Inhibition of HIV-1 gp41 expression with hammerhead ribozymes

2015 ◽  
Vol 471 (1) ◽  
pp. 53-66 ◽  
Author(s):  
Agnieszka Fedoruk-Wyszomirska ◽  
Maciej Szymański ◽  
Paweł Głodowicz ◽  
Marta Gabryelska ◽  
Eliza Wyszko ◽  
...  
Keyword(s):  
Hammerhead Ribozyme ◽  
In Vitro Studies ◽  
Hammerhead Ribozymes ◽  
Intracellular Activity ◽  
Highly Effective ◽  
Hiv 1

We propose a new highly effective hammerhead ribozyme against HIV-1 gp41 that can be potentially used as a basis in HIV-1 therapy. We demonstrate that the ribozyme's intracellular activity cannot be inferred solely from in vitro studies.

Focus on the therapeutic efficacy of 3BNC117 against HIV-1: In vitro studies, in vivo studies, clinical trials and challenges

2017 ◽  
Vol 52 ◽  
pp. 44-50 ◽  
Author(s):  
Zhi-Jun Liu ◽  
Jing Bai ◽  
Feng-Li Liu ◽  
Xiang-Yang Zhang ◽  
Jing-Zhang Wang
Keyword(s):  
Clinical Trials ◽  
Therapeutic Efficacy ◽  
In Vitro Studies ◽  
In Vivo Studies ◽  
Hiv 1

In vitro studies of HIV-1 expression in thymocytes from infants and children

AIDS ◽  
1992 ◽  
Vol 6 (3) ◽  
pp. 265-272 ◽  
Author(s):  
Esther F. Hays ◽  
Christel H. Uittenbogaart ◽  
John C. Brewer ◽  
Leanne W. Vollger ◽  
Jerome A. Zack
Keyword(s):  
In Vitro Studies ◽  
Infants And Children ◽  
Hiv 1

In Vitro Studies of HIV-1 Infection in Thymic Lymphocytes: A Putative Role of the Thymus in AIDS Pathogenesis

1990 ◽  
Vol 6 (3) ◽  
pp. 287-298 ◽  
Author(s):  
ANITA De ROSSI ◽  
MARIA LUISA CALABRO ◽  
MARINA PANOZZO ◽  
DANIELE BERNARDI ◽  
BEATRICE CARUSO ◽  
...  
Keyword(s):  
In Vitro Studies ◽  
Putative Role ◽  
Hiv 1

scholarly journals Plectasin Shows Intracellular Activity against Staphylococcus aureus in Human THP-1 Monocytes and in a Mouse Peritonitis Model

2009 ◽  
Vol 53 (11) ◽  
pp. 4801-4808 ◽  
Author(s):  
Karoline Sidelmann Brinch ◽  
Anne Sandberg ◽  
Pierre Baudoux ◽  
Françoise Van Bambeke ◽  
Paul M. Tulkens ◽  
...  
Keyword(s):  
Staphylococcus Aureus ◽  
Animal Studies ◽  
Kinetic Studies ◽  
Response To Therapy ◽  
In Vitro Studies ◽  
In Vivo Model ◽  
Intracellular Activity ◽  
Peritonitis Model

ABSTRACT Antimicrobial therapy of infections with Staphylococcus aureus can pose a challenge due to slow response to therapy and recurrence of infection. These treatment difficulties can partly be explained by intracellular survival of staphylococci, which is why the intracellular activity of antistaphylococcal compounds has received increased attention within recent years. The intracellular activity of plectasin, an antimicrobial peptide, against S. aureus was determined both in vitro and in vivo. In vitro studies using THP-1 monocytes showed that some intracellular antibacterial activity of plectasin was maintained (maximal relative efficacy [E max], 1.0- to 1.3-log reduction in CFU) even though efficacy was inferior to that of extracellular killing (E max, >4.5-log CFU reduction). Animal studies included a novel use of the mouse peritonitis model, exploiting extra- and intracellular differentiation assays, and assessment of the correlations between activity and pharmacokinetic (PK) parameters. The intracellular activity of plectasin was in accordance with the in vitro studies, with an E max of a 1.1-log CFU reduction. The parameter most important for activity was fC peak/MIC, where fC peak is the free peak concentration. These findings stress the importance of performing studies of extra- and intracellular activity since these features cannot be predicted from traditional MIC and killing kinetic studies. Application of both the THP-1 and the mouse peritonitis models showed that the in vitro results were similar to findings in the in vivo model with respect to demonstration of intracellular activity. Therefore the in vitro model was a good screening model for intracellular activity. However, animal models should be applied if further information on activity, PK/pharmacodynamic parameters, and optimal dosing regimens is required.

scholarly journals HIV gene expression deactivates redox-sensitive stress response program in mouse tubular cells both in vitro and in vivo

