PubMed Health⌕ Search

Biomedical subjects

G F Springer

Publications and source records attributed to G F Springer.

At least 37 records · Page 2Linked to original sources
PubMed1989

Carcinoma autoantigens T and Tn and their cleavage products interact with Gal/GalNAc-specific receptors on rat Kupffer cells and hepatocytes.

We studied interactions of isolated Thomsen-Friedenreich (T)- and Tn-specific glycoproteins with the Gal/GalNAc-specific receptors on rat Kupffer cells and compared them to those with rat hepatocytes. Immunoreactive T and Tn are specific pancarcinoma epitopes. Electron microscopy of gold-labelled T and Tn antigens revealed their specific binding to Kupffer cells, followed by their uptake via the coated pit/vesicle pathway of receptor-mediated endocytosis. Preincubation of Kupffer cells with GalNAc and GalNAc-BSA, but not GlcNAc or GlcNAc-BSA specifically inhibited binding of the T and Tn glycoproteins. Desialylated, isologous erythrocytes (T RBC) are known to bind to the Gal/GalNAc receptors of rat Kupffer cells and hepatocytes. This attachment was specifically inhibited by T and Tn in a concentration-dependent manner: 50% T RBC-Kupffer cell contacts were inhibited at 8.5.10(-6) mM T and 8.5.10(-5) mM Tn antigen concentrations, respectively. The corresponding figures for hepatocytes were 6.10(-6) mM T and 1.2.10(-6) mM Tn antigen. Amino-terminal cleavage products of the T glycoprotein, possessing clusters terminating in non-reducing Gal/GalNAc, inhibited T RBC binding to Kupffer cells and hepatocytes usually at 10(-2) to 10(-5) mM concentrations, whereas GalNAc, galactose and galactose glycosides inhibited at millimolar concentrations. Galactose-unrelated carbohydrates were inactive at concentrations greater than or equal to 50 mM.

Animals↗
PubMed1989

Are pancarcinoma T and Tn differentiation antigens?

Human fetal tissues were studied for presence of immunoreactive Thomsen-Friedenreich (T) and Tn epitopes (EPs) using well-defined anti-T and anti-Tn rodent monoclonal antibodies. T and Tn are universal (pan) carcinoma (CA) markers that are occluded in normal postfetal tissues except in some immunoprivileged enclaves. Immunohistochemical methods using avidin-biotin immunoperoxidase for staining were employed. Tissues between 45 and 117 days after ovulation were studied. In most instances, anti-T and anti-Tn antibodies showed similar immunoreactivity as demonstrated by positive immunohistochemical staining. The most intense staining was in epithelial and mesothelial components; the mesenchyme stained more faintly. All human sera have anti-T and anti-Tn antibodies, stimulated largely by intestinal flora. The presence of immunoreactive T and Tn during an early phase of fetal development, as shown here, and their known absence in noncarcinomatous postfetal tissues suggests that T and Tn, in addition to their association with CA, are stage-specific oncofetal antigens in pretolerogenic differentiation phases.

Antigens, Differentiation↗
PubMed1989

Tn epitope (N-acetyl-D-galactosamine alpha-O-serine/threonine) density in primary breast carcinoma: a functional predictor of aggressiveness.

This interpretive review attempts to dovetail advanced work by different groups of investigators on blood group and carcinoma (CA) glycoconjugates that have terminal, immunoreactive Tn epitopes (GalNAc alpha-O-Ser/Thr), and on the interaction of those structures with complementary antibodies and lectins. Fenlon et al. (1987) and Leathem and Brooks (1987) found a positive correlation between primary breast CA aggressiveness and its affinity for Helix pomatia (HPA) lectin. This phenomenon was used successfully to accurately predict, in studies on 305 breast CA patients, early or late CA recurrence and patient survival time. The innate specificity of the large HPA combining groove (aside from its avid reactivity with appropriately spaced GalNAc alpha-O-) remains obscure, despite careful investigation for more than a decade (Baker et al., 1983). Leathem and Brooks presumed that HPA recognizes a hitherto "undefined biological marker" that indicates a breast CA's aggressiveness. Our own work has shown that the chemically fully defined Tn epitope, as measured with human polyclonal and murine monoclonal anti-Tn antibodies, occurs in immunoreactive form in approximately 90% of all breast and lung adenoCAs studied. Tn is occluded and non-reactive in healthy and non-CA-diseased tissues. We found that CA-associated Tn is an adhesion molecule in attachment to healthy cells; an increase in its density on breast CA cell membranes parallels greater aggressiveness of breast tumors in both humans and mice (the only species studied). Thus, Tn may be all or a major part of the postulated "as yet undefined biological marker" associated with high breast CA aggressiveness. Besides being helpful in the elucidation of some aspects of breast CA pathogenesis, these findings on primary breast CA have clinical implications in that they should facilitate stratification of breast CA patients for adjuvant treatment.

