مطالعه کاهش حدت نئوسپورا کنینوم بعد از پاساژهای متوالی روی رده سلولی J774

نوع مقاله: مقاله کامل

نویسندگان

1 گروه بیوشیمی دانشگاه آزاد اسلامی واحد شیراز

2 موسسه تحقیقات سرم و واکسن سازی رازی شعبه شیراز، سازمان تحقیقات، آموزش و ترویج کشاورزی

چکیده

هدف از این مطالعه، ایجاد تخفیف حدت در تک یاخته‌ی نئوسپورا کانینوم (Neospora caninum) از طریق پاساژ متوالی روی رده سلولی J774 و ارزیابی میزان حدت آن در موش BALB/c بود. ایزوله شماره یک نئوسپورا کانینوم بر روی رده سلولی J774، 90 مرتبه پاساژ داده شد تا سوش تخفیف حدت‌یافته تولید شود؛ سپس 35 سر موش BALB/c ماده با متوسط سن شش تا هشت هفته به صورت تصادفی به هفت گروه پنج‌تایی تقسیم شدند. به گروه اول به عنوان شاهد فقط محیط کشت، به گروه دوم و سوم و چهارم سوش حاد Nc1 با دوزهای 106×5، 106×10 و106×20 و به گروه پنجم، ششم و هفتم سوش تخفیف حدت یافته Nc1 با دوزهای106×5، 106×10 و106×20 به صورت زیر جلدی تزریق شد. تمامی موش‌ها به صورت روزانه بررسی و نشانه‌های بیماری و تلفات احتمالی موش‌ها ثبت گردید. در این مطالعه ایمنی سلولی با آزمایش جلدی و ایمنی هومورال با آزمایش الایزا بررسی شد. نتایج این مطالعه نشان داد موش‌هایی که سوش تخفیف حدت یافته را دریافت کرده‌ بودند، نه تنها تلف نشدند بلکه علائم بیماری را نیز نشان ندادند. به علاوه پاسخ ایمنی هومورال و سلولی نیز در آن ها نسبت به گروه‌های دریافت کننده سوش حاد به طور معنی داری بالاتر بود. در جمع بندی نهایی این مطالعه نشان داد که با استفاده از رده سلولی J774 که یک سلول دفاعی می‌باشد، می‌توان به طور معنی داری بیماری‌زایی سوش حادNc1 را کاهش داد؛ لذا این روش می‌تواند روش مناسبی برای تحقیقات آتی در راستای دستیابی به واکسن زنده تخفیف حدت یافته باشد.

کلیدواژه‌ها


عنوان مقاله [English]

Study on the attenuation of Neospora caninum after continuous passage on J774 cell line

نویسندگان [English]

  • P. Beigi 1
  • Mehdi Namavari 2
  • N. Razmi 1
1 Biochemistry Group, Islamic Azad University Branch Shiraz.
2 Vaccine and Serum Research Institute, Shiraz Branch, Agricultural Research, Education and Extension Organisation (AREEO), Tehran, Iran.
چکیده [English]

The aim of this study was to establish attenuated Neospora caninum (NC-1) tachyzoites through serial passage by J774 cell line. The virulence of attenuated tachyzoites for Balb/c mice was also evaluated. The NC-1 isolates were passaged 90 times in J774 cell line to produce a high-passage or attenuated strain. A total of 35 female BALB/c mice with an average age of six to eight weeks were randomly separated into seven groups. The first group as a control was inoculated subcutaneously with cell culture media and the second, third and fourth groups received low passage or acute NC-1 strain with different doses (5×106, 10×106, 20×106) of tachyzoites. The other three groups were inoculated with high-passage of NC-1 tachyzoites with similar doses. The mortality rate and pathological changes in all groups were noted daily. The cellular and humoral immunity were assessed by Skin test and ELISA, respectively. The results of the present study showed that the groups that received high-passage strain of Nc1 not only survived, but did not show pathological changes of neosporosis. In addition, cellular and humoral immune responses to NC-1 were significantly higher in mice that received high-passage tachyzoites of NC-1. These findings indicated that the serial passages of NC-1 in J774 cell line significantly led to decrease the virulence of NC-1. This attenuated tachyzoite could be used as a candidate for further research to develop live vaccines against Neospora caninum.

کلیدواژه‌ها [English]

  • Neospora caninum
  • J774
  • BALB/c

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