[Heterogeneity of clonal cell lines from a spontaneous osteosarcoma].
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Biomedical subjects
Publications and source records attributed to V Erfle.
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This study analyzed the transforming potential of murine viruses in organ cultures of mouse fetal condylar cartilage: Finkel-Biskis-Jinkins-Murine sarcoma virus (FBJ-MuSV) and Finkel-Biskis-Reilly-Murine sarcoma virus (FBR-MuSV). It was only the FBR-MuSV isolated from a radiation-induced osteosarcoma, that induced morphological changes as early as 24 hours following the infection. The latter manifested itself by a marked enlargement of the number of progenitor cells concomitant with an accumulation of spindle-like cells, giant cells, and pleomorphic cells along with large bone spicules and heavy mineralization of the remaining cartilage. The newly formed tissue synthesized type I collagen and revealed profound invasive characteristics. By day 7, FBR-MuSV-infected cultures acquired the appearance of an osteosarcomatouslike lesion. Electron microscopy examinations revealed that both the matrix and the osteogenic cells in the induced tumors differed markedly from that encountered in normal mammalian osseous tissue. To determine which cells within the condylar tissue served as the target for the FBR-MuSV, we used antibodies against viral P30 protein for an indirect immunoperoxidase reaction. The chondroprogenitor cells were the only ones that reacted positively for the virus-specific protein. Further, the in vitro-induced tumor was tumorigenic in syngeneic mice, hence, brought about the development of osteo-fibrosarcoma subcutaneously. By contrast to FBR-MuSV, the FBJ-MuSV did not elicit similar transformative effects in vitro. Since both viruses possess the fos oncogene, it has been suggested that the unique tumorigenic potential of the FBR-MuSV may be linked to structural alterations in the fos oncogene product.
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Newborn female strain NMRI mice were injected with a mouse retrovirus (OA MuLV) known to induce osteopetrosis. Primary skeletoblast cell cultures were established from humeri and calvaria of 3-day-old, 7-day-old, and 28-day-old animals. Infectious ecotropic MuLV was found in all humerus cultures from infected animals and in 7-day and 28-day calvaria cell cultures. Levels of alkaline phosphatase activity were markedly higher in cultures of calvaria and humeri from infected mice than in those from controls. In vitro infection of undifferentiated periosteal cells was followed by a decrease in cell growth and an increase in alkaline phosphatase activity. In contrast, differentiated osteoblast-like cells were barely susceptible to OA MuLV infection, and the virus did not influence their cell growth or differentiation. Electron-microscopic studies of skeletal tissue from infected old osteopetrotic mice showed virus particles associated with and budding from osteocytes and accumulated in devitalized osteocyte lacunae. The results indicate that progenitor cells of the osteoblastic lineage represent the target cells for OA MuLV in bone tissue, that virus infection induces an increase in osteoblastic activity, and that infected cells produce virus until full development of the disease.
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The expression of 7 protooncogenes (c-sis, c-abl, c-mos, c-bas, c-Ki-ras, c-fos, c-myc) was examined in transplants and established cell lines from spontaneous and radiation-induced murine osteosarcomas. The transplant tumors were compared with different tissues, particularly skeletal tissue (sternum), and the osteosarcoma cell lines with fibroblast lines from the same mouse strains. C-sis was expressed above the level of controls in 2 osteosarcomas (TV, Os5). Three osteosarcomas showed over-expression of c-abl (TVK, DOS, Os5), c-bas (DOS, Os5 and V893) and c-fos (TVK, DOS, Os5), and 4 osteosarcomas showed over-expression of c-Ki-ras (TVK, DOS, Os5, Os16) and c-myc (TVK, DOS, TV, Os5). C-mos expression was not observed under the conditions used. One cell line (Os5) showed an altered transcript (1 kb transcript of c-fos). Apart from the relatively frequent increase in expression of the c-myc and c-ras-family, there was no indication that any particular protooncogene or combination of protooncogenes was associated with murine osteosarcomas.
An N-ecotropic murine leukemia virus (OA MuLV), originally isolated from spontaneous osteomas of strain 101 mice, was molecularly cloned. The virus induces osteomas, osteopetrosis, and malignant lymphomas in NMRI mice. The cloned virus was analyzed by heteroduplex analysis, restriction enzyme mapping, and oligonucleotide mapping. The data show a very close relationship to the endogenous Akv prototype virus with some differences in the gag and the env region. The nucleotide sequence of the U3 region of OA MuLV LTR revealed a structure within the presumable enhancer region very similar to the U3 sequences of the FBJ murine sarcoma virus and its associated helper virus. The significance of these specific structures for the oncogenicity of the virus and the development of the typical disease pattern is discussed.
Antibodies against HTLV-III, neopterin levels in blood and urine, TH/TS ratio and hepatitis marker were determined in 34 clinically symptom-free persons known to be intravenous drug abusers. 15 persons were positive in the ELISA and Western-blot tests. There was a strong reaction to protein p24 compared with that to protein p41. In 12 of 14 persons who were antibody-positive the neopterin level in morning urine was elevated; an abnormal TH/TS ratio was present in nine of 13 persons. In future, determination of neopterin and of antibodies against certain proteins of HTLV-III may make it possible to provide a simple way of prognosticating on the course of an HTLV-III infection.
