Ex vivo culture of human colorectal tissue for the evaluation of candidate microbicides
| Autor: | Peter A. Anton, Julie Elliott, Ian McGowan, Patricia S. Fletcher, Robin J. Shattock, Jean-Charles Grivel, Leonid Margolis |
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| Rok vydání: | 2006 |
| Předmět: |
Sexually transmitted disease
Rectal microbicide Sexual transmission Colon Administration Topical Immunology HIV Infections Cell Separation Biology Pharmacology Virus Replication Antiviral Agents Tissue Culture Techniques Tissue culture Leukocytes medicine Humans Immunology and Allergy Reverse-transcriptase inhibitor Rectum Flow Cytometry Virology Microbicides for sexually transmitted diseases Phenotype Infectious Diseases HIV-1 Drug Evaluation Gels Ex vivo medicine.drug Explant culture |
| Zdroj: | AIDS. 20:1237-1245 |
| ISSN: | 0269-9370 |
| Popis: | Objectives Establishment of an in vitro model to evaluate rectal safety and the efficacy of microbicide candidates. Design An investigation and characterization of human colorectal explant culture for screening candidate microbicides to prevent rectal transmission of HIV-1 infection. Methods Human colorectal explants were cultured at the liquid-air interface on gelfoam rafts. Phenotypic characterization of HIV-1 target cells was performed by fluorescence-activated cell sorter analysis. HIV-1 infection was determined by the measurement of p24 antigen release, viral RNA, and proviral DNA accumulation. Results Colorectal explant CD4 T cells expressed higher CCR5 and CXCR4 levels compared with blood. Minor differences between the rectal and sigmoid colon were observed with a trend for slightly higher CCR5 and HLA-DR expression in cells from the sigmoid colon. Favourable culture conditions were established for colorectal tissue. Although tissue structure degenerated with time, CD4: CD8 cell ratios remained constant, and tissue supported productive HIV-1 infection. The ability of candidate microbicides to inhibit R5 HIV-1 infection was evaluated. Polyanion candidates, PRO2000 and dextrin sulphate, provided 99% protection at 1 microg/ml and 1 mg/ml, respectively, equivalent to 1/5000 and 1/40 of the vaginal formulations. The nucleotide reverse transcriptase inhibitor (NRTI) 9-[2-(phosphonomethoxy)propyl]adenine (PMPA) provided protection at concentrations 1000-fold lower (10 microg/ml) than the proposed vaginal formulation (1%). Furthermore, non-NRTI UC-781 and TMC-120 provided greater than 99% inhibition at 3.3 or 0.33 microg/ml, respectively. No products demonstrated toxicity to rectal mucosa at inhibitory concentrations. Conclusion Colorectal explant culture was shown to be a useful tool for the preclinical evaluation of potential microbicides. The data suggest that rectally applied microbicides might provide protection from HIV-1 transmission. |
| Databáze: | OpenAIRE |
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