Isolation, Characterization and Implications of Anti-TF (Thomsen-Friedenreich) Agglutinins from Different Sources

Kania, Jürgen; Uhlenbruck, G.; Janssen, Eveline; Klein, P. J.; Vierbuchen, M.

doi:10.1016/s0171-2985(80)80097-0

Cited by 15 publications

(7 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…On the other hand, the determination of fine specificity with complete glycoconjugates is more rele vant. The isolation of human serum anti bodies on acid-activated Sepharose was demonstrated recently [16]. The specificity of the isolated antibodies were found to be directed to gal-P(l-3)galNAc and galactose and is confirmed and extended by our find ings.…”

Section: Detection O F Exposed Tf Antigens On Human Breast Cancer Celsupporting

confidence: 80%

“…Anti-TF antibodies eluted by this procedure were named anti-TF2. Isolation of anti-TF antibodies by chromatography on acid-activated Sepharose 4B was performed as described [16]. Briefly, Sepharose 4B (Pharmacia, Uppsala, Sweden) was ac tivated by incubation for 3 h at 50 °C with 0.2 M HC1, neutralized and washed.…”

Section: Methodsmentioning

confidence: 99%

“…However, the molecules re sponsible for these mechanisms are not known. The naturally occurring TF anti bodies in human serum on the other hand were found to be heterogeneous regarding their immunoglobulin classes [2], their tem perature sensitivity [16], complement Fixing properties [17], and their carbohydrate specificities. These parameters varied de pending on the isolation procedure used.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Characterization of Thomsen-Friedenreich Antibody Subpopulations from Normal Human Serum

Wolf

Koerner²,

Klumpp³

et al. 1987

Tumor Biol

View full text Add to dashboard Cite

Thomsen-Friedenreich (TF) antibodies were prepared from human serum by different enrichment procedures. This resulted in three antibody preparations all of which agglutinated neuraminidase-treated erythrocytes. On the other hand, each of the three antibody populations showed a distinct specificity pattern. Anti-TF1 antibodies could be inhibited in the hemagglutination inhibition assay by asialofetuin, asialotransferrin, asialoglyco-phorin and asialomucin. The sialylated form of these glycoproteins showed no inhibition. No significant inhibition could be achieved with several mono- or disaccharides. This suggests that anti-TF1 recognizes common structures on glycoproteins normally hidden by sialic acid. Anti-TF2 antibodies showed specificity for asialofetuin, bovine submaxillary mucin, asialomucin, asialoglycophorin, the disaccharide gal-β(l-3)N-acetyl-galactosamine (galNAc) and nitrophenyl-β-galactoside. Because asialotransferrin or unbound lactosamine were not inhibitory, we suppose that the residual common structure of the inhibitors is (gal)-galNAc-O-Ser/Thr, which is present in high amounts in submaxillary mucin. Anti-TF3 antibodies were inhibited by asialoglycophorin but not by asialomucin or asialofetuin. Strong saccharide inhibitors were gal-β(l-3)galNAc, nitrophenyl-β-galactoside, as well as galactose. Therefore, both of the antibody preparations, anti-TF2 and anti-TF3, could be inhibited by gal-β-( 1– 3)galNAc, but showed preference to one or the other sugar component of the disaccharide resulting in a differential recognition of glycoconjugate inhibitors. Anti-TF2 and anti-TF3 seem to recognize the carbohydrates in the context of a protein backbone, because gal-β-(l-3)galNAc in connection with a ceramide backbone (GMj) was not inhibitory. When tested on three human breast cancer cell lines, only anti-TF2 recognized epitopes exposed on the cell surface. We, therefore, conclude that human serum contains at least three subpopulations of TF antibodies with distinct specificities. Only anti-TF2 can detect cryptic eryth-rocyte epitopes which are also exposed on human breast cancer cell lines.

show abstract

Section: Detection O F Exposed Tf Antigens On Human Breast Cancer Celsupporting

confidence: 80%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Characterization of Thomsen-Friedenreich Antibody Subpopulations from Normal Human Serum

Wolf

Koerner²,

Klumpp³

et al. 1987

Tumor Biol

View full text Add to dashboard Cite

show abstract

“…They obtained two fractions with different specificities, in which the hemagglutination by one of them was not inhibited by the TF disaccharide. Kania et al [21] prepared anti-TF antibodies from human serum using chromatography on acid-activated sepharose. The specificity and concentration of the antibodies were estimated by hemagglutination.…”

Section: Discussionmentioning

confidence: 99%

Isolation and Characterization of Thomsen-Friedenreich-Specific Antibodies from Human Serum

