000101279 001__ 101279
000101279 005__ 20181203020749.0
000101279 0247_ $$2doi$$a10.1034/j.1399-3089.2003.00110.x
000101279 022__ $$a0908-665X
000101279 02470 $$2ISI$$a000182262300002
000101279 02470 $$2DAR$$a3529
000101279 037__ $$aARTICLE
000101279 245__ $$aPresence of Gal-alpha1,3Gal epitope on xenogeneic lines: implications for cellular gene therapy based on the encapsulation technology
000101279 260__ $$c2003
000101279 269__ $$a2003
000101279 336__ $$aJournal Articles
000101279 500__ $$aGene Therapy Center and Division of Surgical Research, Lausanne University Medical School, Lausanne, Switzerland. nicole.deglon@epfl.ch
000101279 520__ $$aExposure to human serum induces the lysis of xenogeneic cells through natural antibodies and complement activation. The carbohydrate Galactose-alpha1,3-Galactose (Gal-alpha1,3-Gal) epitope, has been shown to be the principal antigenic determinant on target cells. This reaction is, therefore, particularly important for xenogeneic cell-based therapy. As a first step toward the evaluation of the impact of this phenomenon for encapsulated xenogeneic cells, we have evaluated the presence of the Gal-alpha1,3Gal epitope on two cell lines currently being used for the systemic delivery of protein in the periphery or the treatment of neurodegenerative diseases. In the second part of the study, we have tested and compared human serum and cerebrospinal fluid (CSF) for the presence of xenoreactive natural antibodies (XNAs) and their potential impact on the survival of xenogeneic cells. Fluorescence-activated cell sorting analysis indicated that baby hamster kidney (BHK) cells expressed low levels of the alpha-Gal epitope, whereas mouse myoblast C2C12 cells were extensively stained with the specific IB4-lectin. There was a direct correlation between serum killing and the level of Gal-alpha1,3-Gal epitope expression on these cells. Importantly, we showed that CSF did not lyse BHK and C2C12 cells as determined by cytotoxic crossmatch assays. The reaction was specific as the addition of soluble Gal-alpha1,3-Gal sugar to human serum effectively reduced cell killing, and the overproduction of alpha-1,3-galactosyltransferase in BHK cells significantly increased inactivation by human serum. To interfere with this antibody-antigen reaction and develop cell lines particularly suitable for cell-based therapy, we either selected C2C12 clones expressing low levels of Gal-alpha1,3-Gal or high levels of alpha-1,2-fucosyltransferase. These cells were found to be resistant to complement-mediated cytolysis. These strategies may, therefore, protect encapsulated xenogeneic cells transplanted in the periphery or the central nervous system even in an unlikely event of a blood-brain barrier breakage and the post-transplantation development of an antibody response.
000101279 6531_ $$aAnimals
000101279 6531_ $$aBase Sequence
000101279 6531_ $$aCHO Cells
000101279 6531_ $$aCell Line
000101279 6531_ $$aCricetinae
000101279 6531_ $$aCytotoxicity
000101279 6531_ $$aImmunologic
000101279 6531_ $$aDNA Primers
000101279 6531_ $$aDNA
000101279 6531_ $$aComplementary/genetics
000101279 6531_ $$aDisaccharides/ immunology
000101279 6531_ $$aEpitopes/immunology
000101279 6531_ $$aFlow Cytometry
000101279 6531_ $$aFucosyltransferases/genetics/metabolism
000101279 6531_ $$aHumans
000101279 6531_ $$aLymphocytes/ immunology
000101279 6531_ $$aMice
000101279 6531_ $$aMice
000101279 6531_ $$aInbred C3H
000101279 6531_ $$aRecombinant Proteins/metabolism
000101279 6531_ $$aSpleen/immunology
000101279 6531_ $$aTransfection
000101279 6531_ $$aTransplantation
000101279 6531_ $$aHeterologous/ immunology
000101279 6531_ $$aMice
000101279 700__ $$0241685$$aDeglon, N.$$g150324
000101279 700__ $$aAubert, V.
000101279 700__ $$aSpertini, F.
000101279 700__ $$0241461$$aWinkel, L.$$g150356
000101279 700__ $$0240206$$aAebischer, P.$$g104359
000101279 773__ $$j10$$k3$$q204-13$$tXenotransplantation
000101279 909C0 $$0252067$$pLEN$$xU10457
000101279 909CO $$ooai:infoscience.tind.io:101279$$pSV$$particle
000101279 937__ $$aLEN-ARTICLE-2003-011
000101279 970__ $$a43/LEN
000101279 973__ $$aEPFL$$rREVIEWED$$sPUBLISHED
000101279 980__ $$aARTICLE