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البحوث المنشورة
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البحث (1):
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عنوان البحث:
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Genetically engineered
two-warhead evasins provide a method to achieve precision targeting of
disease-relevant chemokine subsets
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رابط إلى البحث:
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Click Here
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تاريخ النشر:
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20/04/2018
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موجز عن البحث:
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Both CC and CXC-class chemokines
drive inflammatory disease. Tick salivary chemokine-binding proteins (CKBPs),
or evasins, specifically bind subsets of CC- or CXC-chemokines, and could
precisely target disease-relevant chemokines. Here we have used yeast surface
display to identify two tick evasins: a CC-CKBP, P1243 from Amblyomma
americanum and a CXC-CKBP, P1156 from Ixodes ricinus. P1243 binds 11
CC-chemokines with Kd < 10 nM, and 10 CC-chemokines with Kd between 10 and
100 nM. P1156 binds two ELR + CXC-chemokines with Kd < 10 nM, and four ELR
+ CXC-chemokines with Kd between 10 and 100 nM. Both CKBPs neutralize
chemokine activity with IC50 < 10 nM in cell migration assays. As both CC-
and CXC-CKBP activities are desirable in a single agent, we have engineered
“two-warhead” CKBPs to create single agents that bind and neutralize subsets
of CC and CXC chemokines. These results show that tick evasins can be linked
to create non-natural proteins that target subsets of CC and CXC chemokines.
We suggest that “two-warhead” evasins, designed by matching the activities of
parental evasins to CC and CXC chemokines expressed in disease, would achieve
precision targeting of inflammatory disease-relevant chemokines by a single
agent.
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البحث (2):
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عنوان البحث:
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The N-terminal domain of a tick
evasin is critical for chemokine binding and neutralization and confers
specific binding activity to other evasins
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رابط إلى البحث:
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Click
Here
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تاريخ النشر:
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27/02/2018
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موجز عن البحث:
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Tick
chemokine-binding proteins (evasins) are an emerging class of biologicals
that target multiple chemokines and show anti-inflammatory activities in
preclinical disease models. Using yeast surface display, we identified a
CCL8-binding evasin, P672, from the tick Rhipicephalus pulchellus We found
that P672 binds CCL8 and eight other CC-class chemokines with a Kd < 10 nm
and four other CC chemokines with a Kd between 10 and 100 nm and neutralizes
CCL3, CCL3L1, and CCL8 with an IC50 < 10 nm The CC chemokine-binding
profile was distinct from that of evasin 1 (EVA1), which does not bind CCL8.
We also show that P672’s binding activity can be markedly modulated by the
location of a StrepII-His purification tag. Combining native MS and bottom-up
proteomics, we further demonstrated that P672 is glycosylated and forms a 1:1
complex with CCL8, disrupting CCL8 homodimerization. Homology modeling of
P672 using the crystal structure of the EVA1 and CCL3 complex as template
suggested that 44 N-terminal residues of P672 form most of the contacts with
CCL8. Replacing the 29 N-terminal residues of EVA1 with the 44 N-terminal
residues of P672 enabled this hybrid evasin to bind and neutralize CCL8,
indicating that the CCL8-binding properties of P672 reside, in part, in its
N-terminal residues. This study shows that the function of certain tick
evasins can be manipulated simply by adding a tag. We conclude that homology
modeling helps identify regions with transportable chemokine-binding
functions within evasins, which can be used to construct hybrid evasins with
altered properties.
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البحث (3):
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عنوان البحث:
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Yeast surface display identifies
a family of evasins from ticks with novel polyvalent CC chemokine-binding
activities
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رابط إلى البحث:
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Click Here
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تاريخ النشر:
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27/06/2017
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موجز عن البحث:
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Chemokines function via
G-protein coupled receptors in a robust network to recruit immune cells to
sites of inflammation. Due to the complexity of this network, targeting
single chemokines or receptors has not been successful in inflammatory
disease. Dog tick saliva contains polyvalent CC-chemokine binding peptides
termed evasins 1 and 4, that efficiently disrupt the chemokine network in
models of inflammatory disease. Here we develop yeast surface display as a
tool for functionally identifying evasins, and use it to identify 10 novel
polyvalent CC-chemokine binding evasin-like peptides from salivary
transcriptomes of eight tick species in Rhipicephalus and Amblyomma genera.
These evasins have unique binding profiles compared to evasins 1 and 4,
targeting CCL2 and CCL13 in addition to other CC-chemokines. Evasin binding
leads to neutralisation of chemokine function including that of complex
chemokine mixtures, suggesting therapeutic efficacy in inflammatory disease.
We propose that yeast surface display is a powerful approach to mine
potential therapeutics from inter-species protein interactions that have
arisen during evolution of parasitism in ticks.
