The online version of this article (doi:10.1186/1750-1326-9-58) contains supplementary material, which is available to authorized users.
Michael D Boska, Prasanta K Dash contributed equally to this work.
The authors declare that they have no competing interests.
MDB was responsible for the neuroimaging studies, provided all experimental design advice for each of the sections with HEG and participated directly in manuscript preparation and editing. PKD participated in the animal infections, blood sampling, virology, immunology and histology tested. He participated in behavioral testing and supported the manuscript preparation and editing. JK participated in histology and blood work analyses for the animals. AAE was involved in the neuroimaging and behavioral testing of animals. SPA was involved in histology and blood work for the animals. NF participated in histologic and immune testing of the animals. RH was responsible for design and performance of statistical analyses, EM participated in the histology, viral and immune studies for the animals. SB participated in the design and interpretation of behavioral tests and manuscript preparation, HAG was involved in experimental design and manuscript preparation and editing, LYP participated in experimental design, interpretation, and manuscript preparation, HEG designed the overall experimental schemes with MDB and participated in each part of the experimental design, interpretation, and manuscript preparation and editing, and SG was involve in mouse model preparation, experimental design and manuscript preparation and editing. All authors read and approved the final manuscript.
Host-species specificity of the human immunodeficiency virus (HIV) limits pathobiologic, diagnostic and therapeutic research investigations to humans and non-human primates. The emergence of humanized mice as a model for viral infection of the nervous system has overcome such restrictions enabling research for HIV-associated end organ disease including behavioral, cognitive and neuropathologic deficits reflective of neuroAIDS. Chronic HIV-1 infection of NOD/scid-IL-2Rgc null mice transplanted with human CD34+ hematopoietic stem cells (CD34-NSG) leads to persistent viremia, profound CD4+ T lymphocyte loss and infection of human monocyte-macrophages in the meninges and perivascular spaces. Murine cells are not infected with virus.
Changes in mouse behavior were measured, starting at 8 weeks after viral infection. These were recorded coordinate with magnetic resonance spectroscopy metabolites including N-acetylaspartate (NAA), creatine and choline. Diffusion tensor magnetic resonance imaging (DTI) was recorded against multispectral immunohistochemical staining for neuronal markers that included microtubule associated protein-2 (MAP2), neurofilament (NF) and synaptophysin (SYN); for astrocyte glial fibrillary acidic protein (GFAP); and for microglial ionized calcium binding adaptor molecule 1 (Iba-1). Oligodendrocyte numbers and integrity were measured for myelin associated glycoprotein (MAG) and myelin oligodendrocyte glycoprotein (MOG) antigens.
Behavioral abnormalities were readily observed in HIV-1 infected mice. Longitudinal open field activity tests demonstrated lack of habituation indicating potential for memory loss and persistent anxiety in HIV-1 infected mice compared to uninfected controls. End-point NAA and creatine in the cerebral cortex increased with decreased MAG. NAA and glutamate decreased with decreased SYN and MAG. Robust inflammation reflected GFAP and Iba-1 staining intensities. DTI metrics were coordinate with deregulation of NF, Iba-1, MOG and MAG levels in the whisker barrel and MAP2, NF, MAG, MOG and SYN in the corpus callosum.
The findings are consistent with some of the clinical, biochemical and pathobiologic features of human HIV-1 nervous system infections. This model will prove useful towards investigating the mechanisms of HIV-1 induced neuropathology and in developing novel biomarkers and therapeutic strategies for disease.
