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Biomedical subjects

C M Croce

Publications and source records attributed to C M Croce.

At least 433 records · Page 24Linked to original sources

Selective suppression of the transcription of ribosomal genes in mouse-human hybrid cells.

Mouse-human hybrid cells that preferentially segregate either mouse or human chromosomes were analyzed for their relative content of mouse and human rRNA genes and for their capacity to transcribe these genes. A distinctive Hind III restriction fragment containing 28S rRNA sequences was used to distinguish between mouse and human rDNA and a set of distinctive loop structures in the 45S pre-rRNA was used to distinguish between mouse and human gene transcripts. Our results indicate that the genes of only one species are transcriptionally active in these hybrid cells, even though both sets of genes are present.

Animals↗

Expression of malignancy in hybrids between normal and malignant cells.

Somatic cell hybrids between either normal human fibroblasts, phenotypically normal mouse fibroblasts or mouse peritoneal macrophages and HT1080 human diploid fibrosarcoma cells were studied for their ability to form tumors in nude mice. The results of this study indicate that tumorigenic behavior is expressed as a dominant trait in both human-human and mouse-human hybrid cells.

Animals↗

Somatic cell hybrids between totipotent mouse teratocarcinoma and rat hepatoma cells.

We have produced somatic cell hybrids between totipotent mouse teratocarcinoma cells and rat hepatoma cells. These hybrids were tested for the expression of liver specific functions expressed in the hepatoma cell parent and for their ability to differentiate when injected into nude mice. The results of this study indicate that hybrid cell clones do not resemble either of the parental cells, since they do not produce albumin and tyrosine aminotransferase that are expressed in the rat hepatoma parent, and are incapable of forming either teratocarcinomas or hepatomas when injected in experimental animals.

Albumins↗

Segregation of rat chromosomes in somatic cell hybrids between rat cells and HT 1080 human fibrosarcoma cells.

We produced somatic cell hybrids between HT 1080-6TG human fibrosarcoma cells and either rat white blood cells (WBC) or cells directly derived from rat spleen. Karyologic and isozyme analyses of hybrid cells indicated that they preferentially lose rat chromosomes. Hypoxanthine-aminopterine thymidine-selected hybrid clones expressing rat hypoxanthine phosphoribosyltransferase (HPRT), glucose-6-phosphate dehydrogenase (G6PD), and phosphoglycerate kinase (PGK) and containing the rat X chromosome were counterselected in a medium containing 30 micrograms/ml of 6-thioguanine. Concordant loss of the rat X chromosome and of the expression of rat HPRT and G6PD was observed in the hybrid clones.

Animals↗

Expression of late viral functions in SV40-infected rat-monkey somatic cell hybrids.

Somatic cell hybrids between a rat hepatoma cell line and an SV40-transformed African green monkey kidney cell line (T22 TK-) have been isolated and their response to infection by SV40 virus characterized. The parental rat cells are nonpermissive to productive infection by SV40, while the parental monkey cells, which are T-antigen positive and capsid antigen negative, are permissive to productive infection. Hybrid clones, which had segregated monkey chromosomes, were infected with SV40 virus, and some clones were found to be negative for SV40 capsid antigen production while other clones were positive. Capsid antigen-positive clones production while other clones were positive. Capsid antigen-positive clones produced little or no form I SV40 DNA, no infectious virus, and were not capable of rescuing SV40 from SV40-transformed mouse cells. Such hybrids, which produce late viral proteins in the absence of mature progeny viral DNA may be useful in the study of control of SV40 RNA transcription.

Animals↗

Xenogeneic gene expression in chimeric mice derived from rat--mouse hybrid cells.

