Engineering an in vitro human immune system for rapid vaccine evaluation

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Engineering an in vitro human immune system for rapid vaccine evaluation

William Warren, Ph.D.VaxDesign CorporationOrlando, FL 32826

[email protected]

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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

MIMIC™ Technology Overview

MIMIC: an in vitro biomimetic of the human immune response:

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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

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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

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Peripheral Tissue Module ReplicatesInnate Immune Response

• Adjuvants and immunopotentiators

– “Alum”, MF59, CpG, poly I:C, etc.

• Innate response profiles of vaccines

– DTaP, Influenza, Hep B, Yellow Fever, Rabies, etc.

• Innate response to biologics

– TNFα, CD154, OKT3, Avastin

• Immunosuppressant effects

– Dexamethasone, Cyclosporine, Hydrocortisone, etc.

• Cytokine profiles of pathogens

– Viruses, bacteria

++++

++++

‐/0

DC Maturation

++++++++CpG

0/+++++MF59

++0/+Alum

Cytokine Production

DC Development

Adjuvant

With Jeff Ulmer & Andreas Wack, Novartis Vaccines

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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, 4781–4789 (2006).Harari, A., Petitpierre, S., Vallelian, F. & Pantaleo, G.. Blood 103, 966–972 (2004).Younes, S. A. et al. J. Exp. Med. 198, 1909–1922 (2003)Robert A. Seder, Patricia A. Darrah, Mario Roederer, Nat. Rev. Immunology 8, 257 (2008)

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Magnitude of Primary In Vitro CTL Responses to Yellow FeverCD8 T cells recognize “naïve” antigen

0 102

103

104

105

<PE‐Cy7‐A>: IFN‐g

0

102

103

104

105

<APC

‐Alexa750‐A>: CD107a

3.98 1.73

0.793.6

0 102

103

104

105

<PE‐Cy7‐A>: IFNg

0.33 0.0056

0.00699.7

0 102 103 104 105

<PE‐Cy7‐A>: IFN‐g

0

102

103

104

105

<APC

‐Alexa750‐A>: CD107a

2.97 5.14

5.4586.5

0

102

103

104

105

<APC

‐Alexa750‐A>: 107a

0 102 103 104 105

<PE‐Cy7‐A>: IFNg

0

102

103

104

105

<APC

‐Alexa750‐A>: 107a

0.57 0.019

0.02699.4

Yellow Feverno Antigen

PrimaryStimulation(7 days)

IFNγ

CD10

7a

SecondaryStimulation (12 days)

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Quality of CD8 T Cell Responses: In Vitro Yellow Fever Vaccination

0.0%

0.1%

0.2%

0.3%

0.4%

CGIT CGI CGT GIT CIT CI CT CG IT GI GT G C I T

Freq

uenc

y

2.58% 7.24%

32.53%57.65%

QuadrupleTripleDoubleSingle

Day 7

Day 15 Re‐stimulation

0%

2%

4%

6%

CGIT CGI CGT GIT CIT CI CT CG IT GI GT G C I T

Freq

uenc

y

1.04%14.87%

28.91%55.17%

QuadrupleTripleDoubleSingle

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Quantity: Pre/Post Influenza Vaccine Stimulation:Anti‐Fluvirin IgG

Antigen stimulation & vaccination boosted MIMIC anti‐Fluvirin IgG

Anti‐FluvirinIgG (n

g/ml)

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Quality of Ab Responses: Avidity Indices and Cross ReactivityLast year’s flu vaccine, esp. Solomon Island H1 component, is associated with

reduced Ab avidity and reduced cross‐reactivity

Stronger NC memory, high avidity, response in pre‐vaccinated donors

Stronger SI, low avidity, response in post‐vaccinated donors

Pre‐vaccination Post‐vaccination

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Rapid T and B Cell Immunogen Screening

ControlControl

Ag1Ag1

Ag2Ag2

Ag3Ag3Don

or 1

Don

or 1

Don

or 2

Don

or 2

Don

or 3

Don

or 3

Don

or 4

Don

or 4

Don

or 5

Don

or 5

Don

or 6

Don

or 6

● 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

● Permits the evaluation of primary and recall responses

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AutomationManual

1 hr2.5 hrsBlood Processing

Labor Hours

3 hrs40 hrsELISAHours per 36 plates

10 hrs40 hrsELISPOTHours per 32 plates

24 hrs 6 hrsVaccination SiteHours per 32 plates

Significant Time/Cost Savings via Automation

Automated ELISA Results

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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

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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

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Engineering an in vitro human immune system for rapid vaccine evaluation