تاثیر رویکرداستفاده از اجوانت ها بر میزان کارآیی و ایمنی زایی واکسن ضد اسهال اشریشیا کلی O157:H7
محورهای موضوعی : میکروب شناسی مولکولی
ناهید حیدری
1
,
یحیی تهمتن
2
,
هاجر مولایی
3
1 - دانش آموخته میکروب شناسی، دانشگاه آزاد اسلامی واحد شیراز
2 - بخش باکتری شناسی موسسه تحقیقات واکسن و سرم سازی رازی شیراز
3 - بخش میکروب شناسی، مؤسسه تحقیقات واکسن و سرم سازی رازی شعبه شیراز، سازمان تحقیق، آموزش و ترویج کشاورزی، شیراز، ایران.
کلید واژه: واکسن, اسهال, اجوانت, اشریشیاکلی بیماری زا,
چکیده مقاله :
سابقه و هدف: باکتری اشریشیا کلی انترو هموراژیک از مهمترین عوامل ایجاد اسهال می باشد. با وجود پیشرفت های اخیر، درمان مؤثری برای عفونت ایجاد شده با این باکتری وجود ندارد. توسعه واکسن ساز و کار اولیه ایجاد مقاومت در برابر باکتری محسوب می شود. این پژوهش با هدف ارزیابی نقش اجوانت های مختلف به منظور تهیه واکسن این باکتری انجام شد. مواد و روش ها: ابتدا کشت باکتری محیط اختصاصی انجام شد و سپس به منظور تهیه آنتی ژن و تزریق به موش، با افزودن فرمالین غیر فعال گردید. چهار گروه مختلف باکتری غیر فعال شده، همراه اجوانت آلوم(AO)، اجوانت مونتاناید(MO)، بدون اجوانت(O) و کنترل (C) به صورت زیر جلدی به موش آزمایشگاهی ماده Balb/C تزریق گردید. از آزمون الیزا به منظور سنجش عیار آنتی بادی استفاده شد. 28 روز پس از ایمنی زایی، موش ها به چالش کشیده شدند.یافته ها: بیشترین میزان عیار آنتی بادی از تزریق باکتری با اجوانت آلوم به دست آمد، اما اجوانت مونتاناید در ماه اول تیتر آنتی بادی بالاتری داشت. چالش تایید کرد که گروه AO توانایی ایجاد محافظت در 100 درصد موش ها مورد بررسی را دارد.نتیجه گیری: نتایج نشان داد که پادتن تولید شده با اجوانت آلوم ایمنی در حیوان را برای طولانی مدت نگه می دارد. از آنجا که طراحی واکسن برای جلوگیری از بیماری های عفونی نیاز به سیستم تحویل آنتی ژن کارآمد دارد، این تحقیق می تواند راهبرد جدیدی برای تولید واکسن علیه اشریشیا کلی باشد .
Background & Objectives: Enterohemorrhagic Escherichia coli is one of the major reasons for the bacterial diarrhea. Despite the vast development, there is still no comprehensive treatment for this infection. Vaccine development is the first strategy against bacterial resistance. Due to the pathogenicity of Escherichia coli, the present study was aimed to investigate the role of different adjuvants for vaccination against this bacteria.Materials & Methods: The bacteria were cultured and inactivated by formaldehyde for antigen preparation and injection into the mice. Female Balb/c mice were injected subcutaneously with four different bacterial groups including, bacteria with alum adjuvant (AO), montanide adjuvant (MO), non-adjuvant bacteria (O) and control group. Following, the reminder dose was injected after 14 days and blood samples were collected during 7 weeks. The Elisa test was used for evaluation of antibody titer. 28 days after post immunization, the mice were challenged and were evaluated for 72 h. Statistical analysis was accomplished using ANOVA and SPSS software.Results: The results suggested that while the highest antibody titer was obtained for the AO group, but the MO group has more antibody titer in the first month. Also, challenge results exposed that the AO group protected 100 % of the challenge group mice. Conclusion: consequently, the produced antibody of the alum adjuvant keep higher the animal’s immune system for a longer period. Since vaccine design to prevent infectious diseases requires effective antigen delivery system, this study could be a new strategy to produce the E. Coli O157:H7 vaccine.
