human bacterial challenge experiments as an alternative
DESCRIPTION
Professor Stephen Gordon's presentation at Meningitis Research Foundation's 2013 Conference, Meningitis & Septicaemia in Children & AdultsTRANSCRIPT
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November 2013 MRF Annual Conference, London
Experimental Human Infection with Pneumococcus
Stephen Gordon
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2© The Liverpool School of Tropical Medicine
Pathogenesis of pneumococcal disease
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Outline
•Previous studies of experimental pneumococcal carriage–Jeff Weiser early work–Our first experiments
•How we do our EHPC model–Method, safety issues
•Results–Challenge / re-challenge–Humoral response–Cellular response
•Where next?–Vaccine testing–Vaccine discovery–Host immunity–Microbial ecology
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Previous studies of experimental carriage
•Jeff Weiser (U Penn, Philadelphia)
•JEM 2002 and I&I 2003
•Type 23F 6/14 colonised
•Type 6B 6/8 colonised
•Association noted with IgG to 22kD protein in serum
•Did not particularly comment on dose dependency
•3 washes prior to inoculation
•F/up twice weekly and then weekly
Type cfu/naris colonised
23F 5000 0 (4)
23F 7000 3 (6)
23F 17000 3 (4)
6B 22000-39000
6 (8)
J Expt Med 2002;195:359-362 and I&I 2003;71:5724-32
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Experimental Human Pneumococcal Carriage Early Studies
Study number Description Dates n Carriers
1 23F Bronchoscopy Nov 09-Feb 10 9 1 =11%
2 23F No Bronchoscopy Mar 10-Apr 10 6 0
3 6B No Bronchoscopy Oct 2010 7 5= 71%
4 6B Bronchoscopy Jan 11-Mar 11 9 0
5 6B No Bronchoscopy Mar 11-Aug 11 9 2 =22%
6 6B Re-challenge Jun 11-Sep 11 3 0
7 6B No Bronchoscopy-inocula test Sep 2011 5 1=20%
8 23F No Bronchoscopy-inocula test Sep 2011 4 2 =50%
9 Gates EHPC –dose ranging 6B
Nov 11-Feb 12 50 21=42%
10 Gates EHPC –dose ranging 23F
Feb 12- 60 3=6.0%
11 Gates EHPC - 6B re-challenge Mar 12-May 12 7 0
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6© The Liverpool School of Tropical Medicine
Outline
•Previous studies of experimental pneumococcal carriage–Jeff Weiser early work–Our first experiments
•How we do our EHPC model–Method, safety issues
•Results–Challenge / re-challenge–Humoral response–Cellular response
•Where next?–Vaccine testing–Vaccine discovery–Host immunity–Microbial ecology
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7© The Liverpool School of Tropical Medicine
Method
Watch the video
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8© The Liverpool School of Tropical Medicine
Experimental Human Pneumococcal Carriage
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9© The Liverpool School of Tropical Medicine
Outline
•Previous studies of experimental pneumococcal carriage–Jeff Weiser early work–Our first experiments
•How we do our EHPC model–Method, safety issues
•Results–Dose ranging and challenge / re-challenge–Humoral response–Cellular response
•Where next?–Vaccine testing–Vaccine discovery–Host immunity–Microbial ecology
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10© The Liverpool School of Tropical Medicine
Dose ranging study
2. Reproducibility study: 42% carriage
1. Dose ranging studies: 6B and 23F
(Gritzfeld, J) in preparation
Over 200 people inoculated with pneumococcus, no SAE
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Re-challenge: Carriage protects against acquisition of carriage
Subjects PS Proteins Neither
s1 ++ +s2 =s3 +s4 =s5 ++s6 + ++s7 +s8 ++s9 + +
s10 +
Ferreira et al, AJRCCM 2013
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Response to carriage- cellular
Note IL-17 response following colonisation
Responses can be compared in non-carriage vs carriage
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12.5ng/ml 125ng/ml 625ng/ml
Increased uptake compared to
vehicle
Decreased uptake compared to
vehicle
p = 0.013
p = 0.004
26% ↑ 37% ↑-Overall % increase in uptake
rhIL-17 dose
Macrophages activated with IL17 kill pneumococci
Percentage change in cfu recovered compared to media
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14© The Liverpool School of Tropical Medicine
Outline
•Previous studies of experimental pneumococcal carriage–Jeff Weiser early work–Our first experiments
•How we do our EHPC model–Method, safety issues
•Results–Challenge / re-challenge–Humoral response–Cellular response
•Where next?–Vaccine testing–Vaccine discovery–Host immunity–Microbial ecology
