bafilomycin-a1 and Encephalitis--Viral

Autophagy is critical to maintaining cell homeostasis and is implicated in neurodegenerative diseases. This research examined the role of autophagy in human immunodeficiency virus type 1 (HIV-1)-associated encephalitis, the pathologic hallmark of neuroAIDS.. The frontal cortex from 32 HIV-infected persons (12 without evidence HIV-1 encephalitis or clinical signs of central nervous system impairment and 20 with histopathological findings of HIV-1 encephalitis) and 8 persons without HIV infection and any neuropathology were examined postmortem. Green fluorescent protein-labeled (GFP) light chain 3 (LC3)-expressing neuroblastoma SK-N-SH cells treated with gp120 from CXCR4 and CCR5 viruses were also examined. Autophagic markers were assessed by means of Western blot analysis, transmission electron microscopy (TEM), and confocal microscopy.. Autophagic proteins Beclin 1, Autophagy-related gene (Atg)-5, Atg-7, and LC3-II were significantly increased in brains with HIV-1 encephalitis (P < .05). These findings were confirmed by TEM and immunostaining of brain tissue. Additionally, levels of autophagic proteins and autophagosomes were increased in neuronal cells treated with both CXCR4- or CCR5-tropic HIV-1 gp120. No increase in the level of autophagy was observed in the brains of HIV-infected persons without HIV-1 encephalitis compared with the level in brains of HIV-uninfected persons.. Postmortem brains with HIV-1 encephalitis exhibit increased markers of autophagy compared with brains from HIV-infected persons without HIV-1 encephalitis or HIV-uninfected control brains, which suggests that dysregulation of autophagy may be important in the pathogenesis of neuroAIDS.

Topics: Adult; Aged; Autophagy; Autopsy; Blotting, Western; Case-Control Studies; Cell Line, Tumor; Encephalitis, Viral; Female; Frontal Lobe; HIV Envelope Protein gp120; HIV Infections; Humans; Immunohistochemistry; Lysosomal Membrane Proteins; Lysosomes; Macrolides; Male; Microtubule-Associated Proteins; Middle Aged; Phagosomes

We studied the effect of bafilomycin A1 (Baf-A1), a novel and highly specific inhibitor for vacuolar-type proton (V-H+) pump, on the growth of Japanese Encephalitis virus (JEV) in Vero cells. Viral fluorescence microscopic study showed that Baf-A1 induced the complete disappearance of acidified compartments such as endosomes and lysosomes in Vero cells by the treatment with 0.1 microM Baf-A1 for 1 h at 37 degrees C. In proportion to the disappearance of acidified compartments, virus growth was inhibited when Baf-A1 was present from 1 h before infection to the end of incubation in a dose-dependent manner, or added within as early as 5 min after infection. Conversely, the virus growth was recovered in correlation with the reappearance of acidified compartments after removal of Baf-A1. These results suggest that a low pH condition, which is regulated by Baf-A1-sensitive V-H+ pumps, is essential for the early stage of JEV growth.

Topics: Acids; Animals; Anti-Bacterial Agents; Chlorocebus aethiops; Encephalitis Virus, Japanese; Encephalitis, Viral; Endosomes; Enzyme Inhibitors; Macrolides; Proton Pumps; Vacuoles; Vero Cells