Name: humanized nod/scid/il2r&#947;null (hu-nsg) mouse model for hiv replication and latency studies
Uploaded: 2019-01-07T14:00:00
Description: Watch this Scientific Journal Video about Humanized NOD/SCID/IL2rÃŽÂ³null hu-NSG Mouse Model for HIV Replication and Latency Studies at JoVE.com

这项工作得到了国家卫生研究院 [赠款编号 r01ai29329、r01ai42552 和 r01hl07470 至 j. r.) 和国家卫生研究院国家癌症研究所 [赠款编号 p30ca033572 以支持希望城市综合基因组学] 的支持, 分析药理学和分析细胞计芯]。以下试剂是通过国家卫生研究院艾滋病研究和参考试剂方案、艾滋病司、niaid、nih 获得的: hiv bal 病毒。

所有动物护理和程序都是根据希望城市机构动物护理和使用委员会 (iacuc) 审查和批准的协议进行的, 该委员会由这项研究的主要调查员 (john rossi 博士, iacuc #12034) 持有。人类胎儿肝脏组织是根据联邦和州规定从高级生物科学资源 (加州阿拉米达) 获得的, 该资源是一个非营利组织。供应商有自己的机构审查委员会 (irb), 并符合人的主体保护要求。从希望市献血者中心 (加利福尼亚州杜阿尔特) 的匿名健...

