Skip to main content

Isolation and HIV-1 Infection of Primary Human Microglia From Fetal and Adult Tissue

  • Protocol
Human Retrovirus Protocols

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 304))

Summary

Glial inflammation, principally involving astrocytes and microglia, underlies the pathogenesis of a broad range of neurodegenerative disorders, including, most notably, human immunodeficiency virus (HIV-1)-associated dementia. Indeed, for the latter, disease mechanisms are attributed to viral infection and activation of microglia and perivascular macrophages and their resultant neurotoxic activities. Although monocyte-derived macrophages have served as models for microglia, they are limited both qualitatively and quantitatively in their immune responses and susceptibility to viral infection. Thus, the acquisition of primary human microglial cells is critical for laboratory studies of human neurological disease. In this chapter, we provide detailed methods of isolation, cultivation, characterization, HIV-1 infection, and experimental applications of primary human fetal and adult microglial cells, with particular emphasis on studies of HIV-1 neuropathogenesis.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 169.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. del Rio-Hortega, P. (1932) in Cytology and Cellular Pathology of the Nervous System (Penfield, W., ed.), Hoeber, New York: pp. 481–584.

    Google Scholar 

  2. Gehrmann, J., Matsumoto, Y., and Kreutzberg, G. W. (1995) Microglia: intrinsic immuneffector cell of the brain. Brain Res. Brain Res. Rev. 20, 269–287.

    Article  PubMed  CAS  Google Scholar 

  3. Streit, W., Graeber, M., and Kreutzberg, G. (1988) Functional plasticity of microglia: a review. Glia 1, 301–307.

    Article  PubMed  CAS  Google Scholar 

  4. Griffin, W. S., Sheng, J. G., Royston, M. C., Gentleman, S. M., McKenzie, J. E., Graham, D. I., et al. (1998) Glial-neuronal interactions in Alzheimer’s disease: the potential role of a ‘cytokine cycle’ in disease progression. Brain Pathol. 8, 65–72.

    Article  PubMed  CAS  Google Scholar 

  5. Watkins, L. R. and Maier, S. F. (1999) Implications of immune-to-brain communication for sickness and pain Proc. Natl. Acad. Sci. USA 96, 7710–7713.

    Article  PubMed  CAS  Google Scholar 

  6. Raivich, G., Bohatschek, M., Kloss, C. U. A., Werner, A., Jones, L., and Dreutzberg, G. W. (1999) Neuroglial activation repertoire in the injured brain: graded response molecular mechanisms and cues to physiological function. Brain Res. Rev. 30, 77–105.

    Article  PubMed  CAS  Google Scholar 

  7. Williams, K. C. and Hickey, W. F. (2002) Central nervous system damage, monocytes and macrophages, and neurological disorders in AIDS. Annu. Rev. Neurosci. 25, 537–562.

    Article  PubMed  CAS  Google Scholar 

  8. Minagar, A., Shapshak, P., Fujimura, R., Ownby, R., Heyes, M., and Eisdorfer, C. (2002) The role of macrophage/microglia and astrocytes in the pathogenesis of three neurologic disorders: HIV-associated dementia, Alzheimer disease, and multiple sclerosis. J. Neurol. Sci. 202, 13–23.

    Article  PubMed  CAS  Google Scholar 

  9. Gonzalez-Scarano, F. and Baltuch, G. (1999) Microglia as mediators of inflammatory and degenerative diseases. Annu. Rev. Neurosci. 22, 219–240.

    Article  PubMed  CAS  Google Scholar 

  10. Benveniste, E. N. (1997) Role of macrophages/microglia in multiple sclerosis and experimental allergic encephalomyelitis J. Mol. Med. 75, 165–173.

    Article  PubMed  CAS  Google Scholar 

  11. Dickson, D. W., Lee, S. C., Mattiace, L. A., Yen, S. H., and Brosnan, C. (1993) Microglia and cytokines in neurological disease, with special reference to AIDS and Alzheimer’s disease. Glia 7, 75–83.

    Article  PubMed  CAS  Google Scholar 

  12. Ghorpade, A. (1997) in The neurology of AIDS (Gendelman, H. E., Lipton, S. A., Epstein, L., and Swindells, S., eds.), Chapman and Hall, New York, pp. 86–96.

