concanavalin-a and Mesonephroma

In order to differentiate yolk sac-type alpha-fetoprotein (AFP) from hepatic-type AFP, 5 yolk sac tumors (YSTs) and 6 hepatocellular carcinomas (HCCs) were examined immunohistochemically by the peroxidase-antiperoxidase (PAP) method for AFP, and paradoxical concanavalin A (P-Con A) staining, which has been reported to detect glycoprotein including AFP. In all 5 YSTs, AFP was negative for P-Con A staining. On the other hand, AFP was strongly positive for the same staining in all 6 HCCs. A similar staining pattern for AFP was observed in human yolk sac endodermal cells and embryonal hepatocytes. Thus, it was clarified that yolk sac-type AFP was unable to bind with Con A, in contrast with hepatic-type AFP, on tissue sections. It was concluded that the PAP method for AFP and P-Con A staining might facilitate the immunohistochemical differentiation of these two types of AFP, and that it would be useful for clarifying the histogenesis of various AFP-secreting tumors.

Topics: alpha-Fetoproteins; Carcinoma, Hepatocellular; Concanavalin A; Female; Humans; Immunohistochemistry; Liver; Liver Neoplasms; Mesonephroma; Ovarian Neoplasms; Staining and Labeling; Yolk Sac

A sensitive new technique for lectin-affinity immunoelectrophoresis was applied to samples from 28 infants and children in order to distinguish the origin of elevated alpha-fetoprotein (AFP) in sera. This new immunoelectrophoresis was successfully performed within 24 hours in sera with AFP as small as 910 ng/mL. With combined use of concanavalin A (Con A) and lentil agglutinin (LCH) binding tests, AFPs were classified into three subtypes: benign hepatic condition type (six patients), hepatocellular carcinoma type (nine patients) and yolk sac type (12 patients). AFP was of hepatocellular carcinoma type in all seven patients with hepatoblastoma, and of benign hepatic condition type in six of seven patients with elevated AFP due to conditions such as hepatitis, biliary atresia, and normal newborn. The question as to whether AFP produced in "hepatoblastoma" is of benign hepatic condition type or hepatocellular carcinoma type was first answered by the information in this present report. The differentiation between yolk sac and general hepatic AFPs was completed with the Con A binding test.

Topics: Adolescent; alpha-Fetoproteins; Carcinoma, Hepatocellular; Concanavalin A; Female; Hepatitis; Humans; Immunoelectrophoresis; Infant; Lectins; Liver Neoplasms; Mesonephroma; Neoplasms; Ovarian Neoplasms; Plant Lectins

Profiles of concanavalin A (Con A) and lentil agglutinin (LCH) affinity immunoelectrophoresis were compared for serum alpha-fetoprotein (AFP) from patients with yolk sac tumors and carcinomas of the gastrointestinal tract, in order to find some correlations between peaks of AFP subfractions detectable by two different lectins, and to investigate whether or not it is possible to prove that the binding of AFP to LCH is weakened to some extent if a fucosylated sugar chain has, in addition, a bisect N-acetylglucosamine (GlcNAc) attached to the beta-linked mannose. The results obtained with our improved techniques tend to indicate that a Con A-reactive AFP subfraction (peak a) corresponds to an LCH strongly reactive AFP (peak A), while a Con A-nonreactive AFP (peak b) corresponds to an LCH weakly reactive AFP (peak B). the authors consider the present data sufficient to support the above explanation.

Topics: alpha-Fetoproteins; Concanavalin A; Dysgerminoma; Female; Humans; Immunoelectrophoresis; Lectins; Mesonephroma; Ovarian Neoplasms; Pancreatic Neoplasms; Plant Lectins; Stomach Neoplasms

Techniques have been studied which distinguish two variants of human alpha-fetoprotein (AFP) on the basis of characteristics of the carbohydrate moiety of this glycoprotein. AFP in serum samples from six children with tumors of yolk sac origin showed little concanavalin-A (Con A) binding. In contrast, Con A binding of AFP was almost complete in serum samples from 14 other subjects with elevated AFP, including two with liver-cell tumors, eight with neonatal cholestasis, and four normal newborn infants. Differences were confirmed by immunoelectrophoretic studies. Thus, AFP from cells of yolk sac origin can be distinguished from AFP from liver cells or from tumors of hepatic cell origin.

