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Human adenoviruses have been used as gene therapy vectors. Amongst human adenoviruses, the most widely studied and used in gene therapy preclinical studies is human adenovirus 5 (HAdV-5). Nonetheless, use is hampered by several factors such as the high level of pre-existing neutralizing antibodies against HAdV-5 virions in the clinical population and hepatic tropism following intravenous administration, which can lead to acute liver toxicity in humans, non-human primates and rodent models. Mouse Immunoglobulin M (IgM) has a pivotal role in triggering the classical complement pathway in vitro, which can lead to neutralization of adenovirus virions. Assessing the interaction of HAdV-5 with different components of the mouse immune system can help us to better understand these complex mechanisms and provide a theoretical basis when designing gene therapies using HAdV-5 vectors.
SKOV3 cells, HEK-293 cells were cultured under suitable conditions. Viral particles were propagated in HEK-293 cells and purified by CsCl gradient centrifugation. To investigate the mechanism of action between human IgM and other serum components and HAdV-5 viral particles, we performed nanoparticle tracking analysis, neutralization assays, mass spectrometry and in vivo experiments to examine the physical binding of immune components to HAdV-5, respectively.
The data generated suggest that (1) immune response proteins can bind HAdV-5 virions in naïve mice but neutralization only occurs if FX is absent or the virion is unable to bind FX, (2) purified mouse IgM antibodies bind to HAdV-5 only in the presence of complement components and (3) in serum from immunocompetent mice both classical and alternative complement pathways are activated, while in serum from immuno-deficient mice with absent immunoglobulins, the alternative complement pathway mediates partial neutralization. It was ultimately demonstrated that mouse IgM was not critical for HAdV-5 neutralization either in vitro or in vivo.
Fig. 1 Schematic model of the interactions of HAdV-5 with the host immune system.
This study suggest that HAdV-5 neutralization can be mediated by both the classical and alternative pathways and that, in the absence of immunoglobulins, the complement cascade can be activated by direct binding of C3 to the virion.
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