Antibodies, Neoplasm↗
PubMed1989

Coexpression of sialosyl-Tn (NeuAc alpha 2----6GalNAc alpha 1----O-Ser/Thr) and Tn (GalNAc alpha 1----O-Ser/Thr) blood group antigens on Tn erythrocytes.

Polyagglutinable erythrocytes expressing the rare human blood group phenotype Tn were tested for expression of both Tn (GalNAc alpha 1----O-Ser/Thr) and sialosyl-Tn (NeuAc alpha 2----6GalNAc alpha 1----O-Ser/Thr) antigens by agglutination with a panel of well-characterized monoclonal antibodies specifically directed to Tn and sialosyl-Tn antigens, respectively. Tn erythrocytes from 4 patients were strongly agglutinated by both sialosyl-Tn and Tn-specific monoclonal antibodies, indicating coexpression of sialosyl-Tn and Tn antigens on the cell membrane surface of Tn polyagglutinable erythrocytes. Human polyclonal anti-Tn antisera were found to express weak and variable anti-sialosyl-Tn antibodies in conjunction with the anti-Tn antibodies. The Tn phenotype thus involves not only the classically known Tn antigens but also sialosyl-Tn antigens. However, the human anti-Tn antibodies may be directed mainly toward the Tn antigen (GalNAc alpha 1----O-Ser/Thr).

Antibodies, Monoclonal↗
PubMed1988

Blood group Tn-active macromolecules from human carcinomas and erythrocytes: characterization of and specific reactivity with mono- and poly-clonal anti-Tn antibodies induced by various immunogens.

In contrast to healthy and noncarcinoma-diseased tissues, greater than 80% of all carcinomas (CAs) tested express immunoreactive O-(2-acetamido-2-deoxy-alpha-D-galacto-pyranosyl)-(1----3)-serine/threon ine [alpha-D-GalpNAc-(1----3)-Ser/Thr] in their glycoproteins. CA cells shed, into the tumor's environment, Tn, which is involved in cancer pathogenesis as adhesion molecule and as autoimmunogen. An increase in density of Tn on primary CA frequently parallels augmented CA aggressiveness. Tn-Active glycoproteins of culture-grown human breast CA DU 4475 cells were isolated from cytoplasm and from spent growth medium, and erythrocyte (RBC) Tn antigen was prepared by (1----3)-beta-D-galactosidase treatment of isolated human O RBC MN glycoprotein-derived Thomsen-Friedenreich (T) antigen. Immunochemical, serological, physical, and chemical analyses showed close resemblance of CA- and RBC-derived Tn antigens. The preponderant carbohydrate in both Tn glycoproteins is the alpha-D-GalpNAc residue, and the antigens have a qualitatively and quantitatively similar amino acid composition. Highly specific rodent monoclonal (Mo) anti-Tn antibodies (Abs) were elicited with Tn RBC and normal O RBC-derived Tn antigen, and compared with CA-anti-Tn MoAbs unwittingly evoked by others. A sensitive enzyme immunoassay (EIA) with Tn antigen as solid phase was developed. In this system, highly purified, "naturally occurring" anti-Tn antibodies, which all humans possess, were more sensitive in quantitating breast CA Tn structures than the anti-Tn MoAbs induced by Tn RBCs, and by RBC- and CA-derived Tn-active antigens. The sensitivity of anti-Tn MoAbs was higher in detecting RBC-Tn.

