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DeltaRR vaccination protects from KA-induced seizures and neuronal loss through ICP10PK-mediated modulation of the neuronal-microglial axis.

Laing JM, Aurelian L - Genet Vaccines Ther (2008)

Bottom Line: The CM from Delta PK-infected EOC2 and EOC20 cells did not contain IL-10, but it contained TNF-alpha and RANTES.IL-10 neutralization significantly (p < 0.01) decreased, but did not abrogate, the neuroprotective activity of the CM from Delta RR-infected microglial cultures indicating that ICP10PK modulates the neuronal-microglial axis, also through induction of various microglial neuroprotective factors.Protection was associated with a significant (p < 0.001) increase in the numbers of IL-10+ microglia (CD11b+) as compared to Delta PK-treated animals.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pharmacology and Experimental Therapeutics, University of Maryland, School of Medicine, Baltimore, MD 21201, USA. jlain001@umaryland.edu

ABSTRACT
Ischemic brain injury and epilepsy are common neurodegenerative diseases caused by excitotoxicity. Their pathogenesis includes microglial production of inflammatory cytokines. Our studies were designed to examine whether a growth compromised HSV-2 mutant (Delta RR) prevents excitotoxic injury through modulation of microglial responses by the anti-apoptotic HSV-2 protein ICP10PK. EOC2 and EOC20 microglial cells, which are differentially activated, were infected with Delta RR or the ICP10PK deleted virus (Delta PK) and examined for virus-induced neuroprotective activity. Both cell lines were non-permissive for virus growth, but expressed ICP10PK (Delta RR) or the PK deleted ICP10 protein p95 (Delta PK). Conditioned medium (CM) from Delta RR-, but not Delta PK-infected cells prevented N-methyl-D-aspartate (NMDA)-induced apoptosis of primary hippocampal cultures, as determined by TUNEL and caspase-3 activation (76.9 +/- 5.3% neuroprotection). Neuroprotection was associated with inhibition of TNF-alpha and RANTES and production of IL-10. The CM from Delta PK-infected EOC2 and EOC20 cells did not contain IL-10, but it contained TNF-alpha and RANTES. IL-10 neutralization significantly (p < 0.01) decreased, but did not abrogate, the neuroprotective activity of the CM from Delta RR-infected microglial cultures indicating that ICP10PK modulates the neuronal-microglial axis, also through induction of various microglial neuroprotective factors. Rats given Delta RR (but not Delta PK) by intranasal inoculation were protected from kainic acid (KA)-induced seizures and neuronal loss in the CA1 hippocampal fields. Protection was associated with a significant (p < 0.001) increase in the numbers of IL-10+ microglia (CD11b+) as compared to Delta PK-treated animals. Delta RR is a promising vaccination/therapy platform for neurodegeneration through its pro-survival functions in neurons as well as microglia modulation.

No MeSH data available.


Related in: MedlinePlus

RANTES production is inhibited in ΔRR-infected microglia. EOC2 and EOC20 cells were mock-infected with PBS, or infected with ΔRR, ΔPK (moi = 5) or mock-infected with PBS and culture supernatants collected 1–72 hrs p.i. were assayed for RANTES by ELISA, as described in Materials and Methods. Results are the mean of three independent experiments ± SD. (***p < 0.001 relative to ΔRR-infected).
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Figure 6: RANTES production is inhibited in ΔRR-infected microglia. EOC2 and EOC20 cells were mock-infected with PBS, or infected with ΔRR, ΔPK (moi = 5) or mock-infected with PBS and culture supernatants collected 1–72 hrs p.i. were assayed for RANTES by ELISA, as described in Materials and Methods. Results are the mean of three independent experiments ± SD. (***p < 0.001 relative to ΔRR-infected).

Mentions: RANTES/CCL5 is a member of the C-C (β) chemokine family, which is believed to contribute to the recruitment of T cells and monocytes from the periphery into the CNS. RANTES is produced by microglia in response to pro-inflammatory stimuli [32]. Having seen that TNF-α production is inhibited in ΔRR-, but not ΔPK-infected EOC20 cells, we wanted to know whether this is also true for RANTES. Duplicate samples of the CM from the mock- or virus-infected EOC2 and EOC20 cells were assayed for RANTES by ELISA. RANTES was produced in both EOC2 and EOC20 cells infected with ΔPK. Its levels were significantly (2-fold) higher in EOC2 than EOC20 cells (Fig. 6), suggesting that its regulation is distinct from that of TNF-α. Significantly, however, RANTES was not seen in CM from ΔRR-infected EOC2 or EOC20 cells (Fig. 6), indicating that ICP10PK inhibits its production, independent of the cell activation state.


