Virus Expression Database

GSE11238

Vaccinia E3L mutant virus infected HeLa cell lines (langl-affy-human-215499)

Submitted by Winnie Liang (Translational Genomics, USA) on Apr 22 2008

Platform: microarray – [HG-U133_Plus_2] Affymetrix Human Genome U133 Plus 2.0 Array

Pubmed: None listed

Summary Smallpox is a highly communicable, often fatal diseae. There is currently no licensed treatment for smallpox and vaccinia virus (VV) is currently used for immunization. While immunization with VV can provide good protection against exposure to the smallpox virus, the current vaccine is far from optimal. Complications occur in 1/1,000-1/10,000 vaccinees, with at least one death per million vaccinees. We have constructed recombinant VV strains which are less pathogenic, yet provide a protective immune response. These viruses contain various mutations in the E3L which is known to block the host antiviral response. Identifying the host genes involved in producing a strong protective immunological response to these attenuated viruses would not only increase our understanding of the proteins and pathways involved in effective smallpox vaccination, but aid in the development of alternative vaccine strains which enhance these specific immune responses.
We will determine gene expression patterns in HeLa cells at various times following infection with wtVV and several VV constructs containing mutations in the E3L gene.
The VV E3L gene product blocks the host antiviral response by sequestering viral danger signals, including double-stranded RNA and Z-DNA. VV constructs containing mutations in E3L which allow host cell recognition of either of these danger signals leads to a decrease in viral pathogensis. In this project we will dissect the cellular inflammatory response to infection with wtVV in comparison to VV containing mutations in the E3L gene. By understanding why certain strains of VV are non-pathogenic, yet highly immunogenic, it is possible to gain a better understanding on the mechanisms of poxvirus pathogenesis and the host response.
We will examine three times points following infection with VV: 2 HPI, 6 HPI and 9 HPI. These times points represent keys points in the virus replication cycle. Several VV constructs will be used which contain mutations in the E3L gene. These constructs alter the ability of E3L to sequester double-stranded RNA and/or Z-DNA and therefore have a direct effect on viral pathogenesis. Fourteen constructs will be used including: mock, wtVV, VVdelE3L, VVE3Ldel83N, VVE3Ldel37N, VVE3Ldel26C, VVE3Ldel7C, VVE3L Y48A, VVE3L P63A, VVE3L K167T, VV-ATV, VV-ADAR/E3L, VVdelK3L, VVdelK3L-E3Ldel37N. Cells will be infected at an MOI of 5 to allow infection of all cells. At each time point, cells will be harvested by scraping. RNA will be isolated using a Trizol RNA extraction protocol (Invitrogen) followed by RNA purification using the RNeasy cleanup kit available from Qiagen.
Keywords: time-course

