secreting glucagon or PP separately. Double immunogold labeling performed on both surfaces of the thin tissue sections allowed differentiation between the glucagon/PP cells and the single-labeled glucagon or PP cells at the ultra-structural level. The pancreatic glucagon/PP cells displayed the morphological characteristics of either A cells or PP cells. Both peptides were found in the same secretory granules of the glucagon/PP cells and, in the human pancreas, the glucagon/PP cells displayed secretory granules with a dense core in which both peptides were rather concentrated. The coexistence of glucagon and PP is assumed to originate from the simultaneous expression of the two different genes in the same cell and suggests that the cellular processing through the rough en-doplasmic reticulum-Golgi apparatusgranule secretory pathway for both peptides takes place in parallel.     

Obese mice and the satiety effects of cholecystokinin, bombesin and pancreatic polypeptide

Cholecystokinin (CCK-8), bombesin (BBS) and pancreatic polypeptide (PP) are gastrointestinal hormones which are released during a meal and which decrease food intake when administered exogenously. The satiety effects of CCK-8, BBS and PP were measured in weanling and adult Bar Harbor obese and lean mice. Weanling mice fasted 5.5 hr were less sensitive to both CCK-8 and BBS at 5–6 weeks of age, when body weights of the obese were 20% greater than lean. The obese were equally sensitive as lean mice to CCK-8 and BBS at 7–8 weeks of age when the obese were 50% heavier. After a 3.0 hr fast adult obese mice, weighing 100% more than lean mice, were less sensitive to satiety effects of lower doses of CCK-8 (1.0 μg/kg) and PP (8 μg/kg) but equally sensitive to higher doses of CCK-8 (2.0 μg/kg), PP (16 μg/kg) and all doses of BBS (2.0, 4.0 and 8.0 μg/kg). After a 6-hr fast, 16 μg/kg PP did not affect food intake in obese or lean mice, whereas 32 μg/kg PP decreased food intake more in obese than lean mice. Thus both weanling and adult obese mice, as obese rats, are less sensitive to the putative satiety agent CCK-8.     

Defective induction of antigen-reactive proliferating T cells in B cell-deprived mice. II. Anti-mu treatment affects the initiation and recruitment of T cells

Mice injected from day of birth onwards with rabbit anti-mouse IgM (antim-mu) antibodies were found to be B cell-deficient and defective for the induction of antigen-reactive proliferating T cells (TPRLF). This defective induction was not due to the absence of circulating antigen-specific antibodies since the daily injections of such antibodies during exposure to antigen did not restore the ability of anti-IgM treated animals to generate TPRLF. Analyzing the cellular events implicated in the induction of virgin antigen-reactive T cells, anti-mu-treated mice manifested impairment of the three interacting cell types involved in the induction of TPRLF. Thus, peritoneal and splenic antigen-presenting cells from such animals were impaired in their capacity to signal a primary antigen-specific T cell reaction. Their splenic lymphocytes could not function as initiator cells in transferring immunogenic signals to recruit TPRLF in normal recipients. Potent antigen-specific splenic initiator cells failed to induce the recruitment of specific TPRLF in anti-mu-treated mice. The defective induction of TPRLF in anti-mu-treated mice may be due to a functional impairment of cells expressing membrane-bound IgM molecules which seemingly play a central role in the transfer of immunogenic signals for the recruitment of antigen-specific circulating T cells. We suggest that splenic B cells function as initiators in the transfer of antigen-induced signals from peritoneal antigen-presenting cells to T cells. These seems to be the primary targets of anti-mu treatment.     

Immunolocalization of islet amyloid polypeptide (IAPP) in pancreatic beta cells by means of peroxidase-antiperoxidase (PAP) and protein A-gold techniques.

A novel putative polypeptide hormone identified as islet amyloid polypeptide (IAPP) was recently purified from islet amyloid (IA) of diabetic humans and cats, and also from amyloid of a human insulinoma. Although the function of IAPP is yet unknown, its occurrence in pancreatic endocrine tissue and its partial amino acid sequence identity with calcitonin gene-related peptide (CGRP) suggests an endocrine regulatory effect. In the present investigation, the authors utilized antisera to insulin, glucagon, somatostatin, pancreatic polypeptide, synthetic human CGRP, and a synthetic human IAPP (7-17) undecapeptide to immunohistochemically (PAP technique) document the presence of IAPP immunoreactive cells in the islets of the cat, dog, mouse, and rat, but not in the islets of the horse or calf. In serial sections of islets from these species it was shown that IAPP immunoreactivity occurred in insulin-reactive beta cells. This observation was confirmed immunocytochemically in cat islets by means of protein A-gold probes. With protein A-gold labeling techniques, IAPP immunoreactivity was localized to the outer lucent compartment of the beta cell secretory granule, whereas insulin immunoreactivity was associated with the electron-dense core. These findings provide strong evidence that IAPP or an IAPP precursor is synthesized by beta cells and is stored in beta cell granules for subsequent co-secretion with insulin. The conservation of IAPP in humans and multiple animal species and the localization of IAPP to pancreatic beta cells provide further evidence that IAPP has an important endocrine regulatory function. The propensity of IAPP to polymerize and form IA fibrils in diabetes associated with aging may indicate that IAPP is in some way also linked to the development of Type 2 diabetes.     

Human islet amyloid polypeptide expression in COS-1 cells. A model of intracellular amyloidogenesis.