2. Jay CM, Bhaskaran S, Rathore S, Waghela SD. Enterotoxigenic 99+ Escherichia coli
attachment to host cell receptors inhibited by recombinant pili protein. Vet Microbiol. 2004;
101: 153-160.
3. Clarke SC. Diarrhoeagenic Escherichia coli an emerging problem. Diagn Microbiol Infect Dis.
2001; 41: 93-98.
4. Heuvelink A, Bleumink B, Van den Biggelaar F, Te Giffel M, Beumer R, De Boer E.
Occurrence and survival of verocytotoxin-producing Escherichia coli O157 in raw cow's milk
in The Netherlands. J Food Prot. 1998; 61: 1597-1601.
5. Walker T. Microbiology. Saunders Company, Philadelphia. 1998.
6. armali MA. Infection by verocytotoxin-producing Escherichia coli. Clin Microbiol Rev. 1989;
2: 15-38.
7. Hara- udo J. Flourogenic and chromogenic media, the rapid technique for isolation of V.
cholera. J. Appl. And Eviron. Mic. 2003; 67: 5819-5823.
8. Schroeder CM, Cuiwei Z, Chitrita D, Jocelyn T, Shaohua Z, White DG. Antimicrobial
Resistance of Escherichia coli O157 Isolated from Humans, Cattle, Swine, and Food. Appl
Environ Microbiol. 2002; 68(2): 576–581.
9. Salyers AA. Bacterial pathogenesis: a molecular approach, ASM press Washington. 1994.
10. Serra-Moreno R, Jofre J, Muniesa M. The CI Repressors of Shiga Toxin-Converting
Prophages Are Involved in Coinfection of Escherichia coli Strains, Which Causes a Down
Regulation in the Production of Shiga Toxin. J Bacteriol. 2008; 190(13): 4722–4735.
11. Maleki H, Salmanian AH, Amani J, ordenaiej A, Jafari M. Production of Bivalent Protein
EspA-Tir from Escherichia coli O157:H7 in Tobacco (Nicotiana tobbacum). Crop Biotech.
2013; 5: 1-9. [in Persian]
12. Lima M, dos Santos SA, Rodrigues JM, Silva CL. Vaccine adjuvant: it makes the difference.
Vaccine. 2004; 22: 2374-2379.
13. Aguilar J. Vaccine adjuvants revisited. Vaccine. 2007; 25: 3752-3762.
14. Mac ichan ML, O'Hagan DT, Singh M. Recent developments in adjuvants for vaccines
against infectious diseases. Biomol Eng. 2001; 18: 69-85.
15. Alving CR. Design and selection of vaccine adjuvants: animal models and human trials.
Vaccine. 2002; 20: 56-64.
16. Rojo-Montejo S, Collantes-Fernández S, Regidor-Cerrillo E, Rodríguez-Bertos J, Prenafeta A,
Gomez-Bautista A. Influence of adjuvant and antigen dose on protection induced by an
inactivated whole vaccine against Neospora caninum infection in mice. Vet Parasitol. 2001;
175: 220-229.
17. Doavi T, Mousavi SL, amali M, Amani J, Ramandi MF. Chitosan-based intranasal vaccine
against Escherichia coli O157: H7. IBj. 2016; 20(2): 97. [in Persian]
18. Meng J., Zhao Sh, Doyle MP, Mitchell SE, resovich S. Polymerase chain reaction for
detecting Escherichia coli O157:H7. Int J Food Microbiol. 1996; 32: 103-113.
19. Namdar N, argar M, Tahamtan Y, Hosseini MH. Antagonistic effects of bifidobacterium spp.
in decrease of cytopathic effects of Verotoxin-producing Escherichia coli. JMW. 2013; 5:
94-104. [In Persian].
20. Tahamtan Y, Alimohamadi N. Evaluation of three dilutions of E. coli antigen in
immunogenicity in balb/c mice. Vet Res Biol. 2017; 117: 40-49. [In Persian].
21. Allison AC, Byars NE. Immunological adjuvants: desirable properties and side-effects. Mol
Immunol. 1991; 28: 279-284.
