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The Influence of the Express of TLR9 and IRF7 Imposed by HBcAg Recombinant Adenovirus’s Transfection of Human Peripheral Blood Mononuclear Cells

Author: GaoZhen
Tutor: ZhangLunLi
School: Nanchang University
Course: Infectious Diseases
Keywords: Recombinant adenovirus Hepatitis B core antigen (HBcAg) Transfection Peripheral blood mononuclear cells (PBMCs) TLR9 IRF7 IFN-α
CLC: R363
Type: Master's thesis
Year: 2011
Downloads: 12
Quote: 0
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Abstract


Objective: adenovirus the carrier HBcAg gene the load normal people and chronic HBV carriers in peripheral blood mononuclear cells (PBMCs) detection TLR9, IRF7 expression before and after transfection, IFN-a produces a horizontal, in order to understand the chronic HBV infection, immune identifying the state change. Method: build carry HBcAg gene replication defective recombinant adenovirus transfection healthy subjects and chronic HBV carriers in PBMCs, I grouped by high copy serum HBV-DNA and (or) CpG-ODN2006 stimulation, RT-PCR was used to detect detection TLR9 of TLR9, IRF7mRNA expression before and after transfection, Western-Blot method IRF7 protein expression the Elisa was detected in the culture medium level of IFN-a. Results: ① RT-PCR results before transfection chronic HBV carriers TLR9 the expression of IRF7mRNA lower than normal (P lt; 0.05), which, before transfection to CpG-ODN2006 stimulate HBV carriers and healthy normal TLR9 , IRF7mRNA expression has not increased significantly (P gt; 0.05); transfection I of CpG - ODN2006 stimulate HBV carriers and healthy normal than those transfected TLR9 IRF7mRNA expression significantly increased, and the difference was statistically significant (P lt ; 0.05) ② and TLR9 IRF7mRNA expression consistent with Western-Blot results showed that transfection health than normal to each stimulus before transfection TLR9, IRF7 protein expression enhanced, and the difference was statistically significant (P lt; 0.05), chronic after transfection of HBV carriers than before transfection TLR9, IRF7 protein expression enhancement, the difference was statistically significant (P lt; 0.05). ③ Elisa results show that chronic HBV carriers after transfection IFN-α production level is lower than the healthy control group (P lt; 0.05, P lt; 0.05), in normal health and chronic HBV carriers after transfection, culture supernatants produce IFN-a level than before transfection significantly enhanced, and the difference was statistically significant (P lt; 0.05). Conclusion: ① transfection before chronic HBV infection in PBMCs TLR9, IRF7mRNA and protein expression, IFN-a produce levels significantly lower than normal, suggesting that chronic HBV infection of PBMCs to identify HBV nucleic acid DNA decreased ability can not effectively activate IRF7 not secrete sufficient quantities of IFN-α rid of the virus infection may cause chronic persistent; ② Ad-HBcAg in chronic HBV carriers transfected PBMCs after produce IFN-a, IFN-a level lower than the healthy control group after transfection to generate level, suggesting that persistent HBV infection and PBMCs decreased immune function related; (3) Ad-HBcAg in transfected PBMCs TLR9 IRF7mRNA and protein expression and secretion of IFN-α level than that before transfection enhanced confirmed TLR9 are intracellular receptors to intracellular stimulation can produce strong anti-HBV immunity; ④ transfected with CpG-ODN can activate the expression of chronic HBV carriers PBMCs TLR9, IRF7, while enhancing the secretion of IFN-α, tips to play through the use of CpG-ODN body's innate immune against HBV infection.

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