G F Springer

Publications of G F Springer

• Specific role of T and Tn tumor-associated antigens in adhesion between a human breast carcinoma cell line and a normal human breast epithelial cell line.

  Authors: T Kishikawa, M Ghazizadeh, Y Sasaki, G F Springer

  Japanese journal of cancer research : Gann. 03/1999; 90(3):326-32.

  The possibility that tumor-associated antigens T and Tn act as adhesion molecules between normal and malignant breast epithelial cells at the early stages of recognition in the metastatic pathway wasThe possibility that tumor-associated antigens T and Tn act as adhesion molecules between normal and malignant breast epithelial cells at the early stages of recognition in the metastatic pathway was examined in vitro. The adhesive specificity of the antigens was assessed by means of in vitro adhesion tests between a carcinomatous breast cancer cell line (ZR75-30) and a normal epithelial breast cell line (HLB100) using both monoclonal antibodies and lectins specific as well as nonspecific for each antigen. Adhesion assay was performed using monolayers of the normal cell line prepared on plastic culture plates and the tumor cell line labeled with a fluorescent dye as a probe. The adhesion between the two cell types occurred with significant specificity via T and Tn antigens (P<0.001), and was temperature-dependent. The results suggest that at the early stages of recognition by tumor cells in the metastatic process, T and Tn antigens play a role as adhesion molecules between the tumor cells and adjacent normal cells.

• T (Thomsen-Friedenreich) and Tn epitope location and their spatial relations to adhesion plaques on human breast carcinoma cells: immunogold-silver staining studies at scanning electron microscopic level.

  Authors: B L Wang, G F Springer, L C Harwick

  Journal of submicroscopic cytology and pathology. 11/1998; 30(4):503-9.

  T and Tn are pancarcinoma epitopes (EPs) which can be immunodetected in about 90% of adenocarcinomas (CAs). To study the location of T and Tn EPs and their relations to adhesion plaques on CA cells,T and Tn are pancarcinoma epitopes (EPs) which can be immunodetected in about 90% of adenocarcinomas (CAs). To study the location of T and Tn EPs and their relations to adhesion plaques on CA cells, immunogold-silver staining method was employed at scanning electron microscope (SEM) level. Human breast CA cells grown on coverslips were fixed in paraformaldehyde and glutaraldehyde, reacted with cocktails of monoclonal antibodies against T or Tn EPs, followed by incubation with 10 nm gold conjugated goat anti-mouse immuno-globulins. The positive gold particle labelling was amplified with silver enhancement solution, and the specimens were then routinely critical point dried, sputter-coated with palladium and observed under SEM. The studies show that T and Tn EPs are not randomly distributed on CA cell surface; they are aggregated at the adhesion plaque area. These results confirm that T and Tn EPs play roles in CA cell adhesion, and suggest that they may represent the initial points of contact by immuno-effectors and therefore can be utilized in immunointervention and anti-adhesion therapy against CA.

• Quantitative computerized image analysis of Tn and T (Thomsen-Friedenreich) epitopes in prognostication of human breast carcinoma.

  Authors: B L Wang, G F Springer, S C Carlstedt

  The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society. 11/1997; 45(10):1393-400.

  The precursors of the blood group N and M-immunodominant structures, Tn and T (Thomsen-Friedenreich) epitopes (EPs) occur in approximately 90% of carcinomas (CAs) but are masked in benign-diseasedThe precursors of the blood group N and M-immunodominant structures, Tn and T (Thomsen-Friedenreich) epitopes (EPs) occur in approximately 90% of carcinomas (CAs) but are masked in benign-diseased and healthy tissues. We determined quantitatively on 55 primary invasive ductal breast CAs, stages I to IV, the prognostic value of extent of Tn and T EP expression over an observation period exceeding 5 years postoperatively. Classical, established pathological and histological prognostic characteristic indicators associated with survival were subdivided by standard criteria into favorable and unfavorable categories. Tissue sections were reacted with monoclonal anti-Tn and -T antibodies, followed by the streptavidin-biotin-peroxidase-DAB procedure; counterstain was methyl green. Tn and T EPs were then quantitated by computerized image analysis. Of the 55 CAs, 51 clearly expressed Tn and T, and four had traces. Strong Tn EP expression was statistically significantly associated with shortened 5-year disease-free interval, increasing pTNM stages, positive lymph node status, and increasing combined histological grades. T EPs were usually well expressed but showed no significant association with prognostic factors. Our results suggest that quantitative immunohistochemistry-image analysis of Tn EPs of primary breast CAs may add new parameters to prognostication.

