Sensitive Detection Of Measles Virus Infection In The Blood And Tissues Of Humanized Mouse By One-Step Quantitative RT-PCR

Tuesday, 12th of November 2013

[source]Frontiers in Microbiology[|source]

Currently, humanized mouse systems are widely used as alternatives to non-human primate models, especially for the study of human-tropic infectious diseases such as HIV, human T cell leukemia virus, dengue virus, and Epstein Bar virus. Of the different humanized mice models, the Bone Marrow/Liver/Thymus transplanted mouse, which is transplanted with human fetal liver and thymus tissue in addition to hematopoietic stem cells, is recognized as the model that most closely mimics the human immune response. To study measles virus (MV) infection in humanized mice, the authors infected an MV vaccine strain expressing enhanced green fluorescent protein and analyzed MV-infected cells by flow cytometry. This report documents a humanized mouse model that is expected to be a useful tool for studying the pathogenesis of measles virus infection and vaccine efficacy in the days towards elimination. More details are available at: 

http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3795360/

Abstract

Live attenuated measles virus (MV) has long been recognized as a safe and effective vaccine, and it has served as the basis for development of various MV-based vaccines. However, because MV is a human-tropic virus, the evaluation of MV-based vaccines has been hampered by the lack of a small-animal model. The humanized mouse, a recently developed system in which an immunodeficient mouse is transplanted with human fetal tissues or hematopoietic stem cells, may represent a suitable model. Here, we developed a sensitive one-step quantitative reverse transcription (qRT)- PCR that simultaneously measures nucleocapsid (N) and human RNase P mRNA levels. The results can be used to monitor MV infection in a humanized mouse model. Using this method, we elucidated the replication kinetics of MV expressing enhanced green fluorescent protein both in vitro and in humanized mice in parallel with flow-cytometric analysis. Because our qRT-PCR system was sensitive enough to detect MV expression using RNA extracted from a small number of cells, it can be used to monitor MV infection in humanized mice by sequential blood sampling.