Cloning and expression of the VP1 major capsid protein of diabetogenic encephalomyocarditis (EMC) virus and prevention of EMC virus-induced diabetes by immunization with the recombinant VP1 protein.
The development of diabetes in mice induced by encephalomyocarditis (EMC) virus provides the best experimental evidence that viruses have an aetiological role in the pathogenesis of this disease. The major capsid protein (VP1) of EMC virus is important for both the attachment of the virus to pancreatic beta cells and for the determination of antigenicity. This experiment was initiated to clone the gene for the major capsid protein, VP1, of the diabetogenic EMC (EMC-D) virus, express the VP1 protein, and test whether the recombinant VP1 protein can prevent development of EMC-D virus-induced diabetes in mice. We successfully cloned the VP1 gene of the EMC-D virus in the expression vector pRSET and subsequently expressed the protein in Escherichia coli. The recombinant VP1 protein was then purified by affinity chromatography. Five- to six-week-old male SJL/J mice were immunized intraperitoneally with purified VP1 protein and then challenged after various intervals with highly diabetogenic EMC-D virus. None of the VP1-immunized mice developed diabetes, irrespective of the interval between immunization and virus challenge, whereas 80 to 95% of the EMC-D virus-infected control mice did develop diabetes. All of the VP1-immunized mice showed intact pancreatic islet architecture, whereas most of the infected control mice showed severe beta cell necrosis and lymphocytic infiltration of their pancreatic islets. On the basis of these observations, we conclude that the recombinant VP1 protein of EMC-D virus can completely prevent the development of EMC-D virus-induced diabetes in mice.
Jun
H S
HS
Department of Microbiology and Infectious Diseases, Faculty of Medicine, University of Calgary, Alberta, Canada.
Yoon
S W
SW
Kang
Y
Y
Pak
C Y
CY
Lee
M C
MC
Yoon
J W
JW
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D002213:Capsid / Q000737:chemistry / Q000235:genetics* / Q000276:immunology*; D036022:Capsid Proteins; D018188:Cardiovirus Infections / Q000150:complications* / Q000821:virology; D003921:Diabetes Mellitus, Experimental / Q000517:prevention & control* / Q000821:virology; D003922:Diabetes Mellitus, Type 1 / Q000517:prevention & control / Q000821:virology; D004686:Encephalomyocarditis virus / Q000235:genetics / Q000276:immunology*; D005814:Genes, Viral / Q000235:genetics*; D007328:Insulin / Q000032:analysis; D007515:Islets of Langerhans / Q000473:pathology; D008297:Male; D051379:Mice; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000276:immunology; D017422:Sequence Analysis, DNA; D014611:Vaccination; D015678:Viral Structural Proteins / Q000235:genetics*; D014765:Viral Vaccines / Q000276:immunology
D036022:Capsid Proteins; D007328:Insulin; D011993:Recombinant Fusion Proteins; C096256:VP1 protein, encephalomyocarditis virus; D015678:Viral Structural Proteins; D014765:Viral Vaccines