Synthesis in animal cells of hepatitis B surface antigen particles carrying a receptor for polymerized human serum albumin.

Michel, Marie‐Louise; Pontisso, Patrizia; Sobczak, Eliane; Malpiece, Yves; Streeck, Rolf E.; Tiollais, Pierre

doi:10.1073/pnas.81.24.7708

Cited by 173 publications

(85 citation statements)

References 36 publications

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“…On the other hand, antibodies against the pHSA receptor which have been shown in serum during acute hepatitis B may have an important role for viral clearance (Alberti et al, 1984;Pontisso et al, 1983b). As a result, it would be useful to use HBsAg particles carrying the pHSA receptor in vaccination programs (Michel et al, 1984). The recombinant adenovirus which is capable of eliciting in vivo both anti-HBs and anti-pHSA receptor antibodies would constitute the basis for an efficient live vaccine.…”

Section: Discussionmentioning

confidence: 99%

“…The use of recombinant DNA technology which allows the production of 22 nm HBsAg particles in eucaryotic cells is, therefore, needed. Presently, HBsAg particles are produced in vitro in animal cells (Michel et al, 1984;Patzer et al, 1984) or yeast (Miyanohara et al, 1983;Valenzuela et al, 1982) and in vivo through the vaccinia virus used as a vector (Smith et al, 1983). Chimpanzees infected 3864 with a vaccinia virus-HBV recombinant were found to be protected against a further HBV challenge (Moss et al, 1984).…”

Section: Discussionmentioning

confidence: 99%

“…Antibody levels were expressed as International Units [3.5 RIA units are equivalent to 1 milliintemational unit (mIIJ)]. Anti-polymerized human serum albumin receptor antibodies (anti-pHSA receptor) were detected by hemagglutination inhibition experiments according to Michel et al (1984). For immunoglobulin class characterization of anti-HBs antibodies, IgG were obtained by ion-exchange chromatography on DE-52 columns (Whatman).…”

Section: Purification Of Hbsagmentioning

confidence: 99%

“…HBsAg was purified from pooled supernatants according to Michel et al (1984). Tris-HCI (pH 7.4) was added for 1 h at 4°C.…”

Section: Purification Of Hbsagmentioning

confidence: 99%

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In vitro and in vivo synthesis of the hepatitis B virus surface antigen and of the receptor for polymerized human serum albumin from recombinant human adenoviruses.

Ballay¹,

Levrero²,

Buendía³

et al. 1985

The EMBO Journal

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We have developed an adenovirus vector to express foreign proteins under the control of the adenovirus Ela promoter. Two recombinant plasmids, harbouring either the S gene or the pre-S2 region and the S gene of hepatitis B virus under the control of the Ela promoter, were used to construct two recombinant adenoviruses. These two viruses direct the synthesis of hepatitis B virus surface antigen (HBsAg) particles during the time course of an infectious cycle. When the pre-S2 region is present in the constructed virus, the synthesis of particles carrying the receptor for polymerized human serum albumin (pHSA) is observed. Moreover, the inoculation of rabbits with this latter purified recombinant adenovirus elicits the production of antibodies that react with both HBsAg and pHSA receptor.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Purification Of Hbsagmentioning

confidence: 99%

“…HBsAg was purified from pooled supernatants according to Michel et al (1984). Tris-HCI (pH 7.4) was added for 1 h at 4°C.…”

Section: Purification Of Hbsagmentioning

confidence: 99%

See 2 more Smart Citations

In vitro and in vivo synthesis of the hepatitis B virus surface antigen and of the receptor for polymerized human serum albumin from recombinant human adenoviruses.

Ballay¹,

Levrero²,

Buendía³

et al. 1985

The EMBO Journal

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“…Two larger forms containing pre-S sequences are found in minor quantities, whereas the smallest HBsAg is the predominant component. The HBsAg can be integrated into cellular membranes and secreted as 22-nm particles into the culture supematant (Michel et al, 1984). Using the HBsAg reporter system, kinetics of promoter activities could easily be analyzed under a variety of in vitro conditions.…”

Section: Introductionmentioning

confidence: 99%

Hepatitis B virus surface antigen as a reporter of promoter activity

Marschall

Motz

Leser

et al. 1989

Gene

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SUMMARYThe coding sequence for the hepatitis B virus surface antigen (HBsAg) was used as a new reporter gene for studies on eukaryotic promoter activity and upstream regulatory sequences. The data observed in transfection assays were comparable to results obtained with conventional chloramphenicol acetyltransferase (CAT) assays, as was demonstrated using various transcriptional regulation sequences. The expression of HBsAg as a reporter protein offered some advantages:(i) In transient expression assays, a time course of promoter activity depending on variable culture conditions could be monitored over a period of time, since the HBsAg was secreted into the culture supematant.(ii) Evaluation of HBsAg from supematant aliquots and quantification of the corresponding promoter activities could be performed easily, using the very sensitive and readily available diagnostic HBsAg kits.(iii) In contrast to the conventional CAT assay, the cells remained available for further tests, e.g., Western blot, immunofluorescence or transcript analysis.Characteristics of several Epstein-Barr virus (EBV) promoters, depending on the virus state of EBV-positive B-cells (latency, chemical induction, lytic superinfection, truns-activation), were assayed using the HBsAg reporter system.

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