DeltaRR vaccination protects from KA-induced seizures and neuronal loss through ICP10PK-mediated modulation of the neuronal-microglial axis.

Laing JM, Aurelian L - Genet Vaccines Ther (2008)

RANTES production is inhibited in ΔRR-infected microglia. EOC2 and EOC20 cells were mock-infected with PBS, or infected with ΔRR, ΔPK (moi = 5) or mock-infected with PBS and culture supernatants collected 1–72 hrs p.i. were assayed for RANTES by ELISA, as described in Materials and Methods. Results are the mean of three independent experiments ± SD. (***p < 0.001 relative to ΔRR-infected).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2268933&req=5

Figure 6: RANTES production is inhibited in ΔRR-infected microglia. EOC2 and EOC20 cells were mock-infected with PBS, or infected with ΔRR, ΔPK (moi = 5) or mock-infected with PBS and culture supernatants collected 1–72 hrs p.i. were assayed for RANTES by ELISA, as described in Materials and Methods. Results are the mean of three independent experiments ± SD. (***p < 0.001 relative to ΔRR-infected).
Mentions: RANTES/CCL5 is a member of the C-C (β) chemokine family, which is believed to contribute to the recruitment of T cells and monocytes from the periphery into the CNS. RANTES is produced by microglia in response to pro-inflammatory stimuli [32]. Having seen that TNF-α production is inhibited in ΔRR-, but not ΔPK-infected EOC20 cells, we wanted to know whether this is also true for RANTES. Duplicate samples of the CM from the mock- or virus-infected EOC2 and EOC20 cells were assayed for RANTES by ELISA. RANTES was produced in both EOC2 and EOC20 cells infected with ΔPK. Its levels were significantly (2-fold) higher in EOC2 than EOC20 cells (Fig. 6), suggesting that its regulation is distinct from that of TNF-α. Significantly, however, RANTES was not seen in CM from ΔRR-infected EOC2 or EOC20 cells (Fig. 6), indicating that ICP10PK inhibits its production, independent of the cell activation state.

Bottom Line: The CM from Delta PK-infected EOC2 and EOC20 cells did not contain IL-10, but it contained TNF-alpha and RANTES.IL-10 neutralization significantly (p < 0.01) decreased, but did not abrogate, the neuroprotective activity of the CM from Delta RR-infected microglial cultures indicating that ICP10PK modulates the neuronal-microglial axis, also through induction of various microglial neuroprotective factors.Protection was associated with a significant (p < 0.001) increase in the numbers of IL-10+ microglia (CD11b+) as compared to Delta PK-treated animals.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Pharmacology and Experimental Therapeutics, University of Maryland, School of Medicine, Baltimore, MD 21201, USA. jlain001@umaryland.edu

ABSTRACT
Ischemic brain injury and epilepsy are common neurodegenerative diseases caused by excitotoxicity. Their pathogenesis includes microglial production of inflammatory cytokines. Our studies were designed to examine whether a growth compromised HSV-2 mutant (Delta RR) prevents excitotoxic injury through modulation of microglial responses by the anti-apoptotic HSV-2 protein ICP10PK. EOC2 and EOC20 microglial cells, which are differentially activated, were infected with Delta RR or the ICP10PK deleted virus (Delta PK) and examined for virus-induced neuroprotective activity. Both cell lines were non-permissive for virus growth, but expressed ICP10PK (Delta RR) or the PK deleted ICP10 protein p95 (Delta PK). Conditioned medium (CM) from Delta RR-, but not Delta PK-infected cells prevented N-methyl-D-aspartate (NMDA)-induced apoptosis of primary hippocampal cultures, as determined by TUNEL and caspase-3 activation (76.9 +/- 5.3% neuroprotection). Neuroprotection was associated with inhibition of TNF-alpha and RANTES and production of IL-10. The CM from Delta PK-infected EOC2 and EOC20 cells did not contain IL-10, but it contained TNF-alpha and RANTES. IL-10 neutralization significantly (p < 0.01) decreased, but did not abrogate, the neuroprotective activity of the CM from Delta RR-infected microglial cultures indicating that ICP10PK modulates the neuronal-microglial axis, also through induction of various microglial neuroprotective factors. Rats given Delta RR (but not Delta PK) by intranasal inoculation were protected from kainic acid (KA)-induced seizures and neuronal loss in the CA1 hippocampal fields. Protection was associated with a significant (p < 0.001) increase in the numbers of IL-10+ microglia (CD11b+) as compared to Delta PK-treated animals. Delta RR is a promising vaccination/therapy platform for neurodegeneration through its pro-survival functions in neurons as well as microglia modulation.

No MeSH data available.


Related in: MedlinePlus