Tumour specific cell surface antigen and Forssman antigen associated with rat and hamster cells transformed with different strains of avian sarcoma virus.
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Alpharetrovirus↗
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Distribution of Forssman antigen in chickens.
The distribution of Forssman antigen in various tissues of chickens was studied by the indirect and direct immunofluorescence (IF) tests using rabbit immune serum against purified Forssman glycosphingolipid (GSL) [GalNAc (alpha, 1-3)GalNAc(beta, 1-3)Gal(alpha, 1-4)Gal(beta, 1-4)Glc-ceramide]. A general finding in all tissue sections was the presence of Forssman antigen in vascular endothelium and perivascular connective tissues. Erythrocytes gave a Forssman positive reaction, but the intensity of fluorescence was slight. Forssman antigen was found in about 20% of all peripheral leukocytes. Fractionation of lymphocytes from peripheral blood into T and B lymphocytes showed that about 30% of the B lymphocytes gave a positive reaction, while only 2-3% of the T lymphocytes gave a positive reaction. About 40% of bone marrow cells expressed Forssman antigens. On the other hand, about 40-50% of adherent cells in vitro cultures from spleen were Forssman-positive. The antigen was also abundant in hematopoietic organs, such as spleen, thymus and the bursa of Fabricius. Positive reactions in the digestive tract, ovary, testis and skin varied from portion to portion of the tissues. In brain and spinal cord, only the vascular endothelium gave a positive reaction. In peripheral nerves, only the connective tissue stroma around the nerve fibers gave a positive reaction. In other tissues, the distribution of the antigen was generally in accord with the results of Tanaka and Leduc (1956). These positive reactions in the IF test were specifically inhibited by addition of purified Forssman GSL. The distributions of Forssman antigen in tissues were found to be the same in chickens of various ages.
Animals↗
Expression of Forssman antigen in human large intestine.
Forssman antigen is a commonly occurring heterophile antigen but is thought not to be present in most humans. Recent biochemical studies, however, have shown the presence of Forssman antigen in several forms of human cancer, including gastric, colon, and lung cancers. Immunohistochemical staining with both monoclonal and polyclonal antibodies has failed to demonstrate this antigen in human tissues. In this study we conclusively demonstrated the presence of Forssman antigen in cytoplasm of colon goblet cells, especially those in the so-called transitional mucosa adjacent to carcinoma. Specimens from 69 of 70 patients with colon cancer contained the antigen in goblet cells in transitional mucosae. The antigen was successfully demonstrated only after removal of sialic acid by alkaline hydrolysis-neuraminidase digestion. Localization of the antigen was quite different from that of Tn and human blood group A, B, and H antigens. This antigen was thought to be associated exclusively with globo-series glycolipids, and its existence in glycoproteins has not been conclusively demonstrated. However, the results presented here, based on proteolytic digestion and lipid extraction studies, strongly suggest that the antigen is also contained in glycoproteins.
Adult↗
Apical and basal Forssman antigen in MDCK II cells: a morphological and quantitative study.
We have studied the surface distribution of a glycosphingolipid (the Forssman antigen) in MDCK II and CCL39 cells. The Forssman antigen is mobile on the surface of both these cell lines. Its surface distribution is homogenous on non-polarized cells. Under conditions where MDCK II cells are well polarized, the Forssman antigen is present in equal amounts on the apical membrane and on the basal membrane and its processes. Very little Forssman antigen can be detected on the lateral membrane. The nature of the mechanism excluding the Forssman antigen from the lateral domain remains to be determined. This surface distribution is established within hours after plating and was observed with cells grown on different types of filters. The surface density of the Forssman antigen on the apical and on the basal domain has been estimated. No involvement of the basal Forssman antigen in cell attachment could be demonstrated. However, the apical Forssman antigen appears to be essential to the establishment of the cells in culture.
Animals↗
Expression of Forssman antigen in the post-implantation mouse embryo.
