Single-chain antibody displayed on a recombinant measles virus confers entry through the tumor-associated carcinoembryonic antigen.
To redirect the tropism of the vaccine strain of measles virus (MV), Edmonston B, to a targeted cell population, we displayed on the viral hemagglutinin (H) a single-chain antibody (scAb) specific for the tumor-associated carcinoembryonic antigen (CEA). We generated H fusion proteins with three forms of the scAb appended, differing in the lengths of the linkers separating the VH and VL domains and thus in the oligomerization states of the scAbs. All proteins were stable, appeared properly folded, and were transported to the cell surface, but only H displaying the long-linker form of scAb was functional in supporting cell-cell fusion. This protein induced extensive syncytia in cells expressing the normal virus receptor CD46 and also in CD46-negative cells expressing the targeted receptor, human CEA. Replication-competent MV with H replaced by H displaying the long-linker form of scAb was recovered and replicated efficiently in both CD46-positive and CD46-negative, CEA-positive cells. Thus, MV not only tolerates the addition of a scAb on its H protein but also infects cells via a novel interaction between the scAb and its targeted receptor.
Hammond
A L
AL
Molecular Medicine Program, Mayo Foundation, Rochester, Minnesota 55905, USA.
Plemper
R K
RK
Zhang
J
J
Schneider
U
U
Russell
S J
SJ
Cattaneo
R
R
D000818:Animals; D000906:Antibodies / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D015703:Antigens, CD / Q000032:analysis; D002272:Carcinoembryonic Antigen / Q000276:immunology* / Q000378:metabolism*; D002460:Cell Line; D005434:Flow Cytometry; D015726:Giant Cells / Q000502:physiology; D006389:Hemagglutinins, Viral / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D006801:Humans; D008459:Measles virus / Q000235:genetics* / Q000276:immunology / Q000502:physiology; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins / Q000032:analysis; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D011995:Recombination, Genetic; D014162:Transfection
D000906:Antibodies; D015703:Antigens, CD; C497857:CD46 protein, human; D002272:Carcinoembryonic Antigen; D006389:Hemagglutinins, Viral; D051916:Membrane Cofactor Protein; D008562:Membrane Glycoproteins; D011993:Recombinant Fusion Proteins; C032397:hemagglutinin protein G, measles virus