Abstract
Adenovirus and adeno-associated virus vector-mediated gene delivery is limited by the induction of a humoral immune response that prevents readministration. To determine whether viral delivery in the “preimmune” fetus would produce dose- or time-dependent tolerance, we evaluated the humoral immune response after sequential pre- and postnatal virus administration. We evaluated six injection route and viral dose combinations of adenovirus (intra-amniotic, intrahepatic, and intramuscular injection at 4 × 108 and 2 × 109 particles/fetus) at d 15 postconception (p.c.); three route and dose combinations at d 13 p.c. (intramuscular injection at 1 × 108, 3 × 108, and 5 × 108 particles/fetus); and one route and dose combination of adeno-associated virus (intramuscular at 2.5 × 1010 genome copies/fetus) at d 15 p.c. In utero injection of either viral vector at any route and dose combination resulted in the production of low titers of neutralizing antivirus and antitransgene (β-galactosidase) antibodies. This primary immune response only partially blocked transgene expression after the readministration of viral vectors postnatally. However, delivery of the virus postnatally triggered an immune response that completely blocked transgene expression after a third viral injection. Together, these results suggest that, for B6/129 F1 mice, in utero injection of recombinant adenovirus or adeno-associated virus between d 13 and 15 p.c. does not induce tolerance to the viral vector or transgene product.
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Abbreviations
- Ad:
-
adenovirus
- AAV:
-
adeno-associated virus
- p.c.:
-
postconception
- i.m.:
-
intramuscular
- i.h.:
-
intrahepatic
- i.a.:
-
intra-amniotic
- Nab:
-
neutralizing antibody
- GC:
-
genome copies
- HSC:
-
hematopoietic stem cells
- Ad.CBlacZ:
-
recombinant adenovirus carrying Escherichia coli lacZ gene controlled by the cytomegalovirus enhancer and chicken β-actin gene promoter
- Ad.CMVhGH:
-
recombinant adenovirus carrying hGH controlled by cytomegalovirus promoter
- AAV.CMVlacZ:
-
recombinant AAV vector carrying E. coli lacZ gene controlled by the cytomegalovirus promoter
- AAV.CMVhGH:
-
recombinant AAV vector carrying hGH controlled by the cytomegalovirus promoter
- hGH:
-
human growth hormone
- lacZ:
-
E. coli lac Z gene encoding β-galactosidase
References
Turkay A, Saunders T, Kurachi K 1999 Intrauterine gene transfer: gestational stage-specific gene delivery in mice. Gene Ther 6: 1685–1694
Sekhon H, Larson J 1995 In utero gene transfer into the pulmonary epithelium. Nat Med 1: 1201–1203
Wang G, Williamson R, Mueller G, Thomas P, Davidson BL, McGray PB 1998 Ultrasound-guided gene transfer to hepatocytes in utero. Fetal Diagn Ther 13: 197–205
Vincent MC, Trapnell BC, Baughman RP, Wert SE, Whitsett JA, Iwamoto HS 1995 Adenovirus-mediated gene transfer to the respiratory tract of fetal sheep in utero. Hum Gene Ther 6: 1019–1020
Mitchell M, Jerebtsova M, Batshaw ML, Newman K, Ye X 2000 Long-term gene transfer to mouse fetuses with recombinant adenovirus adeno-associated virus (AAV) vectors. Gene Ther 7: 1986–1992
Lipshutz G, Gruber CA, Cao Y, Harby J, Contag CH, Gaensler KM 2001 In utero delivery of adeno-associated viral vectors: intraperitonial gene transfer produces long-term expression. Mol Ther 3: 284–292
Chirmule N, Xiao W, Truneh A, Schnell MA, Hughes JV, Zoltick P, Wilson JM 2000 Humoral immunity to adeno-associated virus type 2 vector following administration to murine nonhuman primate muscle. J Virol 74: 2420–2425
Yang Y, Jooss KU, Su Q, Ertl HC, Wilson JM 1996 Immune responses to viral antigens versus transgene product in the elimination of recombinant adenovirus-infected hepatocytes in vivo. Gene Ther 3: 137–144
Yang Y, Li Q, Ertl HC, Wilson JM 1995 Cellular humoral immune responses to viral antigens create barriers to lung-directed gene therapy with recombinant adenoviruses. J Virol 69: 2004–2015
Iwamoto H 1996 The window of opportunity for fetal gene therapy. Mol Hum Reprod 2: 472–474