The expression of Forssman antigen on the surface of cells of post-implantation mouse embryos between 5 and 7 1/2 days old and of cells of the gonads from 10 1/2 days has been followed using the monoclonal antiserum M1/22.25. In the early post-implantation embryo a lineage-related distribution is found. The inner cell mass of the blastocyst was previously shown to be Forssman antigen positive and its derivative tissues the epiblast of the 5-day embryo and the primary embryonic endoderm are also positive. The endoderm cells remain positive both over the embryonic and extraembryonic portions of the embryo but the epiblast becomes Forssman antigen negative as it differentiates into embryonic ectoderm. The extraembryonic ectoderm which is derived from the Forssman negative trophectoderm remains negative throughout. The primordial germ cells are Forssman antigen positive from their first appearance in the germinal ridge until day 14 when they become negative but after that time it is other cells not related by direct lineage which become Forssman antigen positive. These are tentatively identified as Sertoli cells precursors as it is the Sertoli cells which are the antigen-positive population in the adult testis.
Animals↗
Comparative study of rabbit hemolysins to various antigens. II. Hemolysins to the Forssman antigen of guinea pig kidney, human type A red cells and sheep red cells.
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Animals↗
A study of the cellular distribution of Forssman antigen in various species.
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Antigens↗
The presence of Forssman antigen in inbred mice.
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Animals↗
Forssman antigen in BK virus-induced tumor cell lines.
A complement-mediated microcytotoxicity assay was used to detect Forssman antigen in BK virus (BKV)-induced tumor cells. BKT-1B, a transplantable hamster tumor cell line, contained Forssman antigen as did some other hamster cell lines tested for comparison. A rat embryo cell line transformed by BKV in vitro was negative for Forssman antigen. The presence of forssman antigen in hamster cell lines was confirmed by absorption experiments of rabbit forssman antiserum with the tumor cells, although in quantitative terms the results obtained by the two techniques were somewhat different. Immunization of rabbits with tumor cells also induced production of Forssman antibody; a much weaker response was observed in hamsters and rats.
Animals↗
Forssman antigen and phase specific fetal antigens: an evaluation of their role in SV40 tumor immunity.
Forssman heterophile antigen was detected on hamster fetal cells which had been previously shown to be capable of eliciting transplantation resistance to syngeneic hamster SV40 tumors. The expression of Forssman antigen continued throughout fetal development and could be detected postpartum in the tissues of neonate hamsters. In contrast, fetal antigen(s) evoking immunity to SV40 tumors was also present on early fetal cells but, unlike Forssman antigen, was not expressed after the tenth day of gestation. Immunization of hamsters with guinea pig kidney cells or sheep erythrocytes which carry Forssman antigen failed to demonstrate significant protection against SV40 tumor development. Again by contrast, immunization with fetal cells was effective in evoking tumor immunity. Evaluation of serological responses to the FORSSMAN A ANTIGEN IN HAMSTERS INDICATED THAT THE HEMOLYTIC REACTIVITY PRODUCED BY IMMUNIZATION WITH GUINEA PIG KIDNEY CELLS OR SHEEP ERYTHROCYTES WAS ELICITED AGAINST ISOANTIGENS AND NOT THE Forssman antigen. A response to the Forssman determinant could only be detected in the serum from animals receiving exhaustive hyperimmunization with fetal cells or SV40 tumor cells. These data would eliminate a possible role of the Forssman heterophile antigen in the tumor protection evoked by immunization with fetal cells bearing embryonic antigens.
Animals↗
Forssman antigenicity of chemically synthesized globopentaose.
The Forssman antigenicity of a chemically synthesized globopentaose was studied. Globopentaose at 40 ng showed strong inhibitory activity for the formation of a precipitin line between globopentaosylceramide (Forssman glycolipid) and anti-Forssman rabbit antiserum, while much more pentasaccharide (7 and 100 micrograms, respectively) was required to inhibit a 50% quantitative precipitin reaction and a hemolysis reaction. An immune complex of the 3H-labeled globopentaose with anti-Forssman antibody was hardly formed. Thus, the chemically synthesized globopentaose possesses the same antigenic specificity as globopentaosylceramide but it is difficult to achieve a stable complex with Forssman antibody.
