2013 A DNA prime-protein boost vaccination strategy targeting turkey coronavirus spike protein fragment containing neutr

11 Pages·2013·0.74 MB·English

Veterinary
Immunology
and
Immunopathology
152 (2013) 359–
369

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at
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Veterinary
Immunology
and
Immunopathology

j
ourna
l
ho
me
pag
e:
www.elsevier.com/locate/vetimm

Research
paper

A
DNA
prime-protein
boost
vaccination
strategy
targeting
turkey
coronavirus
spike
protein
fragment
containing
neutralizing
epitope
against
infectious
challenge

Yi-Ning
Chen a,1, Ching
Ching
Wu a, Yoon
Yeo b, Peisheng
Xu b,2,
Tsang
Long
Lin a,∗

a Department
of
Comparative
Pathobiology,
Purdue
University
406
South
University
Street,
West
Lafayette,
IN
47907,
USA
b Department
of
Industrial
and
Physical
Pharmacy,
Purdue
University,
575
Stadium
Mall
Drive,
West
Lafayette,
IN
47907,
USA

a r t i c l e
i n
f o

Article
history:
Received

3
April
2012
Received

in
revised
form
10
January
2013
Accepted

12
January
2013

Keywords:
DNA-prime

protein-boost
regimen
Neutralizing

epitopes
Polyethyleneimine
Spike

protein
Turkey

coronavirus

a b
s t r a
c t

The present
study
was undertaken
to determine
immune
response
and protection
efficacy
of

a spike (S) protein
fragment
containing
neutralizing
epitopes
(4F/4R) of turkey
coronavi-
rus
(TCoV) by priming with DNA vaccine
and boosting
with the recombinant
protein from
the

corresponding
DNA vaccine
gene segment.
Turkeys
were
vaccinated
by priming
with
either

one dose (G1-750DP)
or two doses
(G3-750DDP)
of 750 �g DNA vaccine
express-
ing

4F/4R S fragment
and boosting
with one
dose of 200 �g 4F/4R
S fragment.
One dose
of
100
�g DNA vaccine
mixed with
polyethyleneimine
(PEI) and sodium
hyaluronate
(HA)
followed

by one dose of 750 �g
DNA vaccine
and one dose of 200 �g 4F/4R S fragment
were
given to the turkeys
in group G2-100DPH.
After infectious
challenge
by TCoV, clinical
signs

and TCoV detected
by immunofluorescence
antibody
(IFA) assay
were
observed
in
less
number
of turkeys
in vaccination
groups than that
in challenge
control
groups.
TCoV
viral

RNA loads
measured
by quantitative
real-time
reverse transcription-PCR
were lower
in

vaccinated
turkeys
than those in challenge
control turkeys.
The turkeys
in G3-750DDP
produced
the highest
level of TCoV S protein-specific
antibody
and
virus neutralization
(VN)

titer. Comparing
to the turkeys
in G1-750DP,
significantly
less TCoV were
detected
by

IFA in the turkeys
in G2-100DPH
receiving
an extra dose of 100 �g DNA mixed with
PEI
and HA. The results indicated that DNA-prime
protein-boost
DNA vaccination
regimen
targeting

TCoV
S fragment
encompassing
neutralizing
epitopes
induced
humoral
immune
response

and partially
protected
turkeys
against
infectious
challenge
by TCoV.
© 2013 Elsevier B.V. All rights reserved.

Abbreviations:

ANOVA,
one-way
analysis
of
variance;
CFA,
complete
Freund’s
adjuvant;
CLPEI,
disulfide-crosslinked
low
molecular
linear
polyethyleneimine;
CpG,
cytosine-phosphate-guanine;
Ct,
threshold
cycle
value;
DPH,
DNA,
polyethyleneimine,
and
sodium
hyaluronate
complex;
dpi,
days
post
inoculation;
E,
envelope;
E.
coli, Escherichia
coli; EID50, median
embryo
infectious
dose;
GM-CSF,
granulocyte-macrophage
colony-stimulating
factor;

HA,
hyaluronate;
HEPES,
4-(2-hydroxyethyl)-1-piperazineethanesulfonic
acid
buffered
saline;
HRP,
horseradish
peroxidase;
IBDV,
infectious
bursal
disease
virus;
IBV,
infectious
bronchitis
virus;
IFA,
imnunofluorescent
antibody
assay;
Kb,
kilobase;
kDa,
kilodalton;
LB,
lysogeny
broth;
M, membrane;
MHV,

mouse
hepatitis
virus;
N,
nucleocapsid;
NTC,
non-template
control;
OD
450
nm,
optical
density
at
450
nm;
qRT-PCR,
quantitative
real-time
reverse
transcription-polymerase
chain
reaction;
S,
spike;
SARS-CoV,
severe
acute
respiratory
syndrome-coronavirus;
TCoV,
turkey
coronavirus;
VN,
virus
neu-
tralization.
∗ Corresponding

author
at:
406
South
University
Street,
West
Lafayette,
IN
47907,
USA.
Tel.:
+1
765
494
7927;
fax:
+1
765
494
9181.
E-mail
address: