PBMCs with Adenovirus show resistance to HIV-1 infection.txt

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PBMCs with Adenovirus show resistance to HIV-1 infection.txt
2013-04-01 23:41:28

adenovirus pbmc hiv-1 infection
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  1. HIV-infection resistance in PMBC-derived dendritic cells modified with recombinant virus
  2. Aim of Study
    Identify characteristics of PMBC-derived dendritic cells and their RESISTANCE to HIV-1 INFECTION by using adenovirus vector to insert various genes
  3. What is ncluded in adenovirus vector
    • Addn of:
    • 1. a sequence modified CCR5delta32 gene: interferes with CCR5 and CXCR4 fxn
    • 2. transduction of siRNA to block CCR5 expression
    • anti-HIV siRNA genes
    • 3. HIV-1 pol
    • 4. HIV-1 int
    • 1 and 2 role: BLOCK VIRAL ENTRY
    • 3 and 4 role: SUPRESS HIV replication
  4. Results of study
    HIV-1 infection resistance was observed when using cells with these recombinant adenoviral vectors
  5. PBMCs with adenviral vector (Ad-R5delta32siHIV) results
    • CCR5delta32 expression INCREASED
    • CCR5 and CXR4 expression DECREASED: Possibly due to inhibition by CCR5siRNA or delta32
  6. how to monitor HIV-1 viral replication, and results
    • Used p24 ELISAs to monitor replication
    • Results: in UNTRANSFECTED cells, p24 INCREASED vs. Ad-R5delta32siHIV showed NO change of p24.
    • Interpretation: INHIBITION of viral replication seen with Adenoviral infected PBMCs
  7. b-galactosidase
    • Magi cells have b-galac gene under control of HIV LTRs
    • Magi cells are susceptible to HIV strain IIIB infection
    • After transfection with adenovirus, Magi cells did NOT show susceptibility to HIV (Blue from b-galac decreased)
  8. # of INF-alpha-secreting cells
    • When looking at the DCs (Mature dendritic cells) the adenoviral infected cells showed INCREASE in these cells vs control
    • Interpretation: The adenovirally infected cells can INDUCE T-cell IMMUNITY
  9. CD83
    surface marker used as a measure to see if DCs with adenovirus were different than the control DCells