Isoantigen expression in hybrid mouse cells

Klein, George; Gars, Ulla; Harris, Henry

doi:10.1016/0014-4827(79)90516-0

Cited by 44 publications

(15 citation statements)

References 14 publications

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“…Neither hypoxanthine-quanine-phosphoribosyltransferase-deficient cells (A9HT and TA3Bimp-) nor thymidine kinase-deficient cells (B82HT) can survive in HAT medium. The explanted tumor cells therefore died, but after [8][9][10][11][12][13][14] days some HAT-resistant colonies appeared. The cells in these colonies proved to be hybrids.…”

Section: Methodsmentioning

confidence: 99%

“…The TA3Ha tumor is the ascitic form of a spontaneous mammary adenocarcinoma of the A/Sn strain (9 (11).…”

Section: Methodsmentioning

confidence: 99%

“…To measure the concentration on H-2 antigens on the surface of the cell lines, quantitative adsorption tests were done as described previously (11). The ratio of bound to free antibody, (100-P)/P, was calculated by the method of Reif (12).…”

Section: Methodsmentioning

confidence: 99%

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Tumor-Host Cell Hybrids in Radiochimeras

Wiener

Fenyõ

Klein

1974

Proc. Natl. Acad. Sci. U.S.A.

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F1 hybrid mice syngeneic or semiallogeneic with respect to the relevant tumor were lethally irradiated and then reconstituted with hemopoietic cells from strain CBAT6T6 mice. After chimerism had been established, the animals were inoculated with solid or ascites tumors. Tumor-host cell hybrids were selected from enzyme-deficient solid tumors by explanting the tumor cell suspension into hypoxanthine-amethopterin-thymidine containing medium. The selection of hybrid cells from ascites tumors was achieved by exploiting the difference between the ascites tumor cells and hybrid cells in their ability to adhere to the surface of culture vessels. T6T6 chromosomal and H-2 antigenic markers served to distinguish between the hemopoietic cells derived from the donor graft and the cells of the host. All solid tumors tested fused with cells of the irradiated host, whereas ascites tumors fused with repopulating cells ofhemopoietic origin.In two previous publications (1, 2) we have described the successful selective isolation of tumor-host cell hybrids from The type of host cell involved in the fusion is unknown. In order to assess the biological significance of the phenomenon, it was obviously important to define this, at least in outline. Radiation chimeras in which the host and the repopulating cells carried different markers appeared to present a possible approach to this question. MATERIALS AND METHODSThe experimental design was as follows. F1 hybrid mice syngeneic or semiallogeneic with respect to the test tumor were lethally irradiated and then reconstituted with CBAT6T6 hemopoietic cells. After chimerism had been established, the animals were inoculated with solid or ascites tumors. Tumor-host cell hybrids were selected as previously described (1, 2). T6T6 chromosomal and H-2 antigenic markers served to distinguish between the hemopoietic cells derived from the embryonic liver graft and the cells of the host.Production of Chimeras. C3H X DBA/2, C3H X C57B1, and C3H X C57 leaden F. hybrid mice, 6-8 weeks old, were lethally irradiated with 800 R (3). About 5 X 105 hemopoietic cells obtained from the liver of newborn CBAT6T6 mice were suspended in 0.5 ml of sterile Eagle's minimal essential medium and inoculated into the tail vein of an irradiated recipient. The development of chimerism was confirmed by examining metaphase plates of spleen or lymph node cells. As a rule, lymphopoietic and hemopoietic organs were repopulated with T6T6 marker positive cells within 2-3 weeks.Tumors. (a). Solid tumors. A9HT and B82HT are highly malignant variants selected from the L cell sublines, A9 and B82 (4). Like the original in vitro lines from which they were derived, A9HT and B82HT are enzyme-deficient; A9HT lacks hypoxanthinq -guanine -phosphoribosyltransferase (HGPRT), while B82HT lacks thymidine kinase (TK). Due to these enzyme deficiencies both lines are unable to grow on hypoxanthine-amethopterin-thymidine (HAT) medium. With inoculla of 4 X 104 to 4 X 104 cells, both lines grew progressively in newborn x-irradiated (4 J k...

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Section: Methodsmentioning

confidence: 99%

“…The TA3Ha tumor is the ascitic form of a spontaneous mammary adenocarcinoma of the A/Sn strain (9 (11).…”

Section: Methodsmentioning

confidence: 99%

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Tumor-Host Cell Hybrids in Radiochimeras

Wiener

Fenyõ

Klein

1974

Proc. Natl. Acad. Sci. U.S.A.

