A search for the presence of the enteroviral capsid protein VP1 in pancreases of patients with Type 1 (insulin-dependent) diabetes and pancreases and hearts of infants who died of coxsackieviral myocarditis

Abstract

Using an antiserum raised to a recombinant coxsackie virus B 3 capsid protein, VP1, an immunocytochemical technique was developed which was capable of detecting the presence of all coxsackie B viruses in formalin fixed paraffin embedded infected tissue culture cells. This technique was tested on autopsy heart and pancreas from 21 patients who were thought to have died of acute coxsackievirus B myocarditis. Cardiac myocytes were positive for the VP1 protein in 12 of 20 cases where the heart was available for study. Insulitis was present in the pancreas in seven of these cases and in all seven islet endocrine cells containing VP1 were found. VP1 was only rarely found in exocrine pancreas. In heart and pancreas, cells shown to contain VP1 usually showed signs of necrosis. Autopsy pancreases from 88 patients who had died at clinical presentation of Type 1 (insulin-dependent) diabetes mellitus showed no evidence of the presence of VP1. The continuing destruction of insulin-secreting B cells seen at the time of death in the diabetic pancreas is unlikely to be due to a direct cytopathic effect of a coxsackie B virus. However, this study does not exclude the possibility that a persistent infection of B cells by a defective enterovirus may result in their destruction by an autoimmune mechanism.

Analysis of pancreas tissue in a child positive for islet cell antibodies

“…CAR production in islet cells, as observed in this study, confirms a previous report on islet-specific production of this molecule in normal human pancreatic islets [C. Verbeke and G. E. Blair, unpublished results]. This finding fits well with previous observations indicating clear tropism of enterovirus for human islets in patients who died of systemic enterovirus infection or diabetic ketoacidosis; the virus has been repeatedly discovered specifically in the islets, but not in the exocrine pancreas [4,20,24]. In addition, enteroviruses infect human islets in vitro.…”

“…The enterovirus-specific antibody (clone 5-D8/1) has been previously used in studies showing enteroviruses in the pancreas of diabetic patients and in the heart tissue of patients with dilated cardiomyopathy [21,22,24]. Even though its specificity has been demonstrated in these studies, it has also been shown that antibodies against enterovirus VP1 protein may cross-react with heat shock proteins, especially HSP60 [27].…”

Enterovirus infection in human pancreatic islet cells, islet tropism in vivo and receptor involvement in cultured islet beta cells

“…Autopsy pancreases from 12 newborn infants who died of fulminant coxsackievirus infections at the age of 3 days to 10 months [48] were studied for presence of enteroviral ribonucleic acid (RNA) by in situ hybridisation. Most of these patients had coxsackievirus myocarditis and six out of seven enterovirus-positive pancreases showed evidence of insulitis.…”

“…In addition, 40 non-diabetic pancreases from subjects matched for age and sex were included in the study as controls. Autopsy specimens obtained from [48] and the University of Tübingen were treated as anonymous tissue, and studies carried out in accordance with the Declaration of Helsinki of the World Medical Assembly. According to the statement of the ethics committee of the University of Tübingen this kind of historic study can be done without specific permissions.…”

“…Formalin-fixed, paraffin-embedded pancreatic tissue was obtained from newborn infants who died of fulminant coxsackievirus infection [48] and from autopsies of patients suffering from Type 1 diabetes. Control tissue included normal pancreases from newborn infants and adults and pancreas from patients with chronic pancreatitis.…”