Inhibition of rat vascular smooth muscle cell proliferation in vitro and in vivo by recombinant replication-competent herpes simplex virus.
OBJECTIVE
The proliferation of vascular smooth muscle cells (VSMCs) is a common feature associated with vascular proliferative disorders such as atherosclerosis and restenosis after balloon angioplasty. We examined the antiproliferative effects of recombinant replication-competent herpes simplex virus (HSV), hrR3, to proliferative VSMCs both in vitro and in vivo.


METHODS
Early passages of Sprague-Dawley rat VSMCs were infected with hrR3 at a low multiplicity of infection (0.01 to 1.0) to examine the in vitro cytotoxic activity of this recombinant HSV to VSMCs in a proliferative state. Sprague-Dawley rats underwent balloon dilatation injury of the left carotid artery to induce neointimal formation. The injured carotid arteries were infected with hrR3 five days after balloon injury. Two weeks after injury, the left carotid arteries were fixed, and the areas of the neointimal and medial layers were analyzed microscopically. Because the reporter Escherichia coli lacZ gene in hrR3 is expressed only in infected cells in which the virus is actively replicating, virus replication was confirmed by X-gal staining.


RESULTS
A morphometric analysis revealed that there were significant differences in the intima/media ratio between the HSV-treated group and mock-infected group (0. 354+/-0.068 and 1.08+/-0.055, respectively). In the histological study (X-gal staining), positive X-gal staining was observed chiefly in the VSMCs in the medial layer just beneath the internal elastic lamina, indicating active viral replication.


CONCLUSIONS
Virus-mediated cytocidal therapy using recombinant HSV vector is a promising modality for the treatment of the restenosis after balloon angioplasty.
Miyatake
S I
SI
Department of Neurosurgery & Clinical Neuroscience, Kyoto University Graduate School of Medicine, Kyoto, Japan. ichi@kuhp.kyoto-u.ac.jp
Yukawa
H
H
Toda
H
H
Matsuoka
N
N
Takahashi
R
R
Hashimoto
N
N
D000800:Angioplasty, Balloon / Q000009:adverse effects; D000818:Animals; D002339:Carotid Arteries / Q000473:pathology / Q000821:virology; D002340:Carotid Artery Diseases / Q000473:pathology; D002455:Cell Division; D002478:Cells, Cultured; D002863:Chromogenic Compounds; D004547:Elastic Tissue / Q000473:pathology / Q000821:virology; D004730:Endothelium, Vascular / Q000473:pathology / Q000821:virology; D004926:Escherichia coli / Q000235:genetics; D005697:Galactosides; D017930:Genes, Reporter / Q000235:genetics; D006965:Hyperplasia; D007150:Immunohistochemistry; D007211:Indoles; D019394:Ki-67 Antigen / Q000032:analysis; D007763:Lac Operon / Q000235:genetics; D009131:Muscle, Smooth, Vascular / Q000473:pathology / Q000821:virology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012008:Recurrence; D018139:Simplexvirus / Q000235:genetics / Q000502:physiology*; D017539:Tunica Intima / Q000473:pathology / Q000821:virology; D017540:Tunica Media / Q000473:pathology / Q000821:virology; D014779:Virus Replication / Q000235:genetics; D001616:beta-Galactosidase; D014841:von Willebrand Factor / Q000032:analysis
D002863:Chromogenic Compounds; D005697:Galactosides; D007211:Indoles; D019394:Ki-67 Antigen; D014841:von Willebrand Factor; D001616:beta-Galactosidase; C044888:5-bromo-4-chloro-3-indolyl beta-galactoside