Search PubMed⌕ Search

Biomedical subjects

J G Fournier

Publications and source records attributed to J G Fournier.

48 records · Page 3Linked to original sources

On the trail of paramyxoviruses in Paget's disease of bone.

The ultrastructural discovery of microcylindric inclusions in the nuclei and cytoplasm of osteoclasts in tissue affected by Paget's disease of bone has created a new approach. The morphologic similarity of the inclusions to viral structures has stimulated further studies involving immunocytologic techniques and in situ hybridization. Polyclonal antibodies reveal the presence of paramyxovirus antigens and measles virus and respiratory syncytial virus in pagetic osteoclasts. Monoclonal and monospecific polyclonal antibodies demonstrate paramyxovirus antigens of measles virus, simian virus (SV5), and human parainfluenza virus (PF3). In situ hybridization carried out with a 3H-labeled DNA probe, specific for the measles nucleocapsid protein, detects measles virus nucleotide sequences in the nuclei and cytoplasm of pagetic osteoclasts, confirming ultrastructural and immunocytologic findings. Surprisingly, the tritiated probe also hybridizes with a large proportion of mononucleated cells: osteoblasts, osteocytes, fibroblasts, and lymphocytes. This suggests a very wide host cell range for measles virus genomic information which, however, would appear to undergo translation only in osteoclasts. The cause-effect relation between the viral information contained by diseased bone cells and Paget's disease of bone remains to be established.

Antibodies, Monoclonal↗

Subacute sclerosing panencephalitis: detection of measles virus RNA in appendix lymphoid tissue before clinical signs.

An appendix removed 15 days before onset of symptoms of subacute sclerosing panencephalitis was examined retrospectively for measles virus ribonucleic acid (RNA). Tissue sections hybridised in situ to a cloned measles virus probe of deoxyribonucleic acid specific for nucleocapsid protein showed that many cells of the lymphoid tissue contained measles virus RNA. In contrast, only a few infected lymphoid cells were detected in three out of six seropositive controls and none in three seronegative infants. A widespread chronic viral infection of the immune system, established after measles, may promote or even initiate nerve cell infection in subacute sclerosing panencephalitis.

Appendix↗

Measles virus RNA detected in Paget's disease bone tissue by in situ hybridization.

Morphological and immunocytological studies have demonstrated the presence of paramyxovirus antigens in Paget's bone disease tissue and in particular antigens related to measles virus and respiratory syncytial virus. To examine the relationship between measles virus and Paget's bone disease we used in situ hybridization and a cloned measles virus DNA probe specific for the nucleocapsid protein to detect and locate measles virus RNA sequences in Paget's bone tissue. In five patients with the disease, measles virus RNA sequences were detected not only in 80 to 90% of the multinucleated osteoclasts where there is morphological and immunocytological evidence of measles virus activity but also in 30 to 40% of mononucleated bone cells, mainly osteoblasts, osteocytes, fibroblasts and lympho-monocytes. In contrast, no hybridization was observed in bone tissue from three control patients without signs of Paget's bone disease. These results indicate that the host cell range for measles virus in Paget's disease is more widespread than has been supposed. They also demonstrate the usefulness of the in situ hybridization method to detect viral genetic information in cells where viral antigenic activity is not detectable. These observations further support the hypothesis that measles virus is involved in the pathogenesis of Paget's bone disease.

Bone and Bones↗

[Contribution of modern research technics to the identification of a virus in Paget's disease of bone].

The morphological resemblance between osteoclast inclusions in Paget's bone disease and viral structures has encouraged new research with immunocytologic methods and In Situ hybridization. Paramyxovirus antigens of measles virus and respiratory syncitial virus were detected in pagetic osteoclasts using polyclonal antibodies. Paramyxovirus antigens of measles virus, simian virus (SV 5) and human parainfluenza virus (PF 3) were demonstrated using monoclonal and monospecific polyclonal antibodies. Measles virus nucleotide sequences were detected in pagetic osteoclasts by In Situ hybridization with a 3H labelled cDNA probe specific for measles nucleocapsid protein. However, the same probe also hybridized with several types of mononucleated cells suggesting a wide host range for measles virus genomic information. Apparently, this viral information is translated only in the osteoclasts. The link between the presence of viral information in pagetic bone tissue cells and Paget's disease itself still awaits elucidation.