    Google Scholar 

  13. Ghorpade, A., Nukuna, A., Che, M., Haggerty, S., Persidsky, Y., Carter, E., et al. (1998) Human immunodeficiency virus neurotropism: an analysis of viral replication and cytopathicity for divergent strains in monocytes and microglia. J. Virol. 72, 3340–3350.

    PubMed  CAS  Google Scholar 

  14. Ghorpade, A., Xia, M. Q., Hyman, B. T., Persidsky, Y., Nukuna, A., Bock, P., et al. (1998) Role of the beta-chemokine receptors CCR3 and CCR5 in human immunodeficiency virus type 1 infection of monocytes and microglia. J. Virol. 72, 3351–3361.

    PubMed  CAS  Google Scholar 

  15. Ghorpade, A., Persidskaia, R., Suryadevara, R., Che, M., Liu, X. J., Persidsky, Y., et al. (2001) Mononuclear phagocyte differentiation, activation, and viral infection regulate matrix metalloproteinase expression: implications for human immunodeficiency virus type 1-associated dementia. J. Virol. 75, 6572–6583.

    Article  PubMed  CAS  Google Scholar 

  16. Peterson, P. K., Hu, S., Salak-Johnson, J., Molitor, T. W., and Chao, C. C. (1997) Differential production of and migratory response to beta chemokines by human microglia and astrocytes. J. Infect. Dis. 175, 478–481.

    Article  PubMed  CAS  Google Scholar 

  17. Lue, L. F., Brachova, L., Walker, D. G., and Rogers, J. (1996) Characterization of glial cultures from rapid autopsies of Alzheimer’s and control patients. Neurobiol. Aging 17, 421–429.

    Article  PubMed  CAS  Google Scholar 

  18. De Groot, C. J. A., Montagne, L., Janssen, I., Ravid, R., Van Der Valk, P., and Veerhuis, R. (2000) Isolation and characterization of adult microglial cells and oligodensrocytes derived from postmortem human brain tissue. Brain Res. Protocols 5, 85–94.

    Article  Google Scholar 

  19. Yong, V. W. and Antel, J. P. (1997) in Protocols for Neural Cell Culture (Fedoroff, S. and Richardson, A., eds.), Humana Press, Totowa, NJ. pp. 157–172.

    Google Scholar 

  20. Ghorpade, A., Bruch, L., Persidsky, Y., et al. (2005) Development of a rapid autopsy program for studies of brain immunity. J. Neuroimmunol., in press.

    Google Scholar 

  21. Ghorpade, A., Persidsky, Y., Swindells, S., et al. (2005) Investigations of neuroinflammatory responses from microglia recovered from HIV-1-infected and seronegative subjects. J. Neuroimmunol., in press.

    Google Scholar 

  22. Schuenke, K. and Gelman, B. B. (2003) Human microglial cell isolation from adult autopsy brain: brain pH, regional variation, and infection with human immunodeficiency virus type 1. J. Neurovirol. 9, 346–357.

    PubMed  Google Scholar 

  23. Suryadevara, R., Holter, S., Borgmann, K., Persidsky, R., Labenz-Zink, C., Persidsky, Y., et al. (2003) Regulation of tissue inhibitor of metalloproteinase-1 by astrocytes: Links to HIV-1 dementia. Glia 44, 47–56.

    Article  PubMed  Google Scholar 

  24. Zheng, J., Thylin, M., Ghorpade, A., Cotter, R., Persidsky, Y., and Gendelman, H. E. (1998) CXCR4 mediates neuronal dysfunction by HIV-1 infected macrophage secretory product: Importance for HIV-1 associated dementia. Soc. Neurosci. Abs. 24, 776.776 (Abstr).

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2005 Humana Press Inc.

About this protocol

Cite this protocol

Borgmann, K., Gendelman, H.E., Ghorpade, A. (2005). Isolation and HIV-1 Infection of Primary Human Microglia From Fetal and Adult Tissue. In: Zhu, T. (eds) Human Retrovirus Protocols. Methods in Molecular Biology™, vol 304. Humana Press. https://doi.org/10.1385/1-59259-907-9:049

Download citation

  • DOI: https://doi.org/10.1385/1-59259-907-9:049

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-58829-495-1

  • Online ISBN: 978-1-59259-907-3

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics