Induction of neutralizing antibodies against Tityus serrulatus scorpion toxins by immunization with a mixture of defined synthetic epitopes

被引:29
作者
Alvarenga, LM
Diniz, CR
Granier, C
Chávez-Olórtegui, C
机构
[1] Fdn Ezequiel Dias, BR-30550010 Belo Horizonte, MG, Brazil
[2] Univ Montpellier 1, Inst Biotechnol & Pharmacol, Montpellier, France
关键词
D O I
10.1016/S0041-0101(01)00197-0
中图分类号
R9 [药学];
学科分类号
1007 ;
摘要
We have used the Spot method of multiple peptide synthesis to prepare sets of immobilized overlapping peptides of uniform size (15 mer), covering the complete amino acid sequences of TsNTxP a non-toxic and immunogenic protein and TsIV, an alpha -type toxin that is the major lethal component of the venom of scorpion Tityus serrulatus. Anti-TsNTxP antibodies binding to peptides, revealed three antigenic regions, one in the N-terminal, the second in the central part and the other in the C-terminal part of TsNTxP. One peptide epitope in the C-terminal part of TsIV was identified with anti-TsIV neutralizing rabbit antibodies. Anti-peptide antibodies were raised against these four peptides all together covalently coupled to keyhole limpet hemocyanin (KLH) and found to neutralize in vitro the toxic effects of the T serrulatus venom. Quantities of venom equivalent to 13,5 LD50 were effectively neutralized by I ml of the anti-peptide serum. The antigenic specificities of the anti-peptides were compared by an indirect enzyme-linked immunosorbent assay (ELISA) using synthetic peptides and crude venoms from T. serrulatus, T. bahiensis, T. cambridgei, T. stigmurus, Androctonus autralis Hector and Centruroides sculpturatus to coat the microtitration plates. The anti-peptide antibodies had a comparable high reactivity with the crude venom of T. serrulatus, moderate binding to T. bahiensis, T. cambridgei, T. stigmurus and Centruroides sculpturatus venoms but were unable to recognize the venom of Androctonus autralis Hector. These results show that by using peptides derived from the sequence of scorpion toxins, the generation of anti-peptide antibodies able to neutralize the cognate venom appears to be an alternative strategy for the easy preparation of antivenoms. (C) 2001 Published by Elsevier Science Ltd.
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页码:89 / 95
页数:7
相关论文
共 28 条
[11]  
DARBON H, 1983, INT J PEPT PROT RES, V22, P179
[12]   OXIDATION/SULFIDATION OF IRON-ALUMINUM ALLOYS [J].
DEVAN, JH ;
TORTORELLI, PF .
MATERIALS AT HIGH TEMPERATURES, 1993, 11 (1-4) :30-35
[13]  
Diniz CR, 1978, ARTHROPOD VENOMS, P379
[14]   ARCHITECTURE OF SCORPION NEUROTOXINS - A CLASS OF MEMBRANE-BINDING PROTEINS [J].
FONTECILLACAMPS, JC ;
ALMASSY, RJ ;
EALICK, SE ;
SUDDATH, FL ;
WATT, DD ;
FELDMANN, RJ ;
BUGG, CE .
TRENDS IN BIOCHEMICAL SCIENCES, 1981, 6 (11) :291-296
[15]   SPOT-SYNTHESIS - AN EASY TECHNIQUE FOR THE POSITIONALLY ADDRESSABLE, PARALLEL CHEMICAL SYNTHESIS ON A MEMBRANE SUPPORT [J].
FRANK, R .
TETRAHEDRON, 1992, 48 (42) :9217-9232
[16]   Venom variability among several Tityus serrulatus specimens [J].
Kalapothakis, E ;
ChavezOlortegui, C .
TOXICON, 1997, 35 (10) :1523-1529
[17]  
Karber C, 1937, BIOL STANDARDIZATION, P37
[18]   BIOCHEMICAL, PHARMACOLOGICAL AND GENOMIC CHARACTERIZATION OF TS-IV, AN ALPHA-TOXIN FROM THE VENOM OF THE SOUTH-AMERICAN SCORPION TITYUS-SERRULATUS [J].
MARTINEAUCLAIRE, MF ;
CEARD, B ;
RIBEIRO, AM ;
DINIZ, CR ;
ROCHAT, H ;
BOUGIS, PE .
FEBS LETTERS, 1994, 342 (02) :181-184
[19]  
MERY J, 1993, INT J PEPT PROT RES, V42, P44
[20]   PURIFICATION OF ANIMAL NEUROTOXINS - ISOLATION AND CHARACTERIZATION OF 11 NEUROTOXINS FROM VENOMS OF SCORPIONS ANDROCTONUS-AUSTRALIS-HECTOR, BUTHUS-OCCITANUS-TUNETANUS AND LEIURUS-QUINQUESTRIATUS-QUINQUESTRIATUS [J].
MIRANDA, F ;
KUPEYAN, C ;
ROCHAT, H ;
ROCHAT, C ;
LISSITZK.S .
EUROPEAN JOURNAL OF BIOCHEMISTRY, 1970, 16 (03) :514-&