Topics: alpha-Fetoproteins; Bile Ducts; Carcinoma, Hepatocellular; Child; Child, Preschool; Cholestasis; Chromatography, Affinity; Concanavalin A; Female; Hepatitis; Humans; Immunoelectrophoresis, Two-Dimensional; Infant; Infant, Newborn; Jaundice, Neonatal; Liver Neoplasms; Male; Mesonephroma; Pancreatic Neoplasms; Teratoma

Using a modified method of Con A, LCH or PHA-E affinity crossed-line immunoelectrophoresis, we studied AFP subfractions in 78 sera including 58 from patients with primary hepatoma, 11 from patients with hepatic metastasis of gastric cancer and 9 from patients with germ cell tumors of the gonads (yolk sac tumor, immature solid teratoma or mature solid teratoma). It was found that AFP in primary hepatoma, metastatic hepatoma or germ cell tumors of the gonads were differently glycosylated, and different patterns of AFP subfractions identified by Con A, LCH or PHA-E affinity crossed-line immunoelectrophoresis facilitated a differential diagnosis of such AFP related malignancies.

Topics: alpha-Fetoproteins; Carcinoma, Hepatocellular; Concanavalin A; Female; Humans; Immunoelectrophoresis; Immunoelectrophoresis, Two-Dimensional; Lectins; Liver Neoplasms; Male; Mesonephroma; Neoplasms, Germ Cell and Embryonal; Ovarian Neoplasms; Teratoma; Testicular Neoplasms

Major molecular species of human alpha-fetoprotein(AFP), which were separated as single components by serial affinity chromatography with concanavalin A(Con-A) and Lens culinaris agglutinin, were further resolved into several bands by affinity electrophoresis with erythroagglutinating phytohemagglutinin of Phaseolus vulgaris lectin(E-PHA). Among the newly separated main molecular species, both Con-A- and E-PHA-reactive AFP(AFP-1X1) was demonstrated, contrary to the known sugar specificity of Con-A and E-PHA, in addition to molecular species of AFP reacting with Con-A but not with E-PHA(AFP-1X0) and of AFP reacting with E-PHA but not with Con-A(AFP-0X1). AFP-0X1 was formed from AFP-0X0, and AFP-1X1 from AFP-1X0 by neuraminidase treatment; thus, AFP-0X1 and AFP-1X1 represent asialylated and AFP-0X0 and AFP-1X0 sialylated molecular species. AFP-1X1' and AFP-0X0' were present as minor components. AFP-0X0' had no affinity for E-PHA, and the affinity increased in the order of AFP's-0X0(or 0X1), -1X1', -1X1 and -0X1. Proportions of those components varied depending on the pathophysiological conditions of AFP production.

Topics: alpha-Fetoproteins; Asialoglycoproteins; Carcinoma, Hepatocellular; Chromatography, Affinity; Concanavalin A; Electrophoresis; Enzyme-Linked Immunosorbent Assay; Fetuins; Humans; Liver Neoplasms; Mesonephroma; Neoplasms; Neoplasms, Germ Cell and Embryonal; Phytohemagglutinins; Stomach Neoplasms; Substrate Specificity

We used affinity chromatography on concanavalin A Sepharose to study the serum alpha-fetoprotein of 10 patients with histologically proven germ-cell tumors and 12 patients with primary liver cancer. Less than 50% of the fetoprotein from germ-cell tumors bound to concanavalin A, as compared with more than 80% of the alpha-fetoprotein from primary liver cancers.

Topics: Adult; Aged; alpha-Fetoproteins; Carcinoma, Hepatocellular; Chromatography, Affinity; Concanavalin A; Female; Humans; Liver Neoplasms; Male; Mesonephroma; Middle Aged; Ovarian Neoplasms; Sepharose; Testicular Neoplasms

Topics: Adenocarcinoma; alpha-Fetoproteins; Animals; Carcinoma, Hepatocellular; Concanavalin A; Diagnosis, Differential; Humans; Isoenzymes; L-Lactate Dehydrogenase; Liver Neoplasms; Lung Neoplasms; Mediastinal Neoplasms; Mesonephroma; Mice; Protein Binding

