Biology Reference
In-Depth Information
Chapter 10
The Analysis of Innate Immune Response to Adenovirus
Using Antibody Arrays
Nelson C. Di Paolo and Dmitry M. Shayakhmetov
Abstract
Even though natural infections with adenovirus (Ad) are largely harmless in humans, an intravenous Ad
vector administration for gene delivery purposes, especially at high doses, stimulates strong innate and
adaptive immune responses, and can be fatal to the host. In animal models, intravenous Ad administration
has been shown to induce transcription and release in the serum of a great number of pro-infl ammatory
cytokines and chemokines. Macrophages, including tissue residential macrophages (e.g., Kupffer cells in
the liver), and dendritic cells throughout the body are considered to be the primary source of these
pro-infl ammatory mediators following their transduction with Ads. Here, we provide an overview and
methodology for the qualitative and quantitative analyses of pro-infl ammatory cytokine and chemokine
expression in the spleen and their release into the bloodstream after intravenous Ad delivery using anti-
body arrays.
Key words
Infl ammation, Innate immunity, Adenovirus vectors, Systemic administration
1
Introduction
To date, accumulating evidence suggests that Ad triggers highly
complex multifaceted innate immune and infl ammatory responses,
which are refl ected at a clinical level in cytokinemia, thrombocyto-
penia, complement activation, disseminated intravascular coagula-
tion, and multiple organ failure due to (at least in part) collateral
damage from infi ltrating pro-infl ammatory leukocytes. Despite con-
siderable recent progress in defi ning early mediators of Ad-induced
infl ammation [
1
-
6
], the unifying mechanistic description of sequen-
tial steps of events that lead from early Ad-host cell interactions to
clinical signs of Ad-triggered systemic toxicity remains illusive.
Within minutes after intravenous administration, Ad particles
are trapped by tissue residential macrophages that trigger activa-
tion and release on numerous pro-infl ammatory cytokines and
chemokines that orchestrate host responses targeted at recruitment
of polymorphonuclear leukocytes, destruction of Ad particles, and
