IAS 2015 Towards an HIV Cure symposium Vancouver Immune recognition following latency reversal
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1 IAS 2015 Towards an HIV Cure symposium Vancouver Immune recognition following latency reversal Marcus Altfeld Professor of Medicine
2 Outline Immune recognition of HIV-1-infected cells Kinetics of antigen expression and immune recognition following latency reversal NK cell-mediated recognition of latently HIV-1-infected cells
3 Immune recognition of HIV-1-infected cells B cell ADCC
4 Immune recognition of HIV-1-infected cells Latently HIV-1-infected cells are invisible to the immune system
5 Archin et al, Nature 2012; Deeks, Nature 2012
6 Challenges for immune recognition Tissue localization of latent HIV-1 reservoir vs immune effector cells No / incomplete latency reversal Impaired antiviral immune responses Viral diversification / immune escape ect
7 Challenges for immune recognition Tissue localization of latent HIV-1 reservoir vs immune effector cells
8 Tissue sanctuaries
9 Tissue sanctuaries Immunotherapeutic approaches to access tissue sanctuaries: Specific targeting of those immune effector mechanisms present in tissue sanctuaries Modulation of immune effector cell trafficking Disruption of GCs Adoptive immunotherapy..
10 Challenges for immune recognition Tissue localization of latent reservoir vs effector cells No / incomplete latency reversal
11 Incomplete HIV-1 latency reversal
12 Immune recognition following latency reversal neutralizing antibodies binding antibodies /ADCC adapted from Deeks, Nature 2012
13 Immune recognition following latency reversal HLA class I presented viral peptides can be recognized by HIV-1-specific CD8+ T cellls adapted from Deeks, Nature 2012
14 Immune recognition following latency reversal MIC-A, MIC-B, others Viral oligonucleotides trigger the expression of stress molecules that serve as ligands for activating NK cell receptors adapted from Deeks, Nature 2012
15 Immune recognition following latency reversal neutralizing antibodies binding antibodies /ADCC adapted from Deeks, Nature 2012
16 Therapeutic use of neutralizing antibodies Barouch et al., Nature 2012
17 Therapeutic use of neutralizing antibodies Applications in HIV-1-infected individuals on HAART in combination with latency reversal Caskey, Klein et al., Nature 2015
18 Therapeutic use of neutralizing antibodies Picker & Deeks, Nature 2013
19 Immune recognition following latency reversal HLA class I presented viral peptides can be recognized by HIV-1-specific CD8+ T cellls
20 CTL recognition following latency reversal HIV-1-specific CTLs can recognize and kill latently infected cells following reactivation (Shan, Immunity 2012, Sung, JID 2015), but: in vitro studies only following in vitro stimulation of CTLs only if viruses have not already escaped (Deng et al., Nature 2015)
21 CTL recognition following latency reversal The latent HIV reservoir will contain CTL escape variants selected during the acute /primary phase of infection Deng et al, Nature 2015; Picker & Lifson, Nature 2015
22 CTL recognition following latency reversal Requirement for therapeutic immunizations to target non-escaped CTL epitopes Picker & Lifson, Nature 2015
23 CTL recognition following latency reversal Requirement for therapeutic immunizations to target non-escaped CTL epitopes
24 Challenges for immune recognition Tissue localization of latent HIV-1 reservoir vs immune effector cells No / incomplete latency reversal Impaired antiviral immune responses Viral diversification / immune escape
25 Challenges for immune recognition Tissue localization of latent HIV-1 reservoir vs immune effector cells No / incomplete latency reversal Impaired antiviral immune responses additional immunotherapies (anti-pd1/pd-1l, cytokines, ect)
26 Immune recognition following latency reversal HIV-1 RNA+ cells might be recognized by NK cells
27 NK cell-mediated recognition of infected cells NK cells can recognize virusinfected cells through several different pathways Altfeld et al NRI 2011
28 Hypothesis Induction of HIV RNA transcription in latently infected cells is sufficient to trigger cellular stress and stress receptor expression How can we distinguish latently infected cells that express only HIV RNA from those that also harbor HIV proteins?? vs
29 Identification of HIV RNA+ vs protein+ cells PrimeFlow TM technique Gloria Martrus, P.15
30 Identification of HIV RNA+ vs protein+ cells HIV p24 Gag RNA J89 cells HIV p24 Gag protein Gloria Martrus, P.15
31 Gloria Martrus, P.15 Identification of HIV RNA+ vs protein+ cells
32 Can NK cells kill HIV RNA+ cells expressing stress receptors? MIC-A, MIC-B, others HIV Nef downregulates NKG2D ligands
33 Can NK cells kill HIV RNA+ cells expressing stress receptors? HIV-1 induces NKG2D ligand expression NKG2D ligand ACH-2, 6 hrs, PB P24 antigen NKG2D ligand HIV-1 Nef downregulates NKG2D ligand expression Eileen Scully, CNIHR workshop 2015 % NKG2DL+ of p24+ NL43 NL43 delta Nef
34 Nef inhibition to enhance NK cell killing 1. Reactivation (HDACi) 3. Nef blockade, enhancing upregulation of NKG2D ligands 2. Viral nucleic acid transcription and production of viral proteins c 4. NKG2D receptor engagement and NK cell degranulation 5. Target cell death Eileen Scully, P.32
35 Nef inhibition enhances NKG2D-L expression ACH-2 system after induction with 20nM Panobinostat mcherry Neffin HIV p24 antigen Eileen Scully, P.32
36 Nef inhibition enhances NKG2D-L expression ACH-2 system after induction with 20nM Panobinostat 2500 mcherry Neffin MFI of NKG2D ligand p24+neffin+ p24+neffin- HIV p24 antigen 0 no stim 2.5h PB 5h PB 24h PB Eileen Scully, P.32
37 Nef inhibition might enhance NK cell killing 10:1 mcherry - Neffin 5:1 NO NK CTRL Eileen Scully, CNIHR workshop 2015
38 Conclusions HIV-1 latency reversal will lead to different stages of reactivation in different cells no reactivation Viral RNA Viral proteins Virus production
39 Future Approaches Combined approaches: therapeutic immunization + reactivation immune modulators + reactivation adoptive transfer of ABs + reactivation adoptive transfer of CTLs or NK cells + reactivation gene therapy + reactivation /TI
40 Acknowledgments Gloria Martrus Eileen Scully Members of HPI Research Unit Virus Immunology Collab: D Kuritzkes, J Hauber, F Kirchhoff, S Benichou
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