global dengue & aedes-transmitted diseases consortium … · 2019-09-13 · dengue and other...
TRANSCRIPT
New discoveries about the molecular specificity of
the human antibody response to dengue and Zika
viruses
Aravinda de Silva
University of North Carolina School of Medicine
PDC Workshop on New and Innovative Approaches to Laboratory Diagnosis of Zika,
Dengue and other Arboviruses, Annecy, France (05/2017)
Outline
• The specificity of Ab responses to primary and secondary
DENV infections
• Recent data on the specificity and kinetics of the Ab
response to Zika
• Implications for Laboratory-based and POC diagnostics
Conventional View: The human Ab response to
flaviviruses is dominated by cross-reactivity.
New Discoveries: While cross-reactive Abs are
present, people also develop robust flavivirus
type-specific Abs.
- POC tests
- Surveillance tools (vaccine trials)
- Past exposure history
5/5/2017 3
Dissecting Antibody Neutralization Capacity
using FACS-based DENV Infection Assay
DENV1
DENV2DENV3DENV4
Reciprocal Neut50 Titer
DENV
1
DENV
2
DENV
3
DENV
4
Baselin
e
<40 <40 <40 <40
Fo
llo
w-U
p
153 <40 <40 <40
Baselin
e
<40 <40 298 <40
Fo
llo
w-U
p
127 142 503 <40
Representative 2° Infection
log1/Dilution
-4.1 -3.6 -3.1 -2.6 -2.1 -1.6-10
0
10
20
30
40
50
60
70
80
90
100
% N
eu
tra
liza
tio
n
Baseline Follow-Up
log1/Dilution
-4.1 -3.6 -3.1 -2.6 -2.1 -1.6-10
0
10
20
30
40
50
60
70
80
90
100
Representative 1° Infection
-4.1 -3.6 -3.1 -2.6 -2.1 -1.6-10
0
10
20
30
40
50
60
70
80
90
100
log1/Dilution
% N
eu
tra
liza
tio
n
Baseline Follow-Up
-4.1 -3.6 -3.1 -2.6 -2.1 -1.6-10
0
10
20
30
40
50
60
70
80
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100
log1/Dilution
Depletion of dengue virus reactive antibody from human sera
Obtain highly pure
dengue virus
Immobilize virus
on to beads
Virus conjugated Beads + Human Sera
Incubated at 37 °C for 2 hrs (3x)
Depleted human sera
Envelope
preM
Capsid
M
Levels of TS and CR Abs after primary and secondary DENV infections
5/5/2017 6
Primary
DENV
infections
Secondary
DENV
infections
Patel et al PLOS NTD In Press.
5/5/2017 7
DENV type-specific neutralizing antibodies reflect past infection history
Patel et al PLOS NTD In Press.Collaboration with
Eva Harris and Magelda Montoya
5/5/2017 8Patel et al PLOS NTD In Press.
Model for antibody responses after primary and secondary DENV infections
Dengue envelope glycoprotein
I IIpr M I IIII II TM
E protein
EDIII
EDIII
EDI
EDIEDII
EDII
Kuhn, RJ et al. Cell, 2002
DENV2 neutralizing human MAbs target quaternary
structure epitopes
2D22 1L12 3F9 + -0.00
0.25
0.50
0.75
1.00
1.25
MAb
Bin
din
g (
OD
405)
DENV2 Virus
2D22 1L12 3F9 + -0.00
0.25
0.50
0.75
1.00
MAb
Bin
din
g (
OD
405)
DENV2 rE
2D22 1L12 3F9 + -0.00
0.25
0.50
0.75
1.00
MAb
Bin
din
g (
OD
405)
DENV2 rEDIII
Emily Gallichotte
The Epitope of DENV2-specific hMAb 2D22
Fibriansha et al Science 2015: Vol. 349 no. 6243 pp. 88-91
Location of quaternary epitopes defined with type-specific neutralizing hMAbs (MBC)
DENV2 hMAb 2D22 6 FAbs per raft
DENV1hMAb 1F4, 14C10 4 FAbs per raft
DENV3hMAb 5J72 FAbs per raft
DENV4 epitope mapping inprogress
Location of epitopes defined with cross-reactive neutralizing hMAbs (MBC)
EDE1 and EDE2hMAbs
Fusion loop/BC Loop
Rouvinski et al. Nature 2015
Dejnirattisai et al. Nat Med 2015
Tsai et al. JV 2014
Smith mBio 2014
Summary
• Primary Infections
» Durable serotype-specific Ab responses
» Tertiary and quaternary structure epitopes
» Abundance of cross-reactive poorly neutralizing Abs
• Secondary Infections
» Dominance of cross reactive neutralizing Abs
» Type-specific antibodies are maintained
» Tertiary and quaternary (EDE) structure epitopes (?)
» NAb response becomes more cross reactive with each new
exposure to DENV
5/5/2017 14
Is Zika a 5th DENV serotype?
•No !
5/5/2017 15
Collins MH, McGowan E, Jadi R, Young E, Lopez CA, Baric RS, et al.
