Exploitation of Herpesviral Transactivation Allows Quantitative Reporter Gene-Based Assessment of Virus Entry and Neutralization
نویسندگان
چکیده
Herpesviral entry is a highly elaborated process requiring many proteins to act in precise conjunction. Neutralizing antibodies interfere with this process to abrogate viral infection. Based on promoter transactivation of a reporter gene we established a novel method to quantify herpesvirus entry and neutralization by antibodies. Following infection with mouse and human cytomegalovirus and Herpes simplex virus 1 we observed promoter transactivation resulting in substantial luciferase expression (>1000-fold). No induction was elicited by UV-inactivated viruses. The response was MOI-dependent and immunoblots confirmed a correlation between luciferase induction and pp72-IE1 expression. Monoclonal antibodies, immune sera and purified immunoglobulin preparations decreased virus-dependent luciferase induction dose-dependently, qualifying this approach as surrogate virus neutralization test. Besides the reduced hands-on time, this assay allows analysis of herpesvirus entry in semi-permissive and non-adherent cells, which were previously non-assessable but play significant roles in herpesvirus pathology.
منابع مشابه
Human papillomavirus genotype 16 pseudovirus production and purification in HEK-293FT cells
Introduction: Human papillomavirus (HPV) is the main causative agent of cervical cancer worldwide leading to a big health problem, especially in the developing countries. Among 14 common high-risk genotypes, HPV16 accounts for more than 50% of all cervical cancers. The current prophylactic vaccines against HPV infection are based on L1 protein. Due to some drawbacks in the current vaccines such...
متن کاملAn Enterovirus-Like RNA Construct for Colon Cancer Suicide Gene Therapy
Background: In gene therapy, the use of RNA molecules as therapeutic agents has shown advantages over plasmid DNA, including higher levels of safety. However, transient nature of RNA has been a major obstacle in application of RNA in gene therapy. Methods: Here, we used the internal ribosomal entry site of encephalomyocarditis virus and the 3’ non-translated region of Poliovirus to design an en...
متن کاملQuantitative Studies on the Neutralization Reaction between African Horse-sickness Virus and Antiserum
متن کامل
ساخت وکتور ریپورتر القایی لوسیفراز برای ردیابی فعالیت ویروس HTLV-1
Background: HTLV-1 (Human T_cell lymphotropic virus type), with about 20 million individuals infected worldwide, is a global health problem and is endemic in certain area such as Japan and Khorasan province of Iran. HTLV-1 is the causative agent of progressive diseases, Adult T cell Leukemia (ATL), and HTLV-I Associated Myelopathy/Tropical Spastic Paraparesis (HAM/TSP) which have no ...
متن کاملتولید هورمون رشد انسانی نوترکیب توسط سلول تخمدان هامستر چینی و بررسی فعالیت زیستی آن به روش سنجش گزارشگر ژنی
Background: Cultivated mammalian cells, because of their capacity for proper protein folding, assembly and post–translational modification, have become the dominant system for production of recombinant proteins in clinical application. Therefore, the quality and efficacy of protein can be superior when expressed in mammalian cells compared to other hosts such as bacteria. Gene reporte...
متن کاملذخیره در منابع من
با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید
عنوان ژورنال:
دوره 6 شماره
صفحات -
تاریخ انتشار 2011