Amino Acids↗
PubMed1988

Comparison by leukocyte adherence inhibition of human immune response to cancer-associated immunogens, Thomsen-Friedenreich (T) and Tn, myelin basic protein, and organ-specific cancer neoantigens.

Peripheral blood leukocytes from cancer patients exhibit nonadherence to glass as an index of antigen recognition when incubated individually with four distinct, soluble tumor-related substances. Crude cancer extracts, purified antigens, T and Tn, myelin basic protein (MBP), and organ-specific cancer neoantigens (OSN), all elicited narrow dose-dependent leukocyte adherence inhibition (LAI) response curves. The present study focused on the reasons for the narrow antigen dose-LAI response relationship. Between 9 and 20 pmol of antigens elicited the maximum number of nonadherent leukocytes; cleavage products of T antigen and the nonapeptide (T18) of MBP required about a 10-fold increase in molar concentration for the same LAI response. When crude cancer extracts were combined with pure antigen or the pure antigens were combined at concentrations shown to give maximum LAI responses, the positive LAI responses were negated. The chemoattractant LTB4 at 10(-11) M triggered maximum LAI. But when MBP was added with the LTB4 at progressively increasing concentrations, there was dose-dependent negation of LAI. The magnitude of LAI depended on the total amount of mediator released rather than the rate of release. When leukocytes from cancer patients were incubated with optimum to high concentrations of MBP, the supernatants contained a mediator that gave similar bell-shaped dose-LAI responses on control leukocytes indicating that leukocytes from cancer patients react to a much broader range of antigen concentration than indicated by the LAI assay alone. High antigen dose negated LAI because of excess mediator production. Antigen-generated mediators had a biphasic effect inducing nonadherence and then adherence of leukocytes.

Antigens, Neoplasm↗
PubMed1985

Tn, a carcinoma-associated antigen, reacts with anti-Tn of normal human sera.

Tn antigen is the immediate precursor of the carcinoma (CA)-associated T antigen; both are masked in non-CA tissues. Tn antigen was detected by absorption of human anti-Tn antibody in 46 of 50 primary breast CAs and in all 6 metastases originating from Tn-positive primary CAs. Thirteen of 25 (52%) anaplastic CAs, but only 2 of 15 (13%) well differentiated CAs had more Tn than T; 1 anaplastic CA had neither antigen. Eighteen of 20 benign breast lesions had no Tn; the 2 positive lesions were premalignant. All 19 breast CAs, studied immunohistochemically, reacted strongly with human polyclonal anti-Tn; benign or normal glandular tissues had minimal or no reactivity. Among live cancer cell lines, the most malignant sublines had more Tn than T on their cell surfaces. Preliminary studies with rodent monoclonal anti-Tn and anti-T antibodies gave immunohistochemical reactivity patterns similar to those of the polyclonal antibodies, but the former were less sensitive in absorption tests. Tn is a CA marker that promises to be useful in tumor detection.

Animals↗
PubMed1985

Tn epitopes, immunoreactive with ordinary anti-Tn antibodies, on normal, desialylated human erythrocytes and on Thomsen-Friedenreich antigen isolated therefrom.

