ارزیابی مقایسه‌ای پاسخ ایمنی‌زایی نانوآنتی‌ژن‌ بتا کلســتریدیوم پرفرنجنس پوشش یافته با غشاء گویچه قرمز و بدون پوشش در مدل حیوانی خرگوش

نوع مقاله : مقاله کامل

نویسندگان

1 گروه میکروبیولوژی، واحد کرمان، دانشگاه آزاد اسلامی، کرمان، ایران

2 بخش تحقیقات و فناوری، شعبه کرمان، موسسه تحقیقات واکسن و سرم سازی رازی، سازمان تحقیقات، آموزش و ترویج کشاورزی، کرمان، ایران

3 بخش تحقیق و توسعه، شعبه کرمان، موسسه تحقیقات واکسن و سرم سازی رازی، سازمان تحقیقات، آموزش و ترویج کشاورزی، کرمان، ایران

چکیده

کلستریدیوم پرفرنجنس تیپ C عامل انتروتوکسمی در دام‌های جوان و بالغ و از عوامل مهم خسارات اقتصادی به صنعت دامپروری می‌باشد. مهم‌ترین عامل ویرولانس در این تیپ، توکسین بتا است. در دهه‌های اخیر استفاده از نانوذرات برای بهبود اثر واکسن‌ها موردتوجه قرار گرفته است. جهت افزایش بقاء نانوذرات، از غشاء سلول‌هایی همچون گویچه قرمز برای پوشش استفاده می‌گردد، لذا تحقیق حاضر باهدف ارزیابی مقایسه‌ای پاسخ‌ ایمنی‌زایی نانوآنتی‌ژن‌ بتا کلســتریدیوم پرفرنجنس پوشش داده شده با غشاء گویچه قرمز و بدون پوشش انجام گرفت. ابتدا نانوآنتی‌ژن‌های محبوس در غشاء گویچه‌های قرمز آماده و پس از انجام آزمون‌های استرلیتی و عدم سمیت، به همراه نانوآنتی‌ژن‌های بدون پوشش طبق برنامه ایمن‌سازی و در بازه زمانی ۱۵۰ روز، به دو گروه خرگوش تزریق شدند. جهت ارزیابی تیتر آنتی‌بادی از روش الیزای غیرمستقیم استفاده شد. جهت نشان‌دادن قابلیـت ایمنی‌زایی نانوذرات پوشش یافته با غشاء گویچه قرمز، چالش و تست همولیزین انجام شد. تعیین پاسخ آنتی‌بادی، تحریک ایمنی‌زایی بالا و افزایش پایدار تیتر آنتی‌بادی در گروه‌ ایمن با نانوآنتی‌ژن‌های محبوس در غشاء گویچه‌ قرمز را نشان داد. ارزیابی چالش و تست همولیزین بیانگر فعالیت مثبت نانوآنتی‌ژن‌ بتا پوشش یافته با غشاء گویچه‌ قرمز در تحریک سیستم ایمنی خرگوش‌ بود. نتایج مطالعه نشان داد که غشاءهای گویچه قرمز یک لایه ایمن برای مهار نانوتوکسین فراهم می‌کنند و سبب رهایش آهسته و تدریجی آنتی‌ژن‌های کپسوله می‌شوند.

کلیدواژه‌ها


عنوان مقاله [English]

Comparative evaluation of the immunogenic response of Clostridium perfringens beta-antigens coated with RBC membrane and uncoated in rabbit as an animal model

نویسندگان [English]

  • Sima Pooyan 1
  • Mehrdad Shamsaddini Bafti 2
  • Nadia Kazemipour 1
  • Mojtaba Ali Mollai 3
  • Farokha Rokhbakhsh-Zamin 1
1 Department of Microbiology, Kerman Branch, Islamic Azad University, Kerman, Iran.
2 Department of Research and Technology, Kerman Branch, Razi Vaccine & Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Kerman, Iran
3 Department of Research and Development, Kerman Branch, Razi Vaccine & Serum Research Institute, Agricultural Research, Education and Extension Organization (AREEO), Kerman, Iran
چکیده [English]

Clostridium perfringens (C. perfringens) type C is the cause of enterotoxemia in young and adult cattle and one of the essential factors of economic losses to the animal husbandry industry. The most important virulence factor in this type is beta toxin. In recent decades, the use of nanoparticles to improve the effectiveness of vaccines has been considered. In order to increase the survival of nanoparticles, the membrane of cells such as red blood cells is used for coating. The aim of comparative evaluation of the immunogenic response of C. perfringens beta-antigen coated with RBCMs and uncoated. First, nanoantigens trapped in the membrane of RBC were prepared, and then sterility and residual toxicity tests were carried out. Finally, it was injected into two groups of rabbits along with bare nano antigens according to the immunization schedule and within 150 days. The indirect ELISA method was used to evaluate the antibody titer. In order to demonstrate the immunogenicity of nanoparticles coated with RBCMs, the challenge and hemolysin tests were performed. Determining the antibody response showed high immunogenic stimulation and a stable increase in antibody titer in the immune group with nano antigens trapped in the RBCMs. Challenge evaluation and hemolysin test showed the positive activity of beta nano antigens covered with red RBCMs in stimulating the immune system of rabbits. The study results showed that the RBCMs provide a safe layer to contain the nanotoxin and cause the slow and gradual release of encapsulated antigens.

