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Blood group isoantigen deletion in carcinoma in situ of the urinary bladder.

Blood group isoantigens A and H (O) were measured by the Secific Red Cell Adherence (SRCA) Test in nine radical cystectomy specimens removed from patients with extensive carcinoma in situ of the urinary bladder. All bladders had areas of histologically normal epithelium and areas of epithelial atypia in addition to the carcinoma in situ. In eight cases, tissue-associated blood group isoantigens were deleted in areas showing either atypia or carcinoma in situ. Isoantigens were present in these areas in the ninth case. Blood group isoantigens were absent in approximately half of the sections of histologically normal epithelium. We propose that areas of epithelium which appear benign according to conventional histological criteria but in which the blood group isoantigens are absent may represent areas of low grade carcinoma in situ.

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Isoantigens A, B, H in normal skin and tumors of the epidermal appendages.

Isoantigens A, B, H(O) in biopsy specimens of 26 normal skin and in 35 adnexal tumors were studied by the red cell adherence (RCA) test of Davidsohn. Isoantigens were detected in the stratum corneum, stratum granulosum, stratum spinosum, acrosyringium, keratogenous zone of hair follicle, eccrine duct, and eccrine gland. The apocrine gland and duct consistently had negative test results for isoantigens. Seventy percent of all tumors tested contained isoantigens and included syringoma, hidrocystoma, syringocystadenoma papilliferum, spiradenoma, eccrine poroma, dermal duct tumor, clear cell hidradenoma, and cylindroma. We interpreted the presence of isoantigens in adnexal tumors as being evidence of immunologic maturity with differentiation toward eccrine structures. The RCA test is a new, sensitive, immunologic technique that can complement enzyme histochemical stains and electron microscopy in the study of the histogenesis of adnexal skin tumors.

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Isoantigens A, B, and H in morphologically normal mucosa and in carcinoma of the larynx.

A modified Coombs' mixed cell agglutination reaction was performed on biopsy specimens of 24 patients with squamous cell carcinoma of the larynx. Isoantigens A, B, and H(O) were absent in 15 cases, partially lost in eight, and completely retained in one case. In four cases the initial biopsy specimens were histologically benign yet negative for isoantigens. In subsequent biopsies these showed the presence of carcinoma. The morphologically normal-appearing squamous mucosa adjacent to a carcinoma was found to be devoid of isoantigens in ten cases. The investigation suggested that isoantigenic alteration as detected by mixed cell agglutination reaction was a sensitive sign, since it occurred in squamous mucosa prior to recognizable morphologic change for malignancy. The reaction may represent a functional dedifferentiation in cancerous tissue, analogous to but sometimes preceding morphologic dedifferentiation. The results were comparable to those in previous studies of neoplasms of various other anatomic sites.

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Carcinoembryonic antigen: evidence for multiple antigenic determinants and isoantigens.

Carcinoembryonic antigen (CEA) preparations, from various sources were compared by radioimmunoassay. The preparations studied included four CEA standards (CEA-Roch, CEA-Montreal, CEA-City of Hope, and CEA-British) and CEA from serum and liver metastases of a patient with cancer of the colon who had an extremely high concentration of serum CEA (more than 26,000 ng/ml). The data indicate that the CEA-Roche standard differs significantly from the other three CEA standards tested, and that the serum CEA from the patient was antigenically different from currently available CEA standards as well as from the CEA obtained from the patient's own liver metastases. These antigenic differences were reflected in radioimmunoassay inhibition curves that were different and that were not affected by perchloric acid extraction of CEA. Because of the antigenic variation in the serum CEA, markedly different CEA concentrations (varying by three orders of magnitude) were measurable by two different antisera (Roche and Montreal). All the various CEA standards and samples cochromatographed on columns of Sepharose-6B, despite the large antigenic variation. We postulate that CEA consists of a family of "isoantigens" with multiple antigenic determinants. We identified a serum CEA isoantigen that was different from the currently available standards. Consequently, we believe that results of radioimmunoassays currently used for CEA measurement may not represent absolute concentrations of serum "CEA", but may reflect the binding affinity of different isoantigens to a particular polyvalent CEA antiserum.

Carcinoembryonic Antigen

Isoantigen loss in cervical neoplasia. Demonstration by immunofluorescence and immunoperoxidase techniques.

Tissue sections from normal uterine cervix and from uterine cervices with dysplasia, carcinoma in situ, and invasive cancer were studied in 70 instances using immunofluorescence and immunoperoxidase techniques for demonstrating the presence of isoantigens A and B. Isoantigens were demonstrated in normal squamous epithelium. Antigens were lost in foci of invasive cancer, were considerably reduced or lost in areas of classic carcinoma in situ in all cases, and were reduced in most cases of dysplasia. These observations confirm those obtained with the specific red cell adherence test. The results suggest that immunofluorescence and immunoperoxidase techniques are sensitive, reproducible, easily performed methods for demonstrating the loss of isoantigens in cervical neoplasia.