The clinical, immunological, and serological status of 28 patients with hemophilia A and of 13 patients with hemophilia B was investigated. Thirty-four patients were treated regularly by clotting factor concentrates and 7 patients had been substituted only 1 to 4 times. Almost all patients with severe hemophilia suffered from hepatopathy. No patient had clinical evidence of the acquired immunodeficiency syndrome (AIDS). Asymptomatic hemophiliacs showed a decreased number of T-helper (OKT 4) cells and an increased number of T-suppressor (OKT 8) cells, which resulted in an inversed OKT 4/OKT 8 cell ratio. Natural killer cell activity of all patients was decreased compared to controls. After culture there was no significant difference of NK cell activity between hemophiliacs and controls. This phenomena was interpreted as a possible maturation defect of NK-cells in vivo. No relationship between immunological alterations and hepatopathy, hepatitis markers, CMV antibodies, amount and source of required factor concentrates, and the kind of hemophilia was observed. IgG immunoglobulins were higher and the OKT 4/OKT 8 ratio lower in the eight patients with lymphadenopathy than in patients without lymphadenopathy. The prevalence of antibodies to human T-lymphotropic virus (HTLVIII) was measured in 35 hemophiliacs and in 25 polytransfused patients, most of whom were suffering from acute leukemia. In 8 of 35 hemophiliacs antibodies to HTLVIII virus were detected by an enzyme linked immunosorbent assay (ELISA) and confirmatory tests. All seropositive patients were treated by blood products from the United States. Eight hemophiliacs treated by factor concentrates from German donors only were seronegative. In comparison 2 of 25 examined non-hemophilia patients receiving multiple blood products from local donors were seropositive for HTLVIII. The results show that hemophilia patients treated by imported clotting factor concentrates have a high risk of HTLVIII positivity. Hemophiliacs substituted by blood products obtained by local donor pools have only a small risk of infection. Because non-hemophiliac polytransfused patients had HTLVIII antibodies, there must be asymptomatic virus carriers in the local donor pool. The HTLVIII antibody screening of all donors and the heat treating of factor concentrates will give better therapeutic safety.
We have found human DNA to contain a number of sequences related to simian sarcoma associated virus (SSAV). One of these sequences was isolated from a human genomic library. The molecular clone, termed S71, contains regions homologous to SSAV gag and pol fragments and SSAV LTR. Furthermore, hybridization experiments and DNA sequencing revealed distinct homologies to the reverse transcriptase coding region of several other retroviruses including baboon endogenous virus (BaEV) and murine leukemia viruses (MuLV) as well as retrovirus-like elements. Some sequence homology was also found with the C-type retrovirus-related multicopy human clone 4-1. S71 is present in only one copy per human genome equivalent and exhibits an EcoRI restriction fragment length polymorphism.
The activation of endogenous retroviruses (MuLV) by internal irradiation and the presence of activated retroviruses in radiation-induced murine osteosarcomas as well as their biological properties in vivo and in vitro were studied. Ecotropic and xenotropic MuLV were expressed dependent on the radiation dose in spleen, bone marrow and bone tissues of C57Bl/6 mice after 224Ra treatment. Radiation-induced osteosarcomas of BALB/c, C57Bl/6 and C3H X 101/F1 mice harboured infectious ecotropic and/or xenotropic viruses whereas in osteosarcomas of NMRI mice predominantly virus protein could be detected. In about 50% of the radiation-induced osteosarcomas of BALB/c mice an amplification of ecotropic proviruses could be detected. This was not found in clonally grown cells from non-tumorous tissues. MuLV from radiation-induced osteosarcomas induced osteopetrosis, osteomas and lymphomas after infection of newborn NMRI mice. In differentiating bone tissue the viruses were found to infect predominantly osteoblast precursor cells suggesting that virus infection results in increased growth and metabolic activity of these cells, which may be a possible mechanism for their pathogenic action in bone tissues.
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Mandibular condyles of late embryonic NMRI mice were used to study the effect of the FBR murine osteosarcoma virus in an in vitro tissue culture system. Chondroprogenitor cells and chondroblastic cells present in the condylar tissue normally undergo rapid differentiation in vitro which results in an advanced stage of bone formation. The infection of condyles with FBR murine osteosarcoma virus induced the transformation of bone progenitor cells and the formation of an atypical proliferative osseous lesion. In markedly disorganized tissue many spindle-like cells, giant cells, and pleomorphic cells were seen together with the formation of large bone spicules and the heavy mineralization of osteoid-like material and of the remaining cartilage. Fibroblast-like cells were found to penetrate from the perichondrial zone into the condylar mass and also into the underlying collagen sponge. The in vivo growth characteristics of FBR murine osteosarcoma virus-infected condyles after 3 days in culture were studied via s.c. transplantation into syngeneic mice. Control condyles developed normal trabecular bone, whereas the infected condyles induced a strong cellular response with the presence of atypical cells and newly formed connective tissue and bone in situ. These observations raise the possibility of a novel approach for further investigations related to numerous aspects of virus-induced osteosarcomagenesis.
We report the first case of acquired immunodeficiency-syndrome (AIDS) with Kaposi's sarcoma which at the same time has also a changing biclonal gammopathy (initial: double IgG-paraproteinaemia, later on IgA-IgG double-paraproteinaemia) due to different lymphocyte clones. The relationship of a B-cell neoplasia to a concurrent disease of T-cells is discussed.
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In 1984 10,281 sera were collected in the FRG and examined for antibodies to HTLV-III (LAV) with an enzyme-linked immunosorbent assay and confirmative tests. Of the German AIDS patients 81% have antibodies. Individuals belonging to AIDS risk groups, homosexuals, haemophiliacs and i.v. drug abusers, have antibody frequencies between 25%-72%. The detection of HTLV-III antibodies in blood donors indicates that the virus is being transmitted by blood transfusions.