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See detailIntracellular distribution of the ORF4 gene product of varicella-zoster virus is influenced by the IE62 protein
Defechereux, Patricia; Debrus, Serge; Baudoux, Laurence et al

in Journal of General Virology (1996), 77(Part 7), 1505-1513

Varicella-zoster virus (VZV) open reading frame 4-encoded protein (IE4) possesses transactivating properties for VZV genes as well as for genes of heterologous viruses, The major regulatory immediate ... [more ▼]

Varicella-zoster virus (VZV) open reading frame 4-encoded protein (IE4) possesses transactivating properties for VZV genes as well as for genes of heterologous viruses, The major regulatory immediate-early protein of VZV (IE62) is a transactivator of VZV gene expression, In transfection assays, IE4 has been shown to enhance activation induced by IE62, To investigate the functional interactions underlying this observation, indirect immunofluorescence studies were undertaken to determine whether IE62 could influence IE4 intracellular localization in transfected cells, In single transfections, IE4 was predominantly found in cytoplasm, In cotransfection with IE62, the IE4 localization pattern was altered, with nuclear staining predominating over cytoplasmic staining, This effect was specific to the IE62 protein since the gene products of ORF63 and ORF61, which are also regulatory proteins, did not influence IE4 distribution, The use of IE62 mutants indicated that IE62 influence is independent of its transactivation function and that the integrity of regions 3 and 4 is required, IE62 remained nuclear whether IE4 was present or not, These observations underline differences in the regulation of gene expression between VZV proteins and their herpes simplex virus type 1 homologues, In infected cells, IE4 was only sometimes found to colocalize with IE62 in nuclei, This observation suggests that when all VZV proteins are present, complex interactions probably occur which could diminish the influence of IE62. [less ▲]

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See detailHighly-Expressed P100/P52 (Nfkb2) Sequesters Other Nf-Kappa B-Related Proteins in the Cytoplasm of Human Breast Cancer Cells
Dejardin, Emmanuel ULg; Bonizzi, Giuseppina; Bellahcene, Akeila ULg et al

in Oncogene (1995), 11(9), 1835-41

Several observations have suggested that NF-kappa B transcription factors could be involved in carcinogenesis. To investigate the possibility that members of the NF-kappa B family participate in the ... [more ▼]

Several observations have suggested that NF-kappa B transcription factors could be involved in carcinogenesis. To investigate the possibility that members of the NF-kappa B family participate in the molecular control of the transformed phenotype, we examined the expression of these proteins in human breast cancer cell lines as well as in primary tumors. Western Immunoblots demonstrated high expression of the p52 precursor p100 (NFKB2) in several breast cancer cell lines while human mammary epithelial cells express this protein only faintly. Eighteen primary breast tumors out of 24 displayed significant expression of the p100/p52 protein. In MDA-MB-435 cells, overexpressed p100 and p52 are predominantly cytoplasmic and coimmunoprecipitation experiments demonstrated that p100 sequesters the heterodimer p50/p65 in the cytoplasm. We demonstrate that most p65 protein is complexed with p100 in these cells while it is complexed predominantly with I kappa B-alpha in cell lines expressing less p100. Our data strengthen the hypothesis that NF-kappa B could be involved in carcinogenesis and suggest that the p100/p52 NF-kappa B subunit could play a role in the development of human breast cancers, possibly by sequestering other NF-kappa B-related proteins in the cytoplasm. [less ▲]

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See detailMutational analysis of varicella-zoster virus major immediate-early protein IE62
Baudoux, Laurence; Defechereux, Patricia; Schoonbroodt, Sonia et al

in Nucleic Acids Research (1995), 23(8), 1341-1349

The varicella-zoster virus (VZV) open reading frame 62 encodes an immediate-early protein (IE62) that transactivates expression of various VZV promoters and autoregulates its own expression in transient ... [more ▼]