Carcinoma in Situ

Specific lysis of human colon tumor cells by antibodies to CEA and isoantigen A: dependence on rabbit serum or neuraminidase.

The 51Cr cytotoxicity test was used to measure specific antibody reactions against carcinoembryonic antigen (CEA) and isoantigen A on the surface of human colon tumor cells. When human serum or guinea pig serum was used as a source of complement, no anti-CEA or anti-isoantigen A cytotoxicity was demonstrable despite binding of specific antibodies and activation of complement at least through the C3 component on the cell surface. In contrast, specific anti-CEA and anti-isoantigen A cytotoxicity was demonstrated when rabbit serum was used as a source of complement. Specific antibody-mediated cell lysis was also achieved with guinea pig complement if the cells were treated with neuraminidase before testing. These results support the concept that certain tumor cells have surface properties that render them resistant to immune lysis.

Adenocarcinoma

Isoantigens A, B and H in urinary bladder carcinomas following radiotherapy.

ABH tissue isoantigens were measured by the Specific Red Cell Adherence (SRCA) test in 66 surgical specimens of urinary bladder, including 53 transitional cell carcinomas, 2 squamous cell carcinomas and 11 controls. The SRCA test was strongly positive in 10 of 11 controls. ABH isoantigens were absent or equivocally present in 68 percent of noninvasive carcinomas (stage 0) and in 65 percent of invasive carcinomas. Clinical histories revealed that all patients with invasive carcinoma who had strongly positive SRCA test results had received prior radiotherapy to the bladder region. None of the patients with invasive bladder carcinoma with negative or weakly positive SRCA tests had been radiated. Histopathology of tumors in both groups was similar. Results of this retrospective study support the hypothesis that radiation may induce differentiation in tumors, possibly through an enhancement of Golgi apparatus function. The SRCA test should not be used as a predictor of the biological behavior of future recurrences in patients with bladder carcinoma who have received therapeutic radiation since radiation may produce "false positive" SRCA test results.

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Blood group isoantigens in human benign and malignant vascular tumors.

Paraffin material of 31 benign and malignant vascular tumors was investigated with respect to their blood group isoantigen (BG) content by the mixed cell agglutination reaction (MCAR). In capillary hemangioma, BG was found in endothelial cells as well as in solid buds. Benign hemangioendothelioma found in endothelial cells as well as in solid buds. Benign hemangioendothelioma found in children differed from that found in adults in that in juvenile cases only endothelial cells expressed BG whereas in adult cases BG isoantigenity was present in endothelial cells as well as in intercapillary cellular elements. In pericytomas only endothelial cells were BG positive, whereas the tumor cells lacked BG. Similar results were obtained with glomus tumors. All but one hemangiosarcoma were BG negative. In one case, however, which probably resembled a "true" malignant hemangioendothelioma (Stout and Lattes, 1967) the tumor cells contained BG in conspicuous amounts.

Adult

Tumor-associated antigens in H-2 hemizygous isoantigenic variants of a somatic cell hybrid, derived from the fusion of a 3-methylcholanthrene-induced sarcoma and a mammary carcinoma.

To check the suggestion that 3-methylcholanthrene-induced, sarcoma-associated, tumor-specific transplantation antigens (TSTA) could be modified H-2 antigens, reciprocal isoantigenic variants derived from TA3Ha/MSWBS hybrid cells were examined. This hybrid was produced by the fusion of the TA3Ha ascites carcinoma (H-2a) and the 3-methylcholanthrene-induced MSWBS ascites sarcoma (H-2s). MSWBS expresses a strong TSTA capable of inducing a rejection reaction in the syngeneic A.SW host. The genetic determinants of the H-2 complex are known to be localized on chromosome No. 17. TA3Ha contributes two normal, telocentric chromosomes No. 17 to the hybrid. In contrast, both chromosomes No. 17 of MSWBS are localized on readily identifiable translocations (17/1 and 17/M1). We have previously shown that the chromosomes No. 17 of one parental strain or the other (but not both) can be eliminated from the hybrid upon selective passage in the opposite parental strain. The present studies showed that the two strain A-compatible variants that have lost the sarcoma-derived 17-chromosomes still contained the same TSTA as the two reciprocal strain A.SW-compatible variants that have lost the mammary carcinoma-derived H-2 chromosomes. These findings argue against the possibility that methylcholanthrene-induced TSTA is a modified form of H-2 or that its structural determinants(s) is localized on chromosome 17.

Animals

[Origin of stromal bone marrow mechanocytes according to the results of typing them by isoantigens and chromosomal markers].