<ol>
	<li>Greiner, D. L., Hesselton, R. A., Shultz, L. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=SCID+mouse+models+of+human+stem+cell+engraftment.">SCID mouse models of human stem cell engraftment.</a> Stem cells. 16 (3), 166-177 (1998).</li><li>Rongvaux, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+and+function+of+human+innate+immune+cells+in+a+humanized+mouse+model.">Development and function of human innate immune cells in a humanized mouse model.</a> Nature. 32 (4), 364-372 (2014).</li><li>Walsh, N. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Humanized+Mouse+Models+of+Clinical+Disease.">Humanized Mouse Models of Clinical Disease.</a> Annual review of pathology. 12, 187-215 (2017).</li><li>Shultz, L. D., Brehm, M. A., Garcia-Martinez, J. V., Greiner, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Humanized+mice+for+immune+system+investigation:+progress,+promise+and+challenges.">Humanized mice for immune system investigation: progress, promise and challenges.</a> Nature reviews. Immunology. 12 (11), 786-798 (2012).</li><li>Shultz, L. D., Ishikawa, F., Greiner, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Humanized+mice+in+translational+biomedical+research.">Humanized mice in translational biomedical research.</a> Nature reviews. Immunology. 7, (2007).</li><li>Moir, S., Fauci, A. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=B+cells+in+HIV+infection+and+disease.">B cells in HIV infection and disease.</a> Nature reviews. Immunology. 9 (4), 235-245 (2009).</li><li>McCune, J. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+SCID-hu+mouse:+murine+model+for+the+analysis+of+human+hematolymphoid+differentiation+and+function.">The SCID-hu mouse: murine model for the analysis of human hematolymphoid differentiation and function.</a> Science. 241 (4873), 1632-1639 (1988).</li><li>Mosier, D. E., Gulizia, R. J., Baird, S. M., Wilson, D. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Transfer+of+a+functional+human+immune+system+to+mice+with+severe+combined+immunodeficiency.">Transfer of a functional human immune system to mice with severe combined immunodeficiency.</a> Nature. 335, 256 (1988).</li><li>Shultz, L. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multiple+defects+in+innate+and+adaptive+immunologic+function+in+NOD/LtSz-scid+mice.">Multiple defects in innate and adaptive immunologic function in NOD/LtSz-scid mice.</a> Journal of immunology. 154 (1), 180-191 (1995).</li><li>Ohbo, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulation+of+hematopoiesis+in+mice+with+a+truncated+mutant+of+the+interleukin-2+receptor+gamma+chain.">Modulation of hematopoiesis in mice with a truncated mutant of the interleukin-2 receptor gamma chain.</a> Blood. 87 (3), 956-967 (1996).</li><li>Ito, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=NOD/SCID/gamma(c)(null)+mouse:+an+excellent+recipient+mouse+model+for+engraftment+of+human+cells.">NOD/SCID/gamma(c)(null) mouse: an excellent recipient mouse model for engraftment of human cells.</a> Blood. 100 (9), 3175-3182 (2002).</li><li>Shultz, L. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+lymphoid+and+myeloid+cell+development+in+NOD/LtSz-scid+IL2R+gamma+null+mice+engrafted+with+mobilized+human+hemopoietic+stem+cells.">Human lymphoid and myeloid cell development in NOD/LtSz-scid IL2R gamma null mice engrafted with mobilized human hemopoietic stem cells.</a> Journal of immunology. 174 (10), 6477-6489 (2005).</li><li>Ishikawa, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+functional+human+blood+and+immune+systems+in+NOD/SCID/IL2+receptor+{gamma}+chain(null)+mice.">Development of functional human blood and immune systems in NOD/SCID/IL2 receptor {gamma} chain(null) mice.</a> Blood. 106 (5), 1565-1573 (2005).</li><li>Watanabe, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+analysis+of+the+functions+of+human+B+and+T+cells+in+humanized+NOD/shi-scid/gammac(null)+(NOG)+mice+(hu-HSC+NOG+mice).">The analysis of the functions of human B and T cells in humanized NOD/shi-scid/gammac(null) (NOG) mice (hu-HSC NOG mice).</a> International immunology. 21 (7), 843-858 (2009).</li><li>McDermott, S. P., Eppert, K., Lechman, E. R., Doedens, M., Dick, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+human+cord+blood+engraftment+between+immunocompromised+mouse+strains.">Comparison of human cord blood engraftment between immunocompromised mouse strains.</a> Blood. 116 (2), 193-200 (2010).</li><li>Mazurier, F., Doedens, M., Gan, O. I., Dick, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+myeloerythroid+repopulation+after+intrafemoral+transplantation+of+NOD-SCID+mice+reveals+a+new+class+of+human+stem+cells.">Rapid myeloerythroid repopulation after intrafemoral transplantation of NOD-SCID mice reveals a new class of human stem cells.</a> Nature. 9 (7), 959-963 (2003).</li><li>Melkus, M. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Humanized+mice+mount+specific+adaptive+and+innate+immune+responses+to+EBV+and+TSST-1.">Humanized mice mount specific adaptive and innate immune responses to EBV and TSST-1.</a> Nature medicine. 12, 1316 (2006).</li><li>Lan, P., Tonomura, N., Shimizu, A., Wang, S., Yang, Y. -. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reconstitution+of+a+functional+human+immune+system+in+immunodeficient+mice+through+combined+human+fetal+thymus/liver+and+CD34++cell+transplantation.">Reconstitution of a functional human immune system in immunodeficient mice through combined human fetal thymus/liver and CD34+ cell transplantation.</a> Blood. 108 (2), 487-492 (2006).</li><li>Brehm, M. A., Bortell, R., Verma, M., Shultz, L. D., Greiner, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Humanized+Mice+in+Translational+Immunology.">Humanized Mice in Translational Immunology.</a> Translational Immunology. , 285-326 (2016).</li><li>King, M. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hu-PBL-NOD-scid+IL2rgnull+mouse+model+of+xenogeneic+graft-versus-host-like+disease+and+the+role+of+host+MHC.">Hu-PBL-NOD-scid IL2rgnull mouse model of xenogeneic graft-versus-host-like disease and the role of host MHC.</a> Clinical &amp; Experimental Immunology. 157, 104-118 (2009).</li><li>Halkias, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Conserved+and+divergent+aspects+of+human+T-cell+development+and+migration+in+humanized+mice.">Conserved and divergent aspects of human T-cell development and migration in humanized mice.