Infection of Endothelium With E1 ⁻ E4 ⁺ , but Not E1 ⁻ E4 ⁻ , Adenovirus Gene Transfer Vectors Enhances Leukocyte Adhesion and Migration by Modulation of ICAM-1, VCAM-1, CD34, and Chemokine Expression

Abstract

Abstract-Intravascular introduction of replication-deficient adenoviral vectors (Advectors) provides an ideal model of delivery of transgenes for the treatment of various vascular abnormalities. On the basis of the knowledge that Advectors can induce inflammatory responses after intravascular administration, we speculated that cellular activation by Advector infection could directly modulate the endothelial cell (EC) adhesion molecule/chemokine expression repertoire. Infection of human umbilical vein ECs or bone marrow microvascular ECs with an E1 Ϫ E4 ϩ Advector resulted in the upregulation of intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and CD34, but not E-selectin, P-selectin, CD36, CD13, CD44, HLA-DR or PECAM. Upregulation of ICAM-1, VCAM-1, and CD34 was apparent 12 hours after infection and persisted for weeks after infection. Selective induction of adhesion molecules was mediated by the presence of the E4 gene in the Advector, because infection of ECs with an E1 Ϫ E4Ϫ Advector had no effect on adhesion molecule expression.

Adenoviruses increase endothelial cell proliferation, migration, and tube formation: Partial reversal by the focal adhesion kinase inhibitor, FRNK

“…However, infection of endothelial cells with E1 − E4 + adenoviruses induces the expression of several genes. These genes are involved in intracellular signaling (LBC guanine nucleotide exchange protein, Gα-S, MEK 5) (Ramalingam et al, 1999b), Ca 2+ regulation/cytoskeleton (calpactin p11, calpactin p36, vinculin, SCA1) (Ramalingam et al, 1999a), growth regulation (IGFBP4, TGF-β2) (Ramalingam et al, 1999b), housekeeping (G6PDH) (Ramalingam et al, 1999b), and adhesion (ICAM-1, VCAM, CD34 (Rafii, et al 2001). Likewise, adenoviral infection of human saphenous vein endothelial cells and foreskin micro-vascular endothelial cells caused upregulation of adhesion molecules (CD54, CD62, CD106) and cytokines (Th1, Il-12, IFN-γ), as well as enhanced secretion of cytokines (TNF-α, IL-8, IL-1β, IL-6) (Tan et al, 2005).…”

“…Although adenoviral mediated activation of FAK depends on penton base protein, many of the responses of endothelial cells to infection depend on the presence of the adenoviral E4 region (Ramalingam et al, 1999a;Ramalingam et al, 1999a;Rafii et al, 2001). This region contains seven open reading frames (Shenk, 1996).…”

Chimpanzee-origin adenovirus vectors as vaccine carriers

“…Previous gene therapy trials reported that deletion or functional inactivation of E4 in AdHu5 vectors reduces the inflammatory response to the vector, 34 and decreases AdHu5-mediated upregulation of adhesion molecules in endothelial cells. 35 Such adhesion molecules are crucial for lymphocyte trafficking. Furthermore, E4 has been reported to affect the performance of viral and cellular promoters.…”

Molecular basis of the inflammatory response to adenovirus vectors

“…In epithelial A549 cells and in endothelial cells (human umbilical vein and bone marrow microvascular endothelial cells), adenovirus vectors induced the expression of various leukocyte adhesion molecules such as ICAM-1 and VCAM-1. 36,37 The expression of leukocyte adhesion molecules facilitates leukocyte recruitment to adenovirus vector-transduced tissues. 17 Thus, adenovirus vectors induce a broad array of inflammatory genes in innate and noninnate cells that underlie the inflammatory response to these agents in vivo.…”