• Immunoreactive T and Tn epitopes in cancer diagnosis, prognosis, and immunotherapy.

  Authors: G F Springer

  Journal of molecular medicine (Berlin, Germany). 09/1997; 75(8):594-602.

  Aberrant glycosylation is a hallmark of cancer cells. The blood group precursors T (Thomsen-Friedenreich) and Tn epitopes are shielded in healthy and benign-diseased tissues but uncovered in approx.Aberrant glycosylation is a hallmark of cancer cells. The blood group precursors T (Thomsen-Friedenreich) and Tn epitopes are shielded in healthy and benign-diseased tissues but uncovered in approx. 90% of carcinomas. T and Tn glycoproteins are specific, autoimmunogenic pancarcinoma antigens. These antigens may also be found in neoplastic blood cells (and on LTV-II infected T lymphocytes). Fundamental chemical and physical aspects of these glycoproteins of primary carcinomas are discussed first. Tn and T epitopes are cell and tissue adhesion molecules, essential in invasion by and metastasis of carcinoma which includes adherent and proliferative phases. These properties are then delineated next, followed by consideration of pathophysiological and clinical aspects of these antigens. Immunohistochemical studies of the extent of Tn antigen expression in primary breast carcinoma demonstrate it highly significant correlation with clinicopathological tumor stages, and hence its value as a reliable prognosticator. On the other hand, there is no significant, prognostically useful association between T antigen expression and clinical disease course. Everyone has "preexisting" anticarcinoma anti-Tn and anti-T antibodies, induced predominantly by the intestinal flora, while cellular immune responses to T and Tn epitopes are evoked only by carcinomas and some lymphomas. Carcinoma dedifferentiation leading to predominance of Tn over T epitopes is described, as is the role of Tn and T epitopes in very early, including preclinical, carcinoma detection. The highest sensitivities in carcinoma detection are for preclinical and the earliest clinical stages. Obviously, preclinical carcinoma detection is of practical importance. T/anti-T tests detected preclinical carcinoma in 77% of 48 patients long (mean 6 years) before their biopsy/X-ray results became positive. There were no false predictions of carcinoma in 38 control persons with benign diseases (observation average 4.8 years). These findings open a novel window for both curative approaches and pathophysiological studies. The autoimmunogenicity of carcinoma T/Tn antigen led us more than two decades ago to begin intradermal vaccination of patients with advanced breast carcinoma of stages IV-IIb, predominately after modified radical mastectomy and sometimes lumpectomy plus axillary dissection always followed by adjuvant radio/chemotherapy. The vaccine consists of human group O red blood cell membrane derived, HLA-free T/Tn antigen containing as adjuvant Ca3(PO4)2 plus a trace of phosphoglycolipid A hyperantigen, i.e., S typhi vaccine (USP), which itself has T and Tn specificities. Our efforts have now for up to 20 years remained successful in a large majority of the 32 patients. All 32 patients survived at least 5 years; 10-year survival was statistically highly significantly improved (5-year survival: P < 1 x 10(-7); 10-year survival: P < 1 x 10(-5)) compared to statistics of the United States National Cancer Institute. Because these vaccinations are successful, their extension to large populations with major types of carcinomas should be considered, and even immunological carcinoma prevention may be contemplated.

• Concurrent immunohistochemical staining of tumor-infiltrating lymphocytes and carcinoma-associated T (Thomsen-Friedenreich)/Tn antigens in human breast carcinoma.

  Authors: B L Wang, G F Springer, M W Kaufman

  The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society. 03/1996; 44(2):187-91.

  The composition of tumor-infiltrating lymphocytes (TIL) often reflects the host's immune response to the tumor. To study the relationship of TIL and carcinoma-associated T/Tn antigens in breastThe composition of tumor-infiltrating lymphocytes (TIL) often reflects the host's immune response to the tumor. To study the relationship of TIL and carcinoma-associated T/Tn antigens in breast carcinoma, a straightforward concurrent immunoenzyme staining procedure was developed. Fresh tissue was directly fixed in a zinc-based fixative to preserve lymphocyte markers and then routinely embedded in paraffin. The TIL subtypes in the sections were identified in the first immunostaining cycle by reaction with a monoclonal antibody (MAb) to lymphocyte markers CD3, CD4, CD8, CD19, or CD56, followed by a modified avidin-biotin procedure and diaminobenzidine tetrahydrochloride-H2O2 for color development. This was followed by paraformaldehyde fixation to block antibody crossreactivity. The T and Tn antigens on carcinoma cells were then demonstrated in a second staining cycle by reaction with an MAb against T or Tn antigen, followed by an indirect immunoalkaline phosphatase procedure and corresponding substrate systems for color development. The distinguishable brown color for TIL and blue or red color for T or Tn antigen enabled us to identify the TIL subsets and to describe their relations with T/Tn antigen expression in situ. This approach may contribute to better understanding of the patients' immune defenses against their tumor and aid in prognostication.