</a> Immunology and cell biology. 93 (8), 716-726 (2015).</li><li>Satheesan, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HIV+replication+and+latency+in+a+humanized+NSG+mouse+model+during+suppressive+oral+combinational+ART.">HIV replication and latency in a humanized NSG mouse model during suppressive oral combinational ART.</a> Journal of virology. , (2018).</li><li>Zhou, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Receptor-targeted+aptamer-siRNA+conjugate-directed+transcriptional+regulation+of+HIV-1.">Receptor-targeted aptamer-siRNA conjugate-directed transcriptional regulation of HIV-1.</a> Theranostics. 8 (6), 1575-1590 (2018).</li><li>Zhou, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell-specific+RNA+aptamer+against+human+CCR5+specifically+targets+HIV-1+susceptible+cells+and+inhibits+HIV-1+infectivity.">Cell-specific RNA aptamer against human CCR5 specifically targets HIV-1 susceptible cells and inhibits HIV-1 infectivity.</a> Chemistry &amp; biology. 22 (3), 379-390 (2015).</li><li>Brechtl, J. R., Breitbart, W., Galietta, M., Krivo, S., Rosenfeld, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+highly+active+antiretroviral+therapy+(HAART)+in+patients+with+advanced+HIV+infection:+impact+on+medical,+palliative+care,+and+quality+of+life+outcomes.">The use of highly active antiretroviral therapy (HAART) in patients with advanced HIV infection: impact on medical, palliative care, and quality of life outcomes.</a> Journal of pain and symptom management. 21 (1), 41-51 (2001).</li><li>Richman, D. D., Margolis, D. M., Delaney, M., Greene, W. C., Hazuda, D., Pomerantz, R. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Challenge+of+Finding+a+Cure+for+HIV+Infection.">The Challenge of Finding a Cure for HIV Infection.</a> Science. 323 (5919), 1304-1307 (2009).</li><li>Pace, M. J., Agosto, L., Graf, E. H., O'Doherty, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HIV+reservoirs+and+latency+models.">HIV reservoirs and latency models.</a> Virology. 411 (2), 344-354 (2011).</li><li>Van Herck, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Blood+sampling+from+the+retro-orbital+plexus,+the+saphenous+vein+and+the+tail+vein+in+rats:+comparative+effects+on+selected+behavioural+and+blood+variables.">Blood sampling from the retro-orbital plexus, the saphenous vein and the tail vein in rats: comparative effects on selected behavioural and blood variables.</a> Laboratory animals. 35 (2), 131-139 (2001).</li><li>Autissier, P., Soulas, C., Burdo, T. H., Williams, K. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+a+12-color+flow+cytometry+panel+to+study+lymphocyte,+monocyte,+and+dendritic+cell+subsets+in+humans.">Evaluation of a 12-color flow cytometry panel to study lymphocyte, monocyte, and dendritic cell subsets in humans.</a> Cytometry. 77 (5), 410-419 (2010).</li><li>Lu, W., Mehraj, V., Vyboh, K., Cao, W., Li, T., Routy, J. -. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CD4:CD8+ratio+as+a+frontier+marker+for+clinical+outcome,+immune+dysfunction+and+viral+reservoir+size+in+virologically+suppressed+HIV-positive+patients.">CD4:CD8 ratio as a frontier marker for clinical outcome, immune dysfunction and viral reservoir size in virologically suppressed HIV-positive patients.</a> Journal of the International AIDS Society. 18, 20052 (2015).</li><li>van't Wout, A. B., Schuitemaker, H., Kootstra, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+propagation+of+HIV-1+on+peripheral+blood+mononuclear+cells.">Isolation and propagation of HIV-1 on peripheral blood mononuclear cells.</a> Nature protocols. 3, 363 (2008).</li><li>Reagan-Shaw, S., Nihal, M., Ahmad, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dose+translation+from+animal+to+human+studies+revisited.">Dose translation from animal to human studies revisited.</a> FASEB journal: official publication of the Federation of American Societies for Experimental Biology. 22 (3), 659-661 (2008).</li><li>Han, Y., Wind-Rotolo, M., Yang, H. -. C., Siliciano, J. D., Siliciano, R. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+approaches+to+the+study+of+HIV-1+latency.">Experimental approaches to the study of HIV-1 latency.</a> Nature reviews. Microbiology. 5 (2), 95-106 (2007).</li><li>Marsden, M. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HIV+Latency+in+the+Humanized+BLT+Mouse.">HIV Latency in the Humanized BLT Mouse.</a> Journal of virology. 86 (1), 339-347 (2012).</li><li>Karpel, M. E., Boutwell, C. L., Allen, T. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=BLT+humanized+mice+as+a+small+animal+model+of+HIV+infection.">BLT humanized mice as a small animal model of HIV infection.</a> Current opinion in virology. 13, 75-80 (2015).</li><li>Durand, C. M., Blankson, J. N., Siliciano, R. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Developing+strategies+for+HIV-1+eradication.">Developing strategies for HIV-1 eradication.</a> Trends in immunology. 33 (11), 554-562 (2012).</li><li>Van Lint, C., Bouchat, S., Marcello, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HIV-1+transcription+and+latency:+an+update.">HIV-1 transcription and latency: an update.</a> Retrovirology. 10, 67 (2013).</li><li>Xu, L., Zhang, Y., Luo, G., Li, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+roles+of+stem+cell+memory+T+cells+in+hematological+malignancies.">The roles of stem cell memory T cells in hematological malignancies.</a> Journal of hematology &amp; oncology. 8, 113 (2015).</li><li>Chun, T. -. W., Moir, S., Fauci, A. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HIV+reservoirs+as+obstacles+and+opportunities+for+an+HIV+cure.">HIV reservoirs as obstacles and opportunities for an HIV cure.</a> Nature immunology. 16 (6), 584-589 (2015).</li><li>Redel, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=HIV-1+regulation+of+latency+in+the+monocyte-macrophage+lineage+and+in+CD4++T+lymphocytes.">HIV-1 regulation of latency in the monocyte-macrophage lineage and in CD4+ T lymphocytes.</a> Journal of leukocyte biology. 87 (4), 575-588 (2010).</li><li>Laird, G. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rapid+Quantification+of+the+Latent+Reservoir+for+HIV-1+Using+a+Viral+Outgrowth+Assay.">Rapid Quantification of the Latent Reservoir for HIV-1 Using a Viral Outgrowth Assay.</a> PLoS pathogens. 9 (5), e1003398 (2013).</li></ol>