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15© The Liverpool School of Tropical Medicine
Carriage model to test vaccine efficacy (2010)
N=58
Vaccine 29
Placebo 29
*mock challenge
8
challenge21
*mock challenge
8
challenge21
*Mock challenge to control for natural acquisition
95% 83%
1 1
12 10
T=0 T=4T=1
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16© The Liverpool School of Tropical Medicine
Vaccine (PCV vs Hep A): protection against carriage (2013)
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Vaccine discovery – Me Vaccine
MRC/FAPESP Bilateral AgreementLSTM/ Butantan Institute
Peptide Arrays: linear epitopes
Fragments (conformational epitopes)
Phage display: protective anti-PspA monoclonal antibodies
- IgG - CD4+ Th17
Learning from acquired immunity (EHPC) how to designbetter vaccines – Cross-reactive epitopes of PspA
Me Vaccine to confer serotype-independent protection
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Vaccine discovery - AmiC
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Host immunity – correlates of protection
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Microbial ecology - nasotypes
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Summary of Expt Human Pneumo Carriage project
•Safe inoculation of over 200 volunteers–Strain differences in carriage
•First vaccine study ongoing–Start with PCV–Plan phase 2a in 2014
•Where next?–Vaccine testing – in risk populations–Vaccine discovery – by antigen knockout–Host immunity – role of adjuvant–Microbial ecology – effect of eradication
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Thank YOU!!Thank YOU!!
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Home Team• Daniela M. Ferreira, Adam Wright, Sarah
Glennie, Toni Banyard• Andrea Collins, Sherouk El Batrawy, Jamie
Rylance, Dan Wootton, Ben Morton• Jenna Gritzfeld, Mathieu Bangert, Kondwani
Jambo, Shaun Pennington• Angela Wright, Carole Hancock, Dave
Shaw, Lorna Roche, Jane ArdreyCollaborators• Eddie Ades (CDC)• Jerry Brown (UCL, London)• Geoff Ginsberg (Duke)• David Goldblatt (ICH, London)• Peter Hermans, Amelieke Cremers, Aldert
Zomer (Nijmegen)• Rob Heyderman (Malawi)• Aras Kadioglu (Liverpool)• Keith Klugman, Josh Shak (Emory)• Rick Malley, Mark Lipsitch (Harvard)• Eliane Miyaji, Adriane Moreno (Butantan)Advisors• Jeff Weiser (U Penn), Mark Alderson (PATH)Safety panel• Rob Read, David Lalloo, Brian Faragher...and our willing volunteers
Royal Liverpool and Broad Green University Hospitals NHS Trust
AcknowledgementsAcknowledgements
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24© The Liverpool School of Tropical Medicine
Summary of Expt Human Pneumo Carriage project
•Safe inoculation of over 200 volunteers–Strain differences in carriage
•First vaccine study ongoing–Start with PCV–Plan phase 2a in 2014
•Where next?–Vaccine testing – in risk populations–Vaccine discovery – by antigen knockout–Host immunity – role of adjuvant–Microbial ecology – effect of eradication
Key QuestionsCan we test the model in the elderly? HIV infected?Can we use GMO in the model? Adjuvant?
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Response to carriage - humoral
P=0.01*
PLoS Pathog. 2012 Apr;8(4):e1002622.
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Pre-challenge samples
Post-challenge samples (2 weeks post inoculation)
* ***
*
* * ** **
*** ***** *
Anti-protein IgG - serum
Ab increased 20/27proteins
* P≤0.05 comparing pre vs post samples
Serum response to challenge by MSD
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* *** *
** * * * *
** *
* *** * * * ** * *
***
Pre-challenge samples
Post-challenge samples(2 weeks post inoculation)
*P≤0.05 comparing pre vs post samples
Ab increased 14/27proteins
Ab increased 14/27proteins
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Serum response to challenge by MSD
Mock challenge No carriage found Carriage found
Mock challenge shows lower post inoculation serum levels; carriage alters response to many proteins
Ferreira et al. 2012 Unpub
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Response to carriage- cellular
There are no naive mice in human studies of carriage
We can detect Ag-specific responses in BAL but not NW
Wright et al. 2012 Unpub.
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Colonization density- 6B culture
A higher dose does not equal higher colonization density
Relatively low dose recovery follows inoculation
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A simple model of defence vs pneumococcal disease