کلیدواژه‌ها [English]

  • Clostridium perfringens
  • Beta Toxin
  • Enterotoxemia
  • Biomimetic nanoparticles
  • ELISA
1- Abdolmohammadi Khiav, L. and A. Paradise. 2021. Molecular and Toxigenic Characteristics of Clostridium Perfringens Type B Isolates from Sheep and Lamb. Journal of Veterinary Research 76: 268-276.
2- Abdolmohammadi Khiav, L., A. Paradise and A. Hagh Roosta. 2020. Designing of an Indirect ELISA Method for the Detection of Beta Antitoxin of Clostridium perfringens Type C in Rabbit Serum. Veterinary Researches & Biological Products 33: 21-30.
3- Abdolmohammadi Khiav, L., R. Pilechian Langroodi and A. Paradise. 2021. Recently acquired for NetB and TpeL toxins for vaccine production to protect against avian necrotic enteritis. Veterinary Researches & Biological Products 34: 26-37.
4- Amini, M., M. Shamsaddini Bafti, B. Kheirkhah and F. Rokhbakhsh-Zamin. 2021. Comparison of toxin production power in different types of Clostridium perfringens among sheep and goats isolates by ELISA. Veterinary Researches & Biological Products.
5- Bagheripour, M. J., F. Ebrahimi, A. Hajizadeh, S. Nazarian and M. A. Arefpour. 2016. Preparation of chitosan based botulinum neurotoxin e recombinant nanovaccine and evaluation of its immunogenicity as oral & intradermal route in mice. Journal of Rafsanjan University of Medical Sciences 14: 923-938.
6- Chekman, I. and P. Simonov. 2012. Structure and Function of Biological Membranes: The Impact of Nanoparticles. International Journal of Physiology and Pathophysiology 3: 187-208.
7- Chiu, H. I., N. A. Samad, L. Fang and V. Lim. 2021. Cytotoxicity of targeted PLGA nanoparticles: a systematic review. RSC Advances 11: 9433-9449.
8- Dehaini, D., R. H. Fang and L. Zhang. 2016. Biomimetic strategies for targeted nanoparticle delivery. Bioengineering & translational medicine 1: 30-46.
9- Elmowafy, E. M., M. Tiboni and M. E. Soliman. 2019. Biocompatibility, biodegradation and biomedical applications of poly(lactic acid)/poly(lactic-co-glycolic acid) micro and nanoparticles. Journal of Pharmaceutical Investigation 49: 347-380.
10- Fathi Najafi, M., M. Hemmaty, J. Navidmehr, M. Afsharian, M. Farhoodi and S. Zibaee. 2020. Improvement in the Growth and α-toxin Production of Clostridium septicum by Magnesium Sulfate. Archives of Razi Institute 75: 219-225.
11- Gao, W. and L. Zhang. 2015. Engineering red‐blood‐cell‐membrane–coated nanoparticles for broad biomedical applications. p. 738-746.
12- Golchinfar, F., R. Madani, T. Emami, H. Zoulfagharian, A. Zare and N. Mouhammadpour. 2016. Designing a competitive ELISA for evaluation of anti-snake venom serum potency. Veterinary Researches & Biological Products 29: 9-16.
13- Hu, C.-M. J., R. H. Fang, B. T. Luk and L. Zhang. 2013. Nanoparticle-detained toxins for safe and effective vaccination. Nature Nanotechnology 8: 933-938.
14- Hu, C.-M. J., L. Zhang, S. Aryal, C. Cheung, R. H. Fang and L. Zhang. 2011. Erythrocyte membrane-camouflaged polymeric nanoparticles as a biomimetic delivery platform. Proceedings of the National Academy of Sciences 108: 10980-10985.
15- Izzati Mat Rani, N. N., Z. M. Alzubaidi, H. Azhari, F. Mustapa and M. C. Iqbal Mohd Amin. 2021. Novel engineering: Biomimicking erythrocyte as a revolutionary platform for drugs and vaccines delivery. European Journal of Pharmacology 900: 174009.
16- Kamali, M., M. B. Salehi, M. Tavalaei, G. H. Olad and M. Musavi. 2009. Fast detection of Clostridium perfringens Type A, B, C and D by Multiplex PCR. Veterinary Researches & Biological Products 22: 63-71.