免疫损伤的小鼠与人的细胞组织一起存在着人类的生理特征, 对人类特有疾病的病理、病理生理学和免疫学研究具有巨大的价值。在免疫功能低下的多株小鼠中, nod。cg-prkdc科学il2rgtm1wji (nsg) 模型是最免疫缺陷的, 因为它缺乏先天免疫和适应性免疫, 以及消融的鼠特异性细胞因子信号 3,12, 19.因此, nsg 小鼠?...

由于建立适合人类疾病的动物模型是找到治愈方法的关键, 随着时间的推移, 适当的动物模型早已得到追求和改进。已经开发出多种免疫功能低下的小鼠模型, 允许人体细胞和组织的雕刻, 并随后执行人性化的功能1,2。这种人性化的小鼠模型对于研究人类特有的疾病3、4、5至关重要。
因感染人体免疫机能丧失病毒 (hiv) 而引起的获得性免疫缺陷综合症 (aids) 就是一个例子。在建立人性化的小鼠模型之前, 伦理和技术限制将艾滋病毒/艾滋病临床前动物研究局限于非人类灵长类动物3。然而, 对这种动物进行专门护理的高昂费用和要求阻碍了典型学术环境中的艾滋病毒/艾滋病研究。hiv 主要感染人类 cd4 + t 细胞, 并影响其他人类免疫细胞的发育和免疫反应, 如 b 细胞、巨噬细胞和树突状细胞6;因此, 对移植功能人体免疫系统的小动物模型的需求很大。
1988年取得了突破, 当时 cb17-科学小鼠与 prkdc科学突变的开发, 并显示出成功的人类免疫系统1的雕刻。prkdc科学突变导致小鼠 t 细胞和 b 细胞功能缺陷和消融的适应性免疫系统, 从而能够植入人外周血单个核细胞 (pbmc)、造血干细胞 (hsc) 和胎儿造血组织7,8。尽管如此, 在这种模式中经常观察到低水平的雕刻;可能的原因是 1) 残留的先天免疫活性通过自然杀手 (nk) 细胞调制和 2) 小鼠 t-和 b 细胞 (泄漏)的后期发育。非肥胖糖尿病 (nod)-科学小鼠模型的后续发展实现了 nk 细胞活性的显著降低;因此, 它能够支持更高层次和更可持续的人类免疫系统组成部分9的雕刻。为了进一步抑制或阻碍先天免疫的发展, 建立了小鼠模型, 在 (nod)-科学背景中, 白细胞介素-2 受体γ链 (il2rg) 被截断或完全敲除。il2rg, 也被称为常见的细胞因子受体γ-链, 是各种细胞因子受体 10,11,12,13的不可缺少的组成部分。品种, 如 nod。cg-prkdcscidil2rgtm1wji (nsg) 和 nodshe.cg-prkdcscidil2rgtm1Wji (nog) 对小鼠细胞因子信号转导和 nk 细胞发育的完全消融造成了强大的干扰。此外, 适应性免疫有严重损伤,14, 15,16。
在艾滋病毒/艾滋病研究中经常使用三种具有科学突变和 il2rg 敲除的人的小鼠模型: blt (骨头 marrowyanglemesus) 模型、pbl (外周血白细胞模型) 和 src ( scid 反细胞模型)3. blt 模型是通过在小鼠肾囊下对人的胎儿肝脏和胸腺进行手术移植, 并伴有静脉注射胎儿肝 hsc3,17,18。blt 小鼠模型具有较高的雕刻效果, 在所有血统中开发人类造血细胞, 并建立强大的人体免疫系统;此外, t 细胞在人类自体胸腺中接受教育, 并表现出 hla 受 ha 限制的免疫反应4,5,17,19。然而, 外科手术的要求仍然是 blt 模型的主要缺点。通过与人外周淋巴细胞静脉注射建立 pbl 小鼠模型。pbl 模型提供了方便, 并产生了成功的 t 细胞雕刻, 但其应用有限, 由于不足的 b 细胞和骨髓细胞植皮, 较低的雕刻水平整体, 并开始严重的移植物抗宿主疾病 (gvhd)3 ,20。src 小鼠模型是通过向新生或年轻成人 scid 小鼠注射人 hsc 建立的。它表现出平均雕刻效率超过 25% (被评估为外周血 cd45 百分比), 并支持多系发展的注射 hsc 和制定一个先天的人体免疫系统。然而, src 模型的局限性在于 t 细胞响应是鼠标 h2 受限而不是人类 hla 受限14,21。
src 小鼠模型被认为是临床前艾滋病毒/艾滋病小动物研究的一个简单可靠的模型, 人类免疫系统的一致植入和造血发育的成功就是例证。我们以前报告了核供应国集团 hu-src-scid (hu-nsg) 小鼠模型的建立, 并介绍了其在艾滋病毒复制和延迟研究22、23、24 中的应用。这种 hu-nsg 小鼠模型显示了高水平的骨髓定位, 易感性的艾滋病毒感染, 并概述了 hiv 感染和发病机制。此外, hu-nsg 小鼠模型对组合抗逆转录病毒疗法 (cart) 作出适当反应, 并在 cart 退出后重述血浆病毒反弹, 确认建立了 25, 26 艾滋病毒潜伏期库 ,27岁。从受感染和 cot 治疗的 hu-nsg 小鼠身上分离出的人体外感染病毒的生产进一步证实了这一艾滋病毒潜伏期。
在此, 我们描述了从新生儿 nsg 小鼠建立 hu-nsg 小鼠模型的详细方案, 包括与 hiv 感染和 cart 治疗有关的潜伏期发展的程序。我们期望该协议在 hiv 动物研究中提供一套新的方法, 包括 hiv 病毒学、潜伏期和治疗。

