Characterization of a recombinant herpes simplex virus 1 designed to enter cells via the IL13Ralpha2 receptor of malignant glioma cells.
Malignant glioma tumor cells in situ exhibit on their surfaces the interleukin 13 (IL-13) receptor designated IL13Ralpha2. To target herpes simplex virus 1 to this receptor, we constructed a recombinant virus (R5111) in which the known heparan sulfate binding sites in glycoproteins B and C were deleted and IL-13 was inserted into both glycoproteins C and D. We also transduced a baby hamster kidney cell line lacking the known viral receptors (J1-1) and Vero cells with a plasmid encoding IL13Ralpha2. The J1-1 derivative (J-13R) cell line is susceptible to and replicates the R5111 recombinant virus but not the wild-type parent virus. We report the following. (i) Expression of IL13Ralpha2 was rapidly lost from the surface of transduced cells grown in culture. The loss appeared to be related to ligands present in fetal bovine serum in the medium. None of the malignant glioma cell lines cultivated in vitro and tested to date exhibited the IL13Ralpha2 receptor. (ii) Soluble IL-13 but not IL-4 or IL-2 blocked the replication of R5111 recombinant virus in J-13R cells. (iii) The endocytosis inhibitor PD98059 blocked the replication in J1-1 cells of a mutant lacking glycoprotein D (gD-/-) but not the replication of R5111 in the J-13R cells. We conclude that R5111 enters cells via its interaction with the IL13Ralpha2 receptor in a manner that cannot be differentiated from the interaction of wild-type virus with its receptors.
Zhou
Guoying
G
The Marjorie B. Kovler Viral Oncology Laboratories, The University of Chicago, 910 East 58th Street, Chicago, Il 60637, USA.
Roizman
Bernard
B
D000818:Animals; D002460:Cell Line; D045744:Cell Line, Tumor; D017353:Gene Deletion; D005910:Glioma / Q000378:metabolism*; D006497:Heparan Sulfate / Q000378:metabolism; D018259:Herpesvirus 1, Human / Q000235:genetics / Q000502:physiology*; D006801:Humans; D018793:Interleukin-13 / Q000494:pharmacology / Q000502:physiology*; D053661:Interleukin-13 Receptor alpha1 Subunit; D018123:Receptors, Interleukin / Q000235:genetics / Q000502:physiology*; D053660:Receptors, Interleukin-13; D018124:Receptors, Tumor Necrosis Factor / Q000032:analysis; D053240:Receptors, Tumor Necrosis Factor, Member 14; D011991:Receptors, Virus / Q000032:analysis; D011995:Recombination, Genetic; D014759:Viral Envelope Proteins / Q000235:genetics / Q000378:metabolism; D014779:Virus Replication / Q000187:drug effects
D006497:Heparan Sulfate; D018793:Interleukin-13; D053661:Interleukin-13 Receptor alpha1 Subunit; D018123:Receptors, Interleukin; D053660:Receptors, Interleukin-13; D018124:Receptors, Tumor Necrosis Factor; D053240:Receptors, Tumor Necrosis Factor, Member 14; D011991:Receptors, Virus; D014759:Viral Envelope Proteins; C508181:IL13RA1 protein, human; C506028:TNFRSF14 protein, human; C040495:glycoprotein B, Simplexvirus; C038535:glycoprotein gC, herpes simplex virus type 1