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G F Springer

Publications and source records attributed to G F Springer.

At least 19 recordsLinked to original sources
PubMed1999

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.

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 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.

Antibodies, Monoclonal↗
PubMed1998

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.

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.

Antigens, Tumor-Associated, Carbohydrate↗
PubMed1997

Immunoreactivities of polyclonal and monoclonal anti-T and anti-Tn antibodies with human carcinoma cells, grown in vitro and in a xenograft model.

Human polyclonal, monospecific anti-T and -Tn antibodies were found to be reactive in ELISA tests with human ovarian (IGROV-1, OVCAR-3 and SKOV-3), breast (SKBr-3 and T47D)- and oral (KB)-carcinoma cell lines, but less so or non-reactive with normal epithelia and fibroblasts. The direct binding radioimmunoassay, using 125I-labeled human antibodies, to the IGROV-1 cancer cells was inhibited by homologous unlabeled antibodies of the same concentration, but not by the respective immunodominant haptenic monosaccharides (Gal for T and GalNAc for Tn). Rodent ascitic monoclonal anti-T (Ca3114 and Ca3741) and anti-Tn (Ca3250, Ca3268 and Ca3638) antibodies were also reactive with the ovarian- and breast-cancer cells, as measured by FACS and ELISA tests, but to a lower extent than the polyclonal human antibodies. Both the monoclonal anti-T (Ca3741) and anti-Tn (Ca3250 and Ca3638) antibody-binding reactivities were significantly inhibited by the haptenic free monosaccharides. Addition of the above MAbs to IGROV-1 ovarian-cancer or T47D breast-cancer cells cultured in vitro resulted in significant cytological change and inhibition of the viability of the tumor cells, but not of normal epithelial breast cells. This effect on viability was shown to be complement-independent, yet it was profoundly influenced by the concentration of the serum added to the assay medium. In vivo biodistribution of the anti-T (Ca3114) and anti-Tn (Ca3638) MAbs administered i.p. to athymic IGROV-1 tumor-bearing CD1 female nude mice revealed higher 125I-labeled antibody accumulation in the tumor xenografts and in their lung tissues, as compared with other organs of the same mice tested. The above results thus suggest the feasibility of utilizing these antibodies in immunotherapy and drug targeting.

Animals↗
PubMed1997

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

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.

Antigens, Tumor-Associated, Carbohydrate↗
PubMed1997

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

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-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.

Adult↗
PubMed1996

Flow cytometric analysis of T and Tn epitopes on chronic lymphocytic leukemia cells.

Immunophenotyping of peripheral blood lymphocytes from six patients with B-cell chronic lymphocytic leukemia (B-CLL) and five normal volunteers was done and their T and Tn epitopes analyzed using specific monoclonal antibodies and flow cytometry. Lymphocytes from all patients showed strong Tn expression as compared to normal control lymphocytes. By contrast, T antigen was not expressed, The TN expression may be a useful diagnostic and prognostic marker for B-CLL.

Adult↗
PubMed1996

Thomsen-Friedenreich-related carbohydrate antigens in normal adult human tissues: a systematic and comparative study.

A broad variety of normal human tissues were examined for the expression of Thomsen-Friedenreich (TF)-related histo-blood group antigens, TF (Gal beta 1-3GalNAc alpha 1-R), Tn (TF precursor, GalNAc alpha 1-R), sialosyl-Tn (NeuAc alpha 2-6GalNAc alpha 1-R), considered to be useful in cancer diagnosis and immunotherapy, and sialosyl-TF, the cryptic form of TF. These antigens or, more correctly, glycotopes, were determined by immunohistochemistry with at least two monoclonal antibodies (mAbs) each (except sialosyl-TF) as well as by lectin histochemistry. For a better dissection of sialosyl-TF and TF glycotopes, tissue sections were pretreated with galactose oxidase or the galactose oxidase-Schiff sequence. Staining with mAbs appeared to be more restricted than with the lectins used. Distribution patterns among normal epithelia were different for all four antigens. These antigens were also detected in some non-epithelial tissues. They can be classified in the following sequence according to the frequency of their occurrence in normal tissues: sialosyl-TF > > sialosyl-Tn > Tn > TF. Most of the positively staining sites for TF, Tn, and sialosyl-Tn are located in immunologically privileged areas. The complex results obtained with anti-TF mAbs (after treatment of the tissue sections with sialidase from Vibrio cholerae) and the lectins amaranthin and jacalin revealed a differential distribution of the subtypes of sialosyl-TF [NeuAc alpha 2-3Gal beta 1-3GalNAc alpha 1-R and Gal beta 1-3 (NeuAc alpha 2-6)GalNAc alpha 1-R] in normal human tissues. From our data it can be inferred that TF, Tn, and sialosyl-Tn are promising targets for a cancer vaccine.