Bone and Bones↗

Expression of integrated hepatitis B virus DNA in PLC/PRF/5, Hep 3B, and L6EC3 cell lines detected by in situ hybridisation.

The expression of the DNA of hepatitis B virus (HBV) was analysed in three cell lines (PLC/PRF/5, Hep 3B, L6EC3) which contain the HBV DNA integrated in their genome and release the viral surface antigen (HBsAg) in relation to cell growth. Using the in situ hybridisation technique and a cloned DNA probe specific for hepatitis B virus (PTKH9), the intracellular viral RNA localisation showed that for the three cell lines, HBV RNA are present in the different cell compartments according to the age of the culture. The nucleolar and nuclear localisation are visible in the early stages of the cell growth, whereas in the later stages viral RNA are found in the cytoplasm corresponding to the maximal production of the HBsAg. These observations suggest that the nucleolus is implicated in the expression of the integrated form of HBV genetic information, the regulation of which is linked to cell growth.

Cell Line↗

Detection of measles virus RNA in lymphocytes from peripheral-blood and brain perivascular infiltrates of patients with subacute sclerosing panencephalitis.

To clarify the relation between lymphocytes and measles virus in subacute sclerosing panencephalitis, we used in situ hybridization and a cloned measles virus DNA probe, specific for nucleocapsid protein, to detect measles virus RNA sequences in circulating lymphocytes and brain perivascular cuffs of patients with subacute sclerosing panencephalitis. Seventy to 90 per cent of peripheral mononuclear cells from three such patients were found to contain measles virus RNA sequences. In contrast, only a few infected cells were observed in four seropositive adults (0.1 to 5 per cent) and three age-matched children (10 to 15 per cent) used as controls. In one sample of brain tissue from a patient with subacute sclerosing panencephalitis, viral RNA sequences were also detected in nerve cells and in numerous cells from the perivascular infiltrates. In contrast, no hybridization was observed in brain tissue from a patient with herpetic encephalitis and from a patient with postlymphoma encephalitis. We conclude that measles virus has a strong tropism for lymphocytes and nerve cells in subacute sclerosing panencephalitis and that lymphocytes may be involved in the pathogenesis of the disease.

Adult↗

Localization of measles virus nucleic acid sequences in infected cells by in situ hybridization.

Vero cells were infected with measles virus and hybridized in situ to a cloned DNA fragment containing specific sequences for measles nucleocapsid protein. The DNA was labelled with tritium by nick-translation. The viral RNA were detected in the cytoplasm 21 hrs after infection. In many cells, the probe hybridized to nuclear structures, and in several mitotic cells, to chromosomes. After 36 hrs of infection, hybridization sites were found both in the center and in many nuclei of all the polykaryons. These results indicate that cellular distribution of viral RNA molecules varies in the course of infection. They further suggest that the nucleus plays a more active role than expected in measles virus transcription and replication.

Animals↗

[Application of immunoelectron microscopy to the detection of viruses in a water medium].

Adenoviruses were immersed in demineralyzed and deionized water for 5 days. The water was subsequently analyzed by Immuno-Electron-Microscopy for detection of viral particles. An attempt of quantation was also made on control, untreated particles. Viruses dispersed in water can be detected by the technique employed but quantitation is limited by the heterogeneous dispersion of the particles on the grids.

Adenoviridae↗

Infection of cultivated CNS tissue with herpes virus, HSVII. A reappraisal.

Organized cultures of newborn rat and hamster cerebellum were infected with herpes virus type II, after 7 and 14 days "in vitro". 48 h after the infection, electron microscopic examination of the cultures showed that astrocytes contained numerous intranuclear and intracytoplasmic viral particles, while neurons remained apparently intact. The specificity of the infection for a given cell type is discussed.

Animals↗

Prion diseases: contribution of high-resolution immunomorphology.

The transmisible spongiform encephalopathies or prion diseases are fatal neurological diseases that occur in animals and humans. They are characterized by the accumulation in the cerebral tissue of the abnormal form of prion protein (PrPsc) produced by a post-translational event involving conformational change of its normal cellular counterpart (PrPc). In this short review, we present some results on the biology of prion proteins which have benefited from morphological approaches combining the electron microscopy techniques and the immunodetection methods. We discuss data concerning in particular the physiological function of the normal cellular prion prion (PrPc) which have allowed to open up new vistas on prion diseases, the biogenesis of amyloid plaque and the cellular site involved in the prion protein conversion process.

Animals↗