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Journal of Veterinary Diagnostic
http://vdi.sagepub.com/content/14/5/377
The online version of this article can be found at:
DOI: 10.1177/104063870201400503
2002 14: 377 J VET Diagn Invest
Chang-Won Lee, Corrie Brown and Mark W. Jackwood
Examined by in Situ Hybridization with Antisense Digoxigenin-Labeled Universal Riboprobe
Tissue Distribution of Avian Infectious Bronchitis Virus following in Ovo Inoculation of Chicken Embryos
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What is This?
Gray
JMK
Mass41
Wolgemuth
* 5 positive cells per high-power eld (400); 1 and 5 positive cells per high-power eld; no positive cells.
Figure 2. In situ hybridization using universal riboprobe. Positive staining can be observed as a dark area in the cytoplasm of epithelial
cells. A, Esophagus (20) 2 days postinfection with the Gray strain. B, Shell gland near bursa (40) 5 days postinfection with the JMK
strain. C, Mesentery near kidney (20) 2 days postinfection with the ArkDPI strain. D, Air sac around lung (20) 2 days postinfection
with the Conn strain. All the sections were counterstained with Mayer hematoxylin.
in chicken embryos should not be overlooked in in-
terpreting this result.
In addition to the tissues described above, we fur-
ther identied positively stained epithelial cells in the
air sac, mesentery, shell gland, and esophagus with
certain strains of IBV. Virus replication in the epithe-
lial cells of air sacs has also been demonstrated by
other researchers (Nauwynck H, Pensaert M, 1988,
Abstract In: Proc 1st Int Symp Infect Bronchitis,
Rauischholzhausen, Germany, pp. 113119). In a pre-
vious study,
19
maximum virus isolations were obtained
from the esophagus of chickens infected with an en-
teric isolate of IBV. Extensive staining in the epithelial
cells of the esophagus in our results conrms that IBV
actually multiplies in this tissue. Detection of IBV in
the mesentery and shell gland has been reported pre-
viously.
10
Considering the close proximity of these tis-
sues with visceral organs, these may provide a route
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381 In ovo tissue distribution of IBV
of virus dissemination throughout the chicken embryo.
The results clearly show the strict epitheliotropic na-
ture of IBV. No positive staining in the spleen at any
time further conrms this fact. The receptor on the cell
surface for IBV is known to be 2,3-linked N-acetyl-
neuraminic acid.
11
However, binding of IBV to this
molecule in itself is not likely the basis of viral cell
tropism because sialic acid is a common cell surface
carbohydrate residue. Thus, an additional, more epi-
thelial cell type-specic molecule is probably required
for IBV infection.
The main advantage of the ISH technique is topo-
logic assessment of nucleic acid localization. Seen
against the backdrop of histologic change, ISH can
help dene the role that the virus plays in tissue dam-
age. In addition, ISH can be useful for retrospective
analysis because nucleic acid is indenitely stable in
parafn-embedded tissue.
Sources and manufacturers
a. Charles Rivers Spafas, North Franklin, CT.
b. Fisher Scientic, Pittsburgh, PA.
c. Invitrogen, San Diego, CA.
d. Roche, Indianapolis, IN.
e. ICN, Irvine, CA.
f. Permount, Fisher Scientic, Pittsburg, PA.
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