Adenovirus (Ad)-based vectors are attractive candidates for a variety of gene-transfer applications. In this study, we found that decay-accelerating factor (DAF)–displaying Ads induce significantly decreased cellular immune responses to transgenes expressed from the vectors in both Ad5-naive and Ad5-immune mice. Specifically, we found a diminished ability of splenocytes to secrete interferon-γ after recall exposure to multiple peptides derived from antigens expressed by DAF-displaying Ads. We also confirmed that DAF-displaying Ads induce decreased numbers of antigen-specific, CD8+ effector memory and central memory CD8+ T cells, thereby uncovering a unique role of complement in modulating the induction of robust memory T-cell responses. We also confirmed that DAF-displaying Ads generate significantly reduced titers of Ad capsid–specific neutralizing antibodies after gene transfer in vivo. In conclusion, DAF-displaying Ad5-based vectors exhibit decreased induction of complement-dependent, innate immune responses, resulting in both an improved safety profile and a decreased propensity to induce humoral and cellular adaptive immune responses to Ad capsid proteins and Ad vector-expressed transgene products. This attractive combination of features will be beneficial in a variety of clinically relevant gene-transfer applications.
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References
1.
AbbinkP., LemckertA.A., EwaldB.A.et al.2007. Comparative seroprevalence and immunogenicity of six rare serotype recombinant adenovirus vaccine vectors from subgroups B and D. J. Virol., 81:4654–4663.
2.
AppledornD.M., KiangA., McBrideA.et al.2008a. Wild-type adenoviruses from groups A–F evoke unique innate immune responses, of which HAd3 and SAd23 are partially complement dependent. Gene Ther., 15:885–901.
3.
AppledornD.M., McBrideA., SereginS.et al.2008b. Complex interactions with several arms of the complement system dictate innate and humoral immunity to adenoviral vectors. Gene Ther., 15:1606–1617.
4.
AppledornD.M., AldhamenY.A., DepasW.et al.2010. A new adenovirus based vaccine vector expressing an Eimeria tenella derived TLR agonist improves cellular immune responses to an antigenic target. PLoS One, 5:e9579.
5.
BarryM.E., Pinto-GonzalezD., OrsonF.M.et al.1999. Role of endogenous endonucleases and tissue site in transfection and CpG-mediated immune activation after naked DNA injection. Hum. Gene Ther., 10:2461–2480.
EverettR.S., HodgesB.L., DingE.Y.et al.2003. Liver toxicities typically induced by first-generation adenoviral vectors can be reduced by use of E1, E2b-deleted adenoviral vectors. Hum. Gene Ther., 14:1715–1726.
8.
FangC., MiwaT., ShenH., SongW.C.2007. Complement-dependent enhancement of CD8+ T cell immunity to lymphocytic choriomeningitis virus infection in decay-accelerating factor-deficient mice. J. Immunol., 179:3178–3186.
9.
FischerM.B., MaM., HsuN.C., CarrollM.C.1998. Local synthesis of C3 within the splenic lymphoid compartment can reconstitute the impaired immune response in C3-deficient mice. J. Immunol., 160:2619–2625.
GabitzschE.S., XuY., YoshidaL.H.et al.2009. Novel adenovirus type 5 vaccine platform induces cellular immunity against HIV-1 Gag, Pol, Nef despite the presence of Ad5 immunity. Vaccine, 27:6394–6398.
12.
GasqueP.2004. Complement: a unique innate immune sensor for danger signals. Mol. Immunol., 41:1089–1098.
13.
HartmanZ.C., KiangA., EverettR.S.et al.2007. Adenovirus infection triggers a rapid, MyD88-regulated transcriptome response critical to acute-phase and adaptive immune responses in vivo. J. Virol., 81:1796–1812.
14.
HartmanZ.C., AppledornD.M., AmalfitanoA.2008. Adenovirus vector induced innate immune responses: impact upon efficacy and toxicity in gene therapy and vaccine applications. Virus Res., 132:1–14.
15.
HawlischH., KohlJ.2006. Complement and Toll-like receptors: key regulators of adaptive immune responses. Mol. Immunol., 43:13–21.
16.
HensleyS.E., AmalfitanoA.2007. Toll-like receptors impact on safety and efficacy of gene transfer vectors. Mol. Ther., 15:1417–1422.
17.
HensleyS.E., CunA.S., Giles-DavisW.et al.2007. Type I interferon inhibits antibody responses induced by a chimpanzee adenovirus vector. Mol. Ther., 15:393–403.
18.
HodgesB.L., EvansH.K., EverettR.S.et al.2001. Adenovirus vectors with the 100K gene deleted and their potential for multiple gene therapy applications. J. Virol., 75:5913–5920.
19.
KaufmanD.R., LiuJ., CarvilleA.et al.2008. Trafficking of antigen-specific CD8+ T lymphocytes to mucosal surfaces following intramuscular vaccination. J. Immunol., 181:4188–4198.
20.
KemperC., AtkinsonJ.P.2007. T-cell regulation: with complements from innate immunity. Nat. Rev. Immunol., 7:9–18.
21.
KiangA., HartmanZ.C., EverettR.S.et al.2006. Multiple innate inflammatory responses induced after systemic adenovirus vector delivery depend on a functional complement system. Mol. Ther., 14:588–598.