• Anti-Thomsen-Friedenreich (T) antibody-based ELISA and its application to human breast carcinoma detection.

  Authors: P R Desai, L H Ujjainwala, S C Carlstedt, G F Springer

  Journal of immunological methods. 01/1996; 188(2):175-85.

  An anti-Thomsen-Friedenreich (T) antibody-based enzyme-linked immunosorbent assay (ELISA) with high efficacy in human breast carcinoma detection is described. Immunoreactive T epitopes occur inAn anti-Thomsen-Friedenreich (T) antibody-based enzyme-linked immunosorbent assay (ELISA) with high efficacy in human breast carcinoma detection is described. Immunoreactive T epitopes occur in approximately 90% of all carcinomata; all humans have anti-T antibodies, naturally occurring anti-carcinoma antibodies, induced by their own intestinal flora. Carcinoma patients, but not control subjects, show alterations of serum anti-T hemagglutinin levels. Human anti-T antibodies are predominantly IgM. In the protocol presented here, anti-T IgM antibodies are quantitated by ELISA using Immulon 2 wells coated with human blood group O erythrocyte-derived T antigen as solid phase; in addition, total IgM in each serum is quantitated by ELISA in parallel with the anti-T IgM. Inter-assay coefficient of variation was 2% for both ELISAs. Although anti-T IgM values alone distinguish between carcinoma patients and control subjects, use of the quotient, QMe, which also considers total IgM, increases this distinction. For a given serum, QMe was obtained by the formula: QMe = (100 x (anti-T IgM)2/total IgM). Sera of 242 subjects, 117 breast carcinoma patients, 36 benign breast disease patients and 89 healthy persons were analyzed. QMe identified 88% of the breast carcinoma patients: it all six (100%) in situ, 11/13 (85%) Stage I, 48/58 (83%) Stage II and III and 38/40 (95%) Stage IV patients. Sera from 83% of the 36 benign breast disease patients were negative, i.e. within normal range; five of the six positive sera originated from patients with increased long-term risk of breast carcinoma, while sera from 11 other patients with increased carcinoma risk were negative. Overall, 90% of the 125 non-carcinoma control subjects were negative by both anti-T IgM and QMe. In preliminary studies, the ELISA protocol detected 11/14 (79%) patients with carcinomata other than those of the breast. The identification of all six in situ breast carcinoma patients by QMe points to its usefulness in carcinoma detection, especially early.

• T and Tn pancarcinoma markers: autoantigenic adhesion molecules in pathogenesis, prebiopsy carcinoma-detection, and long-term breast carcinoma immunotherapy.

  Authors: G F Springer

  Critical reviews in oncogenesis. 02/1995; 6(1):57-85.

  Physical and chemical nature of the T and Tn pancarcinoma [CA] glycopeptide epitopes [EPs], which are immediate precursors of the blood group MN EPs, and their role in CA pathogenesis and in clinicalPhysical and chemical nature of the T and Tn pancarcinoma [CA] glycopeptide epitopes [EPs], which are immediate precursors of the blood group MN EPs, and their role in CA pathogenesis and in clinical disease are discussed. T/Tn are immuno-occluded in non-CA diseased and in healthy tissues. Well-differentiated CAs usually express a higher proportion of T than Tn EPs, while Tn predominates in poorly differentiated primary CAs. Measurement of density of T and Tn EP expression on primary breast CA permits disease prognostication. CA-T and -Tn are cell adhesion molecules involved not only in invasion but also in metastasis. Immunological methods readily detect in vivo autoimmune responses to CA-T and -Tn EPs in about 90% of all CA patients from incipience and throughout. Everyone has preexisting anti-T and anti-Tn antibodies [Abs] induced by the intestinal flora. T/anti-T immunoassays are highly efficient in detection of incipient and clinically overt CAs and, importantly, predicted CA in 77% of the patients, months to many years before their biopsy/X-ray turned positive; there were no false immune predictions of CA. Since 1974, we use human O MN red cell-derived T/Tn glycoprotein vaccine plus adjuvants successfully in safe, specific, effective, long-term, active immunotherapy against recurrence of advanced breast CA pTNM Stages IV, III, and II.