<table>
	<tbody>
		<tr>
			<td>CD34 MicroBead Kit, human</td>
			<td>MiltenyiBiotec</td>
			<td>130-046-703</td>
			<td></td>
		</tr>
		<tr>
			<td>CryoStor CS2</td>
			<td>Stemcell Technologies</td>
			<td>07932</td>
			<td></td>
		</tr>
		<tr>
			<td>NOD.Cg-PrkdcscidIl2rgtm1Wji </td>
			<td>The Jackson Laboratory</td>
			<td>005557</td>
			<td>Order breeders instead of experimental mice</td>
		</tr>
		<tr>
			<td>IsoFlo</td>
			<td>Patterson Veterinary</td>
			<td>07-806-3204</td>
			<td>Order through animal facility, restricted item</td>
		</tr>
		<tr>
			<td>Clidox disinfectant</td>
			<td>Fisher Sicentific</td>
			<td>NC9189926</td>
			<td></td>
		</tr>
		<tr>
			<td>Wescodyne</td>
			<td>Fisher Sicentific</td>
			<td>19-818-419</td>
			<td></td>
		</tr>
		<tr>
			<td>Hamilton 80508 syringe/needle</td>
			<td>Hamilton</td>
			<td>80508</td>
			<td>Custom made</td>
		</tr>
		<tr>
			<td>Blood collection tube (K2EDTA)</td>
			<td>BD Bioscience</td>
			<td>367843</td>
			<td></td>
		</tr>
		<tr>
			<td>Blood collection tube (Heparin)</td>
			<td>BD Bioscience</td>
			<td>365965</td>
			<td></td>
		</tr>
		<tr>
			<td>Capillary tube (Heparinized)</td>
			<td>Fisher Sicentific</td>
			<td>22-362574</td>
			<td></td>
		</tr>
		<tr>
			<td>Red Blood Cell Lysis Buffer</td>
			<td>Sigma Aldrich</td>
			<td>11814389001</td>
			<td></td>
		</tr>
		<tr>
			<td>QIAamp Viral RNA mini kit</td>
			<td>Qiagen</td>
			<td>52906</td>
			<td></td>
		</tr>
		<tr>
			<td>TaqMan Fast VIrus 1-step Master Mix</td>
			<td>Thermofisher</td>
			<td>4444434</td>
			<td></td>
		</tr>
		<tr>
			<td>HIV-1 P24 ELISA (5 Plate kit)</td>
			<td>PerkinElmer</td>
			<td>NEK050B001KT</td>
			<td></td>
		</tr>
		<tr>
			<td>IgG from human serum</td>
			<td>Sigma Aldrich</td>
			<td>I4506-100MG</td>
			<td></td>
		</tr>
		<tr>
			<td>IgG from mouse serum</td>
			<td>Sigma Aldrich</td>
			<td>I5381-10MG</td>
			<td></td>
		</tr>
		<tr>
			<td>BB515 Mouse Anti-Human CD45 (clone HI30)</td>
			<td>BD Biosciences</td>
			<td>564586</td>
			<td>RRID: AB_2732068, LOT 6347696</td>
		</tr>
		<tr>
			<td>PE-Cy7 Mouse Anti-Human CD3 (Clone SK7)</td>
			<td>BD Biosciences</td>
			<td>557851</td>
			<td>RRID: AB_396896, LOT 6021877</td>
		</tr>
		<tr>
			<td>Pacific Blue Mouse Anti-Human CD4 (Clone RPA-T4)</td>
			<td>BD Biosciences</td>
			<td>558116</td>
			<td>RRID: AB_397037, LOT 6224744</td>
		</tr>
		<tr>
			<td>BUV395 Mouse Anti-Human CD8 (Clone RPA-T8)</td>
			<td>BD Biosciences</td>
			<td>563795</td>
			<td>RRID: AB_2722501, LOT 6210668</td>
		</tr>
		<tr>
			<td>APC-Alexa Fluor 750 Mouse Anti-Human CD14 (TuK4)</td>
			<td>ThermoFisher</td>
			<td>MHCD1427</td>
			<td>RRID: AB_10373536, LOT 1684947A</td>
		</tr>
		<tr>
			<td>PE Mouse Anti-Human CD19 (SJ25-C1)</td>
			<td>ThermoFisher</td>
			<td>MHCD1904</td>
			<td>RRID: AB_10373382, LOT 1725304B</td>
		</tr>
	</tbody>
</table>

