Haemagglutinin of Measles Virus: Purification and Storage with Preservation of Biological and Immunological Properties

Gerlier, Denis; Garnier, Florence; Forquet, Frédérique

doi:10.1099/0022-1317-69-8-2061

Cited by 20 publications

(14 citation statements)

References 18 publications

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“…The cDNA encoding the sCD46-C4bp␣ protein was also subcloned under the cytomegalovirus promoter into the APEX3 vector (10), which was used to derive human 293EBNA cells expressing the chimeric protein. The secretion of the chimeric protein was determined using a dot blot assay with MCI20.6 anti-CD46 antibody and anti-mouse immunoglobulin (Ig)-alkaline phosphatase conjugate as previously detailed (20). Throughout all the analyses, an immunopurified recombinant monomeric sCD46 previously described (10) was used for comparative studies.…”

Section: Methodsmentioning

confidence: 99%

“…Alternatively, the sCD46-C4bp␣ protein was produced in serum-free medium supplemented with the plant-derived growth factor Prolifix (Biomedia), concentrated on 100-kDa exclusion membrane, and purified by exclusion chromatography on Sephacryl 200 (Pharmacia). Metabolic labeling with Tran 35 S-label (NEN) for 30 min followed by a chase of 2 h and immunoprecipitation of cell extract and cell supernatant using J4-48 antibodies and protein G-Sepharose beads were performed according to published procedures (20,38).…”

Section: Methodsmentioning

confidence: 99%

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Octamerization Enables Soluble CD46 Receptor To Neutralize Measles Virus In Vitro and In Vivo

Christiansen

Devaux

Réveil

et al. 2000

J Virol

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A chimeric fusion protein encompassing the CD46 ectodomain linked to the C-terminal part of the C4b binding protein (C4bp) ␣ chain (sCD46-C4bp␣) was produced in eukaryotic cells. This protein, secreted as a disulfide-linked homo-octamer, was recognized by a panel of anti-CD46 antibodies with varying avidities. Unlike monomeric sCD46, the octameric sCD46-C4bp␣ protein was devoid of complement regulatory activity. However, sCD46-C4bp␣ was able to bind to the measles virus hemagglutinin protein expressed on murine cells with a higher avidity than soluble monomeric sCD46. Moreover, the octameric sCD46-C4bp␣ protein was significantly more efficient than monomeric sCD46 in inhibiting virus binding to CD46, in blocking virus induced cell-cell fusion, and in neutralizing measles virus in vitro. In addition, the octameric sCD46-C4bp␣ protein, but not the monomeric sCD46, fully protected CD46 transgenic mice against a lethal intracranial measles virus challenge.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Octamerization Enables Soluble CD46 Receptor To Neutralize Measles Virus In Vitro and In Vivo

Christiansen

Devaux

Réveil

et al. 2000

J Virol

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“…Measles virus and viral proteins. Measles virus (Halle strain) was produced, purified and inactivated with UV as previously described (Gerlier, et al, 1988, Gerlier et aI., 1994. UVtreatment abolish transcription and replication of the viral genome without inhibiting its binding and fusion properties.…”

Section: Methodsmentioning

confidence: 99%

Efficient MHC Class II‐restricted presentation of measles virus to T cells relies on its targeting to its cellular receptor human CD46 and involves an endosomal pathway

Gerlier¹

1994

Cell Biology International

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The role of the measles virus (MV) receptor, human CD46, in the uptake of MV and antigen presentation by Major Histocompatibility Complex (MHC) class II molecules was investigated. Expression of CD46 in murine B cells resulted in cells highly efficient in capturing UV-inactivated MV particles and presenting both envelope hemagglutinin H and nucleoprotein N to specific T cell hybridomas. Although MV fuse with the plasma membrane of its target cells, presentation of both MV-H and -N was sensitive to inhibition by chloroquine but was not affected by a tripeptide which prevents virus-cell fusion. Whereas 50 microM of chloroquine was required to inhibit presentation of MV-H, purified H or soluble N, only a two-fold lower concentration was required to inhibit that of MV-N. This shows that some CD46-mediated captured MV particles are endocytosed, then disrupted and processed in an endosome/lysosome compartment.

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“…MV (Hallé strain) was produced as previously described [37]. In all experiments, MV was UV irradiated before use by 30 min exposure at 5 cm from a 254-nm UV lamp.…”

Section: And Viral Proteinsmentioning

confidence: 99%

“…In all experiments, MV was UV irradiated before use by 30 min exposure at 5 cm from a 254-nm UV lamp. UV treatment abolished transcription and replication of the viral genome without inhibiting its binding and fusion abilities [37]. Recombinant soluble native H glycoprotein, prepared as described [17], was generously provided by D. Gerlier.…”

Section: And Viral Proteinsmentioning

confidence: 99%

Enhanced MHC class II-restricted presentation of measles virus (MV) hemagglutinin in transgenic mice expressing human MV receptor CD46

et al. 1998

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This study analyzes the role of the measles virus (MV) receptor, i.e. the human CD46 molecule, in the MHC class II-restricted presentation of MV hemagglutinin (H). We generated transgenic mice ubiquitously expressing CD46, with a similar level of transgene expression on the surface of antigen-presenting cells (APC), i.e. B cells, dendritic cells (DC) and macrophages. APC isolated from transgenic mice and nontransgenic controls were tested for their ability to present MV H to H-specific CD4 + I-E d -restricted T cell hybridomas. All three populations of APC were capable of presenting MV to T cell hybridomas, DC being the most efficient. Expression of CD46 on B lymphocytes increased MHC class II-dependent presentation of MV H up to 100-fold, while CD46-transgenic DC stimulated H-specific T cell hybridomas up to 10-fold better than nontransgenic DC. Interestingly, expression of CD46 did not change the presentation efficiency of transgenic macrophages, indicating that CD46-dependent enhancement of antigen presentation depends on the nature of the APC. Furthermore, a single injection of UV-inactivated MV particles into CD46-transgenic mice, but not nontransgenic controls, induced generation of MV-specific T lymphocytes and production of anti-H antibodies, suggesting a role for CD46 in the efficient capture of MV in vivo. These results show for the first time that one ubiquitously expressed cell surface receptor, like CD46, could function in receptor-mediated antigen presentation both in vitro and in vivo and its performance depends on the type of APC which expresses it.

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Haemagglutinin of Measles Virus: Purification and Storage with Preservation of Biological and Immunological Properties

Cited by 20 publications

References 18 publications

Octamerization Enables Soluble CD46 Receptor To Neutralize Measles Virus In Vitro and In Vivo

Octamerization Enables Soluble CD46 Receptor To Neutralize Measles Virus In Vitro and In Vivo

Efficient MHC Class II‐restricted presentation of measles virus to T cells relies on its targeting to its cellular receptor human CD46 and involves an endosomal pathway

Enhanced MHC class II-restricted presentation of measles virus (MV) hemagglutinin in transgenic mice expressing human MV receptor CD46

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