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PVSRIPO directs infection of, non-cytopathogenic vRNA replication in, and IFN-I signaling in the microglial compartment in vivo . (A-C) Flow cytometry analyses of microglia before/after CD11b + -bead isolation from hCD155 -tg mouse (A, C) or <t>human</t> <t>brain</t> <t>tissue</t> (B) . The same kit and identical procedures were used to process human/mouse brain tissue samples (see Materials and methods ). Consistently, ∼90-95% of CD45 hi , CD11b + , MERTK + , Cx3cx1 + (A, B) and Tmem119 + (C) cells were recovered from brain tissue of either species. (D) RT-qPCR analyses of total RNA from hCD155 -tg murine or human microglia as shown. Microglia were either infected ex vivo [multiplicity of infection (MOI) 10; left and middle panels], or CD11b + -bead isolated after infection in vivo [single intracerebral inoculation of PVSRIPO (5 x 10 7 pfu); right panel]. Ipsilateral (to the site of virus inoculation) and contralateral hemispheres were processed separately. vRNA levels in mouse microglia were analyzed by two-tailed Mann-Whitney test on ΔΔCt values at 8 weeks pi (n=5; left panel); the same test was used to analyze vRNA levels in human microglia at 72hpi (n=3; right middle panel). (E) HCR-FISH analyses of vRNA species in human CD11b + -bead isolated microglia infected ex vivo with PVSRIPO (MOI 10). Two individual microglia cells at 24 and 72hpi, each, are shown. (F-G) IF and HCR-FISH analyses in non-tumor-bearing hCD155 -tg mouse brains 48h post single intracerebral PVSRIPO infusion (5 x 10 7 pfu). Tmem119 + microglia stain positive for (-)strand vRNA (F) with widespread ISG15 induction in the microglial compartment (G) .
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PVSRIPO directs infection of, non-cytopathogenic vRNA replication in, and IFN-I signaling in the microglial compartment in vivo . (A-C) Flow cytometry analyses of microglia before/after CD11b + -bead isolation from hCD155 -tg mouse (A, C) or <t>human</t> <t>brain</t> <t>tissue</t> (B) . The same kit and identical procedures were used to process human/mouse brain tissue samples (see Materials and methods ). Consistently, ∼90-95% of CD45 hi , CD11b + , MERTK + , Cx3cx1 + (A, B) and Tmem119 + (C) cells were recovered from brain tissue of either species. (D) RT-qPCR analyses of total RNA from hCD155 -tg murine or human microglia as shown. Microglia were either infected ex vivo [multiplicity of infection (MOI) 10; left and middle panels], or CD11b + -bead isolated after infection in vivo [single intracerebral inoculation of PVSRIPO (5 x 10 7 pfu); right panel]. Ipsilateral (to the site of virus inoculation) and contralateral hemispheres were processed separately. vRNA levels in mouse microglia were analyzed by two-tailed Mann-Whitney test on ΔΔCt values at 8 weeks pi (n=5; left panel); the same test was used to analyze vRNA levels in human microglia at 72hpi (n=3; right middle panel). (E) HCR-FISH analyses of vRNA species in human CD11b + -bead isolated microglia infected ex vivo with PVSRIPO (MOI 10). Two individual microglia cells at 24 and 72hpi, each, are shown. (F-G) IF and HCR-FISH analyses in non-tumor-bearing hCD155 -tg mouse brains 48h post single intracerebral PVSRIPO infusion (5 x 10 7 pfu). Tmem119 + microglia stain positive for (-)strand vRNA (F) with widespread ISG15 induction in the microglial compartment (G) .
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PVSRIPO directs infection of, non-cytopathogenic vRNA replication in, and IFN-I signaling in the microglial compartment in vivo . (A-C) Flow cytometry analyses of microglia before/after CD11b + -bead isolation from hCD155 -tg mouse (A, C) or <t>human</t> <t>brain</t> <t>tissue</t> (B) . The same kit and identical procedures were used to process human/mouse brain tissue samples (see Materials and methods ). Consistently, ∼90-95% of CD45 hi , CD11b + , MERTK + , Cx3cx1 + (A, B) and Tmem119 + (C) cells were recovered from brain tissue of either species. (D) RT-qPCR analyses of total RNA from hCD155 -tg murine or human microglia as shown. Microglia were either infected ex vivo [multiplicity of infection (MOI) 10; left and middle panels], or CD11b + -bead isolated after infection in vivo [single intracerebral inoculation of PVSRIPO (5 x 10 7 pfu); right panel]. Ipsilateral (to the site of virus inoculation) and contralateral hemispheres were processed separately. vRNA levels in mouse microglia were analyzed by two-tailed Mann-Whitney test on ΔΔCt values at 8 weeks pi (n=5; left panel); the same test was used to analyze vRNA levels in human microglia at 72hpi (n=3; right middle panel). (E) HCR-FISH analyses of vRNA species in human CD11b + -bead isolated microglia infected ex vivo with PVSRIPO (MOI 10). Two individual microglia cells at 24 and 72hpi, each, are shown. (F-G) IF and HCR-FISH analyses in non-tumor-bearing hCD155 -tg mouse brains 48h post single intracerebral PVSRIPO infusion (5 x 10 7 pfu). Tmem119 + microglia stain positive for (-)strand vRNA (F) with widespread ISG15 induction in the microglial compartment (G) .