humanized nod/scid/il2r&#947;null (hu-nsg) mouse model for hiv replication and latency studies

人类病理学、免疫学和治疗发展研究的伦理法规和技术挑战, 对小动物模型的需求很高。与人类有着密切的遗传和行为相似, 像老鼠这样的小动物是人类疾病模型的理想选择, 通过这些模型可以重述类似人类的症状和反应。此外, 可以改变小鼠的遗传背景, 以适应不同的需求。nod/scid/il2rγ null (nsg) 小鼠是应用最广泛的免疫功能低下小鼠菌株之一;它允许植入与人类造血干细胞和/或人体组织和随后的功能性人类免疫系统的发展。这是了解艾滋病毒/艾滋病等人类特有疾病的预后和病理生理学以及帮助寻找治愈方法的一个重要里程碑。在此, 我们报告了一个详细的方案, 以产生人性化的 nsg 小鼠模型 (hu-nsg) 造血干细胞移植到辐射条件下的新生儿 nsg 小鼠。hu-nsg 小鼠模型显示了移植的人的干细胞的多谱系发展和对 hiv-1 病毒感染的易感性。它还概述了与组合抗逆转录病毒疗法 (cart) 相响应的关键生物学特征。

流式细胞术分析经常进行, 以验证分离的 hsc 的纯度, 评估雕刻水平, 对病毒感染的特征免疫反应, 并调查相符的疗效。一个典型的抗体面板包含4-6 个荧光标记抗体;因此, 具有多个激光器和多种滤波器的流式细胞仪对于获得准确的结果至关重要。
对于初始雕刻验证, 人类 cd45 + 细胞计数可以从20% 到80% 不等, 人类白细胞的子集应该?...