22. Hasani SH, Hosseini Jazani N, Shahabi Sh, aramati SA. Evalution of the effect of ICI118,
155, A beta adrenoreceptor antagonsit, and allum, as adjuvantes for increasing the protection
of vaccination against almonella typhimurium. Urmia medical journal. 2014; 24: 851-861.
[In Persian].
23. Figueiredo H, Lage AP, Pereira Júnior FN, Leite RC. Passive immunity in cattle against
enterotoxigenic Escherichia coli: serologic evaluation of a bacterin containing 99 and F41
fimbriae in colostrum of vaccinated females and calf serum. Arq Bras Med Vet Zootec. 2004;
56: 425-432.
24. Jazani NH, Parsania S, Sohrabpour M, Mazloomi E, arimzad M, Shahabi Sh. Naloxone and
alum synergistically augment adjuvant activities of each other in a mouse vaccine model of
almonella typhimurium infection. Immunobiology. 2011; 216: 744-751.
25. Raman VS,. Reed SG, Malcolm MS, Christopher B, Matlashewski G. Adjuvants for
eishmania vaccines: from models to clinical application. Front Immunol. 2012; 3: 144.
26. Leotta GA, Miliwebsky ES, Chinen I, Espinosa EM, Azzopardi , Tennant SM.
Characterisation of Shiga toxin-producing Escherichia coli O157 strains isolated from humans
in Argentina. BMC Microbiol. 2008; 8: 46.
27. halesi R, Salimian J, Nazarian SH, Ehsaei Z, Rahimi AA, Amini N. Production and
purification of heat-labile toxin of enterotoxigenic Escherichia coli and its detection by GM1
gangelioside receptor-ELISA based method. AMUJ. 2012; 15: 35-42. [In Persian]
28. Ebadi V, Ebrahimi F, Hajizadeh A, Tarverdizadeh Y, Bakhshi M. Designing a Recombinant
Vaccine containing three bacterial proteins of EHEC, ETEC, and Shigella Dysentery Antigens
in E. coli and Evaluation of its Humoral Immunity in Mic. J Mazandaran Univ Med Sci. 2018;
27: 1 – 16. [in Persian]
_||_
2. Jay CM, Bhaskaran S, Rathore S, Waghela SD. Enterotoxigenic 99+ Escherichia coli
attachment to host cell receptors inhibited by recombinant pili protein. Vet Microbiol. 2004;
101: 153-160.
3. Clarke SC. Diarrhoeagenic Escherichia coli an emerging problem. Diagn Microbiol Infect Dis.
2001; 41: 93-98.
4. Heuvelink A, Bleumink B, Van den Biggelaar F, Te Giffel M, Beumer R, De Boer E.
Occurrence and survival of verocytotoxin-producing Escherichia coli O157 in raw cow's milk
in The Netherlands. J Food Prot. 1998; 61: 1597-1601.
5. Walker T. Microbiology. Saunders Company, Philadelphia. 1998.
6. armali MA. Infection by verocytotoxin-producing Escherichia coli. Clin Microbiol Rev. 1989;
2: 15-38.
7. Hara- udo J. Flourogenic and chromogenic media, the rapid technique for isolation of V.
cholera. J. Appl. And Eviron. Mic. 2003; 67: 5819-5823.
8. Schroeder CM, Cuiwei Z, Chitrita D, Jocelyn T, Shaohua Z, White DG. Antimicrobial
Resistance of Escherichia coli O157 Isolated from Humans, Cattle, Swine, and Food. Appl
Environ Microbiol. 2002; 68(2): 576–581.
9. Salyers AA. Bacterial pathogenesis: a molecular approach, ASM press Washington. 1994.
10. Serra-Moreno R, Jofre J, Muniesa M. The CI Repressors of Shiga Toxin-Converting
Prophages Are Involved in Coinfection of Escherichia coli Strains, Which Causes a Down
Regulation in the Production of Shiga Toxin. J Bacteriol. 2008; 190(13): 4722–4735.
11. Maleki H, Salmanian AH, Amani J, ordenaiej A, Jafari M. Production of Bivalent Protein
EspA-Tir from Escherichia coli O157:H7 in Tobacco (Nicotiana tobbacum). Crop Biotech.
