human t lymphocyte genetic modification with naked dna.

NLM AIDSLINE Human T lymphocyte genetic modification with naked DNA. Jensen MC; Clarke P; Tan G; Wright C; Chung-Chang W; Clark TN; Zhang F;
November 30, 2000

Mol Ther. 2000 Jan;1(1):49-55. Unique Identifier : AIDSLINE MED/20393213 Endowing T lymphocytes with novel functional attributes by genetic modification is under development for a broad range of clinical cellular immunotherapy applications. To circumvent many of the limitations associated with viral vector systems, a plasmid-based electroporation system that reliably generates G418-resistant primary human T lymphocyte clones was developed. TCR alpha/beta+ CD4+CD8-, and CD4-CD8+ T lymphocyte clones can be routinely isolated from OKT3-stimulated peripheral blood mononuclear cells electroporated with linear plasmid DNA in a limiting dilution drug selection format. Fluorescence in situ hybridization (FISH) studies performed on T cell metaphase spreads using a probe specific for plasmid sequence demonstrated a single FISH signal doublet that varied in chromosomal location from clone to clone. Southern blot analysis using a Neo-specific probe verified chromosomal integration of plasmid vector at a single site. Band intensity quantitation of blots developed with a zeta-specific probe capable of annealing to both endogenous TCR-zeta and the introduced chimeric zeta sequence demonstrated that integrated plasmid was present at a single copy number. Expression levels of the CD20-specific chimeric immunoreceptor construct from a CMV immediate/early promoter present in the plasmid vector varied widely from clone to clone but remained stable during ex vivo expansion to cell numbers in excess of 10(10). This T lymphocyte genetic modification strategy is currently being piloted in a FDA-sanctioned adoptive therapy trial for recurrent lymphoma. JOURNAL ARTICLE Cell Line Chimeric Proteins/GENETICS/IMMUNOLOGY Clone Cells CD4-Positive T-Lymphocytes/IMMUNOLOGY CD8-Positive T-Lymphocytes/IMMUNOLOGY Electroporation Gene Therapy/METHODS Genetic Vectors Human Immunotherapy/METHODS In Situ Hybridization, Fluorescence Membrane Proteins/GENETICS/IMMUNOLOGY Plasmids/*GENETICS Receptors, Antigen, T-Cell/GENETICS/IMMUNOLOGY Support, Non-U.S. Gov't Support, U.S. Gov't, P.H.S.

Most Read: Latest Added Related Topics White House LGBT Conference on HIV/AIDS UNITED STATES: White House in Atlanta: LGBT AIDS Conference No more single-theme conferences: global HIV prevention conference planned FHI 360 Senior Scientist Lut Van Damme Receives International Microbicide Conference Lifetime Achievement Award Equatorial Guinea Ministry of Health and Social Welfare and UNICEF Refute Erroneous AIDS Prevalence Figures Photographs from A Day with HIV Now Online Bulgaria urges Libya to investigate AIDS nurses case Engineers enter fight against AIDS in Africa Gilead’s Complera® Non-Inferior to Atripla® Among Treatment-Naive HIV Patients UN chief Ban calls world leaders to end Aids by 2020 HIV messages should include MSM Stock-outs must be top of agenda TEXAS: Free HIV Testing CALIFORNIA: Entertainment AIDS Alliance (EAA) Presents 11th Annual Wine and Wisdom Fundraiser NORTH CAROLINA: ALFA Offers Free Community Health Educator Training in Hickory CALIFORNIA: Over Half of People Living with AIDS in San Francisco Are Aged 50 or Older THAILAND; UNITED STATES: HIV Drug Can Also Protect Injection Drug Users INDONESIA: Growing HIV/AIDS Awareness in Indonesia's Papua Region UNITED STATES: Many Young Americans Unaware They're Infected with HIV Board to Vote on Condoms in Boston Schools New HIV/AIDS and Human Papillomavirus Findings Reported from Stanley Scott Cancer Center Organization of American States adopts bold resolution on HIV and human rights Eminent Judges unite to address HIV, human rights and the law Right to health, right to life: HIV and human rights in Europe Study Results from Eunice Kennedy Shriver National Institute of Child Health and Human Development Provide New Insights into HIV/AIDS Monetising Human Waste and 101 (Slightly) Crazy Other Ideas Inhibikase Therapeutics to present at 5th International Congress on Polyomaviruses and Human Diseases - May 11, 2013 Grand Hotel Bristol, Stresa, Italy Honduras reforms its penal code to end human right violations based on sexual orientation and gender identity Human Rights Activist Pedro Julio Serrano to Receive Distinguished Award at Cielo Latino Gala 2013 Massive development gains in the South is improving the economic, social and health prospects for millions, says Human Development Report
Information in this article was accurate in November 30, 2000. The state of the art may have changed since the publication date. This material is designed to support, not replace, the relationship that exists between you and your doctor. Always discuss treatment options with a doctor who specializes in treating HIV.