Watch this Scientific Journal Video about Humanized NOD/SCID/IL2rÃŽÂ³null hu-NSG Mouse Model for HIV Replication and Latency Studies at JoVE.com

Humanized NOD/SCID/IL2rÃƒÆ’Ã…Â½Ãƒâ€šÃ‚Â³null hu-NSG Mouse Model for HIV Replication and Latency Studies

该方案提供了一种方法, 建立人性化小鼠 (hu-nsg) 通过肝内注射人类造血干细胞到辐射条件下的新生儿 nsg 小鼠。hu-nsg 小鼠容易感染 hiv 感染和组合抗逆转录病毒疗法 (cart), 是治疗艾滋病毒复制和潜伏期调查的合适病理生理模型。

人性化的 nod/scid/il2γ空 (hu-nsg) 小鼠 hiv 复制和延迟研究模型

Ethical regulations and technical challenges for research in human pathology, immunology, and therapeutic development have placed small animal models in ...

这些问题有助于回答艾滋病毒/艾滋病研究中的一些关键问题，例如艾滋病毒感染的分子机制和预后，以及如何以及在什么地点建立延迟储层。这项技术的优点是它是非侵入性的，并建立了hu-NSG小鼠，必须治疗多血统的发展比移植的人类干细胞，是易受HIV感染，并接受cART。首先通过无菌、70微米、尼龙、网状滤网过滤消化的、人、胎、肝组织、浆料样品，获得单细胞悬浮液，并按照制造商的说明，通过磁性活性细胞分选丰富CD34+HSC。计数后，在冰上新鲜 DPBS 中，每毫升浓度以两倍十至七的可行细胞重新悬挂 HSC。接下来，将整个新生儿NO德/SCID/伽马或NSG小鼠的垃圾放入无菌的饼状辐照笼中，用来自200至250摄氏度的伽马射线从137辐射源对动物进行治疗。然后将细胞装入定制的 Hamilton 80508 50 微升注射器中，并配备 30 量针，然后用 5 倍至 10 到 5 的活人 CD34+HSCs 和 25 微升的 DPBS 将每个新生儿动物直接注射到肝脏中。感染后十至十二周，通过离心从每只小鼠身上旋转反轨道、外周、血液样本，将血浆超先输转移到微离心管中，进行80摄氏度的负极性储存，作为实验上适当的。对于 HSC 移植验证，用 200 微升红细胞裂解缓冲液重新悬浮颗粒，并在 1.5 毫米微离离管中将细胞悬浮液拉下，在室温下孵育 10 分钟。然后通过离心收集剩余的白细胞，每次洗涤在0.5毫升新鲜、0.01%牛血清白蛋白或BSA中两次洗涤。第二次洗涤后，用100微升的阻滞鸡尾酒在4摄氏度下阻断20分钟，然后按照每1倍至6次细胞浓度的2微升抗身体鸡尾酒标记适当的抗身体鸡尾酒。在4摄氏度下30分钟后，在新鲜的DPBS加BSA中清洗细胞两次，然后根据标准方案通过流量细胞学评估外周血样，以量化每个样本的人类CD45+CD3+CD14+或CD19+细胞的百分比。然后计算CD4+CD8+T细胞的比例，根据标准、定量、逆转录酶、PCR方案评估血浆样本中的病毒载量。为了评估HIV感染的影响，将HIV包病毒送到麻醉的人类、NSG小鼠，其外周血液中人类CD45+细胞数量超过20%，通过内向管内给皮，每只小鼠浓度为200纳米P24，每两周通过逆轨道出血采集血液样本，并分析CD4-CD8比率和血浆病毒载量，如所证明的。对于病毒抑制验证供应受感染的，人类NSG动物与新鲜，组合，抗逆转录病毒疗法，或cART，每周加糖水4周，收集血液样本通过逆轨道出血每两周，并分析CD4-CD8的比例和血浆病毒载量，如证明。当血浆病毒载量低于检测限值，并恢复CD4-CD8比1.5至2.5之间，在CART提取后每2周通过逆轨道出血收集外周血样时，评估病毒反弹，并评估样本，如所示。在初始移植验证期间，人类CD45+细胞计数应在20%至80%之间，通过流动细胞计量分析，人类白细胞的子集应显示为离散种群。在健康个体中，CD4+-CD8+细胞的比例仍然介于1.5和2.5之间，而在病毒感染时通常观察到显著的CD4+T细胞耗竭。值得注意的是，健康比率恢复，以回应CART治疗。在整个感染过程中，通过定量RT PCR检测血浆病毒载量，并分别绘制并用于评估病毒传递和抗病毒治疗的效率。虽然这种方法可以提供对艾滋病毒/艾滋病病毒学研究和治疗发展的见解，它也可以应用于与人类疾病相关的其他研究，如癌症、糖尿病和自身免疫疾病。hu-NSG小鼠模型的开发为免疫学和肿瘤学的研究人员探索人类新的功能障碍和病理学铺平了道路，特别是针对HIV慢性感染和延迟水库的机械和治疗研究。

