- INTRODUCTION
- MATERIALS AND METHODS
- Chemicals, reagents, and buffers
- Antibodies
- Animals
- Spiders and venoms
- Expression of recombinant protein
- Antisera
- Normal human serum and erythrocytes
- Electrophoresis and Western blotting
- Enzyme-linked immunosorbent assay (ELISA)
- Neutralization of dermonecrotic activity by sera
- Neutralization of hemolytic activity by sera
- Neutralization of SMase activity by sera
- Statistical analysis
- RESULTS
- DISCUSSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1
Futrell JM, 1992. Loxoscelism. Am J Med Sci 304 :261–267.
-
2
Ginsburg CM, Weinberg AG, 1988. Hemolytic anemia and multiorgan failure associated with localized cutaneous lesion. J Pediatr 112 :496–499.
-
3
Gendron BP, 1990. Loxosceles reclusa envenomation. Am J Emerg Med 8 :51–54.
-
4
Bey TA, Walter FG, Lober W, Schmidt J, Spark R, Schlievert PM, 1997. Loxosceles arizonica bite associated with shock. Ann Emerg Med 30 :701–703.
-
5
Ramos-Cerrillo B, Olvera A, Odell GV, Zamudio F, Paniagua-Solis J, Alagon A, Stock RP, 2004. Genetic and enzymatic characterization of sphingomyelinase D isoforms from the North American fidleback spiders Loxosceles boneti and Loxosceles recluse. Toxicon 44 :507–514.
-
6
Newlands G, Isaacson C, Martindale C, 1982. Loxoscelism in the Transvaal, South Africa. Trans R Soc Trop Med Hyg 76 :610–615.
-
7
Denny WF, Dillaha CJ, Morgan PN, 1964. Hemotoxic effect of Loxoscels reclusa venom: in vivo and in vitro studies. J Lab Clin Med 64 :291.
-
8
Schenone H, Saavedra T, Rojas A, Villarroel F, 1998. Loxoscelism in Chile. Epidemiologic, clinical and experimental studies. Rev Inst Med Trop Sao Paulo 31 :403–415.
-
9
Dillaha CJ, Jansen T, Honeycut WM, Hayden CR, 1964. North America Loxoscelism. JAMA 188 :33–36.
-
10
Rees RS, Shack B, Withers F, Madden J, Franklin J, Lynch JB, 1981. Management of the brown recluse spider bite. Plast Reconstr Surg 68 :768–773.
-
11
Koh WL, 1998. When to worry about spiders bite: inaccurate diagnosis can have serious, even fatal, consequences. Postgrad Med 103 :235–250.
-
12
Allen C, 1992. Arachnid envenomation. Emerg Med Clin North Am 10 :269–299.
-
13
Anderson PC, 1997. Spider bites in the United States. Dermathol Clin 5 :307–311.
-
14
Walter FG, Bilden EF, Gibly RL, 1999. Envenomations. Crit Care Clin 15 :627–629.
-
15
Barrett SM, Romine-Jenkins M, Fisher DE, 1994. Dapsone or electric shock treatment of brown recluse spider envenomation. Ann Emerg Med 24 :21–25.
-
16
Bitterman-Deutsch O, Bergman R, Friedman-Birnbaum R, 1990. Brown spider bite. Harefuah 119 :137–139.
-
17
Wille RC, Morrow JD, 1998. Case report: dapsone hypersensitivity syndrome associated with treatment of the bite of brown spider. Am J Med Sci 296 :270–271.
-
18
Mold JW, Thompson DM, 2004. Management of brown recluse spider bites in primary care. J Am Board Fam Pract 1 :347–352.
-
19
Bryant SM, Pittman LM, 2003. Dapsone use in Loxosceles reclusa enevenomation: is there an indication? Am J Emerg Med 21 :89–90.