The varicella-zoster virus (VZV) open reading frame 62 encodes an immediate-early protein (IE62) that transactivates expression of various VZV promoters and autoregulates its own expression in transient expression assays. In Vero cells, IE62 was shown to transactivate the expression of all putative immediate-early (IE) and early (E) genes of VZV with an up-regulating effect at low intracellular concentrations. To define the functional domains involved in the regulatory properties of IE62, a large number of in-frame insertions and deletions were introduced into a plasmid-borne copy of the gene encoding IE62. Studies of the regulatory activities of the resultant mutant polypeptides in transient expression assays allowed to delineate protein regions important for repression of its own promoter and for transactivation of a VZV putative immediate-early gene (ORF61) promoter and an early gene (ORF29) promoter. This mutational analysis resulted in the identification of a new functional domain situated at the border between regions 4 and 5 which plays a crucial role in the IE62 regulatory functions. This domain turned out to be very well conserved amongst homologous alphaherpesvirus regulatory proteins and appeared to be rich in bulky hydrophobic and proline residues, similar to the proline-rich region of the CAAT box binding protein CTF-1. By immunofluorescence, a nuclear localization signal has been mapped in region 3. [less ▲]

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See detailTamoxifen and its active metabolite inhibit growth of estrogen receptor-negative MDA-MB-435 cells
Charlier, Corinne ULg; Chariot, Alain ULg; Antoine, Nadine ULg et al

in Biochemical Pharmacology (1995), 49

Tamoxifen (TAM), the non-steroidal anti-estrogen most widely administered to breast cancer patients, acts, at least in part, by competing with estrogen receptors (ER). However, the existence of an ... [more ▼]

Tamoxifen (TAM), the non-steroidal anti-estrogen most widely administered to breast cancer patients, acts, at least in part, by competing with estrogen receptors (ER). However, the existence of an alternative mechanism of action for this drug is supported by the clinical observations that: (a) 30% of patients with ER-negative cancer cells respond to TAM, and (b) 30% of patients with ER-positive cancer cells are not sensitive to this anti-estrogen. In this study, we observed that growth of the human ER-negative breast cancer cell line MDA-MB-435 was inhibited by TAM and 4-hydroxytamoxifen (4OH-TAM) in a concentration-dependent fashion. Both monoclonal enzymoimmunoassay and Dextran Charcoal Coated Scatchard radioimmunoassay analysis demonstrated that this MDA-MB-435 cell line does not express ER. The absence of ER in MDA-MB-435 cells was also demonstrated at the mRNA level by both northern blot hybridization and reverse transcription-polymerase chain reaction techniques. MDA-MB-435 cell proliferation was not affected by 17 beta-estradiol or by the pure anti-estrogen ICI 164384, further demonstrating that the observed effects of TAM and its active metabolite on the proliferation of MDA-MB-435 cells were due to an ER-independent mechanism, yet to be identified. MDA-MB-435 thus appears to be a promising original model for the study of the alternative ER-independent mechanisms of action of TAM. [less ▲]

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See detailVaricella-zoster virus gene regulation
Piette, Jacques ULg; Defechereux, Patricia; Baudoux, Laurence et al

in Neurology (1995), 45(12, suppl. 8), 23-27

The varicella-zoster virus genome contains 71 open reading frames (ORFs), five of which (ORF62, ORF4, ORF63, ORF61, and ORF10) encode regulatory proteins. ORF62 codes for the major immediate early protein ... [more ▼]

The varicella-zoster virus genome contains 71 open reading frames (ORFs), five of which (ORF62, ORF4, ORF63, ORF61, and ORF10) encode regulatory proteins. ORF62 codes for the major immediate early protein of the virus exhibiting DNA-binding and regulatory functions. This protein, localized in the cell nucleus, is a functional homologue to ICP4 of herpes simplex virus type 1 (HSV-1). It trans-activates several varicella-zoster virus promoters of the various gene classes and autoregulates its own expression. ORF4 protein activates gene promoters provided they have basal activities, but it is not a functional homologue of HSV-1 ICP27. Gene regulation activity appears to be linked to its cysteine-rich C-terminal region. ORF63 codes for an immediate early protein mainly located in the cell nucleus. The regulatory functions it performs are still unclear. ORF61 protein is the functional homologue of HSV-1 ICPO. Its N-terminal region exhibits a RING domain responsible for trans-activating and trans-repressing activities. ORF10 protein exhibits similarities with HSV-1 VP16 and activates the ORF62 promoter. [less ▲]

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See detailExpression of Bone Sialoprotein, a Bone Matrix Protein, in Human Breast Cancer
Bellahcene, Akeila ULg; Merville, Marie-Paule ULg; Castronovo, Vincenzo ULg

in Cancer Research (1994), 54(11), 2823-6

Microcalcifications are often associated with human mammary lesions, particularly with breast carcinomas. To date, the molecular mechanism that leads to the deposition of hydroxyapatite in the mammary ... [more ▼]

