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DC infected by the ANRS MVA HIV vaccine candidate prime NK cells with anti-HIV specific activity through a mechanism involving NKG2D and NKp46 on NK cells and membrane-bound IL-15 on DC. Uriel Y. Moreno- Nieves PhD Student. Introduction. NK cells : Anti-viral activity.

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uriel y moreno nieves phd student

DC infected by the ANRS MVAHIV vaccine candidate prime NK cells with anti-HIV specific activity through a mechanism involving NKG2D and NKp46 on NK cells and membrane-bound IL-15 on DC

Uriel Y. Moreno-Nieves

PhDStudent

introduction
Introduction
  • NK cells :
    • Anti-viral activity.
    • First line of defense against viral pathogens.
    • Increasing evidence of adaptive immune features.
  • Cross-talk NK/DC:
    • Activation of NK cells.
    • Important for the induction of adaptive immunity.
  • MVA : Modified Vaccinia virus Ankara
    • Attenuated, non-replicating poxvirus.
    • MVAHIV : expression of Gag, Pol, Nef peptides.
background and objectives
Background and Objectives
  • NK cells stimulated by MVAHIV-infected DC are able to better control HIV infection in DC.

Objectives:

  • To test control of HIV-1 infection in other cell types by MVAHIV-stimulated NK cells.
  • To investigate the mechanisms of anti-HIV specific priming of NK cells.
material and methods
Material and methods

CD4+ T cells

PBMC

Magnetic

separation

Ficoll

NK cells

Blood from

Healthy donors

Differentiation

7 days

Monocytes

DC

Addition of DC and NK cells

after 18 hours

Control of HIV infection

DC

CD4+ T cells

Analysis of anti-HIV specificity

Infection of DC

by MVA

Priming of NK cells

4 days

control of hiv infection by nk cells is increased after mva hiv priming
Control of HIV infection by NK cells is increased after MVAHIV priming

Control of HIV infection

HIV-infected DC

DC

CD4+ T cell

Analysis of anti-HIV specificity

Priming of NK cells

4 days

CMV-infected DC

HIV-infected CD4+ T cells

Day 6 p.i.

n=10

Day 7 and Day 10 p.i.

n=8 and n=10

priming of nk cells by mva hiv is modulated by nkg2d
Priming of NK cells by MVAHIV is modulated by NKG2D

HIV-infected CD4+ T cells

HIV-infected DC

Day 7 p.i.

n=8

Day 9 p.i.

n=10

nkg2d blockade during mva priming decreases mbil 15 expression on dc
NKG2D blockade during MVA priming decreases mbIL-15 expression on DC

Expression of mbIL-15 on DC

during MVA priming

Modulation of mbIL-15 on DC by NKG2D and NKp46

24, 48 and 72 hours of priming

n=7

72 hours of priming

n=5

il 15 is important for the mva hiv priming
IL-15 is important for the MVAHIV priming

Implication of NKG2D and mbIL-15 during MVA priming

Day 9 p.i.

n=8

conclusions
Conclusions
  • NK cells primed by MVAHIV have increased anti-HIV activity.
    • Better control of HIV infection in DC and CD4+ T cells.
    • MVAHIV priming seems to be anti-HIV-1 specific.
  • NKp46 blockade during MVAHIV priming enhances subsequent control of HIV infection in DC.
  • NKG2D engagement during MVAHIVpriming is important for subsequent control of HIV infection in DC and CD4+T cells.
    • NKG2D regulates mbIL-15 expression on DC.
    • IL-15 is important for MVAHIVpriming.
    • Decreased mbIL-15 expression after NKG2D blockade during MVAHIV priming might be responsible for the decreased control of HIV infection in CD4+ T cells.
    • Possible contribution of NK cells in vaccine efficacy.
acknowledgements
Acknowledgements

Daniel Scott-Algara

Françoise Barré-Sinoussi

Yves Lévy

Jean-Saville Cummings

Vincent Arnold

Adrien Gilbert

Kevin Yarbrough

Céline Didier

Thank you!