Abstract
Viral pathogenesis is studied predominantly in cultures of primary isolated cells or cell lines. Many retroviruses efficiently replicate only in activated cells. Therefore, in order to become efficient viral producers cells should be artificially activated, a procedure which significantly changes cell physiology. However, for many viral diseases, like HIV-1 and other retroviruses’ diseases, critical pathogenic events occur in tissues. Therefore, cell isolation from their native microenvironment prevents single-cell cultures from faithfully reflecting important aspects of cell–cell and cell–pathogen interactions that occur in the context of complex tissue cytoarchitecture. Tissue explants (histocultures) that retain tissue cytoarchitecture and many aspects of cell–cell interactions more faithfully represent in vivo tissue features. Human histocultures constitute an adequate model for studying viral pathogenesis under controlled laboratory conditions. Protocols for various human histocultures as applied to study retroviral pathogenesis, in particular of HIV-1, have been refined by our laboratory and are described in the present publication. Histocultures of human tonsils and lymph nodes, as well as of recto-sigmoid and cervicovaginal tissues can be used to study viral transmission, pathogenesis and as a preclinical platform for antivirals evaluation.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Sebastion A (2001) A dictionary of the history of science. Informa Healthcare, Pearl River, NY
Brown CR, Czapiga M, Kabat J, Dang Q, Ourmanov I, Nishimura Y, Martin MA, Hirsch VM (2007) Unique pathology in simian immunodeficiency virus-infected rapid progressor macaques is consistent with a pathogenesis distinct from that of classical AIDS. J Virol 81:5594–5606
Harrison R (1907) Observations on the living developing nerve fiber. Anat Rec 1
Harrison R (1910) The outgrowth of the nerve fiber as a mode of protoplasmic movement. J Exp Zool 9:787–846
Carrel A (1912) On the permanent life of tissues outside of the organism. J Exp Med 15: 516–528
Leighton J (1951) A sponge matrix method for tissue culture; formation of organized aggregates of cells in vitro. J Natl Cancer Inst 12:545–561
Leighton J (1963) A method for the comparison of the fate of intravascular tumor-cell emboli in vivo and in organ culture. Natl Cancer Inst Monogr 11:157–195
Hoffman RM (1991) Three-dimensional histoculture: origins and applications in cancer research. Cancer Cells 3:86–92
Yang J, Richards J, Bowman P, Guzman R, Enami J, McCormick K, Hamamoto S, Pitelka D, Nandi S (1979) Sustained growth and three-dimensional organization of primary mammary tumor epithelial cells embedded in collagen gels. Proc Natl Acad Sci U S A 76:3401–3405
Strangeways T (1924) Tissue culture in relation to growth and differentiation. W Heffer and Sons Ltd, Cambridge, England
Fell HB, Robison R (1929) The growth, development and phosphatase activity of embryonic avian femora and limb-buds cultivated in vitro. Biochem J 23(767–784):765
Browning TH, Trier JS (1969) Organ culture of mucosal biopsies of human small intestine. J Clin Invest 48:1423–1432
Inch WR, McCredie JA, Sutherland RM (1970) Growth of nodular carcinomas in rodents compared with multi-cell spheroids in tissue culture. Growth 34:271–282
Glushakova S, Baibakov B, Margolis LB, Zimmerberg J (1995) Infection of human tonsil histocultures: a model for HIV pathogenesis. Nat Med 1:1320–1322
Glushakova S, Baibakov B, Zimmerberg J, Margolis L (1997) Experimental HIV infection of human lymphoid tissue: correlation of CD4+ T cell depletion and virus syncytium-inducing/non-syncytium-inducing phenotype in histoculture inoculated with laboratory strains and patient isolates of HIV type 1. AIDS Res Hum Retroviruses 13:461–471
