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Human immunodeficiency virus-1 tat- and tat/nef-defective genomes containing HIV-regulated diphtheria toxin A chain gene inhibit HIV replication. (CROSBI ID 94722)

Prilog u časopisu | izvorni znanstveni rad | međunarodna recenzija

Brdar, Branko ; Matulić, Maja ; Rubelj, Ivica ; Ivanković, Milena ; Reich, Edward Human immunodeficiency virus-1 tat- and tat/nef-defective genomes containing HIV-regulated diphtheria toxin A chain gene inhibit HIV replication. // Croatian medical journal, 43 (2002), 5; 591-597-x

Podaci o odgovornosti

Brdar, Branko ; Matulić, Maja ; Rubelj, Ivica ; Ivanković, Milena ; Reich, Edward

engleski

Human immunodeficiency virus-1 tat- and tat/nef-defective genomes containing HIV-regulated diphtheria toxin A chain gene inhibit HIV replication.

Aim. To assess the ability of human immunodeficiency virus-1 (HIV-1) tat- and tat/nef-defective genomes containing diphtheria toxin A chain gene (DTA) to inhibit replication of HIV in human cells. Methods. Plasmids were constructed to contain the HIV-1 genome disabled by tat and tat/nef deletions, and sequences coding for the A subunit of diphtheria toxin gene were inserted into one of these deletions. An infectious clone of HIV-1 (pBRU-3) was cotransfected into HeLa-CD4 cells, together with plasmids carrying the modified DTA-containing genomes. Cell culture supernatants were collected and titrated for the virus by multinuclear activation of P-galactosidase (MAGI) assay. Results. Each of the DTA-containing plasmids suppressed HIV production by no less than 96%, whereas the defective non-DTA containing plasmids did not interfere with the virus growth. Plasmids containing wild-type DTA inhibited HIV replication slightly more than its moderately attenuated mutant form, probably by limiting the synthesis of viral proteins. These modified DTA-containing HIV constructs gave no evidence of virus growth in HIV susceptible cells that supported the Multiplication of the parent plasmid. None of the modified DTA-containing plasmids was toxic to cells cotransfected with a selectable marker, as shown by the ability of cotransfectants to multiply and form colonies at rates identical to controls exposed to non-specific DNA. This suggested that DTA was probably not expressed in the absence of activating wild-type HIV plasmid. Conclusion. HIV-regulated DTA in the background of a HIV replication and expression of defective provirus may be taken into consideration as a therapy approach to the treatment of HIV infection, based on its selective and specific toxicity only to HIV infected CD4-positive cells. [References: 53]

Acquired immunodeficiency syndrome. Cd4-positive t-lymphocytes. Diphtheria toxin. Gene therapy. Genes; nef. Genes; tat. Genetic vectors. Hela cells. Hiv-1. Trans-activation (genetic). Transfection.Necrosis-factor-alpha. Trans-activator gene. Human t-cells.

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Podaci o izdanju

43 (5)

2002.

591-597-x

objavljeno

0353-9504

Povezanost rada

Biologija

Indeksiranost