A novel replication-competent modified vaccinia Tiantan (MVTT)-based HIV Vaccine
Non-Infectious Papilloma virus–like Particles (VLPs) inhibit HIV-1 replication: Implications for...
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Transcript of Non-Infectious Papilloma virus–like Particles (VLPs) inhibit HIV-1 replication: Implications for...
Non-Infectious Papilloma virus–like Non-Infectious Papilloma virus–like Particles (VLPs) inhibit HIV-1 Particles (VLPs) inhibit HIV-1 replication: Implications for immune replication: Implications for immune control of HIV-1 replication by IL-27.control of HIV-1 replication by IL-27.
Laboratory of Human Retrovirology CSP / NIAID-Frederick
J.Mohamad FakruddinJ.Mohamad Fakruddin
BACKGROUNDBACKGROUNDImmune system uses a range of soluble molecules to suppress viral infections.
Action of non-cytolytic antiviral factors to control HIV-1 replication.
Some viral infections elicit the secretion of soluble HIV-1 suppressor factors.
HPV-like particles(VLPs) bind to dendritic cells and induce the expression of anti-viral cytokines such as IFN-, IL-10 and IFN-.
VLPs have been used as a vaccine to prevent HPV infection.
BACKGROUNDBACKGROUND
VLP StructureVLP Structure
To study the potential ofVLP induced soluble HIV-1 suppressive factors.
AIMAIM
Cells+PHA 3-days
+ HIV-1 X4(NL4.3) or R5(AD8) 2hr/370C/CO2
Cultured with VLP
7 or 10 days
HIV-1 replication evaluated by p24 ELISA
METHODSMETHODS
VLP inhibits HIV-1 replication in PBMCsVLP inhibits HIV-1 replication in PBMCs
X4 virus R5 virus
VLP suppresses HIV-1 replication regardless of the virus tropism
HIV-1 inhibition is Papilloma virus specificHIV-1 inhibition is Papilloma virus specific
SV40 VLP has no effect on replication of HIV
Pretreatment of PBMC with VLP inhibits HIV-1Pretreatment of PBMC with VLP inhibits HIV-1
24 hr pretreatment is sufficient for inhibition
VLP does not alter the expression of HIV receptors
Effect of VLP on HIV-1 receptorsEffect of VLP on HIV-1 receptors
VLP does not inhibits HIV in CD4VLP does not inhibits HIV in CD4++ T cells and CD8 T cells and CD8++ cells not required cells not required
Effect of VLP on subset of PBMCsEffect of VLP on subset of PBMCs
Media
VLP
VLP binds strongly to Monocytes
VLP exerted its inhibitory activity on PBMCs, but not on CD4+ cells.
VLP inhibitory effect is due to anti-HIV soluble factors or by cell to cell interaction.
To study the mechanism- The effect of conditioned medium(CM) derived from VLP treated PBMCs on HIV infected CD4+ T-cells was investigated.
Is there a role for soluble factors?
Effect of VLP treated CM on CD4Effect of VLP treated CM on CD4++ T cells T cells
VLP induces soluble anti-HIV factor(s) from both PBMCs and MDMs
Global gene response by microarrayGlobal gene response by microarray
One point study to examine transcriptional changes.
MDMs and PBMCs at 24h or 36h after exposure to VLP.
Fold Increase
Gene ID Gene name PBMC MDM
24678 IL-27 14.0 20.0
6355 MCP-2 31.0 30.0
10673 TNSF13B 2.6 2.5
3586 IL-10 NS 2.3
8743 TNSF-10 6.4 111.0
3600 IL-15 NS 4.7
6373 CXCL11 85.0 54.0
3627 CXCL10 10.0 58.0
6347 MCP-1 15.0 NS
3469 INF-
143.0 NS
3456 IFN-
135.0 NS
Expression of cytokine genesExpression of cytokine genes
Detection of of anti-HIV cytokines and IL-27 in CMDetection of of anti-HIV cytokines and IL-27 in CM
Significant increase of IL-27 in both PBMCs and MDMs
Blocking the activity of anti-HIV factorsBlocking the activity of anti-HIV factors
VLP stimulated anti-HIV activity is partially blocked by -IL-27
IL-27 expression is specific for HPV-VLPIL-27 expression is specific for HPV-VLP
SV40 VLP has no effect on IL-27 expression
IL-27 is key factor of VLP mediated HIV inhibition
- many fold increase of IL-27 by VLP was observed at both RNA and protein levels.
- in neutralization experiments only IL-27 blocked VLP inhibition.
IL-27 is a member of the IL-6/IL-12 family of cytokines.
IL-27 produced by dendritic cells and monocytes.
Expression of IL-27R can be detected in NK, B cells and T cells.
Nature of the IL-27/IL-27R interaction is pleiotropic.
IL-27 is an immunomodulator as it promotes pro- and anti-inflammatory response.
Biology of IL-27
IL-27 suppresses X4 replication in IL-27 suppresses X4 replication in CD4CD4++ T cells T cells
IL-27 suppresses X4 replication in IL-27 suppresses X4 replication in PBMCsPBMCs
IL-27 suppresses R5 replication in IL-27 suppresses R5 replication in MDMsMDMs
Conclusions Conclusions VLP displayed anti-HIV activity in PBMCs and MDMs.
This inhibitory effect was lost in the enriched CD4+
T-cells.
Soluble factors play a major role in VLP mediated anti-HIV effect.
Neutralization experiments implicated IL-27 to be one of the mediator anti-HIV VLP effect.
Further studies identified IL-27 as a novel anti-HIV cytokine.
Elucidation of the molecular mechanism of action of IL-27 may lead to HIV therapeutics.
ConclusionsConclusions
Tomozumi Imamichi Cliff Lane
Ligia Pinto Alfonso Pineres
Richard Lempicki Joseph Adelsberger
Jun Yang Catherine Watkins
Cathi Yeager
AcknowledgementsAcknowledgements