1 / 34

Rapid and systemic innate immune responses to an adenoviral HIV vaccine

Rapid and systemic innate immune responses to an adenoviral HIV vaccine. Erica Andersen-Nissen McElrath Lab Fred Hutchinson Cancer Research Center. Developing an HIV vaccine. Very challenging task! Two potential scenarios for an HIV vaccine:

red
Télécharger la présentation

Rapid and systemic innate immune responses to an adenoviral HIV vaccine

An Image/Link below is provided (as is) to download presentation Download Policy: Content on the Website is provided to you AS IS for your information and personal use and may not be sold / licensed / shared on other websites without getting consent from its author. Content is provided to you AS IS for your information and personal use only. Download presentation by click this link. While downloading, if for some reason you are not able to download a presentation, the publisher may have deleted the file from their server. During download, if you can't get a presentation, the file might be deleted by the publisher.

E N D

Presentation Transcript

  1. Rapid and systemic innate immune responses to an adenoviral HIV vaccine Erica Andersen-Nissen McElrath Lab Fred Hutchinson Cancer Research Center

  2. Developing an HIV vaccine • Very challenging task! • Two potential scenarios for an HIV vaccine: • Regardless of the type of HIV vaccine developed, we need to better understand how to induce a protective adaptive response • > innate immune response -slow clinical disease progression -reduce transmission DH Barouch Nature 455, 613-619 (2008)

  3. When a microbe invades, the immune system: Recognizes it Mobilizes and eliminates it Remembers it Returns to baseline Arms of the Immune System

  4. Key Players in Innate Immunity Barriers (Skin, epithelia) Enzymes, Anti-microbial peptides Microbial Recognition Receptors Complement proteins IMM/MICROM 441Lecture #4 October 1/08Kaja Page 4 of 26

  5. What influences adaptive immune response to a vaccine? - Innate immune responses shape the quantity, quality and longevity of the adaptive response (Pulendran and Ahmed, Cell, 2006) -Engineer a vaccine to induce desired innate responses Vaccine vector or adjuvant Vaccine antigen http://research.dfci.harvard.edu/innate/images/innate/immunities.gif

  6. Innate Immune Responses to Vaccination • Few studies have examined innate responses in humans after vaccination (Querec et al., NI, 2009; Gaucher et al., JEM, 2008) • We implemented a clinical trial of an adenoviral vector HIV vaccine (Merck Ad5): 1. Which systemic innate immune responses can be measured in blood after vaccination by intramuscular injection? 2. What are the optimal time points to identify these innate responses in humans? 3. Which innate responses correlate with adaptive responses?

  7. Adenoviral vectors for T-cell-based HIV vaccines • Adenoviral vectors containing HIV inserts found to elicit high-magnitude CD8+ T cell responses ~80% of vaccinees demonstrate T cell immunogenicity by ELISpot ~300 SFU/106 PBMC -responses were durable (Priddy et al., CID, 2008) (Corey et al., AIDS 2009) • Step Study

  8. Innate Immune Responses: HVTN 071 (n=35) 1.5 x 1010 genomes MrkAd5 gag/pol/nef Ad5 injection (IM): Sampling points: Day: 0 6h 1 3 7 28 Samples: Whole blood (Flow cytometry) Serum (Luminex) PBMC RNA (Microarray) n=11 PBMC (ICS)

  9. Innate Immune Responses: HVTN 071 1.5 x 1010 genomes MrkAd5 gag/pol/nef Ad5 injection (IM): Sampling points: Day: 0 6h 1 3 7 28 Samples: n=24 (serum only) PBMC (ICS)

  10. Baseline Serum Ad5 Neutralizing Antibody Titers Baseline Ad5 Titer High, >1000 Moderate, 200-999 Low, 19-199 Negative, ≤18 Ad5 serum neutralizing antibody titer PTID: Group A (n=11) Group B (pre-vaccine and day 1) (n=24)

  11. Cells/ml Blood * p<0.05, Timepoint vs. pre-vaccine (0 hrs.), Wilcoxon signed rank test Moderate, 200-999 Low, 19-199 Negative, ≤18 Hours Post Vaccination Rapid Changes in Peripheral Blood Cell Populations after MRKAd5 gag/pol/nef Vaccination Lymphocytes Monocytes * * * Granulocytes Baseline Ad5 Titer *

  12. 14 12 10 8 6 4 2 0 0 24 48 72 168 Multiple Cell Types Exit the Blood CD8 T cells CD4 T cells B cells CD4+ T cells CD19+ Lymphocytes * * * * Myeloid DC NK cells Plasmacytoid DC * Cells/ml Blood Hours Post Vaccination

  13. * p<0.05, Timepoint vs. pre-vaccine (0 hrs.), Wilcoxon signed rank test Moderate, 200-999 Low, 19-199 Negative, ≤18 Serum Proinflammatory Cytokines & Chemokines Increase Rapidly after Vaccination TNF-a IFN-g IL-6 * * * * IP-10/CXCL-10 MCP-1/CCL2 Baseline Ad5 Titer * * * * * Serum pg/ml Hours Post Vaccination

  14. * p<0.05, Timepoint vs. pre-vaccine (0 hrs.), Wilcoxon signed rank test Moderate, 200-999 Low, 19-199 Negative, ≤18 Serum Anti-inflammatory Cytokines Also Increase after MRKAd5 gag/pol/nef Vaccination IL-1Ra IL-10 * * * Serum pg/ml Hours Post Vaccination Baseline Ad5 Titer

