Immunogenicity Test of ESAT-6/CFP-10 Mycobacterium Tuberculosis (Indonesian Strain) Recombinant Protein Fusion: IFN-γ and CD8+ T Cells Expression in PBMC Culture

Authors

  • Anung Sri Handayani Departemen Pulmonologi dan Kedokteran Respirasi, Fakultas Kedokteran Universitas Brawijaya, Malang
  • Tri Wahju Astuti Departemen Pulmonologi dan Kedokteran Respirasi, Fakultas Kedokteran Universitas Brawijaya, Malang
  • Teguh Rahayu Sartono Departemen Pulmonologi dan Kedokteran Respirasi, Fakultas Kedokteran Universitas Brawijaya, Malang
  • Maimun Zulhaidah Arthamin Departemen Patologi Klinik, Fakultas Kedokteran Universitas Brawijaya, Rumah Sakit Umum Saiful Anwar, Malang
  • Fransisca Srioetami Tanoerahardjo Pusat Penelitian dan Pengembangan Kementerian Kesehatan Indonesia

DOI:

https://doi.org/10.36497/jri.v38i4.44

Keywords:

Recombinant protein fusion ESAT-6/CFP-10, Mycobacterium tuberculosis, IFN-γ, CD8 T cells

Abstract

Background: BCG vaccination is one way to control tuberculosis (TB) but still poor in efficacy thus new vaccine development is needed. Immunogenicity test is needed in developing new vaccine. The aim of this study was to understand whether the recombinant protein fusion of ESAT-6/CFP-10 Mycobacterium tuberculosis (M. tuberculosis) can stimulate cellular immune response, especially IFN-γ and CD8 + T cell expression in PBMC cultures. Methods: This study was an experimental laboratory research conducted on PBMC cultures of 3 groups of subjects (TB patients, latent TB patients and healthy subjects) at RSUD Dr. Saiful Anwar in April-July 2017. The sample of each groups was 8 subjects. Each groups induced by recombinant protein fusion ESAT-6/CFP-10 M. tuberculosis as a standard protocol and to establish the immunogenicity status. CD8+ T cells IFN-γ expressed by C8+ were measured by flowcytometry. Result: Recombinant protein fusion ESAT-6/CFP-10 can stimulate CD8+ T cells and IFN-γ expressed by CD8+ T cells in all group. The highest stimulation of CD8+ percentage was found in healthy subject (37.533 ± 7.264) and IFN-γ expressed by CD8+ T cells was found in healthy subject (7.908 ± 4.457); There are increase significantly CD8+ T cells (p=0.001) and IFN-γ expressed by CD8+ T cells (p=0.217) not significantly in healthy subject compared in PPD and without antigen. Conclusion: Recombinant protein fusion ESAT-6/CFP-10 M. tuberculosis can stimulate CD8+ T cells and IFN-γ expressed by CD8+ T cells in healthy subject. Recombinant protein fusion ESAT-6/CFP-10 M. tuberculosis potential as a new vaccine candidate. (J Respir Indo. 2018;38: 210-8)

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References

Tang XL, Zhou YX, Wu SM, Pan Q, Xia B, Zhang XL. CFP10 dan ESAT6 aptamers as effective Mycobacterial antigen diagnostic reagents. J. Infect. 2014;69:569–80

World Health Organization. Global Tuberculosis Report. [Online]. 2016 [Citted 2016 Mar 1]. Available from: http://www.who.int/tb/publications/global_report/gtbr2016_executive_summary.pdf

Kementerian Kesehatan RI. Pedoman Nasional Pengendalian Tuberkulosis. Jakarta: 2014.p.6–7

Kaplan G. Rational vaccine development - A new trend in tuberculosis control. N Engl J Med. 2005;353:16245

Cooper AM. Cell mediated immune responses in tuberculosis. Annu Re Immunol. 2009;27:393–422

Arthamin Z, Didiet T, Fransisca. Uji imunogenitas protein rekombinan fusi 38 kDa Mycobacterium tuberculosis galur Malang pada ekspresi IL-2 dan IL-4 limfosit T CD3+ Pada Kultur PBMC. Indonesian Journal of Clinical Pathology and Medical. 2015;21:244-9

Sastroasmoro S, Ismael S. Dasar-Dasar Metodologi Penelitian Klinis. Edisi 5. Sagung Seto. Jakarta. 2014. hal. 281

