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HIV-1 causes CD4 cell death through DNA-dependent protein kinase during viral integration



Human immunodeficiency virus-1 (HIV-1) has infected more than 60 million people and caused nearly 30 million deaths worldwide1, ultimately the consequence of cytolytic infection of CD4+ T cells. In humans and in macaque models, most of these cells contain viral DNA and are rapidly eliminated at the peak of viraemia2,3,4, yet the mechanism by which HIV-1 induces helper T-cell death has not been defined. Here we show that virus-induced cell killing is triggered by viral integration. Infection by wild-type HIV-1, but not an integrase-deficient mutant, induced the death of activated primary CD4 lymphocytes. Similarly, raltegravir, a pharmacologic integrase inhibitor, abolished HIV-1-induced cell killing both in cell culture and in CD4+ T cells from acutely infected subjects. The mechanism of killing during viral integration involved the activation of DNA-dependent protein kinase (DNA-PK), a central integrator of the DNA damage response, which caused phosphorylation of p53 and histone H2AX. Pharmacological inhibition of DNA-PK abolished cell death during HIV-1 infection in vitro, suggesting that processes which reduce DNA-PK activation in CD4 cells could facilitate the formation of latently infected cells that give rise to reservoirs in vivo. We propose that activation of DNA-PK during viral integration has a central role in CD4+ T-cell depletion, raising the possibility that integrase inhibitors and interventions directed towards DNA-PK may improve T-cell survival and immune function in infected individuals.

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Figure 1: CD4 lymphocytes killed during HIV-1 infection do not express viral gene products.
Figure 2: Dying CD4 lymphocytes lacking viral gene expression had been productively infected prior to cell death.
Figure 3: Proviral DNA integration triggers cell death during HIV-1 infection.
Figure 4: DNA-PK orchestrates a DNA damage response and cell death following proviral DNA integration.


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We thank D. Ambrozak, R. Nguyen, and S. Perfetto for help with cell sorting, U. Olshevsky, J. Casazza, D. Bolton, A. Pegu and M. Louder for discussions and technical help, and A. Tislerics and B. Hartman for manuscript preparation. This research was supported by the Intramural Research Program of the Vaccine Research Center, NIAID, National Institutes of Health. The findings and conclusions in this report are those of the authors and do not necessarily reflect the views of the funding agency.

Author information




A.C. and G.J.N. designed the research studies; A.C., M.G, C.P. and T.Y. performed the research; A.C., M.G. and T.Y. contributed to development and generation of vectors; A.C., C.P., R.A.K. and G.J.N. analysed data; and A.C., R.A.K. and G.J.N. wrote the paper.

Corresponding author

Correspondence to Gary J. Nabel.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figures 1-4 and Supplementary References. These figures show that HIV-1 induced cell death depends on provirus integration and is observed in p24- CD4+ lymphocytes from healthy donors infected in vitro and patient samples ex vivo. Blocking death by DNA-PK inhibitors does not result from diminished viral replication or general resistance to cell death. (PDF 621 kb)

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Cooper, A., García, M., Petrovas, C. et al. HIV-1 causes CD4 cell death through DNA-dependent protein kinase during viral integration. Nature 498, 376–379 (2013).

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