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In JoVE (2)
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Articles by Anne Fiedler in JoVE
Generation of Human CD40-activated B cells
Tanja M. Liebig, Anne Fiedler, Shahram Zoghi, Alexander Shimabukuro-Vornhagen, Michael S. von Bergwelt-Baildon
Laboratory for Tumor and Transplantation Immunology and Stem Cell Transplantation Program, University Hospital of Cologne, Department I of Internal Medicine
In this video we present the ex vivo generation and expansion of human CD40-activated B cells (CD40-B) from peripheral blood mononuclear cells (PBMC) by stimulation with CD40 ligand and interleukin-4.
Murine Model of CD40-activation of B cells
Tanja M. Liebig, Anne Fiedler, Nela Klein-Gonzalez, Alexander Shimabukuro-Vornhagen, Michael von Bergwelt-Baildon
Laboratory for Tumor and Transplantation Immunology, Department I of Internal Medicine, University Hospital of Cologne
In this video, we demonstrate the procedure of CD40-activation and expansion of murine B cells from splenocytes of C57BL/6 mice, which can be used as a model antigen-presenting cell (APC) to study induction of immunity.
Other articles by Anne Fiedler on PubMed
Chemical Conjugation of Linear and Cyclic RGD Moieties to a Recombinant Elastin-mimetic Polypeptide--a Versatile Approach Towards Bioactive Protein Hydrogels
Macromolecular Bioscience. Jun, 2008 | Pubmed ID: 18350537
An elastin-mimetic polypeptide, (EMM)(7), with the amino-acid sequence GRDPSS [VPGVG VPGKG VPGVG VPGVG VPGEG VPGIG](7) was used for chemical conjugation of various integrin ligands (RGD peptides) to prepare bioactive hydrogels. The chemical approach involved (1) chemical protection of lysine residues with Fmoc or Boc groups, (2) chemical ligation of a protected linear or cyclic RGD ligand, with or without a hexanoic-acid spacer to the glutamic acid residue, (3) deprotection of the lysine functionalities and the RGD moieties and (4) cross-linking to form a bioactive hydrogel. (1)H NMR spectroscopy was used to quantify the multiple steps in the reaction. The chemical protection was found to be between 65 and 93% for Fmoc and Boc, respectively. The ligands studied included linear RGD cell-binding [H-FGRGDS-OH (1-l-RGD), H-Ahx--FGRGDS-OH (2-Ahx-FGRGDS) and a cyclic -H(2)N-(CH(2))(6)COHN-cyclo(-RGDfK-) (H-Ahx-c(-RGDfK-)) peptide also with a hexanoic-acid spacer. Cell adhesion with mouse osteoblast cells was dependent on the ligand type, ligand density and the use of a spacer.