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21.
Dimitrios N. Vatakis Gregory C. Bristol Sohn G. Kim Bernard Levin Wei Liu Caius G. Radu Scott G. Kitchen Jerome A. Zack 《Journal of visualized experiments : JoVE》2012,(70)
Small animal models such as mice have been extensively used to study human disease and to develop new therapeutic interventions. Despite the wealth of information gained from these studies, the unique characteristics of mouse immunity as well as the species specificity of viral diseases such as human immunodeficiency virus (HIV) infection led to the development of humanized mouse models. The earlier models involved the use of C. B 17 scid/scid mice and the transplantation of human fetal thymus and fetal liver termed thy/liv (SCID-hu) 1, 2 or the adoptive transfer of human peripheral blood leukocytes (SCID-huPBL) 3. Both models were mainly utilized for the study of HIV infection.One of the main limitations of both of these models was the lack of stable reconstitution of human immune cells in the periphery to make them a more physiologically relevant model to study HIV disease. To this end, the BLT humanized mouse model was developed. BLT stands for bone marrow/liver/thymus. In this model, 6 to 8 week old NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ (NSG) immunocompromised mice receive the thy/liv implant as in the SCID-hu mouse model only to be followed by a second human hematopoietic stem cell transplant 4. The advantage of this system is the full reconstitution of the human immune system in the periphery. This model has been used to study HIV infection and latency 5-8.We have generated a modified version of this model in which we use genetically modified human hematopoietic stem cells (hHSC) to construct the thy/liv implant followed by injection of transduced autologous hHSC 7, 9. This approach results in the generation of genetically modified lineages. More importantly, we adapted this system to examine the potential of generating functional cytotoxic T cells (CTL) expressing a melanoma specific T cell receptor. Using this model we were able to assess the functionality of our transgenic CTL utilizing live positron emission tomography (PET) imaging to determine tumor regression (9).The goal of this protocol is to describe the process of generating these transgenic mice and assessing in vivo efficacy using live PET imaging. As a note, since we use human tissues and lentiviral vectors, our facilities conform to CDC NIH guidelines for Biosafety Level 2 (BSL2) with special precautions (BSL2+). In addition, the NSG mice are severely immunocompromised thus, their housing and maintenance must conform to the highest health standards (http://jaxmice.jax.org/research/immunology/005557-housing.html). 相似文献
22.
Experimental evaluation of sex differences in territory acquisition in a cooperatively breeding bird
Eikenaar Cas; Richardson David S.; Brouwer Lyanne; Bristol Rachel; Komdeur Jan 《Behavioral ecology》2009,20(1):207-214
In many species, territory ownership is a prerequisite for reproduction;consequently, factors that affect success in territory acquisitioncan have a large impact on fitness. When competing for territories,some individuals may have an advantage if, for example, theyare phenotypically superior or more familiar with the site thanothers. The relative importance of the many factors involvedin territory acquisition is, at present, unclear. We studiedpatterns of natural territory acquisition in a closed and saturatedpopulation of Seychelles warblers. Furthermore, by removingbreeders, we experimentally investigated the relative importance,to territory acquisition, of a range of factors and assessedwhether this differed between the sexes. In both sexes, themain route to natural territory acquisition was to dispersefrom the natal territory to immediately claim a vacant dominantposition. Males were older than females when acquiring a territoryfor the first time. In the removal experiment, for both sexes,the proximity of an individual's natal territory to a vacantdominant position was positively related to the individual'schance of claiming the vacancy. Older males were more likelyto gain an experimental vacant dominant position than were youngmales, whereas age did not affect territory acquisition in females.In the Seychelles warbler, the degree of intrasexual competitionfor territory ownership may be stronger for males than for femalesbecause territory ownership is a prerequisite for male reproduction,whereas females can reproduce on their natal territory. In suchcompetition, young males subsequently lose out to older ones. 相似文献
23.