Scott D 1994 The Nature of Immunologic Tolerance. Landes, Austin, TX, p 157
Brusilow S, Horwich A 1995 Urea cycle enzyme. In: Scriver C, Beaudet A, Sly W, Valle D (eds). The Metabolic and Molecular Bases of Inherited Disease, 7th ed. McGraw-Hill, New York, pp 1187–1232
Crombleholme T, Bianchi D 1994 In utero hematopoietic stem cell transplantation gene therapy. Semin Perinatol 18: 376–384
Flake A, Harrison M, Zanjani E 1991 In utero stem cell transplantation. Exp Hematol 19: 1061–1064
Iwamoto HS, Trapnell BC, McConnell CJ, Daugherty C, Whitsett JA 1999 Pulmonary inflammation associated with repeated, prenatal exposure to an E1, E3-deleted adenoviral vector in sheep. Gene Ther 6: 98–106
Yang EY, Kim HB, Shaaban AF, Milner R, Adzick NS, Flake AW 1999 Persistent postnatal transgene expression in both muscle liver after fetal injection of recombinant adenovirus. J Pediatr Surg 34: 766–773
Kozarsky K, Grossman M, Wilson JM 1993 Adenovirus-mediated correction of the genetic defect in hepatocytes from patients with familial hypercholesterolemia. Somat Cell Mol Genet 19: 449–458
Ye X, Robinson MB, Batshaw ML, Furth EE, Smith I, Wilson JM 1996 Prolonged metabolic correction in adult ornitine transcarbamilase-deficient mice with adenoviral vectors. J Biol Chem 271: 3639–3646
Davis A, Wilson J 1996 Adenoviral vectors. In: Dracopoli NC, Haines JL, Korf BR, Morton CC, Seidman CE, Seidman JG, Smith DR (eds) Current Protocols in Human Genetics. Wiley, New York, pp 12.4.7–12.4.15
Xiao W, Chirmule N, Berta SC, McCullough B, Gao G, Wilson JM 1999 Gene therapy vectors based on adeno-associated virus type I. J Virol 73: 3994–4003
Lipshutz G, Flebbe-Rehwaldt L, Gfaensler KM 2000 Reexpression following readministration of an adenoviral vector in adult mice after initial in utero adenoviral administration. Mol Ther 2: 374–380
Billingham R, Brent L, Medawar P 1953 Actively acquired tolerance to foreign cells. Nature 172: 603–605
Carrier E, Lee T, Busch M, Cowan M 1995 Induction of tolerance in nondefective mice after in utero transplantation of major histocompatibility complex-mismatched fetal hematopoietic stem cells. Blood 86: 4681–4690
Fleischman R, Mintz B 1984 Development of adults bone marrow stem cells in H-2 compatible incompatible mouse fetuses. J Exp Med 159: 731–745
Fleischman R, Mintz B 1979 Prevention of genetic anemias in mice by microinjection of normal hematopoietic stem cells into the fetal placenta. Proc Natl Acad Sci USA 76: 5736–5740
Brockstedt D, Podsakoff GM, Fong L, Kurtzman G, Mueller-Ruchholtz W, Engleman EG 1999 Induction of immunity to antigens expressed by recombinant adeno-associated virus depends on the route of administration. Clin Immunol 92: 67–75
Xiao W, Chirmule N, Schnell MA, Tazelaar J, Huges JV, Wilson JM 2000 Route of administration determines induction of T-cell-independent humoral responses to adeno-associated virus vectors. Mol Ther 1: 323–329
Douagi I, Andre I, Ferraz JC, Cumano A 2000 Characterization of T cell precursor activity in the murine fetal thymus: evidence for an input of T cell precursors between days 12 14 gestation. Eur J Immunol 30: 2201–2210
Strasser A, Rolink A, Melchers F 1989 One synchronous wave of B cell development in mouse fetal liver changes at day 16 of gestation from dependence to independence of a stromal cell environment. J Exp Med 170: 1973–1986
Zinkernagel RM 2000 Localization dose time of antigens determine immune reactivity. Semin Immunol 12: 163–171
Fields PA, Armstrong E, Hagstrom JN, Arruda VR, Murphy ML, Farrell JP, High KA, Herzog RW 2001 Intravenous administration of an E1/E3-deleted adenoviral vector induces tolerance to factor IX in C57BL/6 mice. Gene Ther 8: 354–361
Juillard V, Villefroy P, Godfrin D, Pavirani A, Venet A, Guillet JG 1995 Long-term humoral cellular immunity induced by a signal immunization with replication-defective adenovirus recombinant vector. Eur J Immunol 25: 3467–3473