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PVSRIPO directs infection of, non-cytopathogenic vRNA replication in, and IFN-I signaling in the microglial compartment in vivo . (A-C) Flow cytometry analyses of microglia before/after CD11b + -bead isolation from hCD155 -tg mouse (A, C) or <t>human</t> <t>brain</t> <t>tissue</t> (B) . The same kit and identical procedures were used to process human/mouse brain tissue samples (see Materials and methods ). Consistently, ∼90-95% of CD45 hi , CD11b + , MERTK + , Cx3cx1 + (A, B) and Tmem119 + (C) cells were recovered from brain tissue of either species. (D) RT-qPCR analyses of total RNA from hCD155 -tg murine or human microglia as shown. Microglia were either infected ex vivo [multiplicity of infection (MOI) 10; left and middle panels], or CD11b + -bead isolated after infection in vivo [single intracerebral inoculation of PVSRIPO (5 x 10 7 pfu); right panel]. Ipsilateral (to the site of virus inoculation) and contralateral hemispheres were processed separately. vRNA levels in mouse microglia were analyzed by two-tailed Mann-Whitney test on ΔΔCt values at 8 weeks pi (n=5; left panel); the same test was used to analyze vRNA levels in human microglia at 72hpi (n=3; right middle panel). (E) HCR-FISH analyses of vRNA species in human CD11b + -bead isolated microglia infected ex vivo with PVSRIPO (MOI 10). Two individual microglia cells at 24 and 72hpi, each, are shown. (F-G) IF and HCR-FISH analyses in non-tumor-bearing hCD155 -tg mouse brains 48h post single intracerebral PVSRIPO infusion (5 x 10 7 pfu). Tmem119 + microglia stain positive for (-)strand vRNA (F) with widespread ISG15 induction in the microglial compartment (G) .
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PVSRIPO directs infection of, non-cytopathogenic vRNA replication in, and IFN-I signaling in the microglial compartment in vivo . (A-C) Flow cytometry analyses of microglia before/after CD11b + -bead isolation from hCD155 -tg mouse (A, C) or <t>human</t> <t>brain</t> <t>tissue</t> (B) . The same kit and identical procedures were used to process human/mouse brain tissue samples (see Materials and methods ). Consistently, ∼90-95% of CD45 hi , CD11b + , MERTK + , Cx3cx1 + (A, B) and Tmem119 + (C) cells were recovered from brain tissue of either species. (D) RT-qPCR analyses of total RNA from hCD155 -tg murine or human microglia as shown. Microglia were either infected ex vivo [multiplicity of infection (MOI) 10; left and middle panels], or CD11b + -bead isolated after infection in vivo [single intracerebral inoculation of PVSRIPO (5 x 10 7 pfu); right panel]. Ipsilateral (to the site of virus inoculation) and contralateral hemispheres were processed separately. vRNA levels in mouse microglia were analyzed by two-tailed Mann-Whitney test on ΔΔCt values at 8 weeks pi (n=5; left panel); the same test was used to analyze vRNA levels in human microglia at 72hpi (n=3; right middle panel). (E) HCR-FISH analyses of vRNA species in human CD11b + -bead isolated microglia infected ex vivo with PVSRIPO (MOI 10). Two individual microglia cells at 24 and 72hpi, each, are shown. (F-G) IF and HCR-FISH analyses in non-tumor-bearing hCD155 -tg mouse brains 48h post single intracerebral PVSRIPO infusion (5 x 10 7 pfu). Tmem119 + microglia stain positive for (-)strand vRNA (F) with widespread ISG15 induction in the microglial compartment (G) .
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PVSRIPO directs infection of, non-cytopathogenic vRNA replication in, and IFN-I signaling in the microglial compartment in vivo . (A-C) Flow cytometry analyses of microglia before/after CD11b + -bead isolation from hCD155 -tg mouse (A, C) or human brain tissue (B) . The same kit and identical procedures were used to process human/mouse brain tissue samples (see Materials and methods ). Consistently, ∼90-95% of CD45 hi , CD11b + , MERTK + , Cx3cx1 + (A, B) and Tmem119 + (C) cells were recovered from brain tissue of either species. (D) RT-qPCR analyses of total RNA from hCD155 -tg murine or human microglia as shown. Microglia were either infected ex vivo [multiplicity of infection (MOI) 10; left and middle panels], or CD11b + -bead isolated after infection in vivo [single intracerebral inoculation of PVSRIPO (5 x 10 7 pfu); right panel]. Ipsilateral (to the site of virus inoculation) and contralateral hemispheres were processed separately. vRNA levels in mouse microglia were analyzed by two-tailed Mann-Whitney test on ΔΔCt values at 8 weeks pi (n=5; left panel); the same test was used to analyze vRNA levels in human microglia at 72hpi (n=3; right middle panel). (E) HCR-FISH analyses of vRNA species in human CD11b + -bead isolated microglia infected ex vivo with PVSRIPO (MOI 10). Two individual microglia cells at 24 and 72hpi, each, are shown. (F-G) IF and HCR-FISH analyses in non-tumor-bearing hCD155 -tg mouse brains 48h post single intracerebral PVSRIPO infusion (5 x 10 7 pfu). Tmem119 + microglia stain positive for (-)strand vRNA (F) with widespread ISG15 induction in the microglial compartment (G) .