22.
LasaroM.O., ErtlH.C.2009. New insights on adenovirus as vaccine vectors. Mol. Ther., 177:1333–1339.
23.
LiuJ., EwaldB.A., LynchD.M.et al.2008. Magnitude and phenotype of cellular immune responses elicited by recombinant adenovirus vectors and heterologous prime-boost regimens in rhesus monkeys. J. Virol., 82:4844–4852.
24.
LiuJ., O'BrienK.L., LynchD.M.et al.2009. Immune control of an SIV challenge by a T-cell-based vaccine in rhesus monkeys. Nature, 457:87–91.
MehlhopE., DiamondM.S.2006. Protective immune responses against West Nile virus are primed by distinct complement activation pathways. J. Exp. Med., 203:1371–1381.
27.
MehlhopE., WhitbyK., OliphantT.et al.2005. Complement activation is required for induction of a protective antibody response against West Nile virus infection. J. Virol., 79:7466–7477.
28.
MingozziF., HighK.A.2007. Immune responses to AAV in clinical trials. Curr. Gene Ther., 7:316–324.
29.
MollnesT.E., KirschfinkM.2006. Strategies of therapeutic complement inhibition. Mol. Immunol., 43:107–121.
30.
MorganB.P., MarchbankK.J., LonghiM.P.et al.2005. Complement: central to innate immunity and bridging to adaptive responses. Immunol. Lett., 97:171–179.
31.
NgP., GrahamF.L.2002. Construction of first-generation adenoviral vectors. Methods Mol. Med., 69:389–414.
SallustoF., GeginatJ., LanzavecchiaA.2004. Central memory and effector memory T cell subsets: function, generation, and maintenance. Annu. Rev. Immunol., 22:745–763.
34.
SchiednerG., HertelS., KochanekS.2000. Efficient transformation of primary human amniocytes by E1 functions of Ad5: generation of new cell lines for adenoviral vector production. Hum. Gene Ther., 11:2105–2116.
35.
SereginS.S., AmalfitanoA.2009. Overcoming pre-existing adenovirus immunity by genetic engineering of adenovirus-based vectors. Expert Opin. Biol. Ther., 9:1521–1531.
36.
SereginS.S., AmalfitanoA.2010. Improving adenovirus based gene transfer: strategies to accomplish immune evasion. Viruses, 2:2013–2036.
37.
SereginS.S., AldhamenY.A., AppledornD.M.et al.2009a. CR1/2 is an important suppressor of adenovirus-induced innate immune responses and is required for induction of neutralizing antibodies. Gene Ther., 16:1245–1259.
38.
SereginS.S., AppledornD.M., McBrideA.J.et al.2009b. Transient pretreatment with glucocorticoid ablates innate toxicity of systemically delivered adenoviral vectors without reducing efficacy. Mol. Ther., 17:685–696.
39.
SereginS.S., AldhamenY.A., AppledornD.M.et al.2010a. Adenovirus capsid-display of the retro-oriented human complement inhibitor DAF reduces Ad vector-triggered immune responses in vitro and in vivo. Blood, 116:1669–1677.
40.
SereginS.S., HartmanZ.C., AppledornD.M.et al.2010b. Novel adenovirus vectors 'capsid-displaying' a human complement inhibitor. J. Innate Immun., 2:353–359.
41.
SumidaS.M., TruittD.M., KishkoM.G.et al.2004. Neutralizing antibodies and CD8+ T lymphocytes both contribute to immunity to adenovirus serotype 5 vaccine vectors. J. Virol., 78:2666–2673.
42.
SureshM., MolinaH., SalvatoM.S.et al.2003. Complement component 3 is required for optimal expansion of CD8 T cells during a systemic viral infection. J. Immunol., 170:788–794.
43.
TangJ., OliveM., PulmanausahakulR.et al.2006. Human CD8+ cytotoxic T cell responses to adenovirus capsid proteins. Virology, 350:312–322.
44.
TianJ., XuZ., SmithJ.S.et al.2009. Adenovirus activates complement by distinctly different mechanisms in vitro and in vivo: indirect complement activation by virions in vivo. J. Virol., 83:5648–5658.
45.
TripathyS.K., GoldwasserE., LuM.M.et al.1994. Stable delivery of physiologic levels of recombinant erythropoietin to the systemic circulation by intramuscular injection of replication-defective adenovirus. Proc. Natl. Acad. Sci. U.S.A., 91:11557–11561.
46.
TripathyS.K., BlackH.B., GoldwasserE., LeidenJ.M.1996. Immune responses to transgene-encoded proteins limit the stability of gene expression after injection of replication-defective adenovirus vectors. Nat. Med., 2:545–550.
47.
WeaverE.A., BarryM.A.2008. Effects of shielding adenoviral vectors with polyethylene glycol (PEG) on vector-specific and vaccine-mediated immune responses. Hum. Gene Ther., 19:1369–1382.
48.
WolinsN., LozierJ., EggermanT.L.et al.2003. Intravenous administration of replication-incompetent adenovirus to rhesus monkeys induces thrombocytopenia by increasing in vivo platelet clearance. Br. J. Haematol., 123:903–905.
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