Michou AI, Santoro L, Christ M, Julliard V, Pavirani A, Mehtali MV 1997 Adenovirus-mediated gene transfer: influence of transgene, mouse strain type of immune response on persistence of transgene expression. Gene Ther 4: 473–482
DeMatteo RP, Chu G, Ahn M, Chang E, Barker CF, Markmann JF 1997 Long-lasting adenovirus transgene expression in mice through neonatal intrathymic tolerance induction without the use of immunosuppression. J Virol 71: 5330–5335
Camargo F, Huey-Louie D, Sassani A, Finn A, Schneider D, Agah R, Dichek D 2000 Adenovirus-transgenic mice: partial embrionic lethality lack of immunological tolerance to adenoviral vectors. Mol Ther 1: S180–S181( abstr)
Ridge JP, Fuchs EJ, Matzinger P 1996 Neonatal tolerance revisited: turning on newborn T cells with dendritic cells. Science 271: 1726–1728
Gallucci S, Matzinger P 2001 Danger signals: SOS to the immune system. Curr Opin Immunol 13: 114–119
Tran ND, Porada CD, Almeida-Porada G, Glimp, Anderson WF, Zanjani ED 2001 Induction of stable prenatal tolerance to b-galactosidase by in utero gene transfer into preimmune sheep fetuses. Blood 97: 3417–3423
Hackett NR, Kaminsky SM, Sondhi D, Crystal RG 2000 Antivector antitransgene host responses in gene therapy. Curr Opin Mol Ther 2: 376–382
Kearns WG, Afione SA, Fulmer SB, Pang MC, Erikson D, Egan M, Landrum MJ, Flotte TR, Cutting GR 1996 Recombinant adeno-associated virus (AAV-CFTR) vectors do not integrate in a site-specific fashion in an immortalized epithelial cell line. Gene Ther 3: 748–755
Fisher KD, Stallwood Y, Green NK, Ulbrich K, Mautner V, Seymour LW 2001 Polymer-coated adenovirus permits efficient retargeting evades neutralizing antibodies. Gene Ther 8: 341–348
Reynolds PN, Dimitriev I, Curiel DT 1999 Insertion of an RGD motif into the H1 loop of adenovirus fiber protein alters the distribution of transgene expression of the systemically administered vector. Gene Ther 6: 1336–1339
Gu DL, Gonzalez AM, Printz MA, Doukas J, Ying W, D'Andrea M, Hoganson DK, Curiel DT, Douglas JT, Sosnowski BA, Baird A, Aukerman SL, Pierce GF 1999 Fibroblast growth factor 2 retargeted adenovirus has redirected cellular tropism: evidence for reduced toxicity enhanced antitumor activity in mice. Cancer Res 59: 2608–2614
Morral N, O'Neal W, Rice K, Leland M, Kaplan J, Piedra PA, Zhou H, Parks RJ, Velji R, Aguilar-Cordova E, Wadsworth S, Graham FL, Kochanek S, Carey KD, Beaudet AL 1999 Administration of helper-dependent adenoviral vectors sequential delivery of different vector serotype for long-term liver-directed gene transfer in baboons. Proc Natl Acad Sci USA 96: 12816–12821
Parks R, Evelegh C, Graham F 1999 Use of helper-dependent adenoviral vectors of alternative serotypes permits repeat vector administration. Gene Ther 6: 1565–1573
Jooss K, Yang Y, Wilson JM 1996 Cyclophosphamide diminishes inflammation prolong transgene expression following delivery of adenoviral vectors to mouse liver lung. Hum Gene Ther 7: 1555–1566
Filds Pa, Arruda R, Armstrong E, Chu K, Mingozzi F, Hagston JN, Herzog RW, High KA 2001 Risk preventation of anti-factor IX formation in AAV-mediated gene transfer in the context of a large deletion of F9. Mol Ther 4: 201–210
Ye X, Robinson MB, Pabin C, Batshaw ML, Wilson JM 2000 Transient depletion of CD4 lymphocyte improves efficacy of repeted administration of recombinant adenovirus in the ornithine transcarbamylase deficient sparse fur mice. Gene Ther 7: 1761–176
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Supported by National Institutes of Health grant P30HD 40677, and a grant from the Kettering Family Foundation.
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Jerebtsova, M., Batshaw, M. & Ye, X. Humoral Immune Response to Recombinant Adenovirus and Adeno-Associated Virus after In Utero Administration of Viral Vectors in Mice. Pediatr Res 52, 95–104 (2002). https://doi.org/10.1203/00006450-200207000-00018
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DOI: https://doi.org/10.1203/00006450-200207000-00018
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