Lack of Durable Cross-Neutralizing Antibodies Against Zika Virus from
Dengue Virus Infection. Emerg Infect Dis. 2017;23(5):773-781.
https://dx.doi.org/10.3201/eid2305.161630
5/5/2017 16
5/5/2017 17
Primary Zika or DENV exposure does not lead to durable
CR neutralizing antibodies
Collins et al Emerg Infect Dis. 2017;23(5):773-781.
5/5/2017 18
Secondary DENV exposure does not lead to durable
CR neutralizing antibodies
Collins et al. Emerg Infect Dis. 2017;23(5):773-781.
DENV immune individuals exposed to Zika have a ZIKV TS neut Ab response
Deplete sera of DENV2 binding antibodies and then test for
neutralization of DENV and ZIKV
5/5/2017 19
DENV2 neuts ZIKV Neuts
Collins et al Emerg Infect Dis. 2017;23(5):773-781.
Secondary DENV infections, Hospital-based study
Primary DENV infections, Hospital-based study
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DENV1−Primary
Timepoint
NT
50
Acute Conv 3m 6m 12m 18m
<10
10
20
40
80
160
320
640
1280
2560
5120
10240
20480 ZIKV
DENV1
DENV2
DENV3
DENV4
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DENV1−Secondary
Timepoint
NT
50
Acute Conv 3m 6m 12m 18m
<10
10
20
40
80
160
320
640
1280
2560
5120
10240
20480 ZIKV
DENV1
DENV2
DENV3
DENV4
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DENV2−Primary
Timepoint
NT
50
Acute Conv 3m 6m 12m 18m
<10
10
20
40
80
160
320
640
1280
2560
5120
10240
20480 ZIKV
DENV1
DENV2
DENV3
DENV4
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DENV2−Secondary
Timepoint
NT
50
Acute Conv 3m 6m 12m 18m
<10
10
20
40
80
160
320
640
1280
2560
5120
10240
20480 ZIKV
DENV1
DENV2
DENV3
DENV4
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DENV3−Primary
Timepoint
NT
50
Acute Conv 3m 6m 12m 18m
<10
10
20
40
80
160
320
640
1280
2560
5120
10240
20480 ZIKV
DENV1
DENV2
DENV3
DENV4
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DENV3−Secondary
Timepoint
NT
50
Acute Conv 3m 6m 12m 18m
<10
10
20
40
80
160
320
640
1280
2560
5120
10240
20480 ZIKV
DENV1
DENV2
DENV3
DENV4
●
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DENV1−Primary
Timepoint
NT
50
Acute Conv 3m 6m 12m 18m
<10
10
20
40
80
160
320
640
1280
2560
5120
10240
20480 ZIKV
DENV1
DENV2
DENV3
DENV4
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DENV1−Secondary
Timepoint
NT
50
Acute Conv 3m 6m 12m 18m
<10
10
20
40
80
160
320
640
1280
2560
5120
10240
20480 ZIKV
DENV1
DENV2
DENV3
DENV4
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●
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DENV2−Primary
Timepoint
NT
50
Acute Conv 3m 6m 12m 18m
<10
10
20
40
80
160
320
640
1280
2560
5120
10240
20480 ZIKV
DENV1
DENV2
DENV3
DENV4
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DENV2−Secondary
Timepoint
NT
50
Acute Conv 3m 6m 12m 18m
<10
10
20
40
80
160
320
640
1280
2560
5120
10240
20480 ZIKV
DENV1
DENV2
DENV3
DENV4
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DENV3−Primary
Timepoint
NT
50
Acute Conv 3m 6m 12m 18m
<10
10
20
40
80
160
320
640
1280
2560
5120
10240
20480 ZIKV
DENV1
DENV2
DENV3
DENV4
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DENV3−Secondary
Timepoint
NT
50
Acute Conv 3m 6m 12m 18m
<10
10
20
40
80
160
320
640
1280
2560
5120
10240
20480 ZIKV
DENV1
DENV2
DENV3
DENV4
ZIKV neutralization at different times after DENV infection
Data from Eva Harris and Magelda Montoya UC- Berkeley
Summary
• Low cross-neutralization between DENVs and ZIKVs
• People exposed to ZIKV as a primary or secondary
flavivivrus infection develop high levels of TS Nabs
• Mapping of epitopes in progress
5/5/2017 21
Relevance to Diagnostics
• Many candidate recombinant antigens for flavivirus type-
specific POC diagnostics (look beyond virus and whole
protein antigens- Laksmanane presentation)
• The neutralization assay is a valuable tool for surveillance
and for supporting vaccine clinical trials (Include relevant
flaviviruses in circulation and compare levels of Nabs)
5/5/2017 22
• UNC-
» deSilva Lab: Emily Gallichotte, Bhumi Patel, Matthew Collins, Ramesh Jadi, RajendraRaut and Alice Liou
» Ralph Baric and his group
• Vanderbilt University
» James Crowe Lab
• Duke-NUS, Singapore
» Sheemei Lok and Guntur Fibriansah
• UC- Berkeley
» Eva Harris and Magelda Montoya
Grants:
• NIH/NIAID 1-R01-AI125198-01 (de Silva)
• NIH/NIAID 5 R01 AI107731 (de Silva)
• NIH/NIAID P01 AI106695 (Harris)
• ZIKA PLAN (Wilder-Smith)
NEW PARTNERSHIP WITH CDC