Animals↗
Evaluation of the role of the pneumococcal Forssman antigen (F-polysaccharide) in the cross-serotype protection induced by pneumococcal subcellular preparations.
We tested the hypothesis that the capacity of subcellular preparations of rough pneumococci to give cross-serotype protection is due to the presence of the pneumococcal Forssman antigen (F-polysaccharide). We found by hemagglutination inhibition that the Forssman antigen is present in the subcellular extracts. However, we concluded that the Forssman antigen is not the protective immunogen in the pneumococcal subcellular preparation, since absorption with sheep erythrocytes failed to remove the protective capacity from antiserum raised against the vaccine. Other evidence mitigating against the pneumococcal Forssman antigen being the protective immunogen included the absence of a detectable hemolytic titer in protective antiserum raised against the subcellular preparation, the failure of high-titered sheep hemolysin to passively protect mice against pneumococcal infection, and the failure of purified F-polysaccharide to immunize mice against pneumococcal infection.
Animals↗
Interaction of antibody with Forssman antigen in guinea pigs. A mechanism of adaptation to antibody- and complement-mediated injury.
Forssman antigen is a glycosphingolipid with antigenic specificity determined by extra-membrane haptenic sugars similar to blood group antigens and antigens that are the main barrier to xenogeneic organ transplantation. Herein, we describe the localization of Forssman antigen in guinea pig lungs and kidneys and the consequences of its interaction with antibodies in vitro and in vivo (Forssman reaction). Exposure of cultured guinea pig aortic endothelial cells to Forssman antibodies induced rapid redistribution of antigen-antibody complexes at the cell surface, followed by shedding that occurred by blebbing of plasma membrane as vesicles or fragments, and was associated with disappearance of antigen from the cell surface (antigenic modulation). Guinea pigs surviving frequent intravenous infections of increasing amounts of antibodies, for a total of 20 to 40 lethal doses, developed a partial or complete adaptation to generalized Forssman reaction, and adaptation was associated with partial or complete modulation of Forssman antigen at the surface of the pulmonary and, in minor degree, renal endothelial and epithelial cells. These findings support the hypothesis that modulation of endothelial carbohydrate antigens contributes to adaptation of highly vascularized organs exposed to tolerable levels of allo- or xenoantibodies.
Animals↗
[Search for the heterogenetic Forssman antigen in human breast tumor tissue].
The complement fixation test was used to examine 80 human mammary tumors for the presence of heterogeneous Forssman antigen: 62 cases of cancers of different malignancy (adenocarcinoma, solid tumors), 13 cases of fibroadenomas and 5 cases of fibrocystic mastopathies. Subject to study was also normal tissue of the mammary gland. Forssman antigen was not found in all the tumors tested, as it was not detected in tissues of the normal mammary gland. Therefore the process of transformation of normal cells to tumor ones is not accompanied by the occurrence in them of heterogenous Forssman antigen.
Adenocarcinoma↗
Pneumococcal Forssman antigen: enrichment in mesosomal membranes and specific binding to the autolytic enzyme of Streptococcus pneumoniae.
The choline-containing pneumococcal membrane teichoic acid (Forssman antigen) can be isolated with the membrane fractions of the bacteria. The small vesicle (mesosomal) fraction generated during the formation of protoplasts seems to be highly enriched in this material. Forssman antigen was identified in cell fractions on the basis of (i) radioactive choline label, (ii) autolysin-inhibitory activity, and (iii) the sedimentation profile in sucrose density gradients with and without detergent. A membrane teichoic acid could also be isolated from pneumococci grown in medium in which choline was replaced by ethanolamine as the nutritionally required amino alcohol. This material contained radioactive ethanolamine label and behaved similarly to the choline-containing membrane teichoic acid during centrifugation in detergent-containing and detergent-free density gradients. On the other hand, the material had only low autolysin-inhibitory activity. Binding of pure pneumococcal autolysin to micelles of purified Forssman antigen could be demonstrated by mixing these components in vitro and analyzing them by sucrose density gradients and by agarose chromatography. No binding could be observed between the pneumococcal enzyme and the micellar forms of either cardiolipin or polyglycerophosphate-type lipoteichoic acid isolated from Streptococcus lactis.
