Nonreproducibility of “Snap-Frozen” Rectal Biopsies for Later Use in Ex Vivo Explant Infectibility Studies
Publication: AIDS Research and Human Retroviruses
Volume 28, Issue Number 11
Abstract
Sexual transmission accounts for the majority of new HIV infections worldwide with sexually exposed cervicovaginal and colorectal mucosae being primary sites of infection. Two recent Phase 1 rectal microbicide trials included, as an ancillary endpoint, suppression of ex vivo HIV infection of in vivo microbicide-exposed rectal mucosal tissue biopsies. Both trials demonstrated significant suppression of biopsy infectibility in drug-exposed versus placebo-exposed tissue. This potential early biomarker of efficacy has raised the feasibility of utilizing “snap-frozen” tissue samples, acquired at multiple trial sites to be shipped for central processing, providing a mechanism to correlate tissue drug concentrations with a functional index of HIV prevention. While previous reports have indicated acceptable comparability of fresh versus freeze-thawed cervicovaginal tissue samples, no similar evaluations with colorectal tissue biopsies have been done. In this study, rectal biopsies from healthy, HIV-seronegative participants were assessed for structural integrity (histology), viability (MTT assays), and tissue infectibility to compare results from fresh versus combinations of freeze/thaw protocols. Results indicated that while all protocols showed equivalent viability with fresh samples (MTT), histology documented poor preservation of tissue integrity following freezing. Infectibility results from freeze-thawed colorectal tissue were markedly lower (usually<25% of fresh samples) and varied greatly and unpredictably. Centralized colorectal tissue infectibility assays using biopsies from remote trial sites cannot currently be supported under these protocols.
Get full access to this article
View all available purchase options and get full access to this article.
References
1.
Anton PASaunders TElliott J et al. First Phase 1 Double-Blind, Placebo-Controlled, Randomized Rectal Microbicide Trial Using UC781 Gel with a Novel Index of Ex Vivo EfficacyPLoS One201169e23243. 1. Anton PA, Saunders T, Elliott J, et al.: First Phase 1 Double-Blind, Placebo-Controlled, Randomized Rectal Microbicide Trial Using UC781 Gel with a Novel Index of Ex Vivo Efficacy. PLoS One 2011;6(9):e23243.
2.
Herrera CCranage MMcGowan IAnton PShattock RJ. Colorectal microbicide design: Triple combinations of reverse transcriptase inhibitors are optimal against HIV-1 in tissue explantsAIDS201125161971-1979. 2. Herrera C, Cranage M, McGowan I, Anton P, and Shattock RJ: Colorectal microbicide design: Triple combinations of reverse transcriptase inhibitors are optimal against HIV-1 in tissue explants. AIDS 2011;25(16):1971–1979.
3.
Fletcher PSElliott JGrivel JC et al. Ex vivo culture of human colorectal tissue for the evaluation of candidate microbicidesAIDS20062091237-1245. 3. Fletcher PS, Elliott J, Grivel JC, et al.: Ex vivo culture of human colorectal tissue for the evaluation of candidate microbicides. AIDS 2006;20(9):1237–1245.
4.
Anton PCranston RCarballo-Dieguez AKashuba AKhanukhova EElliot J et al. RMP-02/MTN-006: A Phase 1, placebo controlled trial of rectally applied 1% vaginal tenofovir gel with comparison to oral tenofovir disoproxil fumarate18th Conference on Retroviruses and Opportunistic InfectionsBoston, MAFebruary2011Abstract 34LB. 4. Anton P, Cranston R, Carballo-Dieguez A, Kashuba A, Khanukhova E, Elliot J, et al.: RMP-02/MTN-006: A Phase 1, placebo controlled trial of rectally applied 1% vaginal tenofovir gel with comparison to oral tenofovir disoproxil fumarate. 18th Conference on Retroviruses and Opportunistic Infections, Boston, MA, February 2011. Abstract 34LB.
5.
Anton PAElliott JPoles MA et al. Enhanced levels of functional HIV-1 co-receptors on human mucosal T cells demonstrated using intestinal biopsy tissueAIDS200014121761-1765. 5. Anton PA, Elliott J, Poles MA, et al.: Enhanced levels of functional HIV-1 co-receptors on human mucosal T cells demonstrated using intestinal biopsy tissue. AIDS 2000;14(12):1761–1765.
6.
Gupta PRatner DPatterson BK et al. Use of frozen-thawed cervical tissues in the organ culture system to measure anti-HIV activities of candidate microbicidesAIDS Res Hum Retroviruses2006225419-424. 6. Gupta P, Ratner D, Patterson BK, et al.: Use of frozen-thawed cervical tissues in the organ culture system to measure anti-HIV activities of candidate microbicides. AIDS Res Hum Retroviruses 2006;22(5):419–424.
7.