-
20
Elston MD, Miller MS, Young RJ III, Eggers J, McGlasson D, Schmidt WH, Brush A, 2005. Comparation of colchicines, dapsone, triamcinolone and diphenhydramine therapy for the treatment of brown recluse spider envenomation. Arch Dermatol 141 :595–597.
-
21
Smith CW, Micks DW, 1970. The role of polymorphonuclear leukocytes in the lesion caused by the venom of the brown spider, Loxosceles reclusa. Lab Invest 22 :141–144.
-
22
Wilson JR, Hagood CO, Jr Prather ID, 2005. Brown recluse spider bites: a complex problem wound. A brief review and case study. Ostomy Wound Manage 51 :59–66.
-
23
Pizzi T, 1957. A histopathological study on necrotic arachnidism by Loxosceles laeta. Bol Chil Parasitol 30 :34–36.
-
24
Sunderkotter C, Seeliger S, Schonlau F, Roth J, Hallmann R, Luger TA, Sorg C, Kolde G, 2001. Different pathway leading to cutaneous leukocytoclastic vasculitis in mice. Exp Dermatol 10 :391–404.
-
25
Tambourgi DV, Paixão-Cavalcante D, Gonçalves-de-Andrade RM, Fernandes-Pedrosa MF, Magnoli FC, Morgan PB, van den Berg CW, 2005. Loxosceles sphingomyelinase induces complement-dependent dermonecrosis, neutrophil infiltration, and endogenous gelatinase expression. J Invest Dermatol 124 :725–731.
-
26
Ministry of Health, 1998. Manual de Diagnóstico e Tratamento de Acidentes por Animais Peçonhento. Fundação Nacional de Saúde/Coordenação de Controle de Zoonoses e Animais Peçonhentos. Brasília: Centro Neurop.
-
27
Kurpiewski G, Forrester LJ, Barret JT, Campbell BJ, 1981. Platelet aggregation and sphingomyelinase D activity of a purified toxin from the venom of Loxosceles recluse. Biochim Biophys Acta 678 :467–476.
-
28
Tambourgi DV, Magnoli FC, van den Berg CW, Morgan BP, de Araujo PS, Alves EW, Dias da Silva WD, 1998. Sphingomyelinases in the venom of the spider Loxosceles intermedia are responsible for both dermonecrosis and complement-dependent hemolysis. Biochem Biophys Res Commun 251 :366–373.
-
29
Tambourgi DV, Petricevich VL, Magnoli FC, Assaf SL, Jancar S, Dias da Silva W, 1998b. Endotoxemic-like shock induced by Loxosceles spider venoms, pathological changes and putative cytokine mediators. Toxicon 3 :391–403.
-
30
Tambourgi DV, Morgan BP, Gonçalves-de-Andrade RM, Magnoli FC, van den Berg CW, 2000. Loxosceles intermedia spider envenomation induces activation of an endogenous metallo-proteinase, resulting in cleavage of glycophorins from the erythrocyte surface and facilitating complement-mediated lysis. Blood 95 :683–691.
-
31
Tambourgi DV, de Sousa da Silva M, Billington SJ, Goncalves-de-Andrade RM, Magnoli FC, Songer JG, van den Berg CW, 2002. Mechanism of induction of complement susceptibility of erythrocytes by spider and bacterial sphingomyelinases. Immunol 107 :93–101.
-
32
Tambourgi DV, Fernandes-Pedrosa MF, van den Berg CW, Gonçalves-de-Andrade RM, Ferracini M, Paixão-Cavalcante D, Morgan BP, Rushmere NK, 2004. Molecular cloning, expression, function and immunoreactivities of members of a gene family of sphingomyelinases from Loxosceles venom glands. Mol Immunol 41 :831–840.
-
33
Paixão-Cavalcante D, van den Berg CW, Fernandes-Pedrosa MF, Gonçalves-de-Andrade RM, Tambourgi DV, 2006. Role of matrix metalloproteinases in HaCaT keratinocytes apoptosis induced by Loxosceles venom sphingomyelinase D. J Invest Dermatol 126 :61–68.
