Koyunlarda viral hastalık maedi-visna'ya karşı aşı geliştirilmesinde antijenik epitopların keşfi
Küçük Resim Yok
Tarih
2023
Yazarlar
Dergi Başlığı
Dergi ISSN
Cilt Başlığı
Yayıncı
Ege Üniversitesi
Erişim Hakkı
info:eu-repo/semantics/openAccess
Özet
Maedi Visna (MVV), retrovirüsler familyasının Lentivirus alt grubunda yer alan, koyunları enfekte eden virüstür. Hastalığın Maedi formu ilerleyici bir interstisyel pnömonidir, Visna ise bir meningoensefalittir. Hayvan sağlığı ve refahı problemlerinin yanında ciddi ekonomik kayıplara sebep olan bu virüse karşı günümüzde halen etkin bir tedavi yöntemi ve korunmaya yönelik aşı bulunmamaktadır. Öte yandan biyoinformatik alanındaki gelişmeler, aşı çalışmalarının ilk ve en önemli adımını oluşturan antijenik proteinlerin ve epitopların seçimi için büyük olanaklar sağlamaktadır. Buradan yola çıkılarak in silico analizler sonrasında sentezlenen epitopların immünojenitesi in vitro koşullarda ELISA yöntemi ile kategorize edilmiş MVV pozitif koyunlardan toplanan serum örneklerinin kullanımı ile test edilmiştir. Elde edilen sonuçlara göre gag proteininin amino asit sekanslarının daha fazla korunduğu ve daha yüksek antijenite değerine sahip olduğu tespit edilmiştir. Yapılan çalışmada birçok B hücresi ve MHC-I/II epitopu tahmin edilse de bunlardan sadece 19'unun antijenik, alerjik olmayan, toksik olmayan, çözünür ve hemolitik olmayan özelliklere sahip olduğu tespit edilmiştir. Bununla birlikte, multi-epitop aşı konstraktı, toplam 19 epitop ile tasarlanmış ve TLR-2/4 arasında güçlü bir afinite göstermiştir. Yapılan ELISA testlerinden elde edilen absorbans değerlerine göre tüm peptitlerin immünojenik olduğu saptanmıştır. Yapılan ROC analizi sonucunda, sentezlenen peptitlerden gag proteini için 3. peptit' in (CKQGSKE), env proteini için 5. peptit' in (CEHERRTSHRS) gösterdikleri yüksek değerlerdeki sensitivite ve spesifite ile en iyi peptitler olduğu sonucuna varılmıştır. Testin anlamlı olup olmadığını değerlendirmek için yapılan t-testi sonucunda sadece 6. peptit, env epitoplarının karışımı ve env epitoplarının karışımı halinde çalışılan testler p- value değerleri 0,05'den büyük olduğu için anlamsız olarak saptanmıştır. Sonuç olarak, veteriner hekimlikte önemli bir viral hastalık olan MVV'ye karşı peptit aşısı geliştirilmesi sırasında kullanılabilecek immünojenitesi hem in silico hem de in vitro çalışmalar ile gösterilmiş ideal peptitler ilk kez saptanmıştır
Maedi Visna (MVV), is a retrovirus that member of the lentivirus subfamily and it can infect the sheep. The Maedi form of the disease is a progressive interstitial pneumonia, while Visna is a meningoencephalitis. While virus cause serious economic losses as well as animal health and welfare problems, there is no effective treatment and vaccine. For this reason, the need of developing a vaccine against MVV is inevitable. On the other hand, developments in bioinformatics provide great opportunities for the selection of antigenic proteins and epitopes, which constitute the first and most important step of vaccine studies. Moreover, the selected epitopes can be synthesized in a short time, and their immunogenicity can be tested in vitro with the use of serum samples collected from previously infected hosts. From that point, the immunogenicity of the epitopes synthesized after in silico analysis was tested by using serum samples collected from MVV positive sheep, which were categorized by ELISA method under in vitro conditions. According to the obtained data, the amino acid sequence of gag protein was found more conserved and antigenic. Although many B cells and MHC-I/II epitopes were predicted, only 19 of them were found as antigenic, non-allergic, non-toxic, soluble and non-hemolytic properties. However, the multi-epitope vaccine construct was designed with a total of 19 epitopes and showed strong affinity for TLR-2/4. According to the absorbance values obtained from ELISA tests, it was determined as all peptides were immunogenic. As a result of the ROC analysis, it was concluded that the 3. peptide for the gag protein (CKQGSKE) and the 5. peptide for the env protein (CEHERRTSHRS) were best peptides with their high sensitivity and specificity. As a result of the t-test, only the 6. peptide, mixture of gag epitopes and mixture of env epitopes were found to be insignificant because the p-values were greater than 0,05. In conclusion, ideal peptides for the development of a peptide vaccine against MVV have been identified for the first time that demonstrated by both in silico and in vitro studies.
Maedi Visna (MVV), is a retrovirus that member of the lentivirus subfamily and it can infect the sheep. The Maedi form of the disease is a progressive interstitial pneumonia, while Visna is a meningoencephalitis. While virus cause serious economic losses as well as animal health and welfare problems, there is no effective treatment and vaccine. For this reason, the need of developing a vaccine against MVV is inevitable. On the other hand, developments in bioinformatics provide great opportunities for the selection of antigenic proteins and epitopes, which constitute the first and most important step of vaccine studies. Moreover, the selected epitopes can be synthesized in a short time, and their immunogenicity can be tested in vitro with the use of serum samples collected from previously infected hosts. From that point, the immunogenicity of the epitopes synthesized after in silico analysis was tested by using serum samples collected from MVV positive sheep, which were categorized by ELISA method under in vitro conditions. According to the obtained data, the amino acid sequence of gag protein was found more conserved and antigenic. Although many B cells and MHC-I/II epitopes were predicted, only 19 of them were found as antigenic, non-allergic, non-toxic, soluble and non-hemolytic properties. However, the multi-epitope vaccine construct was designed with a total of 19 epitopes and showed strong affinity for TLR-2/4. According to the absorbance values obtained from ELISA tests, it was determined as all peptides were immunogenic. As a result of the ROC analysis, it was concluded that the 3. peptide for the gag protein (CKQGSKE) and the 5. peptide for the env protein (CEHERRTSHRS) were best peptides with their high sensitivity and specificity. As a result of the t-test, only the 6. peptide, mixture of gag epitopes and mixture of env epitopes were found to be insignificant because the p-values were greater than 0,05. In conclusion, ideal peptides for the development of a peptide vaccine against MVV have been identified for the first time that demonstrated by both in silico and in vitro studies.
Açıklama
Anahtar Kelimeler
Biyoteknoloji, Biotechnology
