دورية أكاديمية
Measuring the Uptake and Transactivation Function of HIV-1 Tat Protein in a Trans-cellular Cocultivation Setup.
| العنوان: | Measuring the Uptake and Transactivation Function of HIV-1 Tat Protein in a Trans-cellular Cocultivation Setup. |
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| المؤلفون: | Ruiz AP; Department of Microbiology and Immunology, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, NY, 10461, USA., Prasad VR; Department of Microbiology and Immunology, Albert Einstein College of Medicine, 1300 Morris Park Ave., Bronx, NY, 10461, USA. vinayaka.prasad@einstein.yu.edu. |
| المصدر: | Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2016; Vol. 1354, pp. 353-66. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Humana Press Country of Publication: United States NLM ID: 9214969 Publication Model: Print Cited Medium: Internet ISSN: 1940-6029 (Electronic) Linking ISSN: 10643745 NLM ISO Abbreviation: Methods Mol Biol Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Totowa, NJ : Humana Press Original Publication: Clifton, N.J. : Humana Press, |
| مواضيع طبية MeSH: | Transcriptional Activation*, Coculture Techniques/*methods , HIV Infections/*genetics , HIV Infections/*metabolism , HIV-1/*physiology , tat Gene Products, Human Immunodeficiency Virus/*genetics , tat Gene Products, Human Immunodeficiency Virus/*metabolism, Cell Line ; Cytokines/genetics ; Electrophoresis, Polyacrylamide Gel/methods ; Endocytosis ; Enzyme-Linked Immunosorbent Assay/methods ; Humans ; Transfection/methods |
| مستخلص: | HIV-1 Tat protein is secreted from infected cells and is endocytosed by uninfected bystander cells. Subsequently, Tat is translocated to the nucleus and binds to promoters of host cell genes, increasing the production of inflammatory host cytokines and chemokines. This inflammatory activation of uninfected cells by HIV-1 Tat protein contributes to the overall inflammatory burden in the central nervous system (CNS) that leads to the development of HIV-associated neurocognitive disorders (HAND). Here we describe methods to evaluate the uptake and transcriptional impact of HIV-1 Tat on uninfected cells by using a trans-cellular transactivation system. Cell lines transiently transfected with Tat expression constructs secrete Tat into the culture medium. Trans-cellular uptake and transactivation caused by secreted Tat can be measured by co-culturing LTR-responsive reporter cells with Tat-transfected cells. Such Tat-producer cells can also be co-cultured with immune cell lines, such as monocytic THP-1 cells or lymphocytic Jurkat T-cells, to evaluate transcriptional changes elicited by Tat taken up by the uninfected cells. |
| معلومات مُعتمدة: | R01 MH083579 United States MH NIMH NIH HHS; R37 AI030861 United States AI NIAID NIH HHS |
| فهرسة مساهمة: | Keywords: Chemokine; Cytokine; Endocytosis; HIV-1; Tat; Trans-cellular transactivation; Transcription |
| المشرفين على المادة: | 0 (Cytokines) 0 (tat Gene Products, Human Immunodeficiency Virus) |
| تواريخ الأحداث: | Date Created: 20151231 Date Completed: 20161013 Latest Revision: 20170602 |
| رمز التحديث: | 20221213 |
| DOI: | 10.1007/978-1-4939-3046-3_24 |
| PMID: | 26714724 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 1940-6029 |
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| DOI: | 10.1007/978-1-4939-3046-3_24 |