-
34
van den Berg CW, Gonçalves-de-Andrade RM, Magnoli FC, Marchbank KJ, Tambourgi DV, 2002. Loxosceles spider venom induces metalloproteinases mediated cleavage of MCP/CD46 and MHC I and induces protection against C-mediated lysis. Immunology 107 :102–110.
-
35
Fernandes-Pedrosa MF, Junqueira de Azevedo IL, Gonçalves-de-Andrade RM, van den Berg CW, Ramos CR, Ho PL, Tam-bourgi DV, 2002. Molecular cloning and expression of a functional dermonecrotic and haemolytic factor from Loxosceles laeta venom. Biochem Biophys Res Commun 298 :638–645.
-
36
Bucherl W, 1969. Biology and venoms of the most important South American spiders of the genera Phoneutria, Loxosceles, Lycosa, and Latrodectus. Am Zool 9 :157–159.
-
37
Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC, 1985. Measurement of protein using bicinchronic acid. Anal Biochem 150 :76–85.
-
38
Laemmli UK, 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227 :680–685.
-
39
Towbin H, Staehelin TJ, Gordon J, 1979. Eletrophoretic transfer of proteins from acrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad USA 76 :4350–4354.
-
40
Furlanetto RS, Santos NP, Navas J, 1962. Preparação, purificação e doseamento do sôro antiloxoscélico. Cienc Cult 14 :254.
-
41
Tokomura A, Majima E, Kariya Y, Tominagua K, Kogune K, Ogune K, Yasuda K, Fukuzawa K, 2002. Increased production of bioactive lysophosphatidic acid by serum lysophospholipase D in human pregnancy. J Biol Chem 277 :39436–39442.
-
42
Klinman NR, Rockey JH, Karush F, 1965. Equine antihapten antibody II. The gamma-G (7S-gamma) components and their specific interaction. Int J Cancer Suppl 47 :51–60.
-
43
Tambourgi DV, Magnoli FC, von Eicksted VRD, Benedetti ZC, Petrecevich VL, Dias da Silva W, 1995. Incorporation of a 35 kDa purified protein from Loxosceles intermedia spider venom transforms human erythrocytes into activators of autologous complement alternative pathway. J Immunol 155 :4459–4466.
-
44
Tambourgi DV, Pedrosa MF, de Andrade RM, Billington SJ, Griffiths M, van den Berg CW, 2007. Sphingomyelinases D induce direct association of C1q to the erythrocyte membrane causing complement mediated autologous haemolysis. Mol Immunol 44 :576–582.
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45
Gutierrez JM, Leon G, Lomonte B, 2003. Pharmacokinetic–pharmacodynamic relationships of immunoglobulin therapy for envenomation. Clin Pharmacokinet 42 :721–741.
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46
Scherrmann JM, 1994. Antibody treatment of toxin poisoning: recent advances. J Toxicol Clin Toxicol 32 :363–375.
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47
Fernandes I, Mota I, 1991. Isolation of IgGT from hiperimmune horse anti-snake venom serum: its protective ability. Toxicon 29 :1373–1379.
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48
Fernandes I, Takehara HA, Santos AC, Cormont F, Latinne D, Bazin H, Mota I, 1996. Neutralization of bothropic and crotalic venom toxic activities by IgG(T) and IgGa subclasses isolated from the immune horse serum. Toxicon 35 :931–936.
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49
Saetang T, Treamwattana N, Suttijitpaisal P, Ratanabanangkoon K, 1997. Quantitative comparison on the refinement of horse antivenom by salt fractionation and ion-exchange chromatography. J Chromatogr B 700 :233–239.
