Pressed juices of the aerial parts of Echinacea purpurea are used as non-specific immunostimulants, and arabinogalactan-proteins (AGPs) have been shown to be part of the active principle. Monoclonal antibodies against an AGP from pressed juice of Echinacea purpurea with complement-stimulating activity have been established by means of hybridoma techniques. To test the specificity of the antibodies, several other arabinogalactan-proteins from suspension cultures of Echinacea purpurea, the roots of Echinacea pallida, the aerial parts of Rudbeckia hirta, the roots of Baptisia tinctoria and gum arabic as well as an arabinogalactan from larch wood were tested in a competitive ELISA for cross reactivities. Chemical modifications at the periphery of the AGP molecules either by reduction of uronic acids or by dearabinosylation had no influence on the reactivity of the molecules towards the antibodies. For further characterization of the epitope, different Ara-Gal-oligosaccharides were used as antigens. A hexasaccharide consisting of a backbone of four molecules of 6-linked beta- D-Gal p, the second and the fourth of them branched at O-2 to an alpha- L-Ara f residue showed weak but reproducible cross reactivity, indicating that the antibodies may be at least in part directed to the carbohydrate moiety of the AGP. Testing of anti-AGP antibodies JIM 8 and LM 2 revealed good reactivity of LM 2 with the Echinacea AGP, whereas Jim 8 showed only very weak interaction.

01 Jul 2003·Kidney InternationalQ1 · MEDICINE

Laminin-8/9 is synthesized by rat glomerular mesangial cells and is required for PDGF-induced mesangial cell migration

Q1 · MEDICINE

Article

Author: Christine K. Abrass ; Kim Hansen

BACKGROUNDLaminin (LM), the major glycoprotein component of basement membranes is expressed as multiple isoforms in a developmentally regulated and tissue-specific manner. LM alpha4 has a limited tissue distribution and is highly expressed in the developing glomerulus. In the present study, we investigate the in vivo and in vitro expression and function of LM alpha4 in the glomerulus.METHODSLM alpha4 expression was examined by Northern blot, reverse transcription polymerase chain reaction (RT-PCR), Western blot, and immunofluorescence microscopy. Mesangial cells (MC) were plated on purified LM-1, LM-2, and LM-8/9. Immunofluorescence microscopy was performed to examine the cellular phenotypes induced by LM-1 and LM-8/9. A modified Boyden chamber method was used to assess laminin participation in platelet-derived growth factor (PDGF)-stimulated migration.RESULTSmRNA for LMalpha4 is expressed in cultured rat MC, and isolated rat and mouse glomeruli, but not in cultured rat glomerular epithelial cells or glomerular endothelial cells. Using antibodies specific for LM alpha4, a 240 kD band was detected in MC extract and a slightly smaller band was identified in extracted rat glomeruli. Purified LM-8/9 had MC adhesive activity comparable to LM-1 and LM-2. MC attached to LM-8/9 exhibited a unique phenotype. In contrast to LM-1, attachment of MC to LM-8/9 produced a highly arborized cell morphology with significantly reduced formation of focal contacts or stress fibers. LM alpha4 is utilized by MC during PDGF-stimulated migration.CONCLUSIONLM alpha4 is synthesized by MC and persists in the mature glomerulus. LM-8/9 stimulates a unique cellular morphology, and they are utilized in PDGF-induced migration. These factors suggest that LM alpha4 plays an important role in MC differentiation and in the maintenance of MC phenotype.

13 Dec 1999·The Journal of Cell BiologyQ1 · BIOLOGY

Cell Elongation Induces Laminin α2 Chain Expression in Mouse Embryonic Mesenchymal Cells

Q1 · BIOLOGY

Article

Author: Beqaj, Safedin ; Schuger, Lucia ; Relan, Nand K. ; Miner, Jeffrey H. ; Yang, Yan

Bronchial smooth muscle (SM) mesenchymal cell precursors change their shape from round to spread/elongated while undergoing differentiation. Here we show that this change in cell shape induces the expression of laminin (LM) α2 chain not present in round mesenchymal cells. LM α2 expression is reversible and switched on and off by altering the cell's shape in culture. In comparison, the expression of LM β1 and γ1 remains unchanged. Functional studies showed that mesenchymal cell spreading and further differentiation into SM are inhibited by an antibody against LM α2. Dy/dy mice express very low levels of LM α2 and exhibit congenital muscular dystrophy. Lung SM cells isolated from adult dy/dy mice spread defectively and synthesized less SM α-actin, desmin, and SM-myosin than controls. These deficiencies were completely corrected by exogenous LM-2. On histological examination, dy/dy mouse airways and gastrointestinal tract had shorter SM cells, and lungs from dy/dy mice contained less SM-specific protein. The intestine, however, showed compensatory hyperplasia, perhaps related to its higher contractile activity. This study therefore demonstrated a novel role for the LM α2 chain in SM myogenesis and showed that its decrease in dy/dy mice results in abnormal SM.

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Indication	Highest Phase	Country/Location	Organization	Date
Nonalcoholic Steatohepatitis	Preclinical	CN	CVI Pharmaceuticals (Shanghai) Co Ltd	-
Nonalcoholic Steatohepatitis	Preclinical	US	CVI Pharmaceuticals (Shanghai) Co Ltd	-
Obesity	Preclinical	CN	CVI Pharmaceuticals (Shanghai) Co Ltd	-

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