2012 ◽  
Vol 302 (1) ◽  
pp. F129-F140 ◽  
Author(s):  
Divya Salhan ◽  
Shresh Pathak ◽  
Mohammad Husain ◽  
Pranai Tandon ◽  
Dileep Kumar ◽  
...  
Keyword(s):  
Free Radical ◽  
Stress Response ◽  
Cell Apoptosis ◽  
In Vitro Studies ◽  
Tubular Cell ◽  
Ros Generation ◽  
Tubular Cells ◽  
Hiv 1

Human immunodeficiency virus (HIV)-1 has been reported to cause tubular cell injury both in in vivo and in vitro studies. In the present study, we evaluated the role of oxidative stress in the induction of apoptosis in HIV gene expressing mouse tubular cells in in vivo (Tg26, a transgenic mouse model of HIV-associated nephropathy) and in vitro (tubular cells were transduced with pNL4-3: ΔG/P-GFP, VSV.G psueudo typed virus) studies. Although Tg26 mice showed enhanced tubular cell reactive oxygen species (ROS) generation and apoptosis, renal tissue did not display a robust antioxidant response in the form of enhanced free radical scavenger (MnSOD/catalase) expression. Tg26 mice not only showed enhanced tubular cell expression of phospho-p66ShcA but also displayed nuclear Foxo3a translocation to the cytoplasm. These findings indicated deactivation of tubular cell Foxo3A-dependent redox-sensitive stress response program (RSSRP) in Tg26 mice. In in vitro studies, NL4-3 (pNL4-3: ΔG/P-GFP, VSV.G pseudotyped virus)-transduced mouse proximal tubular cells (NL4-3/MPTEC) displayed enhanced phosphorylation of p66ShcA. NL4-3/MPTECs also displayed greater ( P < 0.01) ROS generation when compared with empty vector-transduced tubular cells; however, both diminution of p66ShcA and N-acetyl cysteine attenuated NL4-3-induced tubular cell ROS generation as well as apoptosis. In addition, both antioxidants and free radical scavengers partially inhibited HIV-induced tubular cell apoptosis. NL4-3/MPTEC displayed deactivation of RSSRP in the form of enhanced phosphorylation of Foxo3A and attenuated expression of superoxide dismutase (SOD) and catalase. Since both SOD and catalase were able to provide protection against HIV-1-induced tubular cell apoptosis, it suggests that HIV-1-induced proapoptotic effect may be a consequence of the deactivated RSSRP.

Discovery of a Novel HIV-1 Integrase/p75 Interacting Inhibitor by Docking Screening, Biochemical Assay, and in Vitro Studies

2017 ◽  
Vol 57 (9) ◽  
pp. 2336-2343 ◽  
Author(s):  
Yan Wang ◽  
Huang-Quan Lin ◽  
Ping Wang ◽  
Jian-Shu Hu ◽  
Tsz-Ming Ip ◽  
...  
Keyword(s):  
In Vitro Studies ◽  
Biochemical Assay ◽  
Hiv 1

scholarly journals Schistosome Satellite DNA Encodes Active Hammerhead Ribozymes

1998 ◽  
Vol 18 (7) ◽  
pp. 3880-3888 ◽  
Author(s):  
Gerardo Ferbeyre ◽  
James M. Smith ◽  
Robert Cedergren
Keyword(s):  
Repetitive Dna ◽  
Satellite Dna ◽  
Schistosoma Haematobium ◽  
Hammerhead Ribozyme ◽  
Unit Length ◽  
Hammerhead Ribozymes ◽  
Gene Coding ◽  
Rna Structural Motifs

ABSTRACT Using a computer program designed to search for RNA structural motifs in sequence databases, we have found a hammerhead ribozyme domain encoded in the Smα repetitive DNA of Schistosoma mansoni. Transcripts of these repeats are expressed as long multimeric precursor RNAs that cleave in vitro and in vivo into unit-length fragments. This RNA domain is able to engage in bothcis and trans cleavage typical of the hammerhead ribozyme. Further computer analysis of S. mansoni DNA identified a potential trans cleavage site in the gene coding for a synaptobrevin-like protein, and RNA transcribed from this gene was efficiently cleaved by the Smα ribozyme in vitro. Similar families of repeats containing the hammerhead domain were found in the closely related Schistosoma haematobium and Schistosomatium douthitti species but were not present in Schistosoma japonicum orHeterobilharzia americana, suggesting that the hammerhead domain was not acquired from a common schistosome ancestor.

scholarly journals Inhibition of human immunodeficiency virus-1 entry using vectors expressing a multimeric hammerhead ribozyme targeting the CCR5 mRNA