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Toro AF, Malta MB, Soares SL, da Rocha GC, da Silva Lira M, De Oliveira TA, Takehara HA, Lopes-Ferreira M, Santoro ML, Guidolin R, Gondo HH, Fernandes I, Bárbaro KC, 2006. Role of IgG(T) and IgGa isotypes obtained from arachnidic antivenom to neutralize toxic activities of Loxosceles gaucho, Phoneutria nigriventer and Tityus serrulatus venoms. Toxicon 48 :649–661.
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51
Fernandes-Pedrosa MF, Junqueira-de-Azevedo IL, Gonçalves-de-Andrade RM, Kobashi LS, Almeida DD, Ho PL, Tam-bourgi DV, 2008. Transcriptome analysis of Loxosceles laeta (Araneae, Sicariidae) spider venomous gland using expressed sequence tags. BMC Genomics 9 :279.
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A New Anti-loxoscelic Serum Produced Against Recombinant Sphingomyelinase D: Results of Preclinical Trials
Daniel Manzoni de Almeida
Daniel Manzoni de Almeida
Laboratório de Imunoquímica, Instituto Butantan, São Paulo, Brazil; Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal do Rio Grande do Norte, Natal, Brazil; Divisão Bioindustrial e Centro de Biotecnologia, Instituto Butantan, Natal, Brazil; Cardiff University, Wales College of Medicine, Cardiff, United Kingdom
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Matheus de F. Fernandes-Pedrosa
Matheus de F. Fernandes-Pedrosa
Laboratório de Imunoquímica, Instituto Butantan, São Paulo, Brazil; Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal do Rio Grande do Norte, Natal, Brazil; Divisão Bioindustrial e Centro de Biotecnologia, Instituto Butantan, Natal, Brazil; Cardiff University, Wales College of Medicine, Cardiff, United Kingdom
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Rute M. Gonçalves de Andrade
Rute M. Gonçalves de Andrade
Laboratório de Imunoquímica, Instituto Butantan, São Paulo, Brazil; Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal do Rio Grande do Norte, Natal, Brazil; Divisão Bioindustrial e Centro de Biotecnologia, Instituto Butantan, Natal, Brazil; Cardiff University, Wales College of Medicine, Cardiff, United Kingdom
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José Roberto Marcelino
José Roberto Marcelino
Laboratório de Imunoquímica, Instituto Butantan, São Paulo, Brazil; Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal do Rio Grande do Norte, Natal, Brazil; Divisão Bioindustrial e Centro de Biotecnologia, Instituto Butantan, Natal, Brazil; Cardiff University, Wales College of Medicine, Cardiff, United Kingdom
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Hisako Gondo-Higashi
Hisako Gondo-Higashi
Laboratório de Imunoquímica, Instituto Butantan, São Paulo, Brazil; Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal do Rio Grande do Norte, Natal, Brazil; Divisão Bioindustrial e Centro de Biotecnologia, Instituto Butantan, Natal, Brazil; Cardiff University, Wales College of Medicine, Cardiff, United Kingdom
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Inácio de L. M. Junqueira de Azevedo
Inácio de L. M. Junqueira de Azevedo
Laboratório de Imunoquímica, Instituto Butantan, São Paulo, Brazil; Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal do Rio Grande do Norte, Natal, Brazil; Divisão Bioindustrial e Centro de Biotecnologia, Instituto Butantan, Natal, Brazil; Cardiff University, Wales College of Medicine, Cardiff, United Kingdom
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Paulo Lee Ho
Paulo Lee Ho
Laboratório de Imunoquímica, Instituto Butantan, São Paulo, Brazil; Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal do Rio Grande do Norte, Natal, Brazil; Divisão Bioindustrial e Centro de Biotecnologia, Instituto Butantan, Natal, Brazil; Cardiff University, Wales College of Medicine, Cardiff, United Kingdom
