Engineering an in vitro human immune system for rapid vaccine evaluation
Transcript of Engineering an in vitro human immune system for rapid vaccine evaluation
1
Engineering an in vitro human immune system for rapid vaccine evaluation
William Warren, Ph.D.VaxDesign CorporationOrlando, FL 32826
2
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:
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
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
5
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
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
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, 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)
8
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)
9
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
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
11
Quantity: Pre/Post Influenza Vaccine Stimulation:Anti‐Fluvirin IgG
Antigen stimulation & vaccination boosted MIMIC anti‐Fluvirin IgG
Anti‐FluvirinIgG (n
g/ml)
12
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
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
14
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
15
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
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