39 Samples

ID Title Cell Type Timepoint Reported Virus Virus Species Exclusion Reason
GSM283458 HeLa Cell Line, unknown: wtVV, 2 HPI cervical adenocarcinoma cell line  [HeLa ] 2 hrs VV Vaccinia virus
GSM283459 HeLa Cell Line, unknown: wtVV, 6 HPI cervical adenocarcinoma cell line  [HeLa ] 6 hrs VV Vaccinia virus
GSM283460 HeLa Cell Line, unknown: wtVV, 9HPI cervical adenocarcinoma cell line  [HeLa ] 9 hrs VV Vaccinia virus
GSM283461 HeLa Cell Line, unknown: VVdelE3L, 2 HPI cervical adenocarcinoma cell line  [HeLa ] 2 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283462 HeLa Cell Line, unknown: VVdelE3L, 6 HPI cervical adenocarcinoma cell line  [HeLa ] 6 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283463 HeLa Cell Line, unknown: VVdelE3L, 9 HPI cervical adenocarcinoma cell line  [HeLa ] 9 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283464 HeLa Cell Line, unknown: VVE3Ldel83N, 2 HPI cervical adenocarcinoma cell line  [HeLa ] 2 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283465 HeLa Cell Line, unknown: VVE3Ldel83N, 6 HPI cervical adenocarcinoma cell line  [HeLa ] 6 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283466 HeLa Cell Line, unknown: VVE3Ldel83N, 9 HPI cervical adenocarcinoma cell line  [HeLa ] 9 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283467 HeLa Cell Line, unknown: VVE3Ldel26C, 2 HPI cervical adenocarcinoma cell line  [HeLa ] 2 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283468 HeLa Cell Line, unknown: VVE3Ldel26C, 6 HPI cervical adenocarcinoma cell line  [HeLa ] 6 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283469 HeLa Cell Line, unknown: VVE3Ldel26C, 9 HPI cervical adenocarcinoma cell line  [HeLa ] 9 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283470 HeLa Cell Line, unknown: VVE3Ldel7C, 2 HPI cervical adenocarcinoma cell line  [HeLa ] 2 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283471 HeLa Cell Line, unknown: VVE3Ldel7C, 6 HPI cervical adenocarcinoma cell line  [HeLa ] 6 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283472 HeLa Cell Line, unknown: VVE3Ldel7C, 9 HPI cervical adenocarcinoma cell line  [HeLa ] 9 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283473 HeLa Cell Line, unknown: VVE3L Y48A, 2 HPI cervical adenocarcinoma cell line  [HeLa ] 2 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283474 HeLa Cell Line, unknown: VVE3L Y48A, 6 HPI cervical adenocarcinoma cell line  [HeLa ] 6 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283475 HeLa Cell Line, unknown: VVE3L Y48A, 9 HPI cervical adenocarcinoma cell line  [HeLa ] 9 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283476 HeLa Cell Line, unknown: VVE3L Y48F, 2 HPI cervical adenocarcinoma cell line  [HeLa ] 2 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283477 HeLa Cell Line, unknown: VVE3L Y48F, 6 HPI cervical adenocarcinoma cell line  [HeLa ] 6 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283478 HeLa Cell Line, unknown: VVE3L Y48F, 9 HPI cervical adenocarcinoma cell line  [HeLa ] 9 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283479 HeLa Cell Line, unknown: VV-ATV, 2 HPI cervical adenocarcinoma cell line  [HeLa ] 2 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283480 HeLa Cell Line, unknown: VV-ATV, 6 HPI cervical adenocarcinoma cell line  [HeLa ] 6 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283481 HeLa Cell Line, unknown: VV-ATV, 9 HPI cervical adenocarcinoma cell line  [HeLa ] 9 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283482 HeLa Cell Line, unknown: VV-ADAR/E3L, 2 HPI cervical adenocarcinoma cell line  [HeLa ] 2 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283483 HeLa Cell Line, unknown: VV-ADAR/E3L, 6 HPI cervical adenocarcinoma cell line  [HeLa ] 6 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283484 HeLa Cell Line, unknown: VV-ADAR/E3L, 9 HPI cervical adenocarcinoma cell line  [HeLa ] 9 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283485 HeLa Cell Line, unknown: VVdel37N, 2 HPI cervical adenocarcinoma cell line  [HeLa ] 2 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283486 HeLa Cell Line, unknown: VVdel37N, 6 HPI cervical adenocarcinoma cell line  [HeLa ] 6 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283487 HeLa Cell Line, unknown: VVdel37N, 9 HPI cervical adenocarcinoma cell line  [HeLa ] 9 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283488 HeLa Cell Line, unknown: VVdelK3L, 2 HPI cervical adenocarcinoma cell line  [HeLa ] 2 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283489 HeLa Cell Line, unknown: VVdelK3L, 6 HPI cervical adenocarcinoma cell line  [HeLa ] 6 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283490 HeLa Cell Line, unknown: VVdelK3L, 9 HPI cervical adenocarcinoma cell line  [HeLa ] 9 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283491 HeLa Cell Line, unknown: VVdelK3Ldel37N, 2HPI cervical adenocarcinoma cell line  [HeLa ] 2 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283492 HeLa Cell Line, unknown: VVdelK3Ldel37N, 6HPI cervical adenocarcinoma cell line  [HeLa ] 6 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283493 HeLa Cell Line, unknown: VVdelK3Ldel37N, 9HPI cervical adenocarcinoma cell line  [HeLa ] 9 hrs VV Vaccinia virus Extra interventions
mutated to avoid response
GSM283494 HeLa Cell Line, unknown: mock infected 6HPI cervical adenocarcinoma cell line  [HeLa ] 2 hrs mock Uninfected
GSM283495 HeLa Cell Line, unknown: mock infected 2HPI cervical adenocarcinoma cell line  [HeLa ] 6 hrs mock Uninfected
GSM283496 HeLa Cell Line, unknown: mock infected 9HPI cervical adenocarcinoma cell line  [HeLa ] 9 hrs mock Uninfected