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Carmen van den Berg
Carmen van den Berg
Laboratório de Imunoquímica, Instituto Butantan, São Paulo, Brazil; Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal do Rio Grande do Norte, Natal, Brazil; Divisão Bioindustrial e Centro de Biotecnologia, Instituto Butantan, Natal, Brazil; Cardiff University, Wales College of Medicine, Cardiff, United Kingdom
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Denise V. Tambourgi
Denise V. Tambourgi
Laboratório de Imunoquímica, Instituto Butantan, São Paulo, Brazil; Programa de Pós-Graduação em Ciências Farmacêuticas, Universidade Federal do Rio Grande do Norte, Natal, Brazil; Divisão Bioindustrial e Centro de Biotecnologia, Instituto Butantan, Natal, Brazil; Cardiff University, Wales College of Medicine, Cardiff, United Kingdom
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Envenomation by Loxosceles species (brown spider) can lead to local dermonecrosis and to serious systemic effects. The main toxic component in the venom of these spiders is sphingomyelinase D (SMase D) and various isoforms of this toxin are present in Loxosceles venoms. We have produced a new anti-loxoscelic serum by immunizing horses with recombinant SMase D. In the present study, we compared the neutralization efficacy of the new anti-loxoscelic serum and anti-arachnidic serum (the latter serum is used for therapy for loxoscelism in Brazil) against the toxic effects of venoms from spiders of the genus Loxosceles. Neutralization tests showed that anti-SMase D serum has a higher activity against toxic effects of L. intermedia and L. laeta venoms and similar or slightly weaker activity against toxic effects of L. gaucho than that of Arachnidic serum. These results demonstrate that recombinant SMase D can replace venom for anti-venom production and therapy.
Author Notes
- INTRODUCTION
- MATERIALS AND METHODS
- Chemicals, reagents, and buffers
- Antibodies
- Animals
- Spiders and venoms
- Expression of recombinant protein
- Antisera
- Normal human serum and erythrocytes
- Electrophoresis and Western blotting
- Enzyme-linked immunosorbent assay (ELISA)
- Neutralization of dermonecrotic activity by sera
- Neutralization of hemolytic activity by sera
- Neutralization of SMase activity by sera
- Statistical analysis
- RESULTS
- DISCUSSION
ProCite
RefWorks
Reference Manager
BibTeX
Zotero
EndNote
-
1
Futrell JM, 1992. Loxoscelism. Am J Med Sci 304 :261–267.
-
2
Ginsburg CM, Weinberg AG, 1988. Hemolytic anemia and multiorgan failure associated with localized cutaneous lesion. J Pediatr 112 :496–499.
-
3
Gendron BP, 1990. Loxosceles reclusa envenomation. Am J Emerg Med 8 :51–54.
-
4
Bey TA, Walter FG, Lober W, Schmidt J, Spark R, Schlievert PM, 1997. Loxosceles arizonica bite associated with shock. Ann Emerg Med 30 :701–703.
-
5
Ramos-Cerrillo B, Olvera A, Odell GV, Zamudio F, Paniagua-Solis J, Alagon A, Stock RP, 2004. Genetic and enzymatic characterization of sphingomyelinase D isoforms from the North American fidleback spiders Loxosceles boneti and Loxosceles recluse. Toxicon 44 :507–514.
-
6
Newlands G, Isaacson C, Martindale C, 1982. Loxoscelism in the Transvaal, South Africa. Trans R Soc Trop Med Hyg 76 :610–615.
-
7
Denny WF, Dillaha CJ, Morgan PN, 1964. Hemotoxic effect of Loxoscels reclusa venom: in vivo and in vitro studies. J Lab Clin Med 64 :291.
-
8
Schenone H, Saavedra T, Rojas A, Villarroel F, 1998. Loxoscelism in Chile. Epidemiologic, clinical and experimental studies. Rev Inst Med Trop Sao Paulo 31 :403–415.
-
9
Dillaha CJ, Jansen T, Honeycut WM, Hayden CR, 1964. North America Loxoscelism. JAMA 188 :33–36.
-
10
Rees RS, Shack B, Withers F, Madden J, Franklin J, Lynch JB, 1981. Management of the brown recluse spider bite. Plast Reconstr Surg 68 :768–773.