Thymidine kinase-deficient OTT6050 mouse teratocarcinoma cells were fused with hypoxanthine phosphoribosyltransferase-deficient Fu5AH rat hepatoma cells by means of inactivated Sendai virus. The resulting hybrid cells, which were selected in hypoxanthine/aminopterin/thymidine medium, retained almost all of the mouse chromosomes and various numbers of rat chromosomes, and showed many chromosomal rearrangements. The hybrid cells, as well as both parental lines, formed tumors after subcutaneous injection into athymic nude mice. Single rat--mouse hybrid cells from a clonally established subline were transplanted into C57BL6/J mouse blastocysts carrying many genetic markers suitable for the detection of hybrid cell-derived tissue contributions. From 144 blastocysts, each of which was injected with a hybrid cell and then surgically transferred to the uterus of a pseudopregnant foster mother, 62 adult mice developed without any visible coat mosaicism. However, three of these mice showed internal hybrid-cell participation in their livers and a limited number of organs of endomesodermal origin. A tumor classifiable as hemangio endothelioma was found in the liver, the only mosaic tissue, of one of the chimeric mice. Nine different rat-specific enzyme variants were detected in the mosaic organs. A considerable number of variations concerning the presence and quantitative activity of the foreign gene products probably resulted from chromosomal segregation, tissue-specific gene activity, or dosage compensation during differentiation in vivo. Our results demonstrate that cultured malignant rat--mouse hybrid cells differentiate normally and become functionally integrated during development. The appearacne in vivo of certain rat-specific gene products that are not found in the hybrid cells under conditions in vitro indicates differential gene expression of the introduced xenogeneic chromosomes.

Alcohol Oxidoreductases↗

Chromosomal location of the genes for human immunoglobulin heavy chains.

We have studied somatic cell hybrids between P3x63Ag8 mouse myeloma cells deficient in hypoxanthine phosphoribosyltransferase (EC 2.4.2.8) and either human peripheral lymphocytes or human lymphoblastoid or myeloma cells for the production of human immunoglobulin chains and for the expression of enzyme markers assigned to each of the different human chromosomes. Human chromosome 14 was the only human chromosome present in all independent hybrids producing mu, gamma, and alpha human heavy chains. In two of the independent hybrids that produced human heavy chains, human chromosome 14 was the only human chromosome present in the hybrid cells. Loss of human chromosome 14 from these hybrids resulted in the concomitant loss of their ability to produce human immunoglobulin heavy chains. In view of these results, we conclude that the genes for human immunoglobulin heavy chains are located on human chromosome 14 in immunoglobulin-producing human cells.

Animals↗

Reactivation of silent rRNA genes by simian virus 40 in human-mouse hybrid cells.

Mouse-human hybrid cells were used to study the ability of simian virus 40 to regulate the expression of rRNA genes in vivo. In these hybrid cells, only the rRNA genes of the dominant species are expressed; the genes for the rRNA of the recessive species are silent. Simian virus 40 infection of these hybrids led to the production of two distinct 28S rRNA species as analyzed by agarose/2.4% polyacrylamide gel electrophoresis. These species were identified as human and mouse rRNAs. This result was confirmed by histochemical studies which indicated that the nucleolus organizer regions of both mouse and human chromosomes were actively synthesizing rRNA in the virus-infected hybrid cells. These results indicate that simian virus 40 infection can induce the expression of otherwise silent rRNA genes.

Animals↗

Genetics of type II glycogenosis: assignment of the human gene for acid alpha-glucosidase to chromosome 17.

We have studied somatic cell hybrids between thymidine kinase (EC 2.7.1.75) deficient mouse cells and human diploid fibroblasts for the expression of human acid alpha-glucosidase (EC 3.2.1.20). A deficiency in this enzyme is associated with the type II glycogenosis or Pompe disease. All 30 somatic cell hybrids selected in hypoxanthine/aminopterin/thymidine medium expressed human acid alpha-glucosidase and galactokinase (EC 2.7.1.6) and retained human chromosome 17; counterselection of the same hybrids in medium containing 5-bromodeoxyuridine resulted in the growth of hybrids that concordantly lost the expression of human acid alpha-glucosidase and galactokinase as well as human chromosome 17. Hybrids between thymidine kinase-deficient mouse cells and fibroblasts from a patient with Pompe disease that contained human chromosome 17 were found not to express human acid alpha-glucosidase. Because we have already shown that hybrids between mouse peritoneal macrophages and GM54VA simian virus 40-transformed human cells selectively retain human chromosome 17 and lose all other human chromosomes, we tested 13 independent mouse macrophage x GM54VA hybrid clones, including two that retained human chromosome 17 and no other human chromosomes, for the expression of human acid alpha-glucosidase and galactokinase. All 13 hybrid clones were found to express these human enzymes. Thus, we conclude that the gene coding for human acid alpha-glucosidase is located on human chromosome 17.

