HIV Protocols pp 367-376 | Cite as

Evaluating the Role of Viral Proteins in HIV-Mediated Neurotoxicity Using Primary Human Neuronal Cultures

  • Vasudev R. Rao
  • Eliseo A. Eugenin
  • Vinayaka R. Prasad
Part of the Methods in Molecular Biology book series (MIMB, volume 1354)

Abstract

Despite the inability of HIV-1 to infect neurons, over half of the HIV-1-infected population in the USA suffers from neurocognitive dysfunction. HIV-infected immune cells in the periphery enter the central nervous system by causing a breach in the blood–brain barrier. The damage to the neurons is mediated by viral and host toxic products released by activated and infected immune and glial cells. To evaluate the toxicity of any viral isolate, viral protein, or host inflammatory protein, we describe a protocol to assess the neuronal apoptosis and synaptic compromise in primary cultures of human neurons and astrocytes.

Key words

HIV-1 Neuronal damage Neuronal apoptosis Neuro-inflammation HIV-associated dementia HAD HIV-associated neurocognitive dysfunction HAND gp120 Tat Cytokines Chemokines 

Notes

Acknowledgements

This work was supported by NIH R01 MH083579 and R21 MH101003 (to V.R.P.). The authors would like to thank Arthur Ruiz for critically reading the manuscript.

References

  1. 1.
    Heaton RK, Franklin DR, Ellis RJ, McCutchan JA, Letendre SL, Leblanc S, Corkran SH, Duarte NA, Clifford DB, Woods SP, Collier AC, Marra CM, Morgello S, Mindt MR, Taylor MJ, Marcotte TD, Atkinson JH, Wolfson T, Gelman BB, McArthur JC, Simpson DM, Abramson I, Gamst A, Fennema-Notestine C, Jernigan TL, Wong J, Grant I, Group C, Group H (2011) HIV-associated neurocognitive disorders before and during the era of combination antiretroviral therapy: differences in rates, nature, and predictors. J Neurovirol 17:3–16. doi: 10.1007/s13365-010-0006-1 CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Resnick L, Berger JR, Shapshak P, Tourtellotte WW (1988) Early penetration of the blood-brain-barrier by HIV. Neurology 38:9–14CrossRefPubMedGoogle Scholar
  3. 3.
    Airoldi M, Bandera A, Trabattoni D, Tagliabue B, Arosio B, Soria A, Rainone V, Lapadula G, Annoni G, Clerici M, Gori A (2012) Neurocognitive impairment in HIV-infected naïve patients with advanced disease: the role of virus and intrathecal immune activation. Clin Dev Immunol 2012:467154. doi: 10.1155/2012/467154 CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    An SF, Groves M, Gray F, Scaravilli F (1999) Early entry and widespread cellular involvement of HIV-1 DNA in brains of HIV-1 positive asymptomatic individuals. J Neuropathol Exp Neurol 58:1156–1162CrossRefPubMedGoogle Scholar
  5. 5.
    Meltzer MS, Skillman DR, Gomatos PJ, Kalter DC, Gendelman HE (1990) Role of mononuclear phagocytes in the pathogenesis of human immunodeficiency virus infection. Annu Rev Immunol 8:169–194. doi: 10.1146/annurev.iy.08.040190.001125 CrossRefPubMedGoogle Scholar
  6. 6.
    Toborek M, Lee YW, Flora G, Pu H, Andras IE, Wylegala E, Hennig B, Nath A (2005) Mechanisms of the blood-brain barrier disruption in HIV-1 infection. Cell Mol Neurobiol 25:181–199CrossRefPubMedGoogle Scholar
  7. 7.
    Rao VR, Ruiz AP, Prasad VR (2014) Viral and cellular factors underlying neuropathogenesis in HIV associated neurocognitive disorders (HAND). AIDS Res Ther 11:13. doi: 10.1186/1742-6405-11-13 CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Rao VR, Neogi U, Eugenin E, Prasad VR (2014) The gp120 protein is a second determinant of decreased neurovirulence of Indian HIV-1C isolates compared to Southern African HIV-1C isolates. PLoS One 9:e107074. doi: 10.1371/journal.pone.0107074 CrossRefPubMedPubMedCentralGoogle Scholar
  9. 9.
    Rao VR, Neogi U, Talboom JS, Padilla L, Rahman M, Fritz-French C, Gonzalez-Ramirez S, Verma A, Wood C, Ruprecht RM, Ranga U, Azim T, Joska J, Eugenin E, Shet A, Bimonte-Nelson H, Tyor WR, Prasad VR (2013) Clade C HIV-1 isolates circulating in Southern Africa exhibit a greater frequency of dicysteine motif-containing Tat variants than those in Southeast Asia and cause increased neurovirulence. Retrovirology 10:61. doi: 10.1186/1742-4690-10-61 CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Vasudev R. Rao
    • 1
  • Eliseo A. Eugenin
    • 2
    • 3
  • Vinayaka R. Prasad
    • 1
  1. 1.Department of Microbiology and ImmunologyAlbert Einstein College of MedicineBronxUSA
  2. 2.Public Health Research Institute (PHRI)Rutgers The State University of New JerseyNewarkUSA
  3. 3.Department of Microbiology and Molecular GeneticsRutgers The State University of New JerseyNewarkUSA

Personalised recommendations