Construction of Human‐<scp>SCID</scp> Chimeric Mice

Roncarolo, Maria Grazia; Carballido, José M.

doi:10.1002/0471142735.im0408s25

Cited by 10 publications

(11 citation statements)

References 32 publications

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“…Hu-BLT mice were generated by following the previously published protocol (22). Six-to eight-week-old NOD.Cg-Prkdc scid Il2rg tm1Wjl /SzJ (NOD/SCID/IL2rγnull, NSG) mice (The Jackson Laboratory) were purchased and maintained in pathogen-free conditions at University of Nebraska-Lincoln Life Sciences Annex.…”

Section: Methodsmentioning

confidence: 99%

Humanized-BLT mouse model of Kaposi’s sarcoma-associated herpesvirus infection

Wang

Kang

Kumar

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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Lack of an effective small-animal model to study the Kaposi's sarcoma-associated herpesvirus (KSHV) infection in vivo has hampered studies on the pathogenesis and transmission of KSHV. The objective of our study was to determine whether the humanized BLT (bone marrow, liver, and thymus) mouse (hu-BLT) model generated from NOD/SCID/IL2rγ mice can be a useful model for studying KSHV infection. We have tested KSHV infection of hu-BLT mice via various routes of infection, including oral and intravaginal routes, to mimic natural routes of transmission, with recombinant KSHV over a 1-or 3-mo period. Infection was determined by measuring viral DNA, latent and lytic viral transcripts and antigens in various tissues by PCR, in situ hybridization, and immunohistochemical staining. KSHV DNA, as well as both latent and lytic viral transcripts and proteins, were detected in various tissues, via various routes of infection. Using double-labeled immune-fluorescence confocal microscopy, we found that KSHV can establish infection in human B cells and macrophages. Our results demonstrate that KSHV can establish a robust infection in the hu-BLT mice, via different routes of infection, including the oral mucosa which is the most common natural route of infection. This hu-BLT mouse not only will be a useful model for studying the pathogenesis of KSHV in vivo but can potentially be used to study the routes and spread of viral infection in the infected host. T he Kaposi's sarcoma (KS)-associated herpesvirus (KSHV), also known as the human herpesvirus 8, was first identified from KS tissues in 1994 (1). It is the etiologic agent for KS and is also associated with primary effusion lymphoma (PEL) and multicentric Castleman's disease (2). More recently it was also found to be associated with KSHV-associated inflammatory cytokine syndrome (3). Although substantial progress has been made in characterizing the virus, there are still many unanswered questions such as how KSHV infection can lead to disease manifestation and whether latent or lytic induction of KSHV are associated with malignancies. One of the reasons is a lack of a good small-animal model to study KSHV infection in vivo, which has hampered studies on how KSHV infects, spreads, and how it interacts with the host and ultimately leads to disease pathogenesis. Moreover, currently there is no vaccine against KSHV infection, and there is need for an effective animal model to evaluate the efficacy of vaccines if they are developed and for the testing of antiviral regimens.An ideal model should have relatively short generation time, reproduce rapidly, be inexpensive to maintain and house, and be easy to manipulate. An example is a rodent model that can be infected by KSHV effectively. Several small-rodent models have been tested for KSHV infection. The models include transplantation with both human KSHV-infected B lymphoma cells and primary human peripheral blood mononuclear cells in the SCID mouse (4), injection of KSHV into the human skin engrafted or the transplant of the SCID mice...

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Section: Methodsmentioning

confidence: 99%

Humanized-BLT mouse model of Kaposi’s sarcoma-associated herpesvirus infection

Wang

Kang

Kumar

et al. 2014

Proc. Natl. Acad. Sci. U.S.A.

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“…In this model, human HSPC transplanted in the fetal tissues engraft and differentiate into T and B mature lymphocytes (Roncarolo and Carballido, 1998). In a first set of experiments, CD34 1 /DNGFR 1 cells transduced and immunoselected using a large-scale procedure (purity 94.5-97.8%) were injected in the subcutaneous graft of SCID- Four experiments were performed with large numbers of CD34 1 cells transduced with different protocols.…”

Section: Resultsmentioning

confidence: 99%

“…In vivo assays were performed in SCID-hu bone/liver/thymus mice constructed as previously described (Roncarolo et al, 1996;Roncarolo and Carballido, 1998). Briefly, C.B-17 scid/scid (SCID) mice or scid/beige mice were anesthetized and implanted subcutaneously with human fetal thymus, bone, and liver prepared as described (Roncarolo and Carballido, 1998) in accordance with the guidelines set forth by the local Ethical Committee. After 10 weeks, the mice received 1.7 Gy irradiation (x-ray source), followed by direct injection of transduced CD34 1 cells into the grafts.…”

Section: Methodsmentioning

confidence: 99%

Mobilized Blood CD34⁺Cells Transduced and Selected with a Clinically Applicable Protocol Reconstitute Lymphopoiesis in SCID-Hu Mice

et al. 2004

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We developed a clinically applicable gene transfer procedure into mobilized peripheral blood (MPB) CD34(+) hematopoietic progenitor cells, based on single viral exposure and selection of engineered cells. CD34(+) cells were transduced with a retroviral vector carrying the truncated form of the nerve growth factor receptor (Delta NGFR) marker gene, and immunoselected for Delta NGFR expression. Optimal time and procedure for viral exposure, length of culture, and transgene expression of MPB CD34(+) cells were determined using in vitro assays. The multipotent capacity of MPB CD34(+)-transduced cells was demonstrated in the SCID-hu bone/liver/thymus mouse model. Transduced Delta NGFR(+) cells retained 50% of long-term culture-colony forming cells (LTC-CFC) compared to unmanipulated CD34(+) cells. In SCID-hu mice, 52% of CD45(+) cells, 27% of CD34(+) cells, 49% of B cells, and more than 50% of T cells were derived from transplanted CD34(+)/Delta NGFR(+) cells. Furthermore, transplantation of purified transduced cells greatly reduced the competition with untransduced progenitors occurring in unselected grafts. These data demonstrate that MPB CD34(+) cells, transduced with a single viral exposure and selected by transgene expression, retain multilineage reconstitution capacity and remarkable transgene expression.