The bone marrow of radiochimaeras and heterotopic bone marrow transplants were used to study the origin of precursors of the fibroblasts growing in the monolayer cultures of hemopoietic tissue. In the bone marrow explants of the (C57BL/6 X CBA) F1 mice, in which the CBA bone marrow was transplanted following the lethal irradiation, the fibroblasts grown in the colonies were of recipient origin judging by isoantigens in the reaction of indirect immunofluorescence with the anti-C57BL/6-serum. At the same time in the bone marrow explants from heterotopic transplants (CBA leads to CBA X C57BL/6) the fibroblasts grown in colonies were of donor origin. The cultures of hemopoietic cells of the bone marrow of females heterotopically transplanted in the singenic male (guinea pigs Huston) contained only fibroblasts which were of donor origin judging by sex chromosomes in the metaphase plates of dividing cells. Hence, the bone marrow precursors of fibroblasts do not depend histogenetically on hemopoietic cells and are not replaced at the expense of repopulating cells of the second partner.

Animals

Isoantigenic expression of Forssman glycolipid in human gastric and colonic mucosa: its possible identity with "A-like antigen" in human cancer.

The heterogenetic Forssman antigen is a glycosphingolipid, a ceramide pentasaccharide with the structure GalNAcalpha1-->3GalNAcbeta1-->3Galalpha1-->4Galbeta1-->4Glc-->ceramide. Forssman-positive animals are capable of synthesizing this compound in tissues or in erythrocytes, in contrast to the Forssman-negative species, including humans, which are incapable of adding the last carbohydrate in the sequence of the Forssman antigen, namely alphaGalNAc. The Forssman glycolipid and its precursor globoside were examined in twenty-one samples of surgically extirpated gastrointestinal mucosa and tumors derived therefrom. The results revealed that a few patients had chemically and immunologically detectable levels of the Forssman glycolipid as a normal component of their gastrointestinal mucosa (F(+) population); in contrast, the majority of patients did not contain this glycolipid in their normal mucosa (F(-) population). Whereas the F(-) population included blood groups A, B, and O, the F(+) population did not correspond to blood group A. The Forssman status in tumors taken from the F(+) or F(-) population showed the following striking features: (i) all tumors derived from F(-) mucosa possessed Forssman glycolipid, whereas (ii) none of the tumors originating in F(+) mucosa contained Forssman glycolipid. Globoside, the immediate precursor of Forssman antigen, was distributed equally among F(+) and F(-) mucosa and the tumors derived therefrom. Thus, the expression of Forssman antigen in gastrointestinal mucosa appears akin to that of an isoantigen. Furthermore, the Forssman antigen that appears in tumors of the F(-) population could represent a human tumor-associated antigen. In view of the strong crossreactivity of Forssman antigen with blood group A determinants, the appearance of Forssman antigen in human tumors could be related to the "A-like antigen" (or "neo-A antigen") of human tumors reported previously [Hakomori, S., Koscielak, J., Black, K. J. & Jeanloz, R. W. (1967) J. Immunol. 98, 31-38; Häkkinen, I. (1970) J. Natl. Cancer Inst. 44, 1183-1193].

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Tumor-associated antigens in isoantigenic variants of a 3-methylcholanthrene-induced sarcoma.

A sarcoma was induced in an (A.CA X A.BY)F1 mouse. Two isoantigenic variants were selected by loss of one H-2 antigen. The tumor-associated transplantation antigens (TATA) of these variants were compared as to their specificities in (A.CA X A.BY)F1 mice. Both transplantation and indirect membrane immunfluorescence tests revealed that TATA of both variants did not cross-react. Thus selecting angainst different H-2 antigens also selected different TATA. Karyotype studies suggested that both variants originated from a unique clone.

Animals

Immunological cross-reactivity between a glycoprotein isolated from Trichophyton mentagrophytes and human isoantigen A.

A glycoprotein isolated from the cell wall of Trichophyton mentagrophytes was assessed for its cross-reaction with human blood group isoantigens. Rabbit antiglycoprotein antibodies agglutinated human erythrocytes of blood groups A1 and A2, and precipitated Blood Group Substance A in agarose gels. Erythrocytes of blood group B were only slightly agglutinated, and O(Rho+) and O(Rho-) erythrocytes were not. Additionally, the glycoprotein was shown to specifically inhibit isoagglutination of erythrocytes of group A. Partial identity between the glycoprotein and a crude extract of the fungus was demonstrated by immunodiffusion. Analyses revealed the glycoprotein to be composed of approximately 17% protein and 80% carbohydrate. The glycoprotein was found by indirect immunofluorescence to be located in the mycelial cell wall. The possibility that cross-reacting antigens may lead to a chronic, spreading infection is discussed.

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