Investigation of Roles of Cellular Prion Protein and Doppel Protein in Pathogenesis of Experimental Toxoplasma gondii Infection in Mice

Abstract

The aim of the present study was to investigate the roles of cellular prion protein PrPc and doppel Dpl protein in experimental infection caused by Toxoplasma gondii in mice. A total of 96 mice Balb/c , 40 of which were control and 56 was treatment group, were used for this purpose in the study. The mice in experiment group were divided into 8 groups, including 7 mice each, according to post inoculation time 6 and 12 hours, 1, 2, 3, 4, 5 and 6 days. These animals were injected with 1 ml inoculum of T. gondii RH strain including 500 tachyzoits/0.1 ml by intraperitoneal route. At the end those times, the mice were euthanased under deep anesthesia and systematically were necropsied. Macroscopically, first lesions were detected after 4 days post inoculation. Especially, multiple gray-whitish foci were detected on the liver and kidneys. In the lung section, the frothy fluid was leaked. At the 6th days post inoculation, three mice were found died; in this group other mice had a lightly cloudy exudate in peritoneal cavity as well as the lesions in the organs mentioned. Microscopically, many organs such as liver, kidneys, lungs and brains included mononuclear cells infiltrations with necrosis. The some brains revealed necrosis and focal gliosis. Immunohistochemical studies showed the presence of T. gondii antigen in many organs excepting with brain tissues. In the experiment groups, PrPc and Dpl expressions increased with postinoculation days, especially in spleen, kidneys, and lungs by immunohistochemistry. In conclusion, PrPc and Dpl showed an increase in the tissues in experimental T. gondii infection in mice, and this study indicated that both proteins would be important in such disease.

Keywords

• Cellular prion protein,Doppel protein,immunohistokimya,mice,Toxoplasma gondii,Western blot.

Farelerde Deneysel Toxoplasma gondii Enfeksiyonunun Patogenezisinde Hücresel Prion Protein ve Doppel Protein’in Rollerinin Araştırılması

Öz

Bu çalışmada farelerde deneysel olarak oluşturulan Toxoplasma gondii enfeksiyonunda hücresel prion protein PrPc ve doppel proteinin Dpl rollerinin araştırılması amaçlanmıştır. Bu amaçla, 56 adet deneme ve 40 adet kontrol olmak üzere toplam 96 adet Balb/c fare kullanıldı. Çalışmada, her grupta 7 farenin bulunduğu 8 deneme grubu ile her deneme grubu için 5 farenin bulunduğu 8 kontrol grubu oluşturuldu. Deneme grubundaki hayvanlara T. gondii RH suşu takizoitleri 500 takizoit/0.1ml dozda, intraperitoneal yolla verildi. Farelerde enfeksiyondan sonraki 6. ve 12. saatlerde; daha sonra 1., 2., 3., 4., 5., 6. günlerde oluşan klinik bulgular kaydedilerek, sistematik nekropsileri yapıldı. Klinik olarak, ilk 3 günde her hangi bir bulgu gözlenmezken, 6. günde üç fare ölü bulundu. Makroskobik olarak, 4. günden itibaren karaciğer ve böbrek üzerinde boz beyaz renkte odaklar ile periton boşluğunda hafif bulanık bir eksudatın varlığı dikkati çekti. Mikroskobik olarak, böbrek, karaciğer, akciğer ve beyinde mononükleer hücre infiltrasyonu ile birlikte, ölenler başta olmak üzere, özellikle 5. ve 6. günlerde beyin ve karaciğerde fokal nekrotik değişiklikler ve mononüklear hücre infiltrasyonları saptandı. İmmunohistokimyasal olarak, T gondii antijeni beyin dışında böbrek, karaciğer, akciğer ve dalakta saptandı. Enfeksiyon dönemlerinde immunohistokimyasal incelemelerde PrPc, kontrollere göre merkezi sinir sistemi, dalak ve karaciğerde artan oranlarda saptanırken, Dpl artışı özellikle dalak, böbrek, karaciğerde daha belirgindi. Sonuç olarak, farelerde T. gondii enfeksiyonunda PrPc ve Dpl’nin dokularda ekspresyonlarının arttığı gözlendi ve böylece farelerde deneysel Toxoplasma gondii enfeksiyonunun patogenezisinde her iki proteinin de önemli olabileceği kanaatine varıldı.

Anahtar Kelimeler

• Doppel protein,fare,hücresel prion protein,immunohistokimya,Toxoplasma gondii,Western blot.

Kaynakça

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