[Effects of adenovirus-mediated gene transfer of ICOSIg fusion protein on experimental autoimmune myocarditis in Lewis rats].
OBJECTIVE
To explore the effects of adenovirus vector-mediated gene transfer of ICOSIg fusion protein on experimental autoimmune myocarditis (EAM) in Lewis rats.


METHODS
Expression vector containing ICOSIg (p-Adeno-ICOSIg) was constructed by fusion of human ICOS and IgGFc segment. Adenovirus vector was digested by PacI enzyme and transfected into HEK 293 cells. Adenovirus expressing ICOSIg was produced. EGFP was constructed into adenovirus vector and used as control. EAM was induced in Lewis rats by injection of porcine cardiac myosin. All immunized Lewis rats were divided into 4 groups. Group A (n = 15) and B (n = 15) received adenovirus containing ICOSIg on day 0 and day 14 respectively to study the effects of costimulatory molecules gene therapy on T cell activation and inflammation; group C (n = 10) and group D (n = 10) received adenovirus containing EGFP on day 0 and day 14 respectively as controls. Group E (n = 10) was normal controls that did not receive immunization. On day 28, all rats were killed after echocardiography examination. Histopathological examination was performed to observe myocardial inflammation. Protein levels of ICOS, ICOSL, B7-1 and B7-2 were detected by Western blot. INF-gamma, IL-2 and IL-4 mRNA were determined by realtime RT-PCR.


RESULTS
On day 28, cardiac function was significantly improved and myocardial inflammation significantly attenuated in group B compared to group A, C and D (all P < 0.05). B7-1 expression at protein level was significantly lower in group B than that of group C (P < 0.05). ICOS and ICOSL expressions at protein level were significantly decreased in both group A and B compared with group C and D (P < 0.05). IFN-gamma mRNA level significantly decreased and IL-4 mRNA significantly increased in group A and B compared to group C and D (P < 0.05).


CONCLUSIONS
Blockade of costimulatory pathway with gene therapy of ICOSIg alleviated autoimmune inflammatory damage and improved cardiac function in Lewis rats with EAM. Down-regulated costimulatory molecules in the myocardium and reduced inflammatory cytokine secretion might be responsible for the beneficial effects of ICOSIg in this model.
Li
Wei-min
WM
Department of Cardiology, First Clinical College of Harbin Medical University, Harbin 150001, China.
Liu
Wei
W
Zhang
Rui-hong
RH
Wang
Zheng
Z
Kong
Yi-hui
YH
Li
Yue
Y
Yang
Shu-sen
SS
Shen
Jing-xia
JX
Yu
Jiang-bo
JB
Fan
Ying
Y
Xue
Jing-yi
JY
Geng
Jian-qiang
JQ
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000945:Antigens, Differentiation, T-Lymphocyte / Q000235:genetics*; D001327:Autoimmune Diseases / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D004195:Disease Models, Animal; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D007141:Immunoglobulin Fc Fragments / Q000235:genetics*; D060889:Inducible T-Cell Co-Stimulator Protein; D008297:Male; D009205:Myocarditis / Q000276:immunology / Q000473:pathology / Q000628:therapy*; D051381:Rats; D011917:Rats, Inbred Lew; D011993:Recombinant Fusion Proteins / Q000235:genetics
D000945:Antigens, Differentiation, T-Lymphocyte; C556692:ICOS protein, human; D007141:Immunoglobulin Fc Fragments; D060889:Inducible T-Cell Co-Stimulator Protein; D011993:Recombinant Fusion Proteins