Engineering an in vitro human immune system for rapid vaccine evaluation
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1 Engineering an in vitro human immune system for rapid vaccine evaluation William Warren, Ph.D. VaxDesign Corporation Orlando, FL
2 MIMIC Technology Overview MIMIC: an in vitro biomimetic of the human immune response: 1. Collect leukocytes from human donors 2. Simulate innate immunity with the Peripheral Tissue (PT) Module 3. Simulate adaptive immunity with the Lymphoid Tissue Equivalent (LTE) Module 4. Measure the effectiveness of the immune response or immune modulator product 2
3 Benefits: Vaccine Trial in a Well Predictive high throughput in vitro immunology to assess novel vaccine candidates Measure immune response in diverse population Faster cycle time for discovery Better selection of vaccine candidates for clinical evaluation New ways to design clinical trials Reduce the time and costs to bring vaccines to the market 3
4 Examples of In Vitro Vaccine Projects 1. Assessing adjuvant responses 2. Immunotoxicology/immunoregulation of biologicals 3. Quantity and Quality of T cell/cmi responses 4. Quantity and Quality of B cell/ab responses 5. Rapid immunogen screen/engineering immunology 4
5 Peripheral Tissue Module Replicates Innate Immune Response Adjuvants and immunopotentiators Adjuvant DC Development DC Maturation Cytokine Production Alum, MF59, CpG, poly I:C, etc. Alum 0/+ /0 ++ Innate response profiles of vaccines MF /+ DTaP, Influenza, Hep B, Yellow Fever, Rabies, etc. Innate response to biologics CpG TNFα, CD154, OKT3, Avastin Immunosuppressant effects Dexamethasone, Cyclosporine, Hydrocortisone, etc. Cytokine profiles of pathogens Viruses, bacteria With Jeff Ulmer & Andreas Wack, Novartis Vaccines 5
6 Examples of In Vitro Vaccine Projects 1. Assessing adjuvant responses 2. Immunotoxicology/immunoregulation of biologicals 3. Quantity and Quality of T cell/cmi responses 4. Quantity and Quality of B cell/ab responses 5. Rapid immunogen screen/engineering immunology 6
7 How Do We Measure a Good T Cell Response to a Vaccine? Magnitude of a T cell response Quantitative measurement of the T cell response, the frequency of CD4+ or CD8+ T cells that are Ag specific. total cytokine secretion, cytolytic activity or proliferation. Quality of the T cell response Combination of T cell functions Proliferation, organization of immune cells, effector cells Multifunctional T cells have been shown to correlate with disease non progression and protection (HIV, EBV, CMV, Influenza, etc.) Betts, M. R. et al. Blood 107, (2006). Harari, A., Petitpierre, S., Vallelian, F. & Pantaleo, G.. Blood 103, (2004). Younes, S. A. et al. J. Exp. Med. 198, (2003) Robert A. Seder, Patricia A. Darrah, Mario Roederer, Nat. Rev. Immunology 8, 257 (2008) 7
8 Magnitude of Primary In Vitro CTL Responses to Yellow Fever CD8 T cells recognize naïve antigen no Antigen Yellow Fever Primary Stimulation (7 days) <APC Alexa750 A>: 107a <APC Alexa750 A>: 107a <PE Cy7 A>: IFNg <PE Cy7 A>: IFNg 0 Secondary Stimulation (12 days) CD107a IFNγ <PE Cy7 A>: IFN g <APC Alexa750 A>: CD107a <PE Cy7 A>: IFN g <APC Alexa750 A>: CD107a
9 Quality of CD8 T Cell Responses: In Vitro Yellow Fever Vaccination Day 7 0.4% 2.58% 7.24% 0.3% Frequency 0.2% 57.65% 32.53% 0.1% 0.0% CGIT CGI CGT GIT CIT CI CT CG IT GI GT G C I T Quadruple Triple Double Single Day 15 Re stimulation 6% 1.04% 14.87% 4% Frequency 55.17% 28.91% 2% Quadruple Triple Double Single 0% CGIT CGI CGT GIT CIT CI CT CG IT GI GT G C I T 9
10 Examples of In Vitro Vaccine Projects 1. Assessing adjuvant responses 2. Immunotoxicology/immunoregulation of biologicals 3. Quantity and Quality of T cell/cmi responses 4. Quantity and Quality of B cell/ab responses 5. Rapid immunogen screen/engineering immunology 10
11 Quantity: Pre/Post Influenza Vaccine Stimulation: Anti Fluvirin IgG Anti Fluvirin IgG (ng/ml) Antigen stimulation & vaccination boosted MIMIC anti Fluvirin IgG 11
12 Quality of Ab Responses: Avidity Indices and Cross Reactivity Last year s flu vaccine, esp. Solomon Island H1 component, is associated with reduced Ab avidity and reduced cross reactivity Pre vaccination Post vaccination Stronger NC memory, high avidity, response in pre vaccinated donors Stronger SI, low avidity, response in post vaccinated donors 12
13 Examples of In Vitro Vaccine Projects 1. Assessing adjuvant responses 2. Immunotoxicology/immunoregulation of biologicals 3. Quantity and Quality of T cell/cmi responses 4. Quantity and Quality of B cell/ab responses 5. Rapid immunogen screen/engineering immunology 13
14 Rapid T and B Cell Immunogen Screening Reduce the complexity of the human immune system into a series of modular bioengineered in vitro constructs Highly sensitive approach for assessing the magnitude and functionality of specific T and B cells Donor 1 Donor 2 Donor 3 Donor 4 Donor 5 Donor 6 Permits the evaluation of primary and recall responses Control Ag1 Ag2 Ag3 14
15 Significant Time/Cost Savings via Automation Manual Automation Blood Processing Labor Hours Vaccination Site Hours per 32 plates ELISA Hours per 36 plates ELISPOT Hours per 32 plates 2.5 hrs 1 hr 24 hrs 6 hrs 40 hrs 3 hrs 40 hrs 10 hrs Automated ELISA Results 15
16 Conclusions 1. MIMIC system appears to replicate human immunity, further validation will improve the situation 2. Data appears encouraging as a biomimetic of the human immune system vaccine responses immunogenicity immunotoxicity in vitro infectious disease models 3. Applications for vaccines: entire drug development timeline preclinical down selection design adaptive clinical trials manufacturing 16
17 Acknowledgements This work is funded by DARPA/DSO in the Rapid Vaccine Assessment Program and by IAVI through it s Innovation Fund To sanofi pasteur for providing the yellow fever vaccine 17
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