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Immunocytochemical Evidence of Amyloid-enhancing Factor (AEF) in Polymorphonuclear Leukocytes
Authors:Tadaaki Yokota  Tokuhiro Ishihara  Hiroo Kawano  Mutsuo Takahashi  Yoshimi Yamashita  Toshikazu Gondo  Yoshihiro Fujinaga  Fumiya Uchino
Affiliation:First Department of Pathology, Yamaguchi University School of Medicine, Ube.
Abstract:Amyloid enhancing factor (AEF) was extracted from spleens of mice that had received amyloidogenic stimulation. Sephacryl S 300 gel filtration of the crude AEF yielded five fractions, among which strong AEF activity was present in the first peak (Fl), and confirmed by an amyloid induction experiment. An anti AEF antiserum was obtained from a rabbit by immunization with Fl. This antibody reacted strongly with splenic polymorphonuclear leukocytes (PML) from mice given amyloidogenic stimulation, and weakly with those from normal untreated mice. Isoelectric focusing (IEF) analysis of both Fl and sera from mice given amyloidogenic stimulation was performed. A single band was observed on IEF analysis of Fl, whereas many bands were seen on IEF analysis of the sera. After the substances in the gel had been transferred to nitrocellulose membranes by capillary blotting, the membranes were made to react with the anti-AEF antiserum. The results suggested that AEF is a high molecular-weight substance derived from PML and increases in the serum at the time of, or shortly prior to, amyloid deposition in the spleen. Acta Pathol Jpn 39: 349∼355, 1989.
Keywords:Amyloid    Amyloid-enhancing factor    Polymorphonuclear leukocyte    Immunocytochemistry    Isoelectric focusing
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