Non-insulin-dependent diabetes mellitus is characterized by concurrent loss of beta-cells and deposition of islet amyloid derived from islet amyloid polypeptide (IAPP). We have previously demonstrated that IAPP-derived amyloid forms intracellularly in humans with chronic excess insulin expression (eg, insulinoma and insulin receptor antibody-induced insulin resistance). To determine whether overexpression of IAPP results in intracellular amyloid in mammalian cells, we transfected COS cells with vectors expressing amyloidogenic human IAPP or non-amyloidogenic rat IAPP. Transfected COS-1 cells secreted comparable amounts of human IAPP and rat IAPP (2.1 to 2.8 nmol/L/48 hours). After 96 hours, 90% of cells expressing human IAPP contained amyloid fibrils and were degenerating or dead, whereas cells transfected with rat IAPP lacked amyloid and were viable. Thus, overexpression of human IAPP can result in intracellular amyloid formation that is associated with cell death, suggesting that intracellular amyloid may play a role in beta-cell loss in non-insulin-dependent diabetes mellitus.     

B lymphocyte development in fetal liver. II. Frequencies of precursor B cells during gestation.

Single cell cultures of fetal liver cells (FLC) from day 13 of gestation onwards until birth develop lipopolysaccharide (LPS)-reactive B cells which can be stimulated by LPS to IgM-secreting plaque-forming cells (PFC). When the cell concentration of FLC is lowered from 4 × 10⁶ cells/ml to 4 × 10^s cells/ml and below, the “in vitro” development to IgM-secreting PFC is lost. Less and less FLC initiate growth to IgM-secreting PFC as the cell concentration is lowered. Addition of 3 × 10⁶ thymus cells/ml from either syngeneic mice or from xenogeneic Lewis rats dramatically increases the efficiency with which FLC develop into LPS-reactive cells and initiate, after stimulation by LPS, clonal growth and maturation to IgM-secreting PFC. Thus, day 13 FLC can be diluted to 1 × 10⁶ cells/culture, day 16 FLC to 1 × 10⁴ cells/culture and day 19 FLC to 3 × 10¹ cells/culture before the responding cells become limiting. The frequencies of precursor B cells developing to LPS reactivity and then to IgM-secreting PFC in culture have been determined by limiting dilution analyses of FLC from day 13 to 19 (birth) of gestation and of liver cells at days 2, 4 and 7 after birth in cultures containing thymus cells and LPS. They are 1 precursor B cell in ～ 1 × 10⁶ FLC at day 13 of gestation, 1 in ～ 1 × 10^s at day 14, 1 in ～ 3 × 10⁴ at day 15, 1 in ～ 1 × 10⁴ at day 16, 1 in ～ 2 × 10³ at day 17, 1 in ～ 1 × 10² at day 18 and 1 in ～ 3 × 10¹ at day 19 of gestation. After birth, these frequencies decline rapidly to 1 in ～ 3 × 10² at day 2, 1 in 3 × 10³ at day 4 and 1 in less than 10^s liver cells at day 7 after birth.     

DeltaNp63 expression in pancreas and pancreatic neoplasia.

DeltaNp63 (DNp63) has become widely used, in particular, for distinguishing invasive carcinomas from noninvasive ducts by highlighting the myoepithelial or basal cells in the breast and prostate, respectively. It is not known whether this marker may have any application in another exocrine organ, the pancreas. As the ductal and intraductal proliferations of this organ become better characterized, the need for markers to distinguish among these processes increases. We investigated immunohistochemical expression of DNP63 in 105 cases. A total of 25 cases were non-neoplastic pancreata, 25 were pancreatic intraepithelial neoplasia (PanIN) of various grades, and 50 were examples of pancreatic ductal adenocarcinoma. Sections of non-neoplastic pancreata included various types of non-neoplastic processes such as squamous/transitional metaplasia (five cases), which can be mistaken for high-grade PanINs, as well as various degrees of reactive ductal atypia and incidental microcysts with attenuated lining (five cases). No DNp63 expression was noted in normal pancreatic ducts. On the other hand, all five foci of squamous/transitional metaplasia were strongly and uniformly positive for this marker. DNp63 labeling was also noted in those incidental microcysts lined by attenuated cells, seen amidst normal pancreatic lobules. All PanINs were negative. Among invasive carcinomas, DNp63 expression was detected only in areas of squamous differentiation and was completely absent in ordinary ductal areas. Based on this observation, five additional cases of adenosquamous/squamous carcinoma was retrieved and stained, and the squamous components of all of these were also positive. In conclusion, (I) DNp63 is a reliable marker of squamous differentiation in the pancreas. It is valuable in distinguishing squamous/transitional metaplasia from PanINs, a distinction of importance for both researchers and diagnosticians. Among invasive carcinomas, it seems to be entirely specific for areas of squamous differentiation. (II) Those incidental microcysts seen in acinar lobules and lined by attenuated cells are also positive for DNp63, which suggests that they may be metaplastic in nature, and that they do not represent neoplastic cells. (III) Unlike the ducts of other exocrine organs, breast and prostate, there are no DNp63-expressing cells in the normal pancreatic ducts, and therefore, this marker cannot be used in distinguishing invasive carcinomas from the non-invasive ducts. (IV) No p63-expressing 'stem' cells are present in the pancreas.