Eighty-four 18-month-old crossbred beef heifers, 3 to 4 months pregnant, were assigned by stratified randomization to either a high or low (control) beta-carotene (B-car) diet to determine the effect of long-term supplementation of B-car on reproductive performance. The heifers were followed through pregnancy, calving and subsequent breeding. The basal diet consisted of barley, canola meal and barley straw. Heifers supplemented by B-car received 625 mg B-car per day in the concentrate. Vitamin A and D complex injections were given monthly to all heifers. Heifers were bred by artificial insemination after Day 60 postpartum. Throughout the study heifers fed the B-car supplement had higher levels of B-car in plasma (> 300 ug/dl) (P < 0.01) than the heifers fed the control diet (< 50 ug/dl). Vitamin A status was satisfactory in all heifers throughout the study. Birth weight of calves, weight gain, and incidence of mortality were not influenced by B-car. For the control and B-car treatments, days postpartum to first normal luteal phase were 67.5 and 62.6 days; days postpartum to first detected estrus were 70.1 and 65.3, and services per conception were 1.24 and 1.29, respectively. Long-term supplementation of B-car increased prepartum plasma progesterone but had no effect on postpartum fertility. 相似文献
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L A Bristol K Sakaguchi E Appella D Doyle L Takács 《Journal of immunology (Baltimore, Md. : 1950)》1992,149(2):367-372
The rat thymocyte costimulating protein appears to be involved in the regulation of CD4-/CD8- thymocyte proliferation in vitro. We show that thymocyte costimulating protein is dipeptidyl peptidase IV (E.C.N.3.4.14.5) also known as CD26. Some bone marrow cells as well as CD4-/CD8- and, and to a lesser extent, more differentiated T cells respond by proliferating to a CD26 specific mAb-mediated costimulus that does not influence dipeptidyl dipeptidase IV enzyme activity. This suggests the existence of a CD26-linked regulatory mechanism of proliferation that is operational on granulocyte- and macrophage-lineage cells and throughout T cell development. 相似文献
28.
健康人口腔中EB病毒排出情况的研究 总被引:1,自引:0,他引:1
本研究对30名血清EB病毒抗体阳性健康人从口腔排出EB病毒的情况,作了15个月的反复观察。发现28人有病毒排出,其中每次检查均为阳性者6人,间断阳性者22人,有2人多次检查均为阴性。这一结果提示,EB病毒可能是通过它在口咽部某些特定细胞内的慢性复制而在活体内持续存在的。 相似文献
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High-efficiency transduction of human lymphoid progenitor cells and expression in differentiated T cells. 总被引:3,自引:2,他引:1
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D S An Y Koyanagi J Q Zhao R Akkina G Bristol N Yamamoto J A Zack I S Chen 《Journal of virology》1997,71(2):1397-1404
Gene therapy strategies for humans have been limited by low transduction efficiencies and poor expression of retroviral vectors in differentiated progeny cells carrying the transduced vector. Here we describe a strategy utilizing a cell surface reporter gene, murine thy-1.2, selectable by fluorescence-activated cell sorting (FACS), to achieve higher gene marking efficiencies. Human CD34-positive cells were transduced by a murine retroviral vector bearing the thy-1.2 marker and pseudotyped with vesicular stomatitis virus G protein, followed by FACS to enrich for CD34-positive cells that express Thy-1.2 on the cell surface. Gene marking and expression after differentiation into thymocytes were assessed in a SCID-hu Thy/Liv mouse model for human lymphoid progenitor cell gene therapy. We found that virtually all of the differentiated T-cell progeny were marked with vector sequences. It is of particular importance that reconstitution with the selected cells resulted in expression of Thy-1.2 in up to 71% of donor-derived thymocytes. It is of note that the donor-derived thymocytes that did not express Thy-1.2 still harbored vector thy-1.2 sequences, suggesting repression of transgene expression in some cells during progenitor cell differentiation into thymocytes. These studies provide a proof of concept for efficient expression of transgenes through T-lymphoid differentiation and a potential basis for utilizing similar strategies in human gene therapy clinical trials. 相似文献