Amidohydrolases↗
Regulation of Forssman antigen expression during maturation of mouse stromal macrophages in haematopoietic foci.
During a trial to develop a monoclonal antibody (mAb) specific to stromal macrophages (M phi) in haematopoietic foci, we have created a mAb, designated F10, that stains the stromal M phi more selectively than any other mAb reported. As F10 was found to react with Forssman glycosphingolipid (GSL) specifically and to give clearer immunostaining than anti-Forssman GSL IgG, we have studied Forssman antigen expression during maturation of the stromal M phi in splenic haematopoietic foci using F10 and a system of allogenic bone marrow transplantation which allows us to know the turnover of the stromal M phi in vivo. C3H/He mice (H-2k) were lethally irradiated and intravenously infused with the bone marrow cells of BALB/c nu-nu mice (H-2d). Splenic frozen-sections and cytocentrifuge preparations of splenic haematopoietic clusters from the recipient mice were stained with F10 and with mAb against major histocompatibility class I antigens. H-2d-type stromal M phi began to appear in the haematopoietic clusters at Week 5 and they gradually replaced H-2k-type stromal M phi. The percentage of Forssman+ stromal M phi gradually decreased and reached a nadir at Week 6, when most stromal M phi were already of the donor type. At Week 8, however, Forssman+ stromal M phi levels returned to normal. The delayed expression of Forssman antigen on the stromal M phi in haematopoietic foci following genotypic conversion suggests that Forssman antigen is regularly expressed on the subpopulation of stromal M phi, which mature well under specific microenvironmental factors in vivo.
Animals↗
Cell mitotic cycle synthesis of NIL hamster glycolipids including the Forssman antigen.
The synthesis of phospholipids and glycolipids during the cell mitotic cycle of an established hamster line, NIL, has been studied. Cells were synchronized with excess thymidine and mitotically harvested by shaking. Cells were radioactively labeled for 4 h with palmitate, glucosamine, or galactose. Lipids were analyzed by thin-layer chromatography. As cells progressed through the mitotic cycle, incorporation into phospholipids increased but the fraction represented by each remained constant. Similarly, ceramide monohexoside, dihexoside, and hematoside were labeled equally in all phases. Ceramide trihexoside and tetrahexoside were labeled only during G(1) and S. Ceramide pentahexoside (the Forssman antigen) shows density-dependent synthesis, accumulation, and reactivity. Ceramide pentahexoside was labeled during all phases of the mitotic cycle but the rate of incorporation decreased in S and G(2). The total amount of lipid assayed immunologically in cell extracts gradually increased. Exposure of the Forssman antigen in untreated or trypsin-treated cells was studied using binding of chemically labeled antiForssman antiserum. The amount of antigen detected in trypsinized cells increased during G(1) and early S but then remained constant. Mitotic cells exposed all detectable antigen. As cells progressed through the mitotic cycle, a large fraction of the Forssman antigen became cryptic.
Animals↗
Isolation, characterization, and biosynthesis of Forssman antigen in human lung and lung carcinoma.
A glycolipid was isolated from normal lung and lung carcinoma tissues. This glycolipid was identified as the Forssman antigen of pentaglycosyl ceramide by means of chemical and immunological methods. The presence of this antigenic glycolipid was observed in all the tissues examined of adult and embryo lungs, and of lung tumors irrespective of histological type. The extracts of human lung and lung tumors were capable of catalyzing the synthesis of Forssman antigen from globoside.
Adenocarcinoma↗