ABO Blood-Group System↗
PubMed1996

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

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 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.

Antigens, CD↗
PubMed1995

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

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 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.

Adult↗
PubMed1995

Expression of Thomsen-Friedenreich-related antigens in primary and metastatic colorectal carcinomas. A reevaluation.

BACKGROUND: Expression of the pancarcinoma Thomsen-Friedenreich (TF) carbohydrate antigen or, more correctly, hapten, in colorectal carcinomas is not generally agreed on. Furthermore, its suggested role in liver metastasis so far has not been substantiated by direct immunohistochemical evidence. METHODS: Cryostat sections from 52 primary tumors (20 with adjacent transitional mucosa), 22 liver metastases of colorectal carcinomas, and 17 samples of normal mucosae were examined immunohistologically with a panel of at least two monoclonal antibodies (mAbs) each to TF, to its precursor, Tn, and to sialosyl-Tn, among them two newly developed anti-TF mAbs. RESULTS: Of the primary colorectal carcinomas, 60% expressed TF. Staining was more intense with TF-alpha/beta-reactive than with exclusively TF-alpha- or TF-beta-reactive mAbs. Normal and transitional mucosae were negative. Liver metastases were positive for TF in a significantly higher percentage of cases (91%) than primary carcinomas. Patients with TF-positive primary tumors had a significantly higher risk to develop liver metastases compared with patients with TF-negative tumors (57% vs. 14%, respectively). Tn and sialosyl-Tn were expressed concomitantly in most primary (85%) and metastatic (95%) colorectal carcinomas. These antigens also were detected in transitional mucosae (Tn in 25%, sialosyl-Tn in 55% of cases). Normal mucosae were negative. CONCLUSIONS: These results prove unequivocally the presence of exposed TF epitopes in a majority of colorectal carcinomas in which both anomers of TF are expressed. These data further suggest that TF favors liver metastasis and that its expression in primary colorectal carcinomas is a significant risk factor for the development of liver metastasis.

Antibodies, Monoclonal↗
PubMed1995

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

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 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.

Antigens, Neoplasm↗
PubMed1995

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

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 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.

Antigens, Tumor-Associated, Carbohydrate↗
PubMed1995

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

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 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.

Adenocarcinoma↗
PubMed1994

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

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 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.

Adult↗
PubMed1993

Idiotypes of pre-existing human anti-carcinoma anti-T and anti-Tn antibodies.

All humans normally possess antibodies, predominantly IgM, that react specifically with the Thomsen-Friedenreich (T) and the Tn antigens which are present in immunoreactive form on > 85% of all human carcinomas, but not in healthy and otherwise diseased tissues. We report here a serological study of idiotype expression and antigen reactivity of the anti-T and anti-Tn antibodies. Idiotypy was analyzed with rabbit antibodies raised against, and made specific for, affinity-purified polyclonal anti-T and anti-Tn antibodies from blood group A1B healthy adult donors. Anti-T and anti-Tn antibodies cross-reacted idiotypically in spite of their distinct epitope specificities. By adsorbing anti-T antibodies on insolubilized synthetic T carbohydrate we could firmly link idiotype expression with antigen reactivity. The relation of idiotype expression to the antigen-binding site of plant seed lectins was also studied; one originated from Arachis hypogaea [peanut agglutinin (PNA)], the other from Artocarpus integrifolia (Jacalin). PNA inhibited only anti-T antibodies. Jacalin inhibited both anti-T and anti-Tn antibodies in a dose-dependent manner. Neither idiotypic nor anti-idiotypic antibodies diminished the binding of lectins to T and Tn epitopes. The shared idiotypes on natural anti-T and anti-Tn antibodies permit consideration of application of their anti-idiotypes in treatment and/or prevention of human carcinoma.

Adult↗
PubMed1990

Immunohistochemical and ultrastructural localization of T antigen in ovarian tumors.

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 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.

Antibodies, Monoclonal↗