Humanized mice mount specific adaptive and innate immune responses to EBV and TSST-1

Abstract

Here we show that transplantation of autologous human hematopoietic fetal liver CD34+ cells into NOD/SCID mice previously implanted with human fetal thymic and liver tissues results in long-term, systemic human T-cell homeostasis. In addition, these mice show systemic repopulation with human B cells, monocytes and macrophages, and dendritic cells (DCs). T cells in these mice generate human major histocompatibility complex class I- and class II-restricted adaptive immune responses to Epstein-Barr virus (EBV) infection and are activated by human DCs to mount a potent T-cell immune response to superantigens. Administration of the superantigen toxic shock syndrome toxin 1 (TSST-1) results in the specific systemic expansion of human Vbeta2+ T cells, release of human proinflammatory cytokines and localized, specific activation and maturation of human CD11c+ dendritic cells. This represents the first demonstration of long-term systemic human T-cell reconstitution in vivo allowing for the manifestation of the differential response by human DCs to TSST-1.

One Percent Tenofovir Applied Topically to Humanized BLT Mice and Used According to the CAPRISA 004 Experimental Design Demonstrates Partial Protection from Vaginal HIV Infection, Validating the BLT Model for Evaluation of New Microbicide Candidates

“…BLT mice were prepared essentially as previously described (6,11,12,24,25,31,41,45). Briefly, thymus/liver-implanted NOD/SCID or NOD/SCID-gamma chain null mice (NSG) (The Jackson Laboratories) were transplanted with autologous human fetal liver CD34 ϩ cells (Advanced Bioscience Resources) and monitored for human reconstitution in peripheral blood by flow cytometry, as we have previously described (11,12,31,45). Mice were maintained either at the Animal Resources Center of UT Southwestern Medical Center at Dallas or with the Division of Laboratory Animal Medicine at the University of North Carolina (UNC) at Chapel Hill in accordance with protocols approved by the each institution's Institutional Animal Care and Use Committee.…”

“…Cells were washed, counted using trypan blue exclusion, and utilized as indicated throughout the text, on figures, and in tables. Other tissues were harvested and then evaluated by molecular, microscopic, and flow cytometric analyses for evidence of HIV infection as we have previously described (11,12,31,45). The flow cytometry antibody panels for analysis of BLT mouse FRT were Analysis of systemic infection was performed on tissues harvested from infected mice or on cells isolated from the tissues indicated throughout the text, on figures, and in tables.…”

“…The flow cytometry antibody panels for analysis of BLT mouse FRT were Analysis of systemic infection was performed on tissues harvested from infected mice or on cells isolated from the tissues indicated throughout the text, on figures, and in tables. Utilizing in situ hybridization, real-time PCR analysis, and coculture with allogeneic human peripheral blood mononuclear cells (PBMC) as previously described (11,12,31,45). An immunofluorescence assay was performed on paraffin-embedded BLT FRT sections.…”

In vivo imaging of malaria parasites in the murine liver

“…Similar approaches should also permit illumination of how protective immunity is achieved and how activated T cells interact with infected hepatocytes, or comparison of immune effectors against parasites in naïve or vaccinated animals. Ultimately, similar techniques might also be applied to study other host-parasite combinations, such as rodent parasites in their natural host (e.g., P. berghei-Grammomys surdaster) or primate systems, or P. falciparum in humanized mice grafted with human cells or tissues 24 . Figure 5 | Release of merosome buds from a hepatic schizont (also see Supplementary Video 3 online).…”

Animal models of Epstein Barr virus infection

“…Animal with HLA transgenes allow for the thymic selection of T cells on these molecules and result in similar epitope specificities as in patients. Regardless of the source, these T cells produce IFN-γ in response to cognate CD8 + T cell epitopes (Strowig et al, 2009) and autologous LCLs (Traggiai et al, 2004;Melkus et al, 2006;Yajima et al, 2008). Moreover, both CD4 + and CD8 + T cell clones that were primed during EBV infection of HIS mice are able to lyse LCLs (Strowig et al, 2009) …”