2008 ◽  
Vol 89 (9) ◽  
pp. 2252-2261 ◽  
Author(s):  
Reza Nazari ◽  
Xue Zhong Ma ◽  
Sadhna Joshi
Keyword(s):  
Human Immunodeficiency Virus ◽  
Viral Entry ◽  
Hammerhead Ribozyme ◽  
Virus Production ◽  
Progeny Virus ◽  
Immunodeficiency Virus ◽  
Infection Inhibition ◽  
Human Ccr5 ◽  
Hiv 1

Rz1–7 is a multimeric hammerhead ribozyme targeting seven unique sites within the human CCR5 mRNA that is active in vitro. Mouse stem cell virus-based MGIN and human immunodeficiency virus (HIV)-1-based HEG1 vectors were used to express Rz1–7 in a human CD4+ T lymphoid cell line. Stable transductants expressed Rz1–7, which was further shown to be active, since CCR5 mRNA and surface CCR5 protein expression levels decreased. High levels of progeny virus were produced when the transduced cells were challenged with an X4-tropic HIV-1 (NL4-3) strain, suggesting that Rz1–7 expression does not affect X4-tropic virus replication. When the transduced cells expressing Rz1–7 were challenged with the R5-tropic HIV-1 (BaL) strain, 99–100 % inhibition of progeny virus production was observed for the duration of the experiment (∼2 months). When the cells were precultured for 2–3 months prior to HIV-1 infection, inhibition was more prominent in cells transduced with MGIN-Rz1–7 than with HEG1-Rz1–7. Inhibition occurred at the level of viral entry, as no HIV-1 DNA could be detected. These results demonstrate that Rz1–7 confers excellent inhibition of R5-tropic HIV-1 replication at the level of entry. Therefore, we anticipate that this multimeric ribozyme will be beneficial for HIV-1 gene therapy.

scholarly journals Inhibition of Hepatitis B Virus Replication and Expression in Vitro and in Vivo by the Hammerhead Ribozymes Targeted Different Sites

2012 ◽  
Vol 1 (4) ◽  
pp. 206-210
Author(s):  
Wei Dai ◽  
Rong Zhou ◽  
Hong Yu ◽  
Xiao-juan Li
Keyword(s):  
Hepatitis B Virus ◽  
Hepatitis B ◽  
Hammerhead Ribozyme ◽  
Hbv Dna ◽  
Hammerhead Ribozymes ◽  
Tail Vein ◽  
B Virus ◽  
Target Sites

Abstract Objective To develop an effective and specific medicine targeting hepatitis B virus (HBV) pregenome. Based on the identified accessible target sites for hammerhead ribozyme in our previous researches, a recombinant hepatitis delta virus (HDV) ribozyme was chosen and used to demonstrate the effective cleavage in vitro and in vivo. Methods Three hammerhead ribozymes for potential target sites (S, X and C genes) and co-expression plasmid (pTr-dB, pTdδ-dB, pTrX-dB and pTrC-dB) as well as four HDV-ribozyme chimera constructs with HBV (pTdXX, pTdXC, pTdSX and pTdSC) were severally chosen to validate the inhibition of the replication and expression of HBV. The co-expression plasmids (pTdδ and pTr-Db) in physiological saline were hydrodynamically injected to mice by tail vein. Results Compared with the group injected with pTr-dB in Huh-7 cell, hepatitis B surface antigen (HBsAg) was reduced by 31% in the group injected with pTdδ-dB, by 54%, 26%, 72% and 97% in the group injected with recombinant-ribozymes pTdSX, pTdSC, pTdXC and pTdXX, respectively. The inhibiting effects of endogenous ribozymes RzX and RzC on the HBsAg expression were 66% and 57%, respectively. Compared with the positive control, the amount of HBsAg was decreased in mice injected with pTdXX through tail vein by 88% and 96% on the second day and the third day, respectively. HBsAg was undetectable on the 6th day and could not primitively be detected on the 9th day in the sera from all mice. HBV DNA was not detected in the sera of BALB/c mice injected with pTdXX-dB, pTrX-dB or replicating-defective plasmid pHBV, while HBV DNA replication in control group could be detected on the 6th day. While HBcAg could not be detected in liver tissues of mice injected with plasmid pTdXX-dB on the 3rd day. Conclusions Encoding regions of HBV S, C and X gene were the effective cleavage sites for hammerhead ribozyme in vitro and in vivo, which provides basis for further construction of therapeutic recombinant HDV and the development of targeting antiviral gene therapy.

Targeted femtosecond laser driven drug delivery within HIV-1 infected cells: in-vitro studies

2017 ◽  
Author(s):  
Charles Maphanga ◽  
Saturnin Ombinda-Lemboumba ◽  
Sello Manoto ◽  
Malik Maaza ◽  
Patience Mthunzi-Kufa
Keyword(s):  
Drug Delivery ◽  
Femtosecond Laser ◽  
In Vitro Studies ◽  
Infected Cells ◽  
Hiv 1
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