Abstract
Humanized mouse models are used as comprehensive small-animal models of EBV infection. Previously, infectious doses of EBV used in vivo have been determined mainly on the basis of TD50 (50% transforming dose), which is a time-consuming process. Here, we determined infectious doses of Akata-EBV-GFP using green Raji units (GRUs), and characterized dose-dependent effects in humanized mice. We defined two outcomes in vivo, including an infection model and a lymphoma model, following inoculation with low or high doses of Akata-EBV-GFP, respectively. Inoculation with a low dose induced primary B cells to become lymphoblastoid cell lines in vitro, and caused latent infection in humanized mice. In contrast, a high dose of Akata-EBV-GFP resulted in primary B cells death in vitro, and fatal B cell lymphomas in vivo. Following infection with high doses, the frequency of CD19+ B cells decreased, whereas the percentage of CD8+ T cells increased in peripheral blood and the spleen. At such doses, a small part of activated CD8+ T cells was EBV-specific CD8+ T cells. Thus, GRUs quantitation of Akata-EBV-GFP is an effective way to quantify infectious doses to study pathologies, immune response, and to assess (in vivo) the neutralizing activity of antibodies raised by immunization against EBV.
Highlights
Epstein-Barr virus (EBV) is a causative agent of infectious mononucleosis (IM), and is associated with a range of human diseases, including malignancies and autoimmune diseases [1,2]
Humanized mouse models have been used as a typical model to analyze virological parameters and immune functions in vivo upon EBV infection [9,11]
green Raji units (GRUs) quantitation was established as a tool to study EBV infection in vitro and in vivo [15,20,28]
Summary
Epstein-Barr virus (EBV) is a causative agent of infectious mononucleosis (IM), and is associated with a range of human diseases, including malignancies (e.g., nasopharyngeal carcinoma, gastric carcinoma, Burkitt lymphoma, and Hodgkin lymphoma) and autoimmune diseases (e.g., rheumatoid arthritis and multiple sclerosis) [1,2]. The development of severely immunodeficient mouse strains, such as NOD/LtSz-scid Il2rg−/ − (NSG), NOD/Shi-scid Il2rgnull (NOG), and Balb/c Rag2−/− IL-2rg−/− (BRG), enabled the in vivo reconstitution of functional human immune system components after transplantation with human hematopoietic stem cells (HSCs) [6,7,8]. These mice are called humanized mice, and have been instrumental to reproduce key features of viral infections that target cells of the human immune system, including human immunodeficiency virus type 1 (HIV-1) and EBV [9,10]
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