Serum alpha-fetoprotein Values in Dogs with Various Hepaic Diseases.

Abstract

ABSTRACT. Serum alpha-fetoprotein (AFP) values were measured in hepatic diseased dogs with or without tumor and non-hepatic tumor bearing dogs by a sandwich ELISA using anti-dog AFP antiserum. Serum AFP values were less than 70 ng/ml in clinically healthy dogs. The values in dogs with hepatocellular carcinoma were higher than 1,400 ng/ml in 7 of 9 dogs, wherever those in two dogs with cholangiocarcinoma were in the normal range. Serum AFP values in hepatic diseased dogs without tumor were also high, however, the values were below 500 ng/ml in 90% of the dogs. In non-hepatic tumor dogs, serum AFP values were less than 500 ng/ml in 76% of the dogs. In the surgically removal cases with hepatocellular carcinoma, serum AFP values rapidly decreased. These results suggested that the sandwich ELISA using anti-dog AFP antiserum was an available method for diagnosis of hepatocellular carcinoma in dogs.-KEY WORDS: alpha-fetoprotein, canine, ELISA, tumor.J. Vet. Med. Sci. 61(6): 657-659, 1999 with hepatocellular carcinoma were also monitored AFP values after surgical removal of masses. Serum: Sera were collected by the usual methods, and kept below 20°C until assay.Preparation of AFP and antibodies: Canine AFP and antidog AFP antiserum were prepared according to the previously described method [17].ELISA for canine AFP: The protein concentration of purified AFP was measured by the Coomassie protein assay methods (Pierce, U.S.A.) using bovine serum albumin (BSA) as a standard. The ELISA for canine AFP was developed by the previously reported avidin-biotin sandwich method with minor modifications [17]. Briefly, the immunoglobulin G (IgG) fraction of anti-dog AFP antiserum from rabbits was coated on 96-well microplates (flat-bottom) at 5 µg per well. The wells were blocked with PBS containing 0.5% BSA, and washed with PBS containing 0.05% Tween 20 (TPBS) by an ELISA plate washer (Immunowash; BioRad, USA). Serum samples were diluted with TPBS at 1:2, 1:10, and 1:50. A standard solution of canine AFP was also diluted at concentrations of 0.25 ng/ml to 600 ng/ml in TPBS. The IgG fraction of anti-dog AFP antiserum was labeled with N-hydroxysuccimidobiotins (Sigma, U.S.A.) as shown by Coligan et al. [3]. The peroxidase-conjugated streptavidin was purchased from Dako (Denmark) and the chromogen 2',2' azino-bis (3-ethylbenzthiazolin-6-sulfonic acid) from Sigma (U.S.A.). Absorption of the samples and standards were measured at 405 nm. Serum AFP was measured in less than 3 weeks after storage, because storage time would affect AFP values [7]. RESULTS Serum AFP values:Serum AFP values in clinically healthy dogs were less than 70 ng/ml. Serum AFP values Serum alpha-fetoprotein (AFP) values have been used for the diagnosis of hepatocellular carcinoma [4,[5][6]14] and also for the understanding pathophysiological states in hepatic failure [8][9] in man. The values in dogs measured by enzyme-linked immunosorbent assay (ELISA) or radio immuno-assay (RIA) using anti-human AFP antibody were also reported [7,9,15], as the incidence of hepato...

Alpha-Fetoprotein in Serum and Tumor Tissues in Dogs with Hepatocellular Carcinoma

“…13 Subsequently, they found that, as in humans, AFP concentrations are elevated in canine tumors, especially in hepatocellular carcinoma (HCC). 12 In those reports, serum AFP was mildly elevated in some dogs with hepatic and nonhepatic diseases, even in those without tumors. A correlation between histopathology, immunohistochemistry, and serum AFP concentration in canine HCC has not been reported.…”

“…It has also been shown that the serum AFP concentration increases when normal hepatocytes are actively dividing, [9][10][11] and a moderate increase in serum AFP concentration is also reported in hepatic dysfunction or tumors other than HCC. 12 Therefore, quantification of serum AFP may not be used per se for a definitive diagnosis of HCC. In a previous study with a small population, 90% of dogs (19/21) with hepatic disease but without tumors were shown to have AFP concentrations of under 500 ng/ml, whereas 92% of tumor-bearing dogs (50/ 54) without hepatic tumors had concentrations of under 1,000 ng/ml.…”

Möglichkeiten der Diagnostik von Tumorerkrankungen am Magen-Darm-Trakt

“…Hierzug ehören α -Fetoprotein, carcinoembryonic antigen (CEA), squamous cellcarcinoma antigen(SCC-antigen), Cytokeratin 19 fragments (CYFRA) und Carbohydrate antigen1 9-9 (CA19-9) (1,8,16). Beim Hund ließ sich bisherlediglichineini-genUntersuchungeneine gewisse Bedeutungvon α -Fetoprotein im Zusammenhangmit demLeberzellkarzinom, Gallengangskarzinom sowiedem malignen Lymphom derLeber nachweisen (13,18,20,33). Dieklinischeund diagnostische Relevanzdieses Tumormarkersi st für denH und jedochb is heute unklaru nd muss erst in weiterenStudienvalidiert werden.…”

Identification of Cell Surface Antigen Expression in Canine Hepatocellular Carcinoma Cell Lines

“…There is a possibility that CD29, CD44, CD133 and Dlk-1 are expressed only in situations where liver regeneration is enhanced, as α-fetoprotein [19], when it might be difficult to distinguish between tumor and normal tissue using only these antigens. In the present study, these surface antigens were not assessed in liver tissue after hepatectomy or hepatitis, and further research is required to determine appropriate cancer cell markers.…”