Lackman-Smith COsterling CLuckenbaugh K et al. Development of a comprehensive human immunodeficiency virus type 1 screening algorithm for discovery and preclinical testing of topical microbicidesAntimicrob Agents Chemother20085251768-1781. 7. Lackman-Smith C, Osterling C, Luckenbaugh K, et al.: Development of a comprehensive human immunodeficiency virus type 1 screening algorithm for discovery and preclinical testing of topical microbicides. Antimicrob Agents Chemother 2008;52(5):1768–1781.
8.
Shanahan F. Intestinal lymphoepithelial communicationAdv Exp Med Biol19994731-9. 8. Shanahan F: Intestinal lymphoepithelial communication. Adv Exp Med Biol 1999;473:1–9.
9.
Griffith TSFerguson TA. The role of FasL-induced apoptosis in immune privilegeImmunol Today1997185240-244. 9. Griffith TS and Ferguson TA: The role of FasL-induced apoptosis in immune privilege. Immunol Today 1997;18(5):240–244.
10.
Isachenko VIsachenko EReinsberg J et al. Cryopreservation of human ovarian tissue: Comparison of rapid and conventional freezingCryobiology2007553261-268. 10. Isachenko V, Isachenko E, Reinsberg J, et al.: Cryopreservation of human ovarian tissue: Comparison of rapid and conventional freezing. Cryobiology 2007;55(3):261–268.
11.
Keros VHultenby KBorgstrom B et al. Methods of cryopreservation of testicular tissue with viable spermatogonia in pre-pubertal boys undergoing gonadotoxic cancer treatmentHum Reprod20072251384-1395. 11. Keros V, Hultenby K, Borgstrom B, et al.: Methods of cryopreservation of testicular tissue with viable spermatogonia in pre-pubertal boys undergoing gonadotoxic cancer treatment. Hum Reprod 2007;22(5):1384–1395.
12.
McGowan IElliott JCortina G et al. Characterization of baseline intestinal mucosal indices of injury and inflammation in men for use in rectal microbicide trials (HIV Prevention Trials Network-056)J Acquir Immune Defic Syndr2007464417-425. 12. McGowan I, Elliott J, Cortina G, et al.: Characterization of baseline intestinal mucosal indices of injury and inflammation in men for use in rectal microbicide trials (HIV Prevention Trials Network-056). J Acquir Immune Defic Syndr 2007;46(4):417–425.
13.
Anton PAIbarrondo FJBoscardin WJ et al. Differential immunogenicity of vaccinia and HIV-1 components of a human recombinant vaccine in mucosal and blood compartmentsVaccine200826354617-4623. 13. Anton PA, Ibarrondo FJ, Boscardin WJ, et al.: Differential immunogenicity of vaccinia and HIV-1 components of a human recombinant vaccine in mucosal and blood compartments. Vaccine 2008;26(35):4617–4623.
14.
Richardson-Harman NLackman-Smith CFletcher PS et al. Multi-site comparison of anti-HIV microbicide activity in explant assays using a novel endpoint analysisJ Clin Microbiol200947113530-3539. 14. Richardson-Harman N, Lackman-Smith C, Fletcher PS, et al.: Multi-site comparison of anti-HIV microbicide activity in explant assays using a novel endpoint analysis. J Clin Microbiol 2009;47(11):3530–3539.
15.
Reed LJMuench H. A simple method of estimating fifty per cent end-pointsAm J Hygiene193827493-497. 15. Reed LJ and Muench H: A simple method of estimating fifty per cent end-points. Am J Hygiene 1938;27:493–497.
16.
Chan DJMcNally LBatterham MSmith DE. Relationship between HIV-RNA load in blood and semen in antiretroviral-naive and experienced men and effect of asymptomatic sexually transmissible infectionsCurr HIV Res200862138-142. 16. Chan DJ, McNally L, Batterham M, and Smith DE: Relationship between HIV-RNA load in blood and semen in antiretroviral-naive and experienced men and effect of asymptomatic sexually transmissible infections. Curr HIV Res 2008;6(2):138–142.
Information & Authors
Information
Published In
AIDS Research and Human Retroviruses
Volume 28 • Issue Number 11 • November 2012
Pages: 1509 - 1512
PubMed: 22831398
Copyright
Copyright 2012, Mary Ann Liebert, Inc.
History
Published online: 5 November 2012
Published in print: November 2012
Published ahead of print: 4 September 2012
Published ahead of production: 25 July 2012
Topics
Authors
Author Disclosure Statement
No competing financial interests exist.
Metrics & Citations
Metrics
Citations
Export Citation
Export citation
Select the format you want to export the citations of this publication.
View Options
Access content
To read the fulltext, please use one of the options below to sign in or purchase access.⚠ Society Access
If you are a member of a society that has access to this content please log in via your society website and then return to this publication.