Grivel JC, Ito Y, Faga G, Santoro F, Shaheen F, Malnati MS, Fitzgerald W, Lusso P, Margolis L (2001) Suppression of CCR5- but not CXCR4-tropic HIV-1 in lymphoid tissue by human herpesvirus 6. Nat Med 7:1232–1235
Condack C, Grivel JC, Devaux P, Margolis L, Cattaneo R (2007) Measles virus vaccine attenuation: suboptimal infection of lymphatic tissue and tropism alteration. J Infect Dis 196:541–549
Grivel JC, Garcia M, Moss W, Margolis L (2005) Measles virus inhibits HIV-1 replication in human lymphoid tissue ex vivo. J Infect Dis 192:71–78
Lisco A, Grivel JC, Biancotto A, Vanpouille C, Origgi F, Malnati MS, Schols D, Lusso P, Margolis LB (2007) Viral interactions in human lymphoid tissue: Human herpesvirus 7 suppresses the replication of CCR5-tropic human immunodeficiency virus type 1 via CD4 modulation. J Virol 81:708–717
Biancotto A, Iglehart SJ, Lisco A, Vanpouille C, Grivel JC, Lurain NS, Reichelderfer PS, Margolis LB (2008) Upregulation of human cytomegalovirus by HIV type 1 in human lymphoid tissue ex vivo. AIDS Res Hum Retroviruses 24:453–462
Lisco A, Vanpouille C, Tchesnokov EP, Grivel JC, Biancotto A, Brichacek B, Elliott J, Fromentin E, Shattock R, Anton P, Gorelick R, Balzarini J, McGuigan C, Derudas M, Gotte M, Schinazi RF, Margolis L (2008) Acyclovir is activated into a HIV-1 reverse transcriptase inhibitor in herpesvirus-infected human tissues. Cell Host Microbe 4:260–270
Vanpouille C, Biancotto A, Lisco A, Brichacek B (2007) Interactions between human immunodeficiency virus type 1 and vaccinia virus in human lymphoid tissue ex vivo. J Virol 81:12458–12464
Saba E, Grivel JC, Vanpouille C, Brichacek B, Fitzgerald W, Margolis L, Lisco A (2010) HIV-1 sexual transmission: early events of HIV-1 infection of human cervico-vaginal tissue in an optimized ex vivo model. Mucosal Immunol 3:280–290
Fletcher PS, Elliott J, Grivel JC, Margolis L, Anton P, McGowan I, Shattock RJ (2006) Ex vivo culture of human colorectal tissue for the evaluation of candidate microbicides. AIDS 20:1237–1245
Glushakova S, Yi Y, Grivel JC, Singh A, Schols D, De Clercq E, Collman RG, Margolis L (1999) Preferential coreceptor utilization and cytopathicity by dual-tropic HIV-1 in human lymphoid tissue ex vivo. J Clin Invest 104:R7–R11
Glushakova S, Grivel JC, Fitzgerald W, Sylwester A, Zimmerberg J, Margolis LB (1998) Evidence for the HIV-1 phenotype switch as a causal factor in acquired immunodeficiency. Nat Med 4:346–349
Boukari H, Brichacek B, Stratton P, Mahoney SF, Lifson JD, Margolis L, Nossal R (2009) Movements of HIV-virions in human cervical mucus. Biomacromolecules 10:2482–2488
Biancotto A, Grivel JC, Lisco A, Vanpouille C, Markham PD, Gallo RC, Margolis LB, Lusso P (2009) Evolution of SIV toward RANTES resistance in macaques rapidly progressing to AIDS upon coinfection with HHV-6A. Retrovirology 6:61
Biancotto A, Brichacek B, Chen SS, Fitzgerald W, Lisco A, Vanpouille C, Margolis L, Grivel JC (2009) A highly sensitive and dynamic immunofluorescent cytometric bead assay for the detection of HIV-1 p24. J Virol Methods 157:98–101
Perfetto SP, Ambrozak D, Nguyen R, Chattopadhyay P, Roederer M (2006) Quality assurance for polychromatic flow cytometry. Nat Protoc 1:1522–1530
Acknowledgments
This work was supported by the NICHD Intramural Program.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2014 Springer Science+Business Media, LLC
About this protocol
Cite this protocol
Arakelyan, A., Fitzgerald, W., Grivel, JC., Vanpouille, C., Margolis, L. (2014). Histocultures (Tissue Explants) in Human Retrovirology. In: Vicenzi, E., Poli, G. (eds) Human Retroviruses. Methods in Molecular Biology, vol 1087. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-670-2_19
Download citation
DOI: https://doi.org/10.1007/978-1-62703-670-2_19
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-669-6
Online ISBN: 978-1-62703-670-2
eBook Packages: Springer Protocols