Journal: bioRxiv

Article Title: Viral Microglia Reprogramming Clears Oligomeric Neurotoxic Debris

doi: 10.64898/2026.04.06.716590

Figure Lengend Snippet: PVSRIPO directs infection of, non-cytopathogenic vRNA replication in, and IFN-I signaling in the microglial compartment in vivo . (A-C) Flow cytometry analyses of microglia before/after CD11b + -bead isolation from hCD155 -tg mouse (A, C) or human brain tissue (B) . The same kit and identical procedures were used to process human/mouse brain tissue samples (see Materials and methods ). Consistently, ∼90-95% of CD45 hi , CD11b + , MERTK + , Cx3cx1 + (A, B) and Tmem119 + (C) cells were recovered from brain tissue of either species. (D) RT-qPCR analyses of total RNA from hCD155 -tg murine or human microglia as shown. Microglia were either infected ex vivo [multiplicity of infection (MOI) 10; left and middle panels], or CD11b + -bead isolated after infection in vivo [single intracerebral inoculation of PVSRIPO (5 x 10 7 pfu); right panel]. Ipsilateral (to the site of virus inoculation) and contralateral hemispheres were processed separately. vRNA levels in mouse microglia were analyzed by two-tailed Mann-Whitney test on ΔΔCt values at 8 weeks pi (n=5; left panel); the same test was used to analyze vRNA levels in human microglia at 72hpi (n=3; right middle panel). (E) HCR-FISH analyses of vRNA species in human CD11b + -bead isolated microglia infected ex vivo with PVSRIPO (MOI 10). Two individual microglia cells at 24 and 72hpi, each, are shown. (F-G) IF and HCR-FISH analyses in non-tumor-bearing hCD155 -tg mouse brains 48h post single intracerebral PVSRIPO infusion (5 x 10 7 pfu). Tmem119 + microglia stain positive for (-)strand vRNA (F) with widespread ISG15 induction in the microglial compartment (G) .

Article Snippet: Microglia were isolated from freshly dissected mouse brains or from epilepsy surgery human brain tissues using a CD11b + -bead isolation kit (Miltenyi MicroBeads, human & mouse; #130-097-142) as described in detail previously ( ).

Techniques: Infection, In Vivo, Flow Cytometry, Isolation, Quantitative RT-PCR, Ex Vivo, Virus, Two Tailed Test, MANN-WHITNEY, Staining