Research

JoVE Journal

Immunology and Infection

Humanized NOD/SCID/IL2r&#947;null (hu-NSG) Mouse Model for HIV Replication and Latency Studies

Genotypic Inference of HIV-1 Tropism Using Population-based Sequencing of V3

V3的HIV - 1取向的基因型推理使用人口为基础的测序

Background: Prior to receiving a drug from CCR5-antagonist class in HIV therapy, a patient must undergo an HIV tropism test to confirm that his or her ...

科研

免疫与感染

Dissecting Host-virus Interaction in Lytic Replication of a Model Herpesvirus

解剖宿主 - 病毒相互作用模型疱疹病毒裂解性复制

In response to viral infection, a host develops various defensive responses, such as activating innate immune signaling pathways that lead to antiviral ...

Using the BLT Humanized Mouse as a Stem Cell based Gene Therapy Tumor Model

BLT人性化的鼠标使用一个基于干细胞的基因治疗肿瘤模型的建立

Small animal models such as mice have been extensively used to study human  disease and to develop new therapeutic interventions. Despite the wealth of  ...

Humanized Mouse Model to Study Bacterial Infections Targeting the Microvasculature

人源化的小鼠模型来研究细菌感染瞄准微血管

Neisseria meningitidis causes a severe, frequently fatal sepsis when it enters the human blood stream. Infection leads to extensive damage of the blood ...

Rapid Screening of HIV Reverse Transcriptase and Integrase Inhibitors

HIV逆转录酶和整合酶抑制剂的快速筛选

Although a number of anti HIV drugs have been approved, there are still problems with toxicity and drug resistance. This demonstrates a need to identify ...

Pairwise Growth Competition Assay for Determining the Replication Fitness of Human Immunodeficiency Viruses

成对增长竞争测定法测定人类免疫缺陷病毒的复制健身

In vitro fitness assays are essential tools for determining viral replication fitness for viruses such as HIV-1. Various measurements have been used to ...

A 3D Human Lung Tissue Model for Functional Studies on Mycobacterium tuberculosis Infection

A 3D Human Lung Tissue Model for Functional Studies on Mycobacterium tuberculosis Infection

关于功能研究的三维人体肺组织模型<em&gt;结核分枝杆菌</em&gt;感染

Tuberculosis (TB) still holds a major threat to the health of people worldwide, and there is a need for cost-efficient but reliable models to help us ...

Measurement of In Vitro Integration Activity of HIV-1 Preintegration Complexes

Measurement of In Vitro Integration Activity of HIV-1 Preintegration Complexes

测量<em&gt;体外</em&gt; HIV-1 Preintegration配合物的整合活动

HIV-1 envelope proteins engage cognate receptors on the target cell surface, which leads to viral-cell membrane fusion followed by the release of the ...

Influenza A Virus Studies in a Mouse Model of Infection

流感病毒在小鼠感染模型中的研究

Influenza viruses cause over 500,000 deaths worldwide1 and are associated with an annual cost of 12 - 14 billion USD in the United States alone ...

Establishment of the Dual Humanized TK-NOG Mouse Model for HIV-associated Liver Pathogenesis

建立与艾滋病毒相关的肝脏发病机制的双人性化TK-NOG小鼠模型

Despite the increased life expectancy of patients infected with human immunodeficiency virus-1 (HIV-1), liver disease has emerged as a common cause of ...