Masruroh, RositaArviana (2014) Isolasi Dan Identifikasi Protein Feline Tetherin/Bst-2 Dari Peripheral Blood Mononuclear Cells (Pbmcs) Yang Diinduksi Interferon-Alpha (Ifn-Α). Sarjana thesis, Universitas Brawijaya.
Abstract
Infeksi enveloped virus diketahui menimbulkan berbagai macam penyakit infeksi serta kematian pada kucing, seperti Feline Immunodeficiency Virus (FIV) dan Feline Leukimia Virus (FELV). Enveloped virus memiliki kemampuan adaptasi yang tinggi pada sel hospes karena mempunyai viral envelope yang menyebabkan mudah untuk menghindar dari sistem pertahanan hospes. Tetherin/BST-2 merupakan protein antiviral yang dihasilkan oleh berbagai jenis sel darah putih, seperti Peripheral Blood Mononuclear Cell (PBMCs) dan diekspresikan sebagai respon terhadap adanya induksi IFN tipe I seperti IFN- Tetherin/BST-2 telah diidentifikasi berperan penting dalam respon innate immunity, salah satunya dapat menghambat pelepasan enveloped virus dari sel hospes terinfeksi. Penelitian ini bertujuan untuk mengetahui ekspresi dan karakteristik protein feline tetherin/BST-2 pada PBMCs yang diinduksi IFN- PBMCs diisolasi dari darah kucing sehat yang berusia lebih dari 1 tahun. PBMCs dibagi menjadi dua kelompok, yaitu kelompok kontrol dan kelompok perlakuan yang diinduksi IFN- dengan dosis 1000 U/ml. Protein total diisolasi dari sel kultur setelah 24 jam inkubasi dan dianalisa dengan teknik SDS-PAGE, dot blot, dan western blotting. Hasil analisis SDS-PAGE menunjukkan adanya pita protein berukuran 35 kDa pada sampel perlakuan induksi IFN-yang diduga merupakan pita protein feline Tetherin. Analisa pengujian dengan dot blot menunjukkan adanya reaksi positif antara antigen protein Tetherin/BST-2 dan antibodi anti BST- 2 berupa dot warna biru keunguan pada sampel perlakuan induksi IFN-. Hasil analisis western blot menunjukkan konfirmasi adanya ekspresi protein feline Tetherin/BST-2 dengan ukuran berat molekul sekitar 35 kDa. Berdasarkan hasil penelitian dapat disimpulkan bahwa pemberian induksi IFN-dengan dosis 1000 U/ml mampu menginduksi ekspresi protein feline Tetherin/BST-2 pada sel kultur PBMC, dan hasil karakterisasi protein feline Tetherin/BST-2 menunjukkan profil protein dengan berat molekul sekitar 35 kDa.
English Abstract
Enveloped virus infection are known caused of various kinds infections and death in cats, such as Feline Immunodeficiency Virus (FIV) and Feline Leukemia Virus (FeLV). Enveloped viruses have high adaptability in host cells because they have viral envelope which facilitated avoiding host defense system. Tetherin/BST-2 is a potential antiviral protein produced by various cell types including Peripheral Blood Mononuclear Cells (PBMCs) and that is expressed in response of IFN type I such as IFN-?? induction. Tetherin/BST-2 has been identified plays an important role in the innate immunity, one of which can inhibits the release of enveloped virus from infected host cells.This study was aimed to characterize the expression of protein feline Tetherin/BST-2 on PBMCs induced by IFN-?. PBMCs were isolated from blood sample of healthy cats with aged more than 1 year. PBMCs were divided into two groups, namely the control group and the treatment group which induced by IFN- ??at dose of 1000 U / ml. Total protein was isolated from cultured cells after 24 hours incubations and analyzed using SDS-PAGE, dot blot, and western blotting. The results of SDSPAGE analysis showed a 35 kDa protein band in samples that were induced by IFN-??which suggested to be the feline tetherin protein bands. The results dot blot analysis was indicate the positive reaction of antigen Tetherin/ BST-2 and antibody anti BST-2 that marked by purplish blue dot also at sample induction of IFN- treatment. The results of western blot analysis showed confirmation of feline tetherin/BST-2 protein expression with a protein band size approximately 35 kDa molecular weight. Based on the results of this study can be concluded that administration of IFN-induction by doses of 1000 U /ml is able to induces the expression of feline tetherin protein/BST-2 in PBMCs cultured cells, and protein characterization results feline Tetherin/BST-2 shows the protein profiles with a molecular weight of about 35 kDa.
| Item Type: | Thesis (Sarjana) |
|---|---|
| Identification Number: | SKR/FKH/2014/14/051401716 |
| Subjects: | 600 Technology (Applied sciences) > 636 Animal husbandry > 636.08 Specific topics in animal husbandry > 636.089 Veterinary medicine |
| Divisions: | Fakultas Kedokteran Hewan > Kedokteran Hewan |
| Depositing User: | Budi Wahyono Wahyono |
| Date Deposited: | 06 Mar 2014 09:38 |
| Last Modified: | 04 Mar 2022 07:17 |
| URI: | http://repository.ub.ac.id/id/eprint/126899 |
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