Dynamics of Compartmentalized HIV-1 Populations in the Central Nervous System Patrick R. Harrington
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1 Dynamics of Compartmentalized HIV-1 Populations in the Central Nervous System Patrick R. Harrington Postdoctoral Fellow Laboratory of Ronald Swanstrom Lineberger Comprehensive Cancer Center University of North Carolina at Chapel Hill Chapel Hill, NC, USA
2 Dynamics of Compartmentalized HIV-1 Populations in the Central Nervous System Kimberly Ritola, 1 Kevin Robertson, 2 Susan A. Fiscus, 1 Colin Hall, 2 and Ronald Swanstrom. 1 Increased HIV-1 env compartmentalization in the presence of HIV-associated dementia. Journal of Virology. In press (Aug. 2005). Patrick R. Harrington, 1 David W. Haas, 3 Kimberly Ritola, 1 and Ronald Swanstrom. 1 Compartmentalized HIV-1 present in cerebrospinal fluid is produced by short-lived cells. Journal of Virology. In press (July 2005). 1 Lineberger Comprehensive Cancer Center, Center for AIDS Research, Department of Biochemistry and Biophysics, and 2 Department of Neurology, University of North Carolina, Chapel Hill, North Carolina, USA 3 Departments of Medicine, Microbiology and Immunology, and Vanderbilt Meharry Center for AIDS Research, Vanderbilt University School of Medicine, Nashville, Tennessee, USA Funding Lineberger Cancer Center, UNC-Chapel Hill NIMH, NINDS, NIH UNC Center for AIDS Research Vanderbilt Meharry Center for AIDS Research
3 Detection of Viral Genetic Populations by HTA
4 Compartmentalization of HIV-1 Genetic Populations in Blood Plasma and CSF Revealed by V1/V2 (env) HTA Similar env populations BP CSF Unique env populations in plasma Unique env populations in CSF Unique env populations in plasma and CSF BP CSF BP CSF BP CSF *single-stranded probe band unique to compartment
5 Increased Compartmentalization Between Plasma and CSF HIV-1 env Populations is Associated with Neurological Status Patients with compartmentalized env sequences Relative Abundance of Unique env Variants in CSF 100% % of Patients 80% 60% 40% 20% Unique Variants in Plasma and CSF Unique Variants in CSF Unique Variants in Plasma All Variants Shared 0% Primary Infection No CNS Disease MCMD HAD HAD No HAD Neurological Status
6 Dynamics of HIV-1 Populations in Blood and CSF of Asymptomatic Subjects During the Initiation of Antiretroviral Therapy
7 At Least Two Phases of Viral Decline in the Periphery During Antiretroviral Therapy Sources of HIV-1 RNA free virus short-lived, productively infected cells (CD4+ T cells) long-lived infected cells (Macrophages, resting T cells) Change in log plasma HIV RNA Limit of detection Time after initiation of antiretroviral therapy (Perelson, et al., Shaw, et al. 1995) Adapted from HIVpositive.com
8 Antiretroviral Therapy and HIV-1 in cerebrospinal fluid Studies of Bulk HIV-1 RNA in CSF Usually a rapid decline in CSF viral load during therapy Slower decline Neurological disease (Ellis, et. al. 2000; Eggers, et. al. 2003) Kinetics of CSF viral decay do not correlate with blood Not known which source is being treated when bulk viral loads decline in CSF Experimental Design: use HTA to measure the decline of individual HIV-1 genetic variants in blood and CSF Periphery CNS CD4 + T cells Macrophages CSF Macrophages Perivascular Microglia Astrocytes Neurons Hypothesis CSF HIV-1 variants produced by cells within the CNS will decline more slowly in response to antiretroviral therapy compared with CSF HIV-1 coming from peripheral sources.
9 Haas Study-4 treatment-naïve, asymptomatic subjects CSF/Plasma Sampling Start CSF/Plasma Sampling End Three-drug therapy Start CSF/Plasma Sampling Start CSF/Plasma Sampling End Baseline hours postinitiation of therapy Time (Hours)
10 HIV-1 Compartmentalization in the CNS BP CSF BP CSF # # # Relative Variant Load (copies/ml) single-stranded probe *unique variant in CSF # enriched variant in CSF
11 Decline of HIV-1 Genetic Variants During Antiretroviral Therapy: Compartmentalized HIV-1 Variants in CSF Decline Rapidly Patient 2 V4/V5 Variants Decline of CSF HIV-1 Variants (h) # Relative Variant Load (copies/ml) Time on therapy (h) CSF HIV-1 variants shared with the periphery Compartmentalized HIV-1 Variants in CSF
12 Decline of HIV-1 Variants in CSF During Antiretroviral Therapy CSF variants shared with blood Compartmentalized CSF variants Subject 1 2, % decrease t 1/2 max (h) % decrease t 1/2 max (h) t 1/2 p value Conclusion A short-lived, HIV-productively infected cell population exists in the central nervous system and contributes to the HIV-1 population in cerebrospinal fluid.
13 Dynamics of HIV-1 Populations in the CNS
14 Dynamics of HIV-1 Populations in the CNS-HAD?
15 Dynamics of Compartmentalized HIV-1 Populations in the Central Nervous System Conclusions 1. The amount of CNS-specific production of HIV-1 in CSF is associated with neurological disease status. 2. In asymptomatic subjects, short-lived cells (perhaps trafficking CD4 + T cells) play an important role in the production and maintenance of locally produced HIV-1 populations in CSF. Grazie molto per la vostra attenzione! (Thank you very much for your attention)
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