  15. Summary of Significant Changes in Serum Cytokines/Chemokines at 24 Hours Other serum analytes that were measured, but did not change at 24 hours: EGF, Eotaxin, Fractalkine, G-CSF, GM-CSF, IFN-a, IL-1a, IL-1b, IL-2, IL-4, IL-5, IL-8, IL-12p40, IL-12p70, IL-13, IL-15, IL-17, MIP-1a, sCD40L, TGF-a, VEGF

  16. Ad5 Negative/Low Titer Individuals Show Significantly Greater Changes in Serum Cytokines/Chemokines at 24 Hours

  17. PBMC Gene Expression PBMC • Discover other innate immune responses to Ad5 vaccination • Identify additional differences in responses between Ad5 naïve and Ad5 immune individuals Isolate RNA Affymetrix human exon arrays (10 people; 5 timepoints Analysis (collaboration with Dan Zak, ISB)

  18. Med Ad5 Titer Low Ad5 Titer Med Ad5 Titer PBMC Microarrays Show Many Genes with Significant* Temporal Expression Responses volunteer ID: timepoint: *p<10-6, 1 way ANOVA Down-regulated Up-regulated

  19. What do the gene expression changes mean? • 2 sources of changes in gene expression: • Changes in the PBMC composition • Changes in the response of cells that are in the blood to the vaccine • Attenuated gene expression changes in Ad5 medium titer subjects • are these individuals effectively exposed to a lower vaccine dose?

  20. 6hrs: 1 gene down-regulated(p<0.001, |log2(FC)| > 0.5) Innate DB: www.innatedb.ca/

  21. 6hrs: 1 gene down-regulated(p<0.001, |log2(FC)| > 0.5) MIP-1a CCL3

  22. 24hrs: 1065 genes up/down-regulated(p<0.001, |log2(FC)| > 0.5, FDR = 0.01)

  23. 24hrs: TLR pathway up-regulated(p<0.001, |log2(FC)| > 0.5, FDR = 0.01)

  24. 24hrs: STAT1 targets/interactors up-regulated(p<0.001, |log2(FC)| > 0.5, FDR = 0.01)

  25. 24hrs: TCR pathway down-regulated(p<0.001, |log2(FC)| > 0.5, FDR = 0.01)

  26. 72hrs: 116 genes up/down-regulated(p<0.001, |log2(FC)| > 0.5, FDR = 0.02)

  27. 72hrs: 116 genes up/down-regulated (p<0.001, |log2(FC)| > 0.5, FDR = 0.02) CSF1R CSF1R

  28. 168hrs: 11 genes up/down-regulated (p<0.001, |log2(FC)| > 0.5, FDR = 0.2) TXNDC5 ENDO-PDI TXNDC5 = EndoPDI: a protein preferentially expressed in endothelial cells that acts as a stress survival factor during hypoxia (Sullivan et al., 2003)

  29. Summary MRK Ad5 gag/pol/nef administered IM induces robust innate immune responses in the blood: rapid influx of monocytes/mDC into the blood and efflux of lymphocytes/pDC from the blood trafficking to and from the site of vaccination? changes in serum chemokines and PBMC gene expression indicate PBMC are the source of many serum factors maximal responses occur at 24hrs post-vaccination Pre-existing neutralizing antibodies to Ad5 attenuate the innate response reduced production of MCP-1, IP-10 and IL-1Ra muted gene expression changes in PBMC Potential correlates for magnitude of CD8 response are being identified

  30. Future Directions • Compare innate responses to Mrk rAd5 vaccine with responses to other vaccines • Other candidate HIV vaccines (e.g. MVA: HVTN 908) • Vaccines currently in use (e.g. YFV) • Evaluate innate immune responses to vaccines at mucosal sites to determine effects at the site of HIV acquisition • Develop an in vitro system to test effects of identified innate response genes on the adaptive response

  31. Julie McElrath Steve De Rosa Eric Peterson Joanne Chang Sam Pine Greg Spies Don Carter Olivier Defawe Mingchao Shen Reneé Ireton Phyllis Stegall Joleen Borgerding Nidhi Kochar Liza Noonan Yunda Huang Li Qin Steve Self Acknowledgements Dan Zak Alan Aderem HVTN Laboratory Program HVTU (Seattle, Rochester, Nashville, Birmingham) Ann Duerr Larry Corey Jim Kublin Volunteers CAVD Investigators Merck Michael Robertson UW STD/AIDS Research Training Fellowship NIAID/DAIDS Margaret Johnston

  32. Innate immune detection of adenovirus (Gilliet et al., 2008)

  33. The Step Study • 3000-person HIV vaccine trial started in late 2004 • Phase IIB test-of-concept study to establish whether MRKAd5 expressing HIV-1 gag/pol/nef could lower: • HIV-1 infection rates • Plasma viremia • Interim analysis performed when 30 per-protocol HIV infections occurred

  34. No effect of the vaccine on viral load Increased HIV infection in vaccinees that had pre-existing immunity to Ad5 All further vaccinations were halted Mechanisms under investigation Step Study Results Overall HIV Incidence (% per year) 0-26 26-52 52-78 Ad5≤18 HIV Incidence (% per year) 0-26 26-52 52-78 Ad5>18 HIV Incidence (% per year) 0-26 26-52 52-78 Time Interval (weeks) (Buchbinder et al., 2008; McElrath et al., 2008) (Buchbinder et al., 2008)

More Related