17- Kheirollahpour, M., M. Mehrabi, N. M. Dounighi, M. Mohammadi and A. Masoudi. 2020. Nanoparticles and Vaccine Development. Pharmaceutical Nanotechnology 8: 6-21.
18- Kroll, A. V., R. H. Fang and L. Zhang. 2017. Biointerfacing and applications of cell membrane-coated nanoparticles. Bioconjugate chemistry 28: 23-32.
19- Liu, J., Z. Liu, Y. Pang and H. Zhou. 2022. The interaction between nanoparticles and immune system: application in the treatment of inflammatory diseases. Journal of Nanobiotechnology 20: 127.
20- Liu, J., A. Stace-Naughton, X. Jiang and C. J. Brinker. 2009. Porous nanoparticle supported lipid bilayers (protocells) as delivery vehicles. Journal of the American Chemical Society 131: 1354-1355.
21- Paradise, A. R. and L. Abdolmohammadi Khiav. 2020. Evaluation of Epsilon Antitoxin of Clostridium Perfringens Type D in The Rabbit Serum By Indirect ELISA. Veterinary Researches & Biological Products 33: 17-30.
22- Pilehchian, L. R., A. R. Jabbari, S. M. Moosawi and A. Pardis. 2015. A Production of pentavalent Clostridial toxoid vaccine and its comparison to conventional bacterin vaccine.
23- Poudineh Morref, M., M. K. Koohi, M. Alimolaei, T. Emami and J. Hassan. 2022. A New Practical Purification Method for Type D Clostridium perfringens Epsilon Toxin by Size-Exclusion Chromatography (SEC) and Ultrafiltration (UF). Iranian Journal of Veterinary Medicine 16: 178-187.
24- Que, X., J. Su, P. Guo, Z. Kamal, E. Xu, S. Liu, J. Chen and M. Qiu. 2019. Study on preparation, characterization and multidrug resistance reversal of red blood cell membrane-camouflaged tetrandrine-loaded PLGA nanoparticles. Drug delivery 26: 199-207.
25- Rahman Mashhadi, M., N. Atyabi, M. Hemmaty, M. Fathi Najafi and M. H. Fallah Mehrabadi. 2018. A comparative hematological, biochemical and immunological factors study following pentavalent toxoid vaccine containing Clostridium novyi with black disease bacterin/toxoid vaccine injection. Veterinary Researches & Biological Products 31: 66-71.
26- Santos, H. A. 2022. Biomimetic platelet membrane-coated nanoparticles for targeted therapy Huijie Han, Raquel Bártolo, Jiachen Li, Mohammad-Ali Shahbazi. European Journal of Pharmaceutics and Biopharmaceutics 172: 1-15.
27- SPSS Statistics for Windows. 2021. Armonk, NY, IBM Corp.
28- Valencia, P. M., P. A. Basto, L. Zhang, M. Rhee, R. Langer, O. C. Farokhzad and R. Karnik. 2010. Single-step assembly of homogenous lipid− polymeric and lipid− quantum dot nanoparticles enabled by microfluidic rapid mixing. ACS nano 4: 1671-1679.
29- Van Schooneveld, M. M., A. Gloter, O. Stephan, L. F. Zagonel, R. Koole, A. Meijerink, W. J. M. Mulder and F. M. F. De Groot. 2010. Imaging and quantifying the morphology of an organic–inorganic nanoparticle at the sub-nanometre level. Nature nanotechnology 5: 538-544.
30- Vasegh, R., M. Ebtekar, M. Shafiee Ardestani and M. Gholamzad. 2018. Comparison of Humoral and Cell-Mediated Immune Response to Tetanustoxin Coated PLGA in Mice. mdrsjrns 22: 7-19.
31- Vasegh, R., M. Ebtekar, M. Shafiee Ardestani and M. Gholamzad. 2018. Comparison of Humoral and Cell-Mediated Immune Response to Tetanustoxin Coated PLGA in Mice. Pathobiology Research 22: 7-19.
32- Wu, P., X. Jiang, S. Yin, Y. Yang, T. Liu and K. Wang. 2021. Biomimetic recombinant of red blood cell membranes for improved photothermal therapy. Journal of nanobiotechnology 19: 1-13.
33- Xia, Q., Y. Zhang, Z. Li, X. Hou and N. Feng. 2019. Red blood cell membrane-camouflaged nanoparticles: a novel drug delivery system for antitumor application. Acta Pharmaceutica Sinica B 9: 675-689.