Islet amyloid polypeptide and its N-terminal and C-terminal flanking peptides' immunoreactivity in islet amyloid of diabetic patients.
We determined immunohistochemically whether the islet amyloid polypeptide (IAPP)/amylin precursor is one component of islet amyloid, using polyclonal antibodies specific for human IAPP8-17 and amino (N)-terminal and carboxy (C)-terminal flanking peptides. To enhance immunostaining of the amyloid, we pretreated the pancreatic tissue sections with 100% formic acid. In three non-diabetic subjects, pancreatic islet cells were immunoreactive to anti-IAPP8-17 and anti-N-terminal and C-terminal flanking peptide antibodies and the reactivity was enhanced with formic acid pretreatment. In six type 2 diabetic subjects and a subject with type A insulin resistance, islet amyloid deposits were reactive to anti-IAPP8-17 antibody, but not to anti-N-terminal and C-terminal flanking peptide antibodies. Formic acid pretreatment markedly enhanced the reactivity to anti-IAPP8-17 antibody; however, it failed to show the reactivity to anti-N-terminal and C-terminal flanking peptide antibodies. Formic acid pretreatment of pancreatic tissue sections prepared for immunostaining is useful for visualization of buried epitopes of mature IAPP and its precursor molecules, either in islet amyloid deposits or in the islet cells. We conclude that the IAPP precursor and N-terminal and C-terminal flanking peptides are not constituents of human islet amyloid.
Kanatsuka
A
A
Second Department of Internal Medicine, Chiba University School of Medicine, Japan.
Makino
H
H
Yagui
K
K
Huang
C I
CI
Taira
M
M
Mikata
A
A
Yoshida
S
S
D000328:Adult; D000368:Aged; D000595:Amino Acid Sequence; D000682:Amyloid / Q000032:analysis* / Q000737:chemistry* / Q000276:immunology / Q000378:metabolism; D003924:Diabetes Mellitus, Type 2 / Q000276:immunology / Q000378:metabolism* / Q000473:pathology; D000939:Epitopes / Q000032:analysis; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D007333:Insulin Resistance / Q000502:physiology; D058228:Islet Amyloid Polypeptide; D007515:Islets of Langerhans / Q000737:chemistry* / Q000378:metabolism / Q000473:pathology; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D010455:Peptides / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D011498:Protein Precursors / Q000032:analysis / Q000276:immunology / Q000378:metabolism
D000682:Amyloid; D000939:Epitopes; D058228:Islet Amyloid Polypeptide; D010455:Peptides; D011498:Protein Precursors