Butschak

Karsten²

2002

Tumor Biol

View full text Add to dashboard Cite

The Thomsen-Friedenreich (TF) disaccharide, galactose (Gal)β1–3GalNAcα-, is a blood group-related oncofetal antigen with remarkable tumor specificity. Postpartum, carbohydrate structures on the cell walls of the gastrointestinal flora evoke natural antibodies of presumed TF specificity. These antibodies may provide an early barrier against TF-carrying tumor cells. Their possible role, however, has been difficult to assess, since so far only a multivalent immunosorbent, asialoglycophorin (aGP), has been employed for their preparation, and therefore their fine specificities have been only insufficiently defined. We have used a novel immunosorbent consisting of synthetic TFα disaccharides (Galβ1–3GalNAcα-) coupled to polyacrylamide (PAA), which itself was covalently bound to cross-linked sepharose. For specificity analyses, aGP and a panel of PAA-conjugated mono- and oligosaccharides were employed. Binding to the PAA moiety was excluded. The affinity-purified anti-TFα antibodies were of the IgM (≧0.5 mg/100 ml of serum) and IgG (approximately 0.05 mg/100 ml of serum) classes. They did partially cross-react with TFβ, although we detected a second group of anti-TFβ antibodies (both IgM and IgG) which did not cross-react with TFα. The affinity-purified TFα antibodies showed only marginal cross-reactivity with the related antigens lactose, Gal, Tn or the Forssman antigen. Besides TF-specific antibodies, we found antibodies to the carbohydrate antigens Tn, Forssman and β-D-Gal as well as to noncarbohydrate epitopes of glycophorin in human serum.

show abstract

“…Two-dimensional gel electrophoresis was performed by a modification of published procedures [30]. Isoelcctric focusing gels contained 8 % polyacrylamide, 9.5 M urea, 4 % Nonidet P40 and 2 "/, ampholytes pH 3.5 ~ 10 (LKB).…”

Section: Methodsmentioning

confidence: 99%

Characterization of the Trimeric, Self‐Recognizing Geodia cydonium Lectin I

Müller

Conrad

Schröder

et al. 1983

European Journal of Biochemistry

View full text Add to dashboard Cite

A D-galactose-specific lectin I was extracted from the sponge Geodia cydonium and purified by affinity chromatography. The molecular weight of Iectin I as determined by high-pressure liquid gel chromatography, was found to be 36500 1300. Disc gel electrophoresis in the presence and in the absence of sodium dodecyl sulfate showed that lectin I is a trimer composed of three different subunits ( M , : 13 800,13 000 and 12200); two of the three subunits are linked by one disulfide bond. Isoelectric focusing gave a PI of 5.6 for the native molecule and a PI of 4.4 and of 7.4 for the subunits. The three subunits carry carbohydrate side chains, composed of D-galactose (94%) and of arabinose (5%). Based on experiments with lectins, the terminal i)-galactose residues are bound by p1+6 and/or fi1-4 glycosidic linkages. The Geodiu lectin I contains, besides two carbohydrate recognition sites, at least one receptor site for a second Iectin I molecule.Lectins are proteins (or glycoproteins) that bind carbohydrates and agglutinate cells; they have been isolated from viruses, bacteria and eukaryotes [l]. Since the initial work of Dodd et al. (21, a series of lectins have also been isolated from the oldest multicellular animals, the sponges [3 -61. In contrast to most other lectins, the biological role(s) of the sponge lectins are known to some extent. It has been demonstrated [7], that the D-galactose-specific lectin from Geodia cydoviium [5] is involved in the sorting-out of cells during reaggregation of allogeneic cells [S]. This lectin controls the activity of the cell membrane-associated antiaggregation receptor [9] by reversible binding to its D-galactose termini. This receptor, in its active state, has been shown to dissociate the complex, formed between the aggregation factor and the aggregation receptor [7, 81. In the xenogeneic system, using cells from the sponge Tethya lyncurium, the Geodia lectin causes cell agglutination [lo]. Later, evidence was presented suggesting that the lectin from Dictyostelium discoideum plays an analogous role in cellcell adhesion of cellular slime molds [II, 121. In previous reports on the properties of the Geodia lectin, two main aspects have been studied; firstly, the relationship between the hemagglutinin and the aggregation factor [5] and secondly, the mitogenic activity of the lectin for human lymphocytes [6]. In the present contribution a detailed physicochemical characterization of the Geodia lectin I (the main lectin in this sponge) is given, demonstrating that it is a trimer of a molecular weight of 36 500, with the property to react with D-galactose termini of its own protein species. These two characteristics have not yet been observed with other lectins. One lectin from rat liver membranes has been described [13], which binds to its own galactose residues after a pretreatment with neuraminidase. MATERIALS AND METHODS Lectin Extraction and PurificationThe siliceous sponge Geodia cydoniurn (Jam.) was collected near Rovinj (Mediterranean Sea) and the lectins were isolatedAhhrev...

show abstract

Isolation, Characterization and Implications of Anti-TF (Thomsen-Friedenreich) Agglutinins from Different Sources

Cited by 15 publications

References 25 publications

Characterization of Thomsen-Friedenreich Antibody Subpopulations from Normal Human Serum

Characterization of Thomsen-Friedenreich Antibody Subpopulations from Normal Human Serum

Isolation and Characterization of Thomsen-Friedenreich-Specific Antibodies from Human Serum

Characterization of the Trimeric, Self‐Recognizing Geodia cydonium Lectin I

Contact Info

Product

Resources

About