Animals↗

Assignment of a gene for uridine diphosphate galactose-4-epimerase to human chromosome 1 by somatic cell hybridization, with evidence for a regional assignment to 1pter yields 1p21.

The presence of human uridine diphosphate galactose-4-epimerase (GALE) was found to correlate with the presence of chromosome 1 in somatic cell hybrids between man and mouse. The gene for GALE can therefore be assigned to human chromosome 1. Using a chromosome 1 rearrangement, we have been able to regionally assign GALE to the pter yields p21 region.

Animals↗

Expression of HLA-A, but not of HLA-B, in mouse-human somatic cell hybrids carrying the region p21 leads to pter of human chromosome 6.

Mouse-human somatic cell hybrids containing human chromosome 17 carrying the region p21 leads to pter of human chromosome 6 and no other human chromosomes, were found to express HLA-A but not HLA-B. Counterselection of the hybrid cells in medium containing 5-bromodeoxyuridine resulted in the growth of hybrid cells that have concordantly lost the expression of HLA-A and the human translocation chromosome.

Animals↗

Studies on the association of the Epstein-Barr virus genome and human chromosomes.

We have used human-mouse somatic cell hybrid to study the association between the EBV genome and the cellular genome. Attempts were made to identify a specific human chromosome(s) with which the EBV genome is associated. Our data suggest that at least in the mouse fibroblast/Burkitt lymphoma hybrid combination studies, the EBV genome is not associated with any specific human chromosome.

Antigens, Viral↗

Species-specific suppression of histone H1 and H2B production in human/mouse hybrids.

Ten human/mouse hybrid cell lines that segregate either human or mouse chromosomes were examined for the expression of human- and mouse-specific histones H1 and H2B. Results of this study indicate that the human and mouse chromosomes in hybrid cells that segregate human chromosomes (M greater than H hybrids) contain only mouse histone H1 and H2B. Chromosomes in hybrid cells that segregate mouse chromosomes (H greater than M hybrids) contain only human H1 and H2B histones. Loss of the ability to produce either human or mouse histones does not seem to be due to the loss of specific human or mouse chromosomes because M greater than H hybrids retaining at least one copy of each human chromosome contain only mouse H1 and H2B and H greater than M hybrids retaining at least one copy of each mouse chromosome contain only human H1 and H2B histones. These results, together with those concerning histone H4 acetylation levels and ratios of variants of histones H3 and H2A that are like those in the dominant parent cell type, indicate that the control mechanisms affecting H1 and H2B expression in H greater than M and in M greater than H hybrid cells affect expression of histones H2A, H3, and H4 genes as well. The present data thus support the hypothesis that none of the histone genes that are active in the recessive parent cell type is expressed in hybrid lines that segregate recessive cell chromosomes.

Cell Line↗

Repertoire of antiviral antibodies expressed by somatic cell hybrids.

Fusion between P3 x 63 Ag8 mouse myeloma cells and spleen cells from BALB/c mice immunized with influenza type A or B or parainfluenza type 1 virus generated reproducibly antiviral antibody-producing somatic cell hybrids (hybridomas). Eleven hybridomas derived from spleen cells of mice immunized with influenza type A virus were directed against the viral hemagglutinin, one reacted with a host component derived from chickens, and one expressed a specificity not further characterized. The hybridoma antibodies tended to be highly specific for the hemagglutinin of the immunizing virus and seemed to express the same repertoire of strain-specific antibody reactivities as splenic precursor B cells, they did not express any of the frequently occurring crossreactive anti-hemagglutinin specificities. Hybridomas producing crossreactive antibodies against hemagglutinin could be obtained if priming and boosting virus were heterologous.

Animals↗