• T/Tn pancarcinoma autoantigens: fundamental, diagnostic, and prognostic aspects.

  Authors: G F Springer, P R Desai, M Ghazizadeh, H Tegtmeyer

  Cancer detection and prevention. 01/1995; 19(2):173-82.

  Pathogenetic aspects of pancarcinoma T/Tn autoantigens were investigated; they are present in approximately 90% of all carcinomas from incipience and throughout. T/Tn are occluded in noncarcinomaPathogenetic aspects of pancarcinoma T/Tn autoantigens were investigated; they are present in approximately 90% of all carcinomas from incipience and throughout. T/Tn are occluded in noncarcinoma (non-CA) diseased and healthy tissues. By serological and immunohistochemical methods, we found that well-differentiated carcinomata express a higher proportion of T than Tn, while in poorly differentiated carcinomata, Tn predominates over T. Tn density of primary carcinomas correlates positively with aggressiveness, early clinical relapse, and early death. Delayed-type skin hypersensitivity to T (DTHR-T) and solid-phase anti-T antibody immunoassay (SPIA-T), respectively, detected 85% of 461 and 88% of 222 carcinoma patients; < 7% of over 450 benign diseased and healthy subjects reacted positively in these assays. T/anti-T assays are highly effective in detecting incipient (TisN0M0 and T1N0M0) carcinomas: DTHR-T-85% of 41, and SPIA-T-96% of 26 patients. Positive anti-T tests predicted CA in 74% of 47 patients months to years before their biopsy/X-ray turned positive.

• T/Tn antigen vaccine is effective and safe in preventing recurrence of advanced breast carcinoma.

  Authors: G F Springer, P R Desai, B D Spencer, H Tegtmeyer, S C Carlstedt, E F Scanlon

  Cancer detection and prevention. 01/1995; 19(4):374-80.

  Since 1974, and as of March, 1993, we have used T/Tn antigen vaccine in safe, specific, effective, long-term intradermal vaccination against recurrence of advanced breast carcinoma (CA). Staging isSince 1974, and as of March, 1993, we have used T/Tn antigen vaccine in safe, specific, effective, long-term intradermal vaccination against recurrence of advanced breast carcinoma (CA). Staging is by the pathologic TNM system. Treatment is ad infinitum. Of 19 consecutive breast carcinoma patients vaccinated, six Stage IV, six Stage III, and seven Stage II all survived > 5 years postoperatively. Three Stage III, three Stage IV, and five Stage II patients (i.e., 11) survived > 10 to > 18 years. Five others are alive but have not reached 10 years; three of them have no evidence of disease (NED). Three patients died of CA before reaching 10 years. An additional three breast CA patients are being treated for > 2 years, but, < 5 years postoperatively, they are NED. The vaccination are presented as a delayed-type hypersensitivity reaction with significant inflammation with increase of helper T lymphocytes and decrease of T suppressor/cytotoxic cell ratio.

• T/Tn antigen vaccine is effective and safe in preventing recurrence of advanced human breast carcinoma.

  Authors: G F Springer, P R Desai, H Tegtmeyer, S C Carlstedt, E F Scanlon

  Cancer biotherapy. 02/1994; 9(1):7-15.

  For nearly 20 yrs, we used T/Tn antigen vaccine in safe, specific, effective, long-term intradermal vaccination against recurrence of advanced breast carcinoma. Treatment is ad infinitum. All 18For nearly 20 yrs, we used T/Tn antigen vaccine in safe, specific, effective, long-term intradermal vaccination against recurrence of advanced breast carcinoma. Treatment is ad infinitum. All 18 breast carcinoma patients treated, pTNM Stages IV (6), III (6), and II (6), survived > 5 yrs postoperatively; 10 survived > 10 to > 18 yrs; of the latter, three patients each are Stages III and IV. Five additional 5 yr survivors have not yet reached 10 yrs. The probability that our survival results are due to chance, with NCI "1991 Standard PDQ Data" as control, for all three stages taken together is: 5-yr survival: p < 1 x 10(-8); 10-yr survival: p < 1 x 10(-5). There were no untoward side effects. The vaccination area presented as a delayed-type hypersensitivity reaction, but at variance with the PPD reaction, with significant inflammation, increase of helper T lymphocytes and decrease of the T suppressor/cytotoxic cell ratio.