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المؤتمرات العلمية:
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المؤتمر (1):
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عنوان المؤتمر:
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2nd European Chemokine and Cell Migration
Conference 2017
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تاريخ الإنعقاد:
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06/09/2017
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مكان
الإنعقاد:
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Cardiff, UK
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طبيعة المشاركة:
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Poster presentation
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عنوان المشاركة:
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Genetically engineered 2-warhead
evasins provide a method for precision targeting of chemokine expression
patterns in disease
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ملخص المشاركة:
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A broad range of diseases are
characterized by inflammation driven by the expression of subsets of CC and
CXC-class chemokines. These form a robust network, mediated by polyvalent
receptor ligand binding, expression of multiple chemokines in disease tissues
and multiple receptors on inflammatory cells, and synergistic interactions
between chemokines. Naturally occurring polyvalent chemokine-binding proteins
(CKBPs) in viruses and helminths polyvalently target both CC and CXC
chemokines but provide no opportunity to selectively target disease relevant
chemokine subsets. Tick salivary CKBPs specifically bind subsets of either
CC- or of CXC-chemokines, and could provide a method to polyvalently but
precisely target disease-relevant chemokines, and such CKBPs have shown
efficacy in pre-clinical disease models. However, both CC- and CXC-CKBP
activities are desirable in a single agent to match the CC and CXC chemokine
expression patterns observed in disease. Here we have used yeast surface
display to identify a CC-CKBP, P1243_AMBAM from the tick Amblyomma americanum
and a CXC-CKBP, P1156_IXORI from the tick Ixodes ricinus. P1243_AMBAM
selectively binds 11 CC-chemokines with Kd <10 nM, and 10 CC-chemokines
with Kd between 10 and 100 nM. P1156_IXORI selectively binds two
CXC-chemokines with Kd <10 nM, and three CXC-chemokines with Kd between 10
and 100 nM. Both CKBPs neutralize chemokine activity in chemotaxis assays
with IC50 in low nM range. We have created “two-warhead” CKBPs by linking
P1243_AMBAM and P1156_IXORI to create single agents that bind and neutralize
subsets of CC and CXC chemokines. These results provide proof-of-concept that
tick CKBPs can be physically linked to create non-natural peptides that
target subsets of CC and CXC chemokines. We suggest that novel “two-warhead”
CKBPs, designed by matching the chemokine-subset binding properties of
parental CKBPs to the CC and CXC chemokines expressed in a particular
inflammatory disease, would achieve precision targeting of disease-relevant
CC and CXC chemokine subsets by a single agent.
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المؤتمر (2):
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عنوان المؤتمر:
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London Inflammation Network 2017
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تاريخ الإنعقاد:
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08/12/2017
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مكان
الإنعقاد:
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London, UK
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طبيعة المشاركة:
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Poster Presentation
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عنوان المشاركة:
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Genetically engineered 2-warhead
evasins provide a method for precision targeting of chemokine expression
patterns in disease
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ملخص المشاركة:
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A broad range of diseases are
characterized by inflammation driven by the expression of subsets of CC and
CXC-class chemokines. These form a robust network, mediated by polyvalent
receptor ligand binding, expression of multiple chemokines in disease tissues
and multiple receptors on inflammatory cells, and synergistic interactions
between chemokines. Naturally occurring polyvalent chemokine-binding proteins
(CKBPs) in viruses and helminths polyvalently target both CC and CXC
chemokines but provide no opportunity to selectively target disease relevant
chemokine subsets. Tick salivary CKBPs specifically bind subsets of either
CC- or of CXC-chemokines, and could provide a method to polyvalently but
precisely target disease-relevant chemokines, and such CKBPs have shown
efficacy in pre-clinical disease models. However, both CC- and CXC-CKBP
activities are desirable in a single agent to match the CC and CXC chemokine
expression patterns observed in disease. Here we have used yeast surface
display to identify a CC-CKBP, P1243_AMBAM from the tick Amblyomma americanum
and a CXC-CKBP, P1156_IXORI from the tick Ixodes ricinus. P1243_AMBAM
selectively binds 11 CC-chemokines with Kd <10 nM, and 10 CC-chemokines
with Kd between 10 and 100 nM. P1156_IXORI selectively binds two
CXC-chemokines with Kd <10 nM, and three CXC-chemokines with Kd between 10
and 100 nM. Both CKBPs neutralize chemokine activity in chemotaxis assays
with IC50 in low nM range. We have created “two-warhead” CKBPs by linking
P1243_AMBAM and P1156_IXORI to create single agents that bind and neutralize
subsets of CC and CXC chemokines. These results provide proof-of-concept that
tick CKBPs can be physically linked to create non-natural peptides that
target subsets of CC and CXC chemokines. We suggest that novel “two-warhead”
CKBPs, designed by matching the chemokine-subset binding properties of
parental CKBPs to the CC and CXC chemokines expressed in a particular
inflammatory disease, would achieve precision targeting of disease-relevant
CC and CXC chemokine subsets by a single agent.
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جوائز التكريم:
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الجائزة (1):
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مسمى الجائزة:
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London
Inflammation Network (LIN) Poster Prize
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الجهة المانحة:
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London
Inflammation Network (LIN) committee
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تاريخ الجائزة:
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08/12/2017
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مجال التكريم:
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Excellent poster titled: Genetically
engineered 2-warhead evasins provide a method for precision targeting of
chemokine expression patterns in disease presented in London Inflammation
Network 2017 .
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