Microcalcifications are often associated with human mammary lesions, particularly with breast carcinomas. To date, the molecular mechanism that leads to the deposition of hydroxyapatite in the mammary tissue has not been elucidated. Bone sialoprotein (BSP) is a glycoprotein the expression of which coincides with the appearance of the first hydroxyapatite crystals during bone development. In this study, we report the observation that BSP, a bone matrix protein, is expressed in human mammary cancer cells. Using an immunoperoxidase technique, we studied the expression of BSP in 79 breast lesions, including 28 benign and 51 malignant specimens. Two polyclonal antibodies, one directed against intact human BSP and the other against a synthetic peptide of BSP (residues 277-294), were used and gave identical results. Normal mammary glands expressed undetectable or barely detectable amounts of BSP, and the majority of the benign lesions examined were generally unstained (0) or weakly stained (1+). Most of the breast carcinoma specimens (around 87%) showed a significant increase (P = 0.0001) in BSP expression. Breast carcinomas with microcalcifications had the highest immunoreactivity (2+ or 3+) to BSP antibodies. This is the first demonstration that BSP expression is significantly increased in breast cancer. Expression of BSP by breast cancer cells could play a major role in the deposition of microcalcifications and in the preferred bone homing of breast cancer cells. [less ▲]

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See detailLa protéine P53, protectrice de l'intégrité du génome: rôle dans la genèse des cancers
Bellahcene, Akeila ULg; Merville, Marie-Paule ULg; Gielen Jacques et al

in Revue Médicale de Liège (1994), 49(5), 274-84

L'homéostasie des tissus de l'organisme est le résultat d'un équilibre dynamique stable entre des facteurs de régulation positifs et négatifs. Ceux-ci contrôlent la prolifération des cellules et ... [more ▼]

L'homéostasie des tissus de l'organisme est le résultat d'un équilibre dynamique stable entre des facteurs de régulation positifs et négatifs. Ceux-ci contrôlent la prolifération des cellules et déterminent leur appartenance tissulaire. Des données récentes indiquent que l'apparition des cancers résulte du déséquilibre de cet état dynamique soit part l'activation de facteurs positifs désignés sous le terme d'oncogènes, soit par l'inactivation de facteurs négatifs qui sont regroupés dans la famille des gènes suppresseurs de tumeurs. Le gène codant pour la protéine p53 est un membre particulièrement important de cette dernière famille. En effet, son inactivation, par mutation et/ou délétion, est l'une des altérations génétiques la plus fréquemment détectée dans les cancers. La fonction principale attribuée à la protéine p53 consiste à préserver le génome des altérations susceptibles d'entraîner, entre autres, la cellule dans un processus de transformation maligne. C'est en permettant la réparation des altérations du DNA survenues lors de sa réplication avant la mitose ou provoquées par des agents extérieurs, que la protéine p53 semble exercer ses fonctions. Des altérations du gène p53 entraînant des perturbations de la fonction de la protéine p53 ont été identifiées au niveau de la plupart des lésions tumorales malignes. A ce titre, cette protéine semble jouer un rôle déterminant au niveau de la genèse des cancers. Aussi le gène p53 fait-il l'objet de recherches intensives qui devraient déboucher sur la mise au point de nouveaux moyens de détection et d'évaluation pronostique des cancers. D'autre part, des expériences visant à restaurer la fonction de la protéine p53 au niveau des cellules cancéreuses ouvrent de nouvelles et séduisantes perspectives thérapeutiques. [less ▲]

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See detailThe Nf-Kappa B Transcription Factor and Cancer: High Expression of Nf-Kappa B- and I Kappa B-Related Proteins in Tumor Cell Lines
Bours, Vincent ULg; Dejardin, Emmanuel ULg; Goujon-Letawe, F. et al

in Biochemical Pharmacology (1994), 47(1), 145-9

NF-kappa B is a pleiotropic transcription factor which controls the expression of many genes and viruses. To date, there is good evidence, but no definitive proof, for its role in tumor formation and ... [more ▼]