-
11
Koh WL, 1998. When to worry about spiders bite: inaccurate diagnosis can have serious, even fatal, consequences. Postgrad Med 103 :235–250.
-
12
Allen C, 1992. Arachnid envenomation. Emerg Med Clin North Am 10 :269–299.
-
13
Anderson PC, 1997. Spider bites in the United States. Dermathol Clin 5 :307–311.
-
14
Walter FG, Bilden EF, Gibly RL, 1999. Envenomations. Crit Care Clin 15 :627–629.
-
15
Barrett SM, Romine-Jenkins M, Fisher DE, 1994. Dapsone or electric shock treatment of brown recluse spider envenomation. Ann Emerg Med 24 :21–25.
-
16
Bitterman-Deutsch O, Bergman R, Friedman-Birnbaum R, 1990. Brown spider bite. Harefuah 119 :137–139.
-
17
Wille RC, Morrow JD, 1998. Case report: dapsone hypersensitivity syndrome associated with treatment of the bite of brown spider. Am J Med Sci 296 :270–271.
-
18
Mold JW, Thompson DM, 2004. Management of brown recluse spider bites in primary care. J Am Board Fam Pract 1 :347–352.
-
19
Bryant SM, Pittman LM, 2003. Dapsone use in Loxosceles reclusa enevenomation: is there an indication? Am J Emerg Med 21 :89–90.
-
20
Elston MD, Miller MS, Young RJ III, Eggers J, McGlasson D, Schmidt WH, Brush A, 2005. Comparation of colchicines, dapsone, triamcinolone and diphenhydramine therapy for the treatment of brown recluse spider envenomation. Arch Dermatol 141 :595–597.
-
21
Smith CW, Micks DW, 1970. The role of polymorphonuclear leukocytes in the lesion caused by the venom of the brown spider, Loxosceles reclusa. Lab Invest 22 :141–144.
-
22
Wilson JR, Hagood CO, Jr Prather ID, 2005. Brown recluse spider bites: a complex problem wound. A brief review and case study. Ostomy Wound Manage 51 :59–66.
-
23
Pizzi T, 1957. A histopathological study on necrotic arachnidism by Loxosceles laeta. Bol Chil Parasitol 30 :34–36.
-
24
Sunderkotter C, Seeliger S, Schonlau F, Roth J, Hallmann R, Luger TA, Sorg C, Kolde G, 2001. Different pathway leading to cutaneous leukocytoclastic vasculitis in mice. Exp Dermatol 10 :391–404.
-
25
Tambourgi DV, Paixão-Cavalcante D, Gonçalves-de-Andrade RM, Fernandes-Pedrosa MF, Magnoli FC, Morgan PB, van den Berg CW, 2005. Loxosceles sphingomyelinase induces complement-dependent dermonecrosis, neutrophil infiltration, and endogenous gelatinase expression. J Invest Dermatol 124 :725–731.
-
26
Ministry of Health, 1998. Manual de Diagnóstico e Tratamento de Acidentes por Animais Peçonhento. Fundação Nacional de Saúde/Coordenação de Controle de Zoonoses e Animais Peçonhentos. Brasília: Centro Neurop.
-
27
Kurpiewski G, Forrester LJ, Barret JT, Campbell BJ, 1981. Platelet aggregation and sphingomyelinase D activity of a purified toxin from the venom of Loxosceles recluse. Biochim Biophys Acta 678 :467–476.
-
28
Tambourgi DV, Magnoli FC, van den Berg CW, Morgan BP, de Araujo PS, Alves EW, Dias da Silva WD, 1998. Sphingomyelinases in the venom of the spider Loxosceles intermedia are responsible for both dermonecrosis and complement-dependent hemolysis. Biochem Biophys Res Commun 251 :366–373.
