Neutralized adenovirus-immune complexes can mediate effective gene transfer via an Fc receptor-dependent infection pathway

Philip L. Leopold, Rebecca L. Wendland, Theresa Vincent, Ronald Crystal

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Neutralization of adenovirus (Ad) by anti-Ad neutralizing antibodies in serum involves formation of Ad-immune complexes that prevent the virus from interacting with target cells. We hypothesized that Ad-immune complexes likely contain viable Ad vectors which, although no longer capable of gaining access to receptors on target cells, may be able to express transgenes in cells bearing Fc receptors for immunoglobulins, i.e., that antibody-based " neutralization" of Ad vectors may be circumvented by the Fc receptor pathway. To test this hypothesis, we expressed the Fcγ receptor IIA (FcγR) in A549 lung epithelial cells or human dermal fibroblasts and evaluated gene transfer in the presence of human neutralizing anti-Ad serum. FcγR-expressing cells bound and internalized copious amounts of Ad, with a distinct population of internalized Ad trafficking to the nucleus. The dose-response curves for inhibition of gene transfer revealed that FcγR-expressing cells required a more-than-10-fold higher concentration of anti-Ad serum to achieve 50% inhibition of Ad-encoded β-galactosidase expression compared with non-FcγR-expressing cells. The discrepancy between neutralization of Ad during infection of FcγR-expressing cells and neutralization of Ad during infection of non-FcγR-expressing cells occurred with either heat-inactivated or non-heat-inactivated sera, was blocked by addition of purified Fc domain protein, and did not require the cytoplasmic domain of FcγR, suggesting that immune complex internalization proceeded via endocytosis rather than phagocytosis. FcγR-mediated infection by Ad-immune complexes did not require expression of the coxsackie virus-Ad receptor (CAR) since similar data were obtained when CAR-deficient human dermal fibroblasts were engineered to express FcγR. However, interaction of the Ad penton base with cell surface integrins contributed to the difference in neutralization between FcγR-expressing and non-FcγR-expressing cells. The data indicate that complexes formed from Ad and anti-Ad neutralizing antibodies, while compromised with respect to infection of non-FcγR- expressing target cells, maintain the potential to transfer genes to FcγR-expressing cells, with consequent expression of the transgene. The formation of Ad-immune complexes that can target viable virus to antigen-presenting cells may account for the success of Ad-based vaccines administered in the presence of low levels of neutralizing anti-Ad antibody.

Original languageEnglish
Pages (from-to)10237-10247
Number of pages11
JournalJournal of Virology
Volume80
Issue number20
DOIs
Publication statusPublished - 1 Oct 2006
Externally publishedYes

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Fc Receptors
Antigen-Antibody Complex
Adenoviridae
Infection
Genes
Adenoviridae Infections
Neutralizing Antibodies
Virus Receptors
Enterovirus
Serum
Transgenes
Adenovirus Vaccines
Coxsackie and Adenovirus Receptor-Like Membrane Protein
Fibroblasts
Galactosidases
Viruses
Immunoglobulin Fc Fragments
Skin
Antigen-Presenting Cells
Endocytosis

ASJC Scopus subject areas

  • Immunology

Cite this

Neutralized adenovirus-immune complexes can mediate effective gene transfer via an Fc receptor-dependent infection pathway. / Leopold, Philip L.; Wendland, Rebecca L.; Vincent, Theresa; Crystal, Ronald.

In: Journal of Virology, Vol. 80, No. 20, 01.10.2006, p. 10237-10247.

Research output: Contribution to journalArticle

Leopold, Philip L. ; Wendland, Rebecca L. ; Vincent, Theresa ; Crystal, Ronald. / Neutralized adenovirus-immune complexes can mediate effective gene transfer via an Fc receptor-dependent infection pathway. In: Journal of Virology. 2006 ; Vol. 80, No. 20. pp. 10237-10247.
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