2013; 5: 1-9. [in Persian]
12. Lima M, dos Santos SA, Rodrigues JM, Silva CL. Vaccine adjuvant: it makes the difference.
Vaccine. 2004; 22: 2374-2379.
13. Aguilar J. Vaccine adjuvants revisited. Vaccine. 2007; 25: 3752-3762.
14. Mac ichan ML, O'Hagan DT, Singh M. Recent developments in adjuvants for vaccines
against infectious diseases. Biomol Eng. 2001; 18: 69-85.
15. Alving CR. Design and selection of vaccine adjuvants: animal models and human trials.
Vaccine. 2002; 20: 56-64.
16. Rojo-Montejo S, Collantes-Fernández S, Regidor-Cerrillo E, Rodríguez-Bertos J, Prenafeta A,
Gomez-Bautista A. Influence of adjuvant and antigen dose on protection induced by an
inactivated whole vaccine against Neospora caninum infection in mice. Vet Parasitol. 2001;
175: 220-229.
17. Doavi T, Mousavi SL, amali M, Amani J, Ramandi MF. Chitosan-based intranasal vaccine
against Escherichia coli O157: H7. IBj. 2016; 20(2): 97. [in Persian]
18. Meng J., Zhao Sh, Doyle MP, Mitchell SE, resovich S. Polymerase chain reaction for
detecting Escherichia coli O157:H7. Int J Food Microbiol. 1996; 32: 103-113.
19. Namdar N, argar M, Tahamtan Y, Hosseini MH. Antagonistic effects of bifidobacterium spp.
in decrease of cytopathic effects of Verotoxin-producing Escherichia coli. JMW. 2013; 5:
94-104. [In Persian].
20. Tahamtan Y, Alimohamadi N. Evaluation of three dilutions of E. coli antigen in
immunogenicity in balb/c mice. Vet Res Biol. 2017; 117: 40-49. [In Persian].
21. Allison AC, Byars NE. Immunological adjuvants: desirable properties and side-effects. Mol
Immunol. 1991; 28: 279-284.
22. Hasani SH, Hosseini Jazani N, Shahabi Sh, aramati SA. Evalution of the effect of ICI118,
155, A beta adrenoreceptor antagonsit, and allum, as adjuvantes for increasing the protection
of vaccination against almonella typhimurium. Urmia medical journal. 2014; 24: 851-861.
[In Persian].
23. Figueiredo H, Lage AP, Pereira Júnior FN, Leite RC. Passive immunity in cattle against
enterotoxigenic Escherichia coli: serologic evaluation of a bacterin containing 99 and F41
fimbriae in colostrum of vaccinated females and calf serum. Arq Bras Med Vet Zootec. 2004;
56: 425-432.
24. Jazani NH, Parsania S, Sohrabpour M, Mazloomi E, arimzad M, Shahabi Sh. Naloxone and
alum synergistically augment adjuvant activities of each other in a mouse vaccine model of
almonella typhimurium infection. Immunobiology. 2011; 216: 744-751.
25. Raman VS,. Reed SG, Malcolm MS, Christopher B, Matlashewski G. Adjuvants for
eishmania vaccines: from models to clinical application. Front Immunol. 2012; 3: 144.
26. Leotta GA, Miliwebsky ES, Chinen I, Espinosa EM, Azzopardi , Tennant SM.
Characterisation of Shiga toxin-producing Escherichia coli O157 strains isolated from humans
in Argentina. BMC Microbiol. 2008; 8: 46.
27. halesi R, Salimian J, Nazarian SH, Ehsaei Z, Rahimi AA, Amini N. Production and
purification of heat-labile toxin of enterotoxigenic Escherichia coli and its detection by GM1
gangelioside receptor-ELISA based method. AMUJ. 2012; 15: 35-42. [In Persian]
28. Ebadi V, Ebrahimi F, Hajizadeh A, Tarverdizadeh Y, Bakhshi M. Designing a Recombinant
Vaccine containing three bacterial proteins of EHEC, ETEC, and Shigella Dysentery Antigens
in E. coli and Evaluation of its Humoral Immunity in Mic. J Mazandaran Univ Med Sci. 2018;
27: 1 – 16. [in Persian]