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“…Hu-BLT mice were generated by following the previously published protocols (19,20). Briefly, 6-to 8-week-old NOD.Cg-Prkdc scid IL2rg tm1Wjl /Szj (NOD/SCID/IL2rgnull, or NSG) mice (The Jackson Laboratory) were purchased and maintained under pathogen-free conditions at the University of Nebraska-Lincoln Life Sciences Annex.…”

Section: Methodsmentioning

confidence: 99%

Topical Tenofovir Disoproxil Fumarate Nanoparticles Prevent HIV-1 Vaginal Transmission in a Humanized Mouse Model

Destache

Mandal

Yuan

et al. 2016

Antimicrob Agents Chemother

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e Preexposure prophylaxis (PrEP) with 1% tenofovir (TFV) vaginal gel has failed in clinical trials. To improve TFV efficacy in vaginal gel, we formulated tenofovir disoproxil fumarate nanoparticles in a thermosensitive (TMS) gel (TDF-NP-TMS gel). TDF-NPs were fabricated using poly(lactic-co-glycolic acid) (PLGA) polymer and an ion-pairing agent by oil-in-water emulsification. The efficacy of TDF-NP-TMS gel was tested in humanized bone marrow-liver-thymus (hu-BLT) mice. Hu-BLT mice in the treatment group (Rx; n ‫؍‬ 15) were administered TDF-NP-TMS gel intravaginally, having TDF at 0.1%, 0.5%, and 1% (wt/vol) concentrations, whereas the control (Ctr; n ‫؍‬ 8) group received a blank TMS gel. All Rx mice ( Presently, a total of 36.9 million people worldwide are living with HIV-1 (1). Topical preexposure prophylaxis (PrEP) currently is a promising preventative strategy (2). The basic idea is to protect the vagina (and/or rectum) from HIV-1 infection by applying gel containing antiretroviral drug(s) around the time of sexual intercourse. This topical preparation is considered a microbicide, inhibiting infection by blocking viral transmission at the mucosal surface. To date, tenofovir (TFV) is the only drug administered locally as a 1% vaginal gel shown to be effective at preventing heterosexual contraction of HIV-1 (3). TFV tissue concentrations indicate a direct relationship between levels of TFV in genitals and protection (4-7). The minimum amount of TFV in cervicovaginal fluid levels when associated with gel that shows protection against HIV-1 infection has been reported to be Ͼ1,000 ng/ml (4). This level is greater than 10 times that seen in patients receiving oral TDF and emtricitabine (4). In female macaques given 1% TFV gel, the intracellular half-life for the active metabolite, tenofovir diphosphate, is significantly shorter (averaging 25 h) in vaginal lymphocytes than peripheral PBMCs (averaging 49 h) (7). A coitally independent strategy using 1% TFV gel has not shown efficacy in several clinical trials (8,9). Based on the dramatic negative results of the Vaginal and Oral Interventions to Control the Epidemic (VOICE) trial, it is important to consider female attitudes and opinions for a vaginal gel-based prevention delivery system. A safe and effective female-controlled, discrete gel-based delivery system has the potential to prevent millions of HIV-1 infections worldwide annually.When designing female-controlled preventative delivery systems, the gel-based system should have features important for the female user. Namely, the delivery system should be (i) easy to administer; (ii) adherent to the mucosal surface once applied vaginally; (iii) low seepage; and (iv) free of side effects or cytotoxicity to the mucosal surfaces of the female genital tract (10). All of these factors, if not optimized, could diminish gel effectiveness or lead to gel aversion. Finally, a long-acting preparation would be highly desirable if it offered long-term protection from HIV-1 (11).Our laboratory has been developing a nanotechn...

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Construction of Human‐SCID Chimeric Mice

Cited by 10 publications

References 32 publications

Humanized-BLT mouse model of Kaposi’s sarcoma-associated herpesvirus infection

Humanized-BLT mouse model of Kaposi’s sarcoma-associated herpesvirus infection

Mobilized Blood CD34⁺Cells Transduced and Selected with a Clinically Applicable Protocol Reconstitute Lymphopoiesis in SCID-Hu Mice

Topical Tenofovir Disoproxil Fumarate Nanoparticles Prevent HIV-1 Vaginal Transmission in a Humanized Mouse Model

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Construction of Human‐SCID Chimeric Mice

Cited by 10 publications

References 32 publications

Humanized-BLT mouse model of Kaposi’s sarcoma-associated herpesvirus infection

Humanized-BLT mouse model of Kaposi’s sarcoma-associated herpesvirus infection

Mobilized Blood CD34+Cells Transduced and Selected with a Clinically Applicable Protocol Reconstitute Lymphopoiesis in SCID-Hu Mice

Topical Tenofovir Disoproxil Fumarate Nanoparticles Prevent HIV-1 Vaginal Transmission in a Humanized Mouse Model

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Mobilized Blood CD34⁺Cells Transduced and Selected with a Clinically Applicable Protocol Reconstitute Lymphopoiesis in SCID-Hu Mice