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“…Ehrlich ascites tumor cells also suppress the antigens of different cells after hybridization (18,19), perhaps by a similar masking mechanism. Fusion between macrophages and embryonic chick fibroblasts brought about a very slow decline in receptor activity.…”

Section: Discussionmentioning

confidence: 99%

Macrophage-Melanoma Cell Heterokaryons

Gordon

Cohn

1971

The Journal of Experimental Medicine

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The plasma membrane of the mouse peritoneal macrophage has specific receptors which enable the cell to ingest antibody-coated sheep red .cells and is also rich in a divalent cation-dependent adenosine triphosphatase (ATPase) activity (1, 2). By fusing macrophages with a strain of mouse melanoma cells which lacks these receptors, one could not only trace the fate of the macrophage membrane in the heterokaryons, but also hope to learn about the expression of cell-specific traits in animal cells (3).Macrophage homokaryons obtained by Sendai virus-induced cell fusion continued to exhibit active phagocytosis of sensitized erythrocytes. The phagocytic receptor could also be detected in heterokaryons shortly after fusion, but was progressively lost over the next 12-24 hr. The ATPase activity disappeared from heterokaryons as well, and in addition, seemed to spread out over the cell surface.Several possible mechanisms could account for these observations. These include mixing of membrane components resulting in dilution and loss of function; endo-or exocytosis without further synthesis; and masking of the macrophage membrane markers, either by serum factors or by the heterokaryon itself. We report experiments to distinguish among these hypotheses. Materials and MethodsCell Culture and Cdl Fusion Technlque.--Carried out as described (3). The macrophage phagocytic receptor was assayed with minor modifications of our previous procedure. Cover slip preparations were exposed at room temperature for 15 rain to a 0.1% suspension of sheep red blood cells (SRBC) 1 which had been coated with a 1/~o00 dilution of a rabbit anti-sheep * This work was partially supported by Grant AI 07012 from the National Institutes of Health.1 Abbreviations used in this paper: AbSRBC, antibody-coated sheep red blood cells; BGG, bovine gamma globulin; BSA, bovine serum albumin; EAT, Ehrlich ascites tumor cells; IgG, immunoglobulin G; IPT-FCS, immunoprecipitin-tested fetal calf serum; NBCS, newborn calf serum; 199 M, medium 199 + 10% NBCS; SRBC, sheep red blood cells; STI, soybean trypsin inhibitor.

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“…However, the hybrids have undergone such extensive chromosome loss that it is possible for some of the steps involved in antigen synthesis or membrane incorporation to be defective. For this reason, antigenic suppression [9,17,18], in which hybrids show an un expected suppression of the antigens contributed by the partner cell, is not a plausible explanation. Some antigen loss may be due to trypsinization of cells, but most antigens usually remain [ 19], Loss of some of the hybrid chromosomes and consequent changes in the expression of surface antigenicity are probably due to prolonged culture.…”

Section: Discussionmentioning

confidence: 99%

Microspectrofluorometric Analysis of Surface Antigens of Murine Melanoma and Hamster Peritoneal Cell Hybrids: Comparisons of Species Antigenicity, Chromosome Number, and Tumorigenicity

Erickson

1979

Oncology

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Somatic cell hybrids from viral fusions of murine melanoma (PAZG) × Chinese hamster peritoneal cells (CH) were compared with respect to surface antigenicity, karyotype and tumorigenicity. One line, F57-(9), which arose from the hybridization of two CH cells and one PAZG cell, had slight (6%) CH chromosome loss but 80% PAZG chromosome loss after 10 months in culture. These cells expressed CH antigens strongly and PAZG antigens weakly. In comparison, another hybrid, F57-(7), formed from one CH and one PAZG cell, lost 20% of its chromosomes after 10 months in vitro. These cells had a stronger expression of PAZG antigens and weaker expression of CH antigens than F57-(9). These findings indicate a direct relationship between chromosome number and antigenicity; tumorigenicity, however, does not appear to depend on the chromosome numbers of the parental cells.

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Isoantigen expression in hybrid mouse cells

Cited by 44 publications

References 14 publications

Tumor-Host Cell Hybrids in Radiochimeras

Tumor-Host Cell Hybrids in Radiochimeras

Macrophage-Melanoma Cell Heterokaryons

Microspectrofluorometric Analysis of Surface Antigens of Murine Melanoma and Hamster Peritoneal Cell Hybrids: Comparisons of Species Antigenicity, Chromosome Number, and Tumorigenicity

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