Lectin affinities of AFP were analyzed using Con A sepharose chromatography and crossed immuno-affino-electrophoresis. With Con A, AFP was divided into three subfractions, nonbound, loosely-bound and tightly-bound by chromatography, or two subfractions, nonbound and bound by electrophoresis. Con A nonbound subfraction was small in percentage in hepatocellular carcinoma (HCC), neonatal hepatitis, congenital biliary atresia (CBA), liver cirrhosis (LC) and cord sera. In contrast with these, the increase of Con A non-bound AFP was observed in yolk sac tumor (YST) and metastatic liver cancer (Meta). With LCA, AFP was divided into three subfractions: nonbound, loosely bound and tightly bound. Loosely bound fraction was very small in every specimen. AFPs from cord sera and LC showed uniform LCA affinity pattern, but AFPs from HCC were not uniform. Our data suggest that the analyses of lectin affinity of AFP serve as a diagnostic tool in differentiating (1) HCC from YST, (2) HCC from Meta, (3) CBA or neonatal hepatitis from YST and (4) LC from some cases of HCC.

Topics: alpha-Fetoproteins; Child; Child, Preschool; Chromatography; Concanavalin A; Diagnosis, Differential; Electrophoresis; Female; Humans; Infant; Infant, Newborn; Lectins; Liver Diseases; Male; Mesonephroma; Plant Lectins; Protein Binding

We determined by affinity chromatography on concanavalin A-Sepharose the carbohydrate variant patterns of alphafetoprotein in the sera of 15 infants and children with endodermal sinus tumors (five cases), a neonatal mature teratoma (one case), hepatoblastomas (two cases), pancreatic carcinoma (one case), biliary atresia (four cases), neonatal hepatitis (one case) and neonatal hyperbilirubinemia (one case), in the sera from four normal neonates, and in the sera from two kinds of nude mice bearing human endodermal sinus tumors. Sera from patients with endodermal sinus tumors and pancreatic carcinoma were found to contain a relatively high proportion (48.4 +/- 4.5 and 52.6%) of alphafetoprotein which did not bind to concanavalin A. Sera from nude mice with human endodermal sinus tumors contained AFP, 96.2% of which did not bind to concanavalin A. Sera from patients with other lesions (nine cases) and from normal neonates, whose AFPs are all presumed to be of hepatic origin, contained much less (5.9 +/- 3.6%) of the concanavalin A non-binding AFP variant. These results indicate that human AFP has three distinct patterns of reactivity with concanavalin A and that studies in xenograft models may give important information relating to the glycosylation and secretion process of AFP.

Topics: Adolescent; Adult; alpha-Fetoproteins; Animals; Biliary Tract Diseases; Carcinoma, Hepatocellular; Child; Child, Preschool; Chromatography, Affinity; Concanavalin A; Female; Humans; Infant; Infant, Newborn; Liver Neoplasms; Male; Mesonephroma; Mice; Mice, Nude; Neoplasm Transplantation; Teratoma

Human AFP purified from fetal serum and amniotic fluid was separated into three different variants by chromatography on concanavalin A insolubilized on Sepharose (Con A--Sepharose). The three variants were indistinguishable in immunodiffusion and radioimmunoassay. Sera from patients with yolk-sac tumor and amniotic fluid from early pregnancy were found to contain a high proportion (15-45%) of AFP which does not bind to Con A, while AFP in fetal and newborn sera, and in amniotic fluid from late pregnancy, contained less (2-6%) of this variant. The use of a large excess of Con A--Sepharose and the fact that the non-bound AFP consistently eluted as non-bound in rechromatography showed that this AFP is non-reactive with Con A. Fractionation of radiolabelled AFP from cord serum in a mixture with amniotic fluid verified the difference in the amount of the Con-A nonreactive variant in AFP from these two sources. These results suggest that AFP synthesized by the yolk-sac tissue and by the liver are glycosylated differently. The variant may prove to be diagnostically useful.

Topics: alpha-Fetoproteins; Amniotic Fluid; Carbohydrates; Chromatography, Affinity; Concanavalin A; Female; Fetal Blood; Humans; Immunodiffusion; Infant, Newborn; Liver; Liver Neoplasms; Mesonephroma; Ovarian Neoplasms; Pregnancy; Pregnancy Trimester, Second; Pregnancy Trimester, Third; Radioimmunoassay; Teratoma