Nofizar D, Nawas A, Burhan E. Identifikasi faktor risiko tuberculosis multidrug resistant (TB-MDR). Maj Kedokt Indon 2010; 60: 537-45

Podewils LJ, Holtz T, Riestina V, Skripeonokav, Zarovska E, Kirvelaite G, et al. Impact of malnutrition on clinical presentation, Clinical course and mortality In MDR TB patients. Epidemiol Infect. 2011; 139: 113-20

Gupta KB, Gupta R, Atreja A, Verma M, Vishkroma S. Tuberculosis and nutrition. Lung India. 2009: 26: 9-16

Pratomo IP, Burhan E, Tambunan V. Malnutrition and tuberculosis. J Indon Med Assoc. 2012; 62:230-7

Bates MN, Khalakdina A, Pel M, Chang L, Lessa F, Smith KR. Risk of tuberculosis from exposure to tobacco smoke. Arch intern Med; 2007; 167: 335-42

Suprayitno B, Rahajoe NN, Rahajoe N, Boediman I, Said M, Setyanto DB. Karakteristik tuberculosis anak dengan biakan positif. Cermin Dunia Kedokteran. 2002;13:22-5

World Health Organization. The immunological basis for immunization series. Module 5: Tuberculosis. WHO Press. 2011. p 1-30

Martın C. 2005. The dream of a vaccine against tuberculosis; new vaccines improving or replacing BCG?. Eur Respir J. 2005;26:162–7

Tiwari S, Amood K, Kapoor SK. Relationship between sputum smear grading and smear conversion rate and treatment outcome in the patient of pulmonary tuberculosis undergoing DOTS - A prospective cohortsStudy. Indian J. Tuberc. 2012;59:132-40

Grippi MA, Elias JA, Fishman JA, Kotloff RM, Pack AI, Senior RM. Fishman’s Pulmonary Diseases and Disorders. 5th Ed. McGraw-Hill

Medical. New York. 2015. p 3971–4010

Perhimpunan Dokter Paru Indonesia. Pedoman Diagnosis dan Penatalaksanaan di Indonesia. Tuberkulosis. 2016. hal 3.

Orme I. Adaptive immunity to mycobacteria. Curr opin immunol. 2004;7:58–61

Hanecom WA, Abel B, Scriba TJ. Immonological protection against tuberculosis. SAMJ. 2007; 97:973-7

Raja A. 2004. Immunology of tuberculosis. Indian J Med Res. 2004;120:213-32

Skold M, Behar SMRole of CD1d restricted NKT cells in microbial immunity. Infect Immun. 2003;71:5447-55

Nagata T, Koide, Y. Immune responses against mycobacterium tuberculosis and the vaccine strategies. In: Cardona, P. J. (ed.) Understanding tuberculosis - analyzing the origin of mycobacterium tuberculosis pathogenicity. Kroasia: InTech. 2012

Eley BS, Beatty DW. The basic immunology of tuberculosis. Saunders; 2009. p. 75-86

Tores M, Herrera T, Villareal H, Rich EA, Sada E. Cytokine profiles for peripheral blood lymphocytes from patients with active pulmonary tuberculosis and healthy household contacts in response to the 30-kilodalton antigen of Mycobacterium tuberculosis. Infect Immun. 1998;66:176-80

Ginsberg AM. . A proposed national strategy for tuberculosis vaccine development. Clin Infect Dis ;30(Suppl 3);S233-42

Samten B, Wang X. Barnes PF. Mycobacterium tuberculosis ESX-1 systemsecreted protein ESAT-6 but not CFP10 inhibits human T-cell immune responses. Tuberculosis. 2009; 89(Suppl 1):S74-S76

Wang X, Barnes PF, Huang F, Alvarez IB, Neuenschwander PF, Sherman DR, et al. Early secreted antigenic target of 6-kDa protein of Mycobacterium tuberculosis primes dendritic cells to stimulate Th17 and inhibit Th1 immune responses. J Immunol. 2012;189: 3092-103

Dawson R, Condos R, Huie ML, Ress S, Tseng CH, Brauns C, et al. Immunomodulation with recombinant interferon-gamma 1b in pulmonary tuberculosis. PLos One. 2009;4: e6984

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Published

2018-10-28

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