ВЫЯВЛЕНИЕ СПЕЦИФИЧЕСКИХ I GE АНТИТЕЛ

, 2009,
8,
1
I GE
FC
,
.
«
»
.
,
Ig.
(Fc RI) –
,
.
Fc RI
-
Dermatophagoides
WRNWDVYK.
pteronyssinus.
.
:
IgE
,
,
Abstract. Such new diagnostic direction as antibody detection with the help
of peptides which are the analogues of Ig-binding site, located in the active
centers of receptors for immunoglobulins is suggested in this article. High affinity
receptor for immunoglobulin E (Fc RI) is a key highly specialized structure on
the surface of a number of immune cells. Fragment of active Fc RI center –
octapeptide WRNWDVYK has been synthesized by means of classical methods
of peptide chemistry. The synthesized octapeptide’s capacity to bind specific IgE
to allergen of mite Dermatophagoides pteronyssinus has been proved with the
help of immunoenzyme test-system. The obtained peptide may potentially be
used for diagnostic purposes.
1
, 2009,
8,
1
Key words: peptide, immunoglobulin E, IgE, high affinity receptor for
immunoglobulin E, Fc RI, bronchial asthma.
:
, 210023, .
,
.
,
27,
,
–
.
.
.
.
[1].
,
.
(Fc RI)
,
–
[2,3],
.
Fc: , , ,
.
Fc,
.
2
Fc RI:
[4,5].
Fc RI (Fc RI )
,
.
1988
Fc R1A
[6].
1
2
, 2009,
8,
,
1
1q23 [7,8].
IgE
Fc RI
.
Fc RI -IgE
2
.
,
Fc RI -IgE
IgE
2
2
IgE.
Fc RI
[9].
Fc RI
IgE
,
IgE.
in
vitro
,
IgE.
WRNWDVYKIgE
,
Fc RI ,
.
9.0
(NCBI) –
2
«
»,
NM_002001,
(PDB) -
1f6a
(
:
S.C.Garman, B.A.Wurzburg, S.S.Tarchevskaya, J.P.Kinet, T.S.Jardetzky)
E (Fc RI ).
,
,
,
2
IgE.
Fc RI
–
WRNWDVYK [10].
IgE
WRNWDVYK
product
(Sigma,
P8375).
WRNWDVYK[11].
3
, 2009,
8,
1
,
A280
,1
= 0,73, A403
,1
,1
= 2,275,
,1
403
= 109 000
-1
-1
0,8
– 5,2
.
,
.
(“
+”,
“
”)
1:2.
+4° .
,
,
WRNWDVYK-
Dermatophagoides pteronyssinus
«
.
IgE
DP0004.
(100 PNU/
)
Dermatophagoides pteronyssinus (5000 PNU/
,
.
»)
0,1
100
Na(Nunc MaxiSort 468667).
18
+4
.
18
3
300
1
,
.
+4°
WRNWDVYKDermatophagoides
pteronyssinus
Anti-Human IgE ( -chain specific) peroxidase
conjugate (Sigma, product
A9667).
Anti-Human IgE ( -chain specific) peroxidase conjugate
(Sigma, product
1
A9667) 2
1:10 PBS-T,
.
-20° .
24
4
, 2009,
8,
1
Dermatophagoides pteronyssinus.
“Global
Intitiative for Asthma. Global Strategy for Asthma Management and Prevention”
[12]
Dermatophagoides pteronyssinus.
DP0004 WRNWDVYK-
1
2
•
2
20 (PBS-T)
180
20
•
,
60
•
3
;
+37° ;
PBS-T;
•
150
WRNWDVYK,
•
PBS-T 1:200;
150
Human IgE
Anti-
,
•
PBS-T 1:200;
60
•
3
•
150
(Sigma, product
•
PBS-T;
3,3'5,5'-Tetramethyl-benzidine (
)
T8665);
60
•
37° ;
+18°
;
100
;
•
,
-01C,
»,
,
450
.
WRNWDVYK-
IgE
.
.
,
.
,
IgE:
Fc-
;
8E/5D4, IT, BSW –
5
E411, XTE4 –
C 2
;
, 2009,
8E/4F4 -
8,
C 3
1
;
8E/Le27 -
8E/5D4.
C 4
,
,
8E/5D4,
C 2
«
»,
IgE
,
-8660
.
-8660 +
•
1
)
WRNWDVYK-
1:100;
,
PBS-T
;
•
2
)
8E/4F4,
PBS-T 1:100;
;
•
3
)
(
,
,
Fc RI )
[13],
PBS-T 1:100;
:
1
- 180
+ 20
;3
- 120
100
;5
+ 140
20
;2
- 160
+ 80
;4
- 80
;7
+ 120
- 40
+ 40
- 100
;6
+ 160
+ 180
•
30
•
3
•
100
+37° ;
PBS-T;
3,3'5,5'-Tetramethyl-benzidine
;
6
+
- 60
;8
-
, 2009,
•
8,
1
30
•
+18°
;
100
;
•
,
-01C,
450
.
