Browsing by Author "Farzani, Touraj Aligholipour"

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    Aşılama stratejierinde bir gen aktarım vektörü olarak BoHV-4 movar suşu kullanımı.BoHV-1 glikoptein D ifade eden BoHV-4(BoHV-1gD) elde edilmesi
    (Ankara Üniversitesi : Bilimsel Araştırma Projeleri, 2020) Dağalp Bilge, Seval; Farzani, Touraj Aligholipour; Doğan, Fırat; Akkutay Yoldar, Ayşe Zeynep; Özkul, Aykut; Alkan, Feray; Dörttaş, Selvi Deniz; Veteriner Fakültesi
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    Aşılama stratejilerinde bir gen aktarım vektörü olarak bohv-4 movar suşu ile bac-bohv-4 elde edilmesi
    (Sağlık Bilimleri Enstitüsü, 2018) Farzani, Touraj Aligholipour; Dağalp, Seval Bilge; Veteriner Hekimliği
    BoHV-4, herpesviridae ailesinin gammaherpesvirinea alt ailesinde ve rhadinovirus cinsinde yer alan bir etkendir. Virus solunum sistemi enfeksiyonunda, vulvovaginitis, endometritis, mastitis, abort, metritis, dermatitis olgularında tespit edilmiştir. Ayrıca sağlıklı görünümlü hayvanların çeşitli materyallerinden de etken izole edilmiştir.Son yıllarda, BoHV-4 farklı patojenlere ait immundominant antijenlerin aktarımında ve kanser terapisinde yeni bir viral vektör olarak sunulmaktadır. BoHV-4'ün biyolojik ve moleküler özellikleri, onu diğer viral vektörlerden daha avantajlı kılmaktadır.Bu çalışmada, BoHV-4'ün Movar suşu kullanılarak bir viral vektör geliştirilmesi planlanmıştır. Rekombinant virus oluşturmak için, virusun ORF2 ve ORF3 gen bölgelerinin arası BAC-EGFP-Neo kasetin klonlanması için tercih edilmiştir.Hücrede gerçekleştirilen homolog rekombinasyondan sonra, rekombinant viruslar yeşil floresan protein ekspresyonuna bağlı olarak bir antibiyotik olan genetisin direncine göre seçilmiştir.Hirt yöntemi kullanılarak elde edilen rekombinant sirküler viruslar DH10B bakterisine transforme edilmiştir. Son basamakta, bakteriden ekstraksiyonu yapılan BoHV-4-BAC-EGFP, MDBK hücresine transfekte edilmiş ve rekombinant virus hazırlanarak stabilitesi DH10B bakterisinde doğrulanmıştır. Elde edilen BoHV-4-BAC viral vektörün, sahip olduğu özelliklere dayanarak, bu bağlamda ileride yapılması planlanan ülkemizde önemli ekonomik kayıpların nedeni ve/veya zoonoz karakterde etkenlere yönelik aşılamalarda ve başta glioma olmak üzere insan ve hayvanlarda birçok kanserin terapisinde kullanımının yararlı olabileceği düşünülmektedir.
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    Bovine Herpesvirus Type 4 (BoHV-4) Vector Delivering Nucleocapsid Protein of Crimean-Congo Hemorrhagic Fever Virus Induces Comparable Protective Immunity against Lethal Challenge in IFNα/β/γR-/- Mice Models
    (2019) Farzani, Touraj Aligholipour; Veteriner Fakültesi
    Crimean-Congo hemorrhagic fever virus (CCHFV) is the causative agent of a tick-borne infection with a significant mortality rate of up to 40% in endemic areas, with evidence of geographical expansion. Due to a lack of effective therapeutics and control measures, the development of a protective CCHFV vaccine remains a crucial public health task. This paper describes, for the first time, a Bovine herpesvirus type 4 (BoHV-4)-based viral vector (BoHV4-∆TK-CCHFV-N) and its immunogenicity in BALB/c and protection potential in IFNα/β/γR-/- mice models in comparison with two routinely used vaccine platforms, namely, Adenovirus type 5 and a DNA vector (pCDNA3.1 myc/His A), expressing the same antigen. All vaccine constructs successfully elicited significantly elevated cytokine levels and specific antibody responses in immunized BALB/c and IFNα/β/γR-/- mice. However, despite highly specific antibody responses in both animal models, the antibodies produced were unable to neutralize the virus in vitro. In the challenge experiment, only the BoHV4-∆TK-CCHFV-N and Ad5-N constructs produced 100% protection against lethal doses of the CCHFV Ank-2 strain in IFNα/β/γR-/- mice. The delivery platforms could not be compared due to similar protection rates in IFNα/β/γR-/- mice. However, during the challenge experiment in the T cell and passive antibody transfer assay, BoHV4-∆TK-CCHFV-N was dominant, with a protection rate of 75% compared to others. In conclusion, vector-based CCHFV N protein expression constitutes an effective approach for vaccine development and BoHV-4 emerged as a strong alternative to previously used viral vectors.
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    Co-delivery effect of CD24 on the immunogenicity and lethal challenge protection of a DNA vector expressing nucleocapsid protein of crimean congo hemorrhagic fever virus
    (2019) Yılmaz, Erkan; http://orcid.org/0000-0001-7355-3878; Dil ve Tarih-Coğrafya Fakültesi; Farzani, Touraj Aligholipour; Hanifehnezhad, Alireza; Földes, Katalin; Ergünay, Koray
    Crimean Congo hemorrhagic fever virus (CCHFV) is the causative agent of a globally-spread tick-borne zoonotic infection, with an eminent risk of fatal human disease. The imminent public health threat posed by the disseminated virus activity and lack of an approved therapeutic make CCHFV an urgent target for vaccine development. We described the construction of a DNA vector expressing a nucleocapsid protein (N) of CCHFV (pV-N13), and investigated its potential to stimulate the cytokine and total/specific antibody responses in BALB/c and a challenge experiment in IFNAR −/− mice. Because of a lack of sufficient antibody stimulation towards the N protein, we have selected cluster of differentiation 24 (CD24) protein as a potential adjuvant, which has a proliferative effect on B and T cells. Overall, our N expressing construct, when administered solely or in combination with the pCD24 vector, elicited significant cellular and humoral responses in BALB/c, despite variations in the particular cytokines and total antibodies. However, the stimulated antibodies produced as a result of the N protein expression have shown no neutralizing ability in the virus neutralization assay. Furthermore, the challenge experiments revealed the protection potential of the N expressing construct in an IFNAR −/− mice model. The cytokine analysis in the IFNAR −/− mice showed an elevation in the IL-6 and TNF-alpha levels. In conclusion, we have shown that targeting the S segment of CCHFV can be considered for a practical way to develop a vaccine against this virus, because of its ability to induce an immune response, which leads to protection in the challenge assays in the interferon (IFN)-gamma defective mice models. Moreover, CD24 has a prominent immunologic effect when it co-delivers with a suitable foreign gene expressing vector. © 2019 by the authors. Licensee MDPI, Basel, Switzerland.