• Pancarcinoma T/Tn antigen detects human carcinoma long before biopsy does and its vaccine prevents breast carcinoma recurrence.

  Authors: G F Springer, P R Desai, H Tegtmeyer, B D Spencer, E F Scanlon

  Annals of the New York Academy of Sciences. 09/1993; 690:355-7.

• Immunohistochemical and ultrastructural localization of T antigen in ovarian tumors.

  Authors: M Ghazizadeh, T Oguro, Y Sasaki, K Aihara, T Araki, G F Springer

  American journal of clinical pathology. 04/1990; 93(3):315-21.

  Thomsen-Friedenreich (T) antigen, the immediate precursor antigen of the human blood group MN system, has been found in many carcinomas, but it is suppressed in normal tissues and nonmalignantThomsen-Friedenreich (T) antigen, the immediate precursor antigen of the human blood group MN system, has been found in many carcinomas, but it is suppressed in normal tissues and nonmalignant diseases. Using a monoclonal antibody specific for the T epitope and an indirect immunoperoxidase technique at light and electron microscopic levels, the authors studied the expression of T antigen and its potential diagnostic value in ovarian tumors. Among 30 serous and mucinous ovarian cystadenocarcinomas, 20 (67%) were positive and 10 (33%) were negative for T antigen. In carcinomas, positive rates increased in parallel with the tumor grade and were 37%, 75% and 80% for grade 1, 2, and 3 tumors, respectively. Of the nine patients with metastasis, seven (78%) had positive and two had negative reactions in their primary and metastatic tumors. T antigen staining was observed at the intraluminal cell surfaces and peripheral cell membranes. The ultrastructural localization of T antigen revealed electron-dense reaction products at the cell surface and microvillous surfaces. Of the ten benign ovarian tumors, three (30%) were weakly positive and seven (70%) were negative for T antigen. These findings indicate a positive correlation between the presence of immunoreactive T antigen and conventional unfavorable prognostic indicators in ovarian carcinoma. The surface location of T antigen suggests that it may have a functional role at the cell membrane and the membrane may be involved in secretion (shedding) of T antigen. Detection of T antigen may be a useful marker of prognosis in ovarian carcinoma.

• Pancarcinoma T and Tn epitopes: autoimmunogens and diagnostic markers that reveal incipient carcinomas and help establish prognosis.

  Authors: G F Springer, P R Desai, W Wise, S C Carlstedt, H Tegtmeyer, R Stein, E F Scanlon

  Immunology series. 02/1990; 53:587-612.

• 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.

  Authors: D M Thomson, G F Springer, P R Desai, R Scanzano, M Gubersky, G Shenouda

  Clinical immunology and immunopathology. 12/1988; 49(2):231-41.

  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.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.

• 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.

  Authors: G F Springer, E V Chandrasekaran, P R Desai, H Tegtmeyer

  Carbohydrate research. 08/1988; 178:271-92.

  In contrast to healthy and noncarcinoma-diseased tissues, greater than 80% of all carcinomas (CAs) tested express immunoreactiveIn 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.

• Intestinal flora, carcinomata and erythrocytes evoke anti-Tn antibodies.

  Authors: P R Desai, H Tegtmeyer, G F Springer, S Metcalfe, R J Svvennsen

  Die Naturwissenschaften. 06/1987; 74(5):247-8.

• The fundamental and diagnostic role of T and Tn antigens in breast carcinoma at the earliest histologic stage and throughout.

  Authors: G F Springer, P R Desai, M K Robinson, H Tegtmeyer, E F Scanlon

  Progress in clinical and biological research. 02/1986; 204:47-70.

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

  Authors: G F Springer, P R Desai

  Molecular immunology. 12/1985; 22(11):1303-10.

  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.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.

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

  Authors: G F Springer, C R Taylor, D R Howard, H Tegtmeyer, P R Desai, S M Murthy, B Felder, E F Scanlon

  Cancer. 03/1985; 55(3):561-9.

  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 50Tn 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.

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

  Authors: G F Springer, W A Fry, P R Desai, R A Semerdjian, H Tegtmeyer, C G Neybert, E F Scanlon

  Cancer detection and prevention. 02/1985; 8(1-2):95-100.

  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 usedWe 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.