NF-kappa B is a pleiotropic transcription factor which controls the expression of many genes and viruses. To date, there is good evidence, but no definitive proof, for its role in tumor formation and development of metastasis. To investigate the possibility that members of the NF-kappa B family could participate in the molecular control of the transformed and invasive phenotype, we examined the expression of these proteins in a variety of human tumor cell lines. The expression of p50, p65, p52 and I kappa B was quantified at the protein level using western immunoblot and mobility shift assay and at the RNA level by northern blot. We observed high expression of the NF-kappa B inhibitor I kappa B in the ovarian carcinoma cell line OVCAR-3 together with constitutive nuclear NF-kappa B activity. We also studied the colon carcinoma cell line HT-29 and its metastatic counterpart HTM-29 and we observed specific expression of the p52 NF-kappa B-related protein in the metastatic cells. Our data confirm that NF-kappa B could be involved in the genesis of a variety of cancers including solid tumors and provide us with interesting models to explore the exact role of these transcription factors in cancer. [less ▲]

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See detailVZV ORF 4 encoded protein is an effective transactivor of gene expression and possesses distinct regulatory properties
Defechereux, Patricia; Baudoux, Laurence; Lambert, Chantal ULg et al

Conference (1994)

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See detailLe tamoxifene dans le traitement du cancer du sein
Charlier, Corinne ULg; Colin, Claude ULg; Merville, Marie-Paule ULg et al

in Journal de Gynécologie, Obstétrique et Biologie de la Reproduction (1994), 23(7), 751-6

Tamoxifen is the most often prescribed non steroidal antioestrogenic agent in the world for breast cancer. Worldwide collaboration. has centralized the results, of different trials throughout the world on ... [more ▼]

Tamoxifen is the most often prescribed non steroidal antioestrogenic agent in the world for breast cancer. Worldwide collaboration. has centralized the results, of different trials throughout the world on oral adjuvant therapy in the early stages of breast cancer. A significative regression of the tumour was observed in most cases. Moreover, recent epidemiological studies suggest that tamoxifen could prevent new contralateral primary tumours. The risk of the disease should thus be reduced by the prophylactic use of antioestrogens such as tamoxifen. Investigations using a variety of models have evaluated the effect of tamoxifen on tumour promotion and cell growth. Tamoxifen-induced growth inhibition is associated with major changes in biochemical events in cultured human breast cancer cells including cell proliferation or growth factor production. Growth inhibition of oestrogen-responsive human breast cancer cells is associated with an induced secretion of autoinhibitory polypeptides (TGF beta) and an antagonistic effect on the synthesis of proliferative proteins (TGF alpha,...). The first step in the mechanism of action of the drug is binding of tamoxifen to the oestrogen receptors. Development of resistance to tamoxifen treatment is a great problem in treatment of breast cancer patients and the mechanism of resistance will require further study: under the influence of the drug, tumours could become remodelled as selected subpopulations emerge resistant-tamoxifen. The fact that some breast cancers which are oestrogen receptor-negative respond to antioestrogen suggests that parallel but separate pathways for oestrogen and antioestrogen action may exist. This paper summarizes the results of the most recent studies concerning this promising drug. [less ▲]

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See detailCharacterization of the regulatory functions of varicella-zoster virus open reading frame-4 gene-product
Defechereux, Patricia; Melen-Lamalle, Laurence ULg; Baudoux, Laurence et al

in Journal of Virology (1993), 67(7), 4379-4385

Varicella-zoster virus (VZV) open reading frame 4 (ORF4) encodes a protein with a predicted molecular weight of 51,540 presenting amino acid sequence homology with the immediate-early regulatory protein ... [more ▼]

Varicella-zoster virus (VZV) open reading frame 4 (ORF4) encodes a protein with a predicted molecular weight of 51,540 presenting amino acid sequence homology with the immediate-early regulatory protein ICP27 of herpes simplex virus type 1. To investigate the regulatory properties of the ORF4 gene product, we performed a series of transient expression assays in Vero cells, using a plasmid expressing ORF4 as effector and several VZV genes and heterologous genes as targets. The VZV target plasmids contained promoter/regulatory regions from genes belonging to the three putative VZV kinetic classes fused to the chloramphenicol acetyltransferase (CAT) gene. The heterologous target plasmids consisted of promoter/regulatory regions of human cytomegalovirus, Rous sarcoma virus, and human immunodeficiency virus type 1 fused to the reporter gene. These experiments demonstrated that the ORF4 gene product activated expression of ORF62 in a dose-dependent fashion but had no effect on the expression of the three other putative immediate-early genes (ORF4, ORF61, and ORF63). When various amounts of ORF4 were transfected in the presence of early gene promoters, dose-dependent transactivation was evidenced with the thymidine kinase gene (ORF36) and the major DNA-binding protein gene (ORF29) promoters; interestingly, little activity was detected with the promoter of the DNA polymerase gene (ORF28). No activation of late gene expression, represented by the glycoprotein I and glycoprotein II genes, was seen even over a wide range of concentrations of input ORF4 plasmid. Expression of pCMVCAT, pRSVCAT, and pHIVCAT was also stimulated by the ORF4 gene product. CAT mRNA analysis showed that activation of VZV target promoters occurs at the transcriptional and/or posttranscriptional level. [less ▲]

