Intracranial administration of P gene siRNA protects mice from lethal Chandipura virus encephalitis.
BACKGROUND: In parts of India, Chandipura Virus (CHPV) has emerged as an encephalitis causing pathogen in both epidemic and sporadic forms. This pediatric disease follows rapid course leading to 55-75% mortality. In the absence of specific treatment, effectiveness of RNA interference (RNAi) was eval...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2010-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC2797643?pdf=render |
| _version_ | 1818017526257287168 |
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| author | Satyendra Kumar Vidya A Arankalle |
| author_facet | Satyendra Kumar Vidya A Arankalle |
| author_sort | Satyendra Kumar |
| collection | DOAJ |
| description | BACKGROUND: In parts of India, Chandipura Virus (CHPV) has emerged as an encephalitis causing pathogen in both epidemic and sporadic forms. This pediatric disease follows rapid course leading to 55-75% mortality. In the absence of specific treatment, effectiveness of RNA interference (RNAi) was evaluated. METHODS AND FINDINGS: Efficacy of synthetic short interfering RNA (siRNA) or short hairpin RNA (shRNA) in protecting mice from CHPV infection was assessed. The target genes were P and M genes primarily because important role of the former in viral replication and lethal nature of the latter. Real time one step RT-PCR and plaque assay were used for the assessment of gene silencing. Using pAcGFP1N1-CHPV-P, we showed that P-2 siRNA was most efficient in reducing the expression of P gene in-vitro. Both quantitative assays documented 2 logs reduction in the virus titer when P-2, M-5 or M-6 siRNAs were transfected 2 hr post infection (PI). Use of these siRNAs in combination did not result in enhanced efficiency. P-2 siRNA was found to tolerate four mismatches in the center. As compared to five different shRNAs, P-2 siRNA was most effective in inhibiting CHPV replication. An extended survival was noted when mice infected intracranially with 100 LD50 CHPV were treated with cationic lipid complexed 5 microg P-2 siRNA simultaneously. Infection with 10LD50 and treatment with two doses of siRNA first, simultaneously and second 24 hr PI, resulted in 70% survival. Surviving mice showed 4 logs less CHPV titers in brain without histopathological changes or antibody response. Gene expression profiles of P-2 siRNA treated mice showed no interferon response. First dose of siRNA at 2 hr or 4 hr PI with second dose at 24hr resulted in 40% and 20% survival respectively suggesting potential application in therapy. CONCLUSIONS: The results highlight therapeutic potential of siRNA in treating rapid and fatal Chandipura encephalitis. |
| first_indexed | 2024-04-14T07:28:08Z |
| format | Article |
| id | doaj.art-505cc4013a564e389fa33bfac2ead114 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-04-14T07:28:08Z |
| publishDate | 2010-01-01 |
| publisher | Public Library of Science (PLoS) |
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| spelling | doaj.art-505cc4013a564e389fa33bfac2ead1142022-12-22T02:05:56ZengPublic Library of Science (PLoS)PLoS ONE1932-62032010-01-0151e861510.1371/journal.pone.0008615Intracranial administration of P gene siRNA protects mice from lethal Chandipura virus encephalitis.Satyendra KumarVidya A ArankalleBACKGROUND: In parts of India, Chandipura Virus (CHPV) has emerged as an encephalitis causing pathogen in both epidemic and sporadic forms. This pediatric disease follows rapid course leading to 55-75% mortality. In the absence of specific treatment, effectiveness of RNA interference (RNAi) was evaluated. METHODS AND FINDINGS: Efficacy of synthetic short interfering RNA (siRNA) or short hairpin RNA (shRNA) in protecting mice from CHPV infection was assessed. The target genes were P and M genes primarily because important role of the former in viral replication and lethal nature of the latter. Real time one step RT-PCR and plaque assay were used for the assessment of gene silencing. Using pAcGFP1N1-CHPV-P, we showed that P-2 siRNA was most efficient in reducing the expression of P gene in-vitro. Both quantitative assays documented 2 logs reduction in the virus titer when P-2, M-5 or M-6 siRNAs were transfected 2 hr post infection (PI). Use of these siRNAs in combination did not result in enhanced efficiency. P-2 siRNA was found to tolerate four mismatches in the center. As compared to five different shRNAs, P-2 siRNA was most effective in inhibiting CHPV replication. An extended survival was noted when mice infected intracranially with 100 LD50 CHPV were treated with cationic lipid complexed 5 microg P-2 siRNA simultaneously. Infection with 10LD50 and treatment with two doses of siRNA first, simultaneously and second 24 hr PI, resulted in 70% survival. Surviving mice showed 4 logs less CHPV titers in brain without histopathological changes or antibody response. Gene expression profiles of P-2 siRNA treated mice showed no interferon response. First dose of siRNA at 2 hr or 4 hr PI with second dose at 24hr resulted in 40% and 20% survival respectively suggesting potential application in therapy. CONCLUSIONS: The results highlight therapeutic potential of siRNA in treating rapid and fatal Chandipura encephalitis.http://europepmc.org/articles/PMC2797643?pdf=render |
| spellingShingle | Satyendra Kumar Vidya A Arankalle Intracranial administration of P gene siRNA protects mice from lethal Chandipura virus encephalitis. PLoS ONE |
| title | Intracranial administration of P gene siRNA protects mice from lethal Chandipura virus encephalitis. |
| title_full | Intracranial administration of P gene siRNA protects mice from lethal Chandipura virus encephalitis. |
| title_fullStr | Intracranial administration of P gene siRNA protects mice from lethal Chandipura virus encephalitis. |
| title_full_unstemmed | Intracranial administration of P gene siRNA protects mice from lethal Chandipura virus encephalitis. |
| title_short | Intracranial administration of P gene siRNA protects mice from lethal Chandipura virus encephalitis. |
| title_sort | intracranial administration of p gene sirna protects mice from lethal chandipura virus encephalitis |
| url | http://europepmc.org/articles/PMC2797643?pdf=render |
| work_keys_str_mv | AT satyendrakumar intracranialadministrationofpgenesirnaprotectsmicefromlethalchandipuravirusencephalitis AT vidyaaarankalle intracranialadministrationofpgenesirnaprotectsmicefromlethalchandipuravirusencephalitis |