Abstract
Replication-defective adenovirus vector without both E1 and E3 is one of the most popular tools in transgenic therapies. However, more attention should be paid to adenovirus vectors mediated-gene modified study on endothelial cells (ECs). To verify the possible danger in that process, we explored the effect of adenovirus on ECs in this study. By using western blot analysis, we showed that the level of both CD40 and CD40L on human umbilical vein endothelial cells (HUVECs) were upregraduated by adenovirus vector infection at 100 multiplicity of infection (MOI). The activation of ECs induced by adenovirus vector infection at MOI 100 can be partly inhibited by a blockade of CD40/CD40L interactions by using the recombinant adenovirus Ad-sCD40LIg or an anti-CD40L monoclonal antibody (mAb) in vitro. On ECs, blockade of CD40/CD40L decreased the expression of IL (interleukin)-6, IL-8 and intercellular adhesion molecule (ICAM) in adenovirus vector-induced cells. In electrophoretic mobility shift assay (EMSA), both Ad-sCD40LIg and anti-CD40L mAb can attenuate the activity of NF-kappaB (NF-κB) pathway contributing to the activation of ECs, which indicated that CD40-CD40L interactions played significant role in the activation of ECs induced by adenovirus vectors via an NF-κB pathway. Our study provide evidences for a supplementary mechanism of the ECs activation induced by adenovirus vector infection and suggests that CD40-CD40L interactions partly participate in the ECs activation induced by adenovirus vectors in an NF-κB-dependent manner. Key words : Adenovirus vector, CD40, CD40L, endothelial cells, NF-kappaB.
Highlights
Replication-defective adenovirus vector without both E1 and E3 is one of the most popular tools in transgenic therapies for its advantages, for instance they can carryAbbreviations: ECs, Endothelial cells; HUVECs, human umbilical vein endothelial cells; MOI, multiplicity of infection; mAb, monoclonal antibody; NF-κB, NF-kappaB.long exogenous DNA fragments, infect variety of dividing and nondividing cells at higher infectious rate and there is less risk of inducing alteration of the host DNA for the infection cannot interfere with the host genome (Kanaya et al, 2003)
In electrophoretic mobility shift assay (EMSA), both Ad-sCD40LIg and anti-CD40 ligand (CD40L) mAb can attenuate the activity of NF-kappaB (NF-κB) pathway contributing to the activation of ECs, which indicated that CD40-CD40L interactions played significant role in the activation of ECs induced by adenovirus vectors via an NF-κB pathway
A concentration-dependent stimulation of ECs by adenovirus vectors was performed and we observed that maximal effect on CD40 and CD40L protein expression was achieved after 24 h of infection with 100 MOI of adenovirus vectors
Summary
Replication-defective adenovirus vector without both E1 and E3 is one of the most popular tools in transgenic therapies for its advantages, for instance they can carry. Long exogenous DNA fragments, infect variety of dividing and nondividing cells at higher infectious rate and there is less risk of inducing alteration of the host DNA for the infection cannot interfere with the host genome (Kanaya et al, 2003). Some problems must be tackled before the vector is applicated for patients. Adenovirus vectors, regardless of containing an exogenous or not, can excite an inflammatory reaction followed by upregulation of the inflammatory genes (Yei et al, 1994; Schaack et al, 2011). Adenovirus vectors can promote the acute inflammation in vivo by inducing chemokines including the C-C chemokine RANTES and IP-10 through
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