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See detailStimulation of the 92-Kd Type Iv Collagenase Promoter and Enzyme Expression in Human Melanoma Cells
Lauricella-Lefebvre, M. A.; Castronovo, Vincenzo ULg; Sato, H. et al

in Invasion & Metastasis (1993), 13(6), 289-300

The 92-kD type IV collagenase is a member of the metalloproteinase family which degrades type IV collagen, a major component of basement membrane and is involved in tumor invasion and metastasis. The ... [more ▼]

The 92-kD type IV collagenase is a member of the metalloproteinase family which degrades type IV collagen, a major component of basement membrane and is involved in tumor invasion and metastasis. The promoter and adjacent regulatory sequences of the 92-kD type IV collagenase have been identified previously and three cis-acting elements homologous to the binding sites for AP-1, NF-KB and SP-1 proteins contributed to induction of the promoter activity by 12-O-tetradecanoylphorbol 13-acetate (TPA) and tumor necrosis factor (TNF-alpha) in HT1080 cells. To date, no direct correlation between promoter activity and expression of the 92-kD type IV collagenase has been reported in normal or cancer cells. In this study, the effects of the transcriptional stimulation of the 92-kD type IV collagenase gene on the expression of the enzyme in human A2058 melanoma cells was analyzed by zymography experiments. Quantitative immunoblots using a monoclonal antibody that recognized specifically and exclusively the 92-kD type IV collagenase, confirmed that the 92-kD gelatinase was 92-kD type IV collagenase. Stimulation of the promoter activity resulted in increased gelatinase activity in the culture medium of A2058 cells. A direct correlation between TPA- and TNF-alpha-mediated promoter stimulation of the 92-kD type IV collagenase gene and its expression was also demonstrated in the human fibrosarcoma HT1080 cells. Interleukin-1 alpha failed to induce 92-kD gene promoter activity and type IV collagenase expression in melanoma and fibrosarcoma cell lines. Our data demonstrated that TPA- and TNF-alpha-induced 92-kD type IV collagenase promoter stimulation leads to a proportional increase of enzyme expression and secretion and thus could contribute to the activation of the invasive phenotype. [less ▲]

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See detailLa régulation de l'expression des gènes du virus de la varicelle et du zona
Piette, Jacques ULg; Defechereux, Patricia; Baudoux, Laurence et al

in Annales de Médecine Vétérinaire (1992), 136(8), 627-635

Varicella-zoster virus (VZV) belongs to the alphaherpesvirus family and shares many important structural and functional similarities with other members of the family such as herpes simplex virus type 1 ... [more ▼]

Varicella-zoster virus (VZV) belongs to the alphaherpesvirus family and shares many important structural and functional similarities with other members of the family such as herpes simplex virus type 1 (HSV-1). VZV is responsible for two different clinical syndromes, varicella which is the result of the primary infection and zoster which is due to virus reactivation remaining latent in the peripheral nervous system. VZV DNA is 124,884 base pair long and encodes four regulatory proteins (IE4, IE61, IE62 and IE63). Using transient expression systems, we have shown that IE4, IE62 and IE63 can regulate the expression of an indicator gene driven by various VZV promoter regions, demonstrating that these proteins play important roles in the infectious cycle. [less ▲]

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See detailAntibodies to varicella-zoster virus modulate antigen distribution but fail to induce viral persistence in vitro.
Sadzot-Delvaux, Catherine ULg; Marc, Philippe; Lebon, Linda et al

in Journal of Virology (1992), 66(12), 7499-504

Varicella-zoster virus (VZV) persists in human sensory ganglia. One of the hypotheses to explain the induction or the maintenance of VZV latency is that it could be promoted by the immune response itself ... [more ▼]