Additional file 1: Figure S1: Identification of HIV-1 infected human cells in spleen and brain. A. Five micron thick paraffin-embedded tissue sections were double florescent stained for human CD14 (macrophages) and HIV-1p24 (viral core protein, top panels) or human CD4 (T lymphocytes) and HIV-1p24 on the bottom panels. Merged pictures show HIV-1 infected (pictured in yellow) human CD14 or CD4 positive cells, original magnification 400×. B. Sections of brain tissues were peroxidase-stained for HLA-DR (human immunocytes) in meninges, brain parenchyma and perivascular spaces. HIV-1p24 cells are pictured on corresponding adjacent sections. Top insets show a magnified view of identified cells (see arrows). Top panel shows a HIV-1p24 cell simultaneously stained for human CD163 (macrophages). C. Sections showing perivascular human CD163 positive macrophages. B and C are at original magnification 200× and insets at 1000×. (DOCX 8 MB)13024_2014_569_MOESM1_ESM.docx
Additional file 2: Figure S2: Complete set of histology results comparing uninfected humanized mice (green, n = 10) to HIV-1 infected controls (red, n = 20) from A: M2 region of the cerebral cortex, B: Whisker barrel region of the cerebral cortex, C: Corpus callosum, D: CA1 region of the hippocampus, E: CA2 region of the hippocampus, F: CA3 region of the hippocampus, G: Dentate gyrus, H: Cerebellum and I: Brainstem. *Significant differences (p < 0.05). (DOCX 728 KB)13024_2014_569_MOESM2_ESM.docx
Additional file 3: Figure S3: Cerebral Cortex Metabolite Levels (Means ± SEM) expressed as a percentage of total signal contribution from 1H MRS scans of (red) HIV-1 infected humanized mice (n = 7) and (black) uninfected humanized mice (n = 7) over time. Time zero in infected mice is preinfection with subsequent spectra acquired every four weeks up to 16 weeks in both infected and uninfected mice. *p < 0.05 control vs infected mice, ^p < 0.05 vs time zero in control mice, (red “^” symbol) p < 0.05 vs preinfection in infected mice. (DOCX 459 KB)13024_2014_569_MOESM3_ESM.docx
Additional file 4: Figure S4: Cerebellum Metabolite Levels (Means ± SEM) expressed as a percentage of total signal contribution from 1H MRS scans of (red) HIV-1 infected humanized mice (n = 7) and (black) uninfected humanized mice (n = 7) over time. Time zero, in infected mice is preinfection with subsequent spectra acquired every four weeks up to 16 weeks in both infected and uninfected mice. *p < 0.05 control vs infected mice, ^p < 0.05 vs time zero in control mice, (red “^” symbol) p < 0.05 vs preinfection in infected mice. (DOCX 457 KB)13024_2014_569_MOESM4_ESM.docx
Additional file 5: Figure S5: Comparison of DTI metrics (mean ± SEM) in CA1, CA2, CA3, and Dentate Gyrus (from left to right) as shown in Figure 7 in uninfected (black, n = 7) and HIV-1 infected (red, n = 8) humanized mice. Shown are (from top to bottom) Mean diffusivity (Dav), Fractional anisotropy (FA), transverse component of diffusivity ((λt), and longitudinal component of diffusivity ((λl). (red “*” symbol) p < 0.05 control vs infected mice, tp < 0.05 vs time zero in control mice, (red “^” symbol) p < 0.05 vs preinfection in infected mice. (DOCX 612 KB)
Additional file 6: Figure S6: Comparison of DTI metrics (mean ± SEM) in frontal cortex, middle cerebral cortex, M2 region of cerebral cortex, and whisker barrels (from left to right) as shown in Figure 7 in uninfected (black, n = 7) and HIV-1 infected (red, n = 8) humanized mice. Shown are (from top to bottom) Mean diffusivity (Dav), Fractional anisotropy (FA), transverse component of diffusivity (λt), and longitudinal component of diffusivity (λl). (red “*” symbol) p < 0.05 control vs infected mice, tp < 0.05 vs time zero in control mice, (red “t” symbol) p < 0.05 vs preinfection in infected mice. (DOCX 685 KB)
Additional file 7: Figure S7: Comparison of DTI metrics over time (mean ± SEM) from left to right in the Genu of the Corpus Callosum (CC) and the Splenium of the CC, as shown in Figure 7. Results are shown from uninfected (black, n = 7) and HIV-1 infected (red, n = 8) humanized mice. Shown are (from top to bottom) Mean diffusivity (Dav), Fractional anisotropy (FA), transverse component of diffusivity (λt), and longitudinal component of diffusivity ((λl). (red “*” symbol) p < 0.05 control vs infected mice, tp < 0.05 vs time zero in control mice, (red “t” symbol) p < 0.05 vs preinfection in infected mice. (DOCX 347 KB)
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- Associations between brain microstructures, metabolites, and cognitive deficits during chronic HIV-1 infection of humanized mice
Michael D Boska
Prasanta K Dash
Adrian A Epstein
Sidra P Akhter
Harris A Gelbard
Larisa Y Poluektova
Howard E Gendelman
- BioMed Central
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