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Poster De Conférence Année : 2017

Association of adenovirus and metallic particles formed during electropulsation promotes virus entry in CAR-negative cells by endocytosis

Résumé

Penetration of large macromolecules (including DNA) across the plasmic membrane during electropulsation is not fully understood. We had previously applied long-lasting electric pulses after infection of CAR-negative cells with an adenovirus vector and shown a large increase in the expression of its reporter gene. This effect was mainly associated to the large production of metallic particles released by the aluminum electrodes of the electroporation cuvettes. We investigated now the contribution of the different experimental factors to this increase and saw if it can be generalized to several conditions and cell lines. The observed effect is due to increased adenovirus entry into cells since no contribution of the further stages of adenovirus cellular trafficking was detected. The largest effect was obtained when the virions associated during the pulses to the metallic particles formed, compared to the results obtained when the virions were mixed to preformed metallic particles or to aluminum hydroxide powder. Analysis of virion/metallic particle aggregates by transmission electromicroscopy (TEM) showed a higher proportion of complexed virions in the former case. We speculate that this complex is the key factor explaining the effect. To confirm this idea, we have studied infected cells by TEM after infection and with different pulse conditions. We also used different endocytosis inhibitors to delineate which endocytosis mechanism(s) is (are) involved in increased adenovirus entry into cells. Hypotheses that may explain this effect will be discussed.

Domaines

Biotechnologies
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Dates et versions

hal-02405042 , version 1 (11-12-2019)

Identifiants

  • HAL Id : hal-02405042 , version 1

Citer

Thierry Ragot, Catherine Pioche-Durieu, Eric Le Cam, Lluis M Mir. Association of adenovirus and metallic particles formed during electropulsation promotes virus entry in CAR-negative cells by endocytosis. XXVth Anniversary Meeting of the European Society of Gene and Cell Therapy, Oct 2017, BERLIN, Germany. Mary Ann Liebert, Inc., Human Gene Therapy, 28, pp.A119, Human Gene Therapy. ⟨hal-02405042⟩
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