Varicella-zoster virus (VZV) persists in human sensory ganglia. One of the hypotheses to explain the induction or the maintenance of VZV latency is that it could be promoted by the immune response itself. It is known that in the case of viruses which bud off the infected cell membrane, virus-specific antibodies can induce antigenic modulation, i.e., spatial redistribution of viral antigens and modulation of their synthesis. To determine whether antigenic modulation occurs during VZV infection in vitro and could possibly be involved in viral persistence, we have grown infected cells in the presence of anti-VZV antibodies either transiently or permanently. The distribution of immune complexes and viral proteins was then analyzed. In transient immunomodulation experiments, the distribution of one or more viral antigens was modified not only in the cytoplasmic membranes but also in the cytoplasm and nucleoplasm of infected cells. When infected cells were kept permanently in the presence of antibodies, the same pattern of redistribution of immune complexes was observed and the localization of internal viral glycoproteins was significantly modified. However, antibodies did not prevent the lytic effect of infection; they altered neither the infectious virus yield nor the Western immunoblot pattern of viral proteins, suggesting that immunomodulation is not the primary effector of viral persistence. [less ▲]

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See detailCharacterization of regulatory functions of the varicella-zoster virus gene-63-encoded protein
Jackers, Pascale ULg; Defechereux, Patricia; Baudoux, Laurence et al

in Journal of Virology (1992), 66(6), 3899-3903

Varicella-zoster virus (VZV) gene 63 encodes a protein (IE63) with a predicted molecular mass of 30.5 kDa which has amino acid similarities to the immediate-early (IE) protein 22 (ICP22) of herpes simplex ... [more ▼]

Varicella-zoster virus (VZV) gene 63 encodes a protein (IE63) with a predicted molecular mass of 30.5 kDa which has amino acid similarities to the immediate-early (IE) protein 22 (ICP22) of herpes simplex virus type 1. ICP22 is a polypeptide synthesized in herpes simplex virus type 1-infected cells, and as is the case for its VZV counterpart, its regulatory functions are unknown. On the basis of the VZV DNA sequence, it has been shown that IE63 exhibits hydrophilic and acidic properties, suggesting that this protein could play a regulatory role during the infectious cycle. We report in this article cotransfection experiments which demonstrate that the VZV gene 63 protein strongly represses, in a dose-dependent manner, the expression of VZV gene 62. On the other hand, transient expression of the VZV gene 63 protein can promote activation of the thymidine kinase gene but cannot affect the expression of the genes encoding glycoproteins I and II. The results of transient expression experiments strongly suggest that the VZV gene 63 protein could play a pivotal role in the repression of IE gene expression as well as in the activation of early gene expression [less ▲]

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See detailVaricella-zoster gene 63 encoded protein is an important regulatory factor
Jackers, Pascale ULg; Defechereux, Patricia; Baudoux, Laurence et al

in Archives Internationales de Physiologie, de Biochimie et de Biophysique (1992), 100

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See detailMolecular characterization of varicella-zoster virus gene expression
Defechereux, Patricia; Baudoux, Laurence; Jackers, Pascale ULg et al

in Archives Internationales de Physiologie, de Biochimie et de Biophysique (1992), 100

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See detailAn in vivo model of varicella-zoster virus latent infection of dorsal root ganglia
Sadzot-Delvaux, Catherine ULg; Merville, Marie-Paule ULg; Delrée, P. et al

in Journal of Neuroscience Research (1990), 26(1), 83-89

We describe here the first in vivo model of varicella-zoster virus (VZV) latent infection in the adult rat peripheral nervous system. Infected Mewo cells were injected subcutaneously along the spine of ... [more ▼]

We describe here the first in vivo model of varicella-zoster virus (VZV) latent infection in the adult rat peripheral nervous system. Infected Mewo cells were injected subcutaneously along the spine of healthy adult rats. No clinical sign of infection was observed even 9 months after inoculation. Humoral immune response to VZV was detected in all infected animals throughout the study (9 months). The presence of viral material in dissociated and cultured dorsal root ganglia (DRG) from inoculated animals was studied by immunoperoxidase and in situ hybridization. When DRGs from infected animals were plated in culture from 1 month and up to 9 months after inoculation, viral nucleic acids and proteins were detected in neurons. Furthermore, trypsinization and subcultivation of infected neurons in culture is needed to reactivate infectious virus at least in some of the neurons. This model provides a useful tool for studying 1) the molecular mechanisms leading to an in vivo latency, 2) the role of the immune system, in particular cellular immunity, on the establishment, maintenance, and reactivation of latency, 3) the neurotropism of mutant viruses, and 4) the effects of antiviral agents. [less ▲]

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