UJI IMUNOGENITAS PROTEIN REKOMBINAN FUSI ESAT-6/CFP-10 Mycobacterium tuberculosis TERHADAP EKSPRESI IFN-γ DAN JUMLAH LIMFOSIT T CD4+ PADA KULTUR PBMC

La’a, dr. Modestus and dr. Maimun Z. Arthamin, M.Kes., Sp.PK. and dr. Agustin Iskandar, M.Kes., Sp.PK. (2019) UJI IMUNOGENITAS PROTEIN REKOMBINAN FUSI ESAT-6/CFP-10 Mycobacterium tuberculosis TERHADAP EKSPRESI IFN-γ DAN JUMLAH LIMFOSIT T CD4+ PADA KULTUR PBMC. Magister thesis, Universitas Brawijaya.

Abstract

Latar Belakang: Vaksinasi merupakan usaha pencegahan dan pemberantasan tuberkulosis (TB) yang efektif dengan menstimulasi respons imun terutama limfosit T CD4+ untuk menghasilkan sitokin-sitokin seperti Interferon gamma (IFN-γ). Interferon gamma memiliki peranan yang sangat penting dalam proteksi terhadap infeksi TB. Protein rekombinan fusi Early Secretory Antigenic Target 6 kDa (ESAT-6) dan Culture Filtrate Protein 10 kDa (CFP-10) mempunyai banyak epitop yang sangat kuat dikenali oleh limfosit T sehingga diharapkan dapat menjadi kandidat vaksin baru. Tujuan penelitian ini adalah untuk menentukan pengaruh pemberian protein rekombinan fusi ESAT-6/CFP-10 M.tb terhadap respons imun seluler pada kultur Peripheral Blood Mononuclear Cell (PBMC) pasien TB, kontak TB dan sehat endemik. Metode Penelitian: Penelitian eksperimental in vitro menggunakan subyek penelitian pasien TB, kontak TB dan sehat endemik masing-masing kelompok 8 orang. Darah vena yang diambil dilakukan isolasi PBMC dan dikultur dengan 3 perlakuan, tidak dipapar protein apapun, dipapar PPD 2 μg dan dipapar protein rekombinan fusi ESAT-6/CFP-10 dosis 2 μg/mL. Pemeriksaan ekspresi IFN-γ dan jumlah limfosit T CD4+ menggunakan metode flowcytometri, kemudian dianalisis dengan Anova atau Kruskal Wallis. Hasil: Rerata ekspresi IFN-γ setelah distimulasi protein rekombinan fusi ESAT-6/CFP-10 pada kelompok pasien [rerata (SD)]: 39,208% (37,431), kontak TB 14,316% (7,821), dan sehat endemik 12,280% (8,376) dengan p=0,679. Rerata jumlah sel T CD4+ setelah distimulasi protein rekombinan fusi ESAT-6/CFP-10 pada kelompok pasien 25,969% (12,169), kontak TB 23,269% (8,709), dan sehat endemik 27,585% (8,085) dengan p=0,264. Kesimpulan: Pemberian protein rekombinan fusi ESAT-6/CFP-10 pada ketiga kelompok subyek penelitian dengan t

English Abstract

Background: Vaccination is effective prevention and eradication program of Tuberculosis (TB) by stimulating the immune response, especially CD4+ T lymphocytes to produce cytokines such as interferon gamma (IFN-γ). Interferon gamma has an important role in protection against TB infection. Fusion recombinant protein called Early Secretory Antigenic Target 6 kDa (ESAT-6) and Culture Filtrate 10 kDa protein (CFP-10) have many epitopes which are very strong to be recognized by T lymphocytes, so they are expected by the researcher to become new vaccine candidates. The aim of this study was to determine the effect of giving ESAT-6/CFP-10 M.tb fusion recombinant protein to the cellular immune response in Peripheral Blood Mononuclear Cell (PBMC) in TB patients, endemic TB and healthy contacts. Research Method: This in vitro experimental study used TB patients, TB contact and healthy endemic contacts as a subject and each group consist of 8 people. Venous blood was isolated from PBMC and cultured with 3 treatments, not exposed to any protein, exposed to 2 μg PPD and ESAT-6/CFP-10 fusion recombinant protein with 2 μg/mL dosage. The flowcytometry method was used to examine IFN-γ expression and CD4+ T lymphocyte count, then these would be analysed by Anova or Kruskal Wallis test. Results: The mean IFN-γ expression after stimulated with ESAT-6/CFP-10 fusion recombinant protein in the patient group was [mean (SD)]: 39,208% (37,431), TB contact 14,316% (7,821), and endemic health 12,280% (8,376) with p=0.679. The mean number of CD4+ T cells after stimulated ESAT-6/CFP-10 fusion recombinant protein in the patient group was 25.969% (12,169), TB contacts 23,269% (8,709), and endemic health 27,585% (8,085) with p=0.264. Conclusion: There was no significant difference in administration of ESAT- 6/CFP-10 fusion recombinant proteins in three research subject groups with three different treatments

Other obstract

Item Type:	Thesis (Magister)
Identification Number:	TES/616.995/LA'A/u/2019/041901637
Uncontrolled Keywords:	Kata kunci : ESAT-6/CFP-10, M.tb, IFN-γ, limfosit T CD4+
Subjects:	600 Technology (Applied sciences) > 616 Diseases > 616.9 Other disease > 616.99 Tumors and miscellaneous communicable diseases > 616.995 Tuberculosis
Divisions:	Profesi Kedokteran > Spesialis Patologi Klinik, Fakultas Kedokteran
Depositing User:	yulia Chasanah
Date Deposited:	26 Aug 2022 07:06
Last Modified:	26 Aug 2022 07:54
URI:	http://repository.ub.ac.id/id/eprint/193645

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