,
WRNWDVYK-
,
,
-8660
8E/5D4
C 2
IgE
.
8E/4F4
(
,
,
Fc RI )
.
(
.1).
1,800
1,600
1,400
1,200
1,000
0,800
WRNWDVYK8E/4F4
0,600
0,400
0,200
0,000
20
40
80
100
120
140
1
.1
,
7
,
160
180
,
, 2009,
8,
1
,
,
,
,
-
.
,
,
.
,
,
,
.
-
.
WRNWDVYK)
8E/5D4.
15-20
,
8
.
,
WRNWDVYK,
.
WRNWDVYK-,
8E/4F4
C 2
.1.
8
IgE (
8E/5D4)
, 2009,
8,
1
1
IgE
Dermatophagoides pteronyssinus
IgE
WRNWDVYR%
WRNWDVY
K
WRNWDVYK-
0,516
0,641
0,624
0,902
0,816
1,061
0,732
0,916
1,288
1,871
1,344
0,657
0,829
0,806
0,864
0,446
0,848
0,965
0,760
1,031
1,229
1,745
1,391
1,435
.
-
%
WRNWDV
YK
WRNWDVY
K-
62,2
79,5
72,2
201,9
96,3
110,0
96,3
88,8
104,8
107,2
96,7
45,8
.
0,887
0,938
1,552
1,050
1,571
1,012
1,216
1,832
1,405
0,711
1,396
1,616
1,386
1,260
1,323
1,155
1,532
1,077
1,285
2,012
1,719
1,177
1,495
1,675
64,0
74,5
117,3
90,9
102,5
94,0
94,7
91,1
81,7
60,4
93,4
96,4
8E/5D4
C 2
IgE
.
,
WRNWDVYK-
,
WRNWDVYKC 2
IgE.
2
8E/4F4
Cε2
IgE (
8E/5D4)
,
-8660
WRNWDVYK8E/4F4) -
IgE
180
160
120
100
80
60
40
20
+ 20
+ 40
+ 80
+ 100
+ 120
+ 140
+ 160
+ 180
.
-
0,375
0,523
0,904
1,191
1,161
1,200
1,728
1,691
.
9
0,069
0,064
0,081
0,064
0,093
0,101
0,207
0,092
0,062
0,058
0,060
0,083
0,073
0,098
0,228
0,228
, 2009,
8,
1
,
IgE
Dermatophagoides
pteronyssinus WRNWDVYK-
IgE
,
.2,
,
WRNWDVYK-
IgE.
IgE
Dermatophagoides pteronyssinus WRNWDVYKIgE
(r = 0,783,
n=24)
.
Dermatophagoides pteronyssinus
,
IgE
pteronyssinus
Dermatophagoides
IgE Anti-Human IgE
WRNWDVYK-
,
(r = -0,093
Anti-Human IgE
r = -0,168
WRNWDVYK-
WRNWDVYK-
).
E
.
WRNWDVYK-
,
,
Dermatophagoides pteronyssinus
.
(r = 0,783, n=24)
WRNWDVYK-
Anti-Human IgE
Dermatophagoides pteronyssinus
.
10
.
, 2009,
8,
1
.
WRNWDVYK
,
,
.
1.
–
/
.:
.
.
.
[
.].
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1353-1365.
3.
, . .
Fc/
. .
,
. – 2006. – . 8,
4.
. .
,
. .
//
2-3. – . 207.
, . .
/ . .
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.:
, 1998. – 252 .
5. Kawakami, T. Regulation of mast-cell and basophil function and survival by
IgE. / T. Kawakami, S. J. Galli // Nat. Rev. Immunol. – 2002. – Vol. 2, N 10. – P.
773-786.
6. Human and rat mast cell high-affinity immunoglobulin E receptors:
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Am. J. Hum. Genet. – 1989. – Vol. 45. – P. 761-765.
8. Le Coniat, M. The human genes for the alpha and gamma subunits of the
mast cell receptor for immunoglobulin E are located on human chromosome band
1q23 / M. Le Coniat, J. P. Kinet, R. Berger // Immunogenetics. – 1990. – Vol. 32,
N 3. – P. 183-186.
11
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9. The structure of the IgE C 2 domain and its role in stabilizing the complex
with its high-affinity receptor Fc RI . / J. M. McDonnell [et al.] // Nature
structural biology. – 2001. – Vol. 8, N 5. – P. 437-441.
10.
study of immunobiological activity of the WRNWDYYK octapeptide /
V. P. Martinovich [et al.] // 30th European Peptide Symposium (30EPS),
Helsinki, Finlandia Hall, 30 Aug.-5 Sept., 2008. – P. 112.
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W. Knapp, K. Holubar, G. Wick. – Elsevier/North Holland Biomedical Press,
Amsterdam, 1978. – P. 215–224.
12. Global Intitiative for Asthma. Global Strategy for Asthma Management
and Prevention. NYLB / WHO Worksop Peport, 1993.
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.] //
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