Hybridoma generation, using specifically, maximally desialylated human blood group O erythrocytes (T RBC) as immunogen, and biochemical studies suggested the presence of immunogenic Tn epitopes. GalNAc alpha-O, on T RBC. We therefore investigated by immunochemical means whether or not Tn-specific epitopes immunoreactive with anti-Tn antibodies present in ordinary human sera occur on T RBC and on Thomsen-Friedenreich (T) antigen prepared from them. We did detect the Tn epitope with such antibodies, in addition to the T epitope, on isolated T antigen. T RBC absorbed specifically, under standard conditions, 25-60% of the heterogeneous anti-Tn antibody populations in ordinary human sera of appropriately adjusted titer score. The anti-Tn eluted from T RBC had scores ranging from 6.5 to 35% of those of the unabsorbed parent sera. The varying fine specificities of eluted anti-Tn were demonstrated by inhibition of Tn RBC agglutination with putative haptens and antigens. Tn-specific haptens and antigens were the most powerful inhibitors. Depending on the serum used to prepare the anti-Tn eluates, the antibodies could be divided into those that were inhibited well exclusively by GalNAc alpha-O derivatives and those that were also inhibited by Gal, notably by Gal alpha-O derivatives and more strongly by GalNAc and Me-alpha-GalNAc. In the two reciprocal hemagglutination inhibition systems used, Tn-specific haptens were considerably more active than the T-hapten Gal beta 1----3GalNAc alpha-O, and desialylated ovine submaxillary mucin (AS-OSM) had higher activity than T antigen. Inhibition of Tn RBC agglutination by haptens was uniformly more efficient than that of T RBC; this is, at least in part, due to the much higher negative charge of Tn as opposed to T RBC. In microprecipitin tests, Helix pomatia lectin was nearly as powerful a precipitin of T antigen as of AS-OSM. The importance of the terminal GalNAc alpha of T antigen for its precipitation with the Helix lectin was demonstrated by the very high and virtually exclusive inhibitory activity of Me-alpha-GalNAc and GalNAc. Our findings may contribute to comprehension of the significance of uncovered Tn in most carcinomas, and the role of anti-Tn as a "natural" anti-carcinoma antibody. They may also help illuminate the rare heterozygous, autosomal, apparently premalignant spot mutation that leads to Tn RBC in vivo.

Antibodies↗
PubMed1985

Further studies on the detection of early lung and breast carcinoma by T antigen.

We measured the cellular and humoral autoimmune response to carcinoma (CA)-associated T antigen in patients with the earliest clinical stages of lung (T1N0M0) and breast (Tis) CA. We used desialylated MN glycoprotein from healthy human erythrocytes in a single measurement of 1) delayed-type skin hypersensitivity response (DTHR-T) and 2) humoral anti-T response with a solid-phase immunoassay (SPIA-T). DTHR-T detected 19/20 lung CA and 10/12 ductal and 7/10 lobular breast CA patients. Sixteen of 18 stage T1N0M0 lung CA patients had a positive SPIA-T as did 7/8 patients with Tis breast CA. Two of 35 patients with benign lung and 11/144 with benign breast disease had a positive DTHR-T. None of 160 persons with either other non-CA diseases or healthy had a positive DTHR-T. Three of 68 such control subjects had a positive SPIA-T. The difference between CA patients and control populations is statistically highly significant.

Antigens, Tumor-Associated, Carbohydrate↗
PubMed1984

T and Tn, general carcinoma autoantigens.

Primary and metastatic carcinomas are epithelial in origin and comprise by far the largest group of malignant tumors in humans. In most of these tumors, T and Tn antigens, whose epitopes have been synthesized, are uncovered and immunoreactive. In all other tissues T and Tn antigens are masked and not accessible to the immune system; they are generally precursors in normal complex carbohydrate chains. Thus, carcinomas have antigens recognized as foreign by the patients' immune system. The expression of T and Tn antigens has pathogenic and clinical consequences, and the antigens themselves are powerful histological markers in carcinoma diagnosis and frequently in prognosis. Most patients distinguish their carcinoma from all other cells, as shown by strong autoimmune responses to T antigen. These responses are readily measured by assays, and they allow detection of carcinomas with greater sensitivity and specificity frequently earlier than previously possible. Moreover, the extent of T and Tn expression often correlates with carcinoma differentiation; on a molecular level, clustered T- and Tn-active structures on carcinoma cell surfaces may be involved in invasion.

Antibody Formation↗
PubMed1983

Lung cancer patients' autoimmune responses to Thomsen-Friedenreich (T) antigen: diagnostic utility.

The usefulness of T antigen in the diagnosis of lung cancer (LCA), including early, was assessed by determining the in vitro delayed type hypersensitivity response to T(DTHR-T), and by measuring with a solid phase immunofluorescent assay the serum anti-T IgM response. Sensitivity of DTHR-T was 89% for 73 patients with LCA including 8/9 with Stage T1N0M0 disease, overall specificity was 95% for 212 healthy persons and those with non-CA disease. The humoral immune assay detected 31/35 (89%) LCA patients including 4/5 Stage T1N0M0 patients. Overall specificity was greater than 90% among 116 persons without CA.

Antibody Formation↗