-
29
Tambourgi DV, Petricevich VL, Magnoli FC, Assaf SL, Jancar S, Dias da Silva W, 1998b. Endotoxemic-like shock induced by Loxosceles spider venoms, pathological changes and putative cytokine mediators. Toxicon 3 :391–403.
-
30
Tambourgi DV, Morgan BP, Gonçalves-de-Andrade RM, Magnoli FC, van den Berg CW, 2000. Loxosceles intermedia spider envenomation induces activation of an endogenous metallo-proteinase, resulting in cleavage of glycophorins from the erythrocyte surface and facilitating complement-mediated lysis. Blood 95 :683–691.
-
31
Tambourgi DV, de Sousa da Silva M, Billington SJ, Goncalves-de-Andrade RM, Magnoli FC, Songer JG, van den Berg CW, 2002. Mechanism of induction of complement susceptibility of erythrocytes by spider and bacterial sphingomyelinases. Immunol 107 :93–101.
-
32
Tambourgi DV, Fernandes-Pedrosa MF, van den Berg CW, Gonçalves-de-Andrade RM, Ferracini M, Paixão-Cavalcante D, Morgan BP, Rushmere NK, 2004. Molecular cloning, expression, function and immunoreactivities of members of a gene family of sphingomyelinases from Loxosceles venom glands. Mol Immunol 41 :831–840.
-
33
Paixão-Cavalcante D, van den Berg CW, Fernandes-Pedrosa MF, Gonçalves-de-Andrade RM, Tambourgi DV, 2006. Role of matrix metalloproteinases in HaCaT keratinocytes apoptosis induced by Loxosceles venom sphingomyelinase D. J Invest Dermatol 126 :61–68.
-
34
van den Berg CW, Gonçalves-de-Andrade RM, Magnoli FC, Marchbank KJ, Tambourgi DV, 2002. Loxosceles spider venom induces metalloproteinases mediated cleavage of MCP/CD46 and MHC I and induces protection against C-mediated lysis. Immunology 107 :102–110.
-
35
Fernandes-Pedrosa MF, Junqueira de Azevedo IL, Gonçalves-de-Andrade RM, van den Berg CW, Ramos CR, Ho PL, Tam-bourgi DV, 2002. Molecular cloning and expression of a functional dermonecrotic and haemolytic factor from Loxosceles laeta venom. Biochem Biophys Res Commun 298 :638–645.
-
36
Bucherl W, 1969. Biology and venoms of the most important South American spiders of the genera Phoneutria, Loxosceles, Lycosa, and Latrodectus. Am Zool 9 :157–159.
-
37
Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC, 1985. Measurement of protein using bicinchronic acid. Anal Biochem 150 :76–85.
-
38
Laemmli UK, 1970. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227 :680–685.
-
39
Towbin H, Staehelin TJ, Gordon J, 1979. Eletrophoretic transfer of proteins from acrylamide gels to nitrocellulose sheets: procedure and some applications. Proc Natl Acad USA 76 :4350–4354.
-
40
Furlanetto RS, Santos NP, Navas J, 1962. Preparação, purificação e doseamento do sôro antiloxoscélico. Cienc Cult 14 :254.
-
41
Tokomura A, Majima E, Kariya Y, Tominagua K, Kogune K, Ogune K, Yasuda K, Fukuzawa K, 2002. Increased production of bioactive lysophosphatidic acid by serum lysophospholipase D in human pregnancy. J Biol Chem 277 :39436–39442.
-
42
Klinman NR, Rockey JH, Karush F, 1965. Equine antihapten antibody II. The gamma-G (7S-gamma) components and their specific interaction. Int J Cancer Suppl 47 :51–60.
-
43
Tambourgi DV, Magnoli FC, von Eicksted VRD, Benedetti ZC, Petrecevich VL, Dias da Silva W, 1995. Incorporation of a 35 kDa purified protein from Loxosceles intermedia spider venom transforms human erythrocytes into activators of autologous complement alternative pathway. J Immunol 155 :4459–4466.
-
44
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