Cloning and expression of pab gene of M. tuberculosis isolated from pulmonary TB patient in E.coli DH5α

Raras, Tri Yudani Mardining; Lyrawati, Diana

doi:10.13181/mji.v20i4.458

Cited by 10 publications

(3 citation statements)

References 6 publications

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“…Recombinant protein Ag38kDa was obtained from the Biomedical Laboratory, Faculty of Medicine, Universitas Brawijaya Malang, Indonesia. The pMB38 plasmid (Raras & Lyrawati, 2011), carrying the pab gene, was expressed in E. coli strain BL21-(DE-3) carrying the Ag38kDa recombinant protein plasmid was purified using Column Nited (Raras, Sholeh, & Lyrawati, 2014).…”

Section: Overexpression Of Ag38kdamentioning

confidence: 99%

Immune Response on the Administration of Recombinant Protein Antibodies Ag-38 Kda Mycobacterium Tuberculosis and Rifampicin Ex-Vivo.

Raras

Fahrinda²,

Yuliati³

et al. 2022

AJID

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Background: Development a granuloma model resembling latent tuberculosis in vitro is needed with a fast and efficient time to be used as an effective therapy. This study aimed to form efficient granulomas, increase cellular immunity and humoral immunity, and evaluate growth on media using recombinant protein antibody Ag38kDa, Rifampicin, and a combination of both. Peripheral Blood Mononuclear Cell (PBMC) in vitro is derived from a healthy individual separated from monocytes and lymphocytes. Materials and methods: Monocytes are matured into macrophages and then combined macrophages and lymphocytes to the Roswell Park Memorial Institute (RPMI) medium. Flow cytometry analysis was used to count the number of cells, and cytokine levels were measured using ELISA. The result from the treatment was planted on the Lowenstein-Jensen medium. Results: Granulomas-like aggregates was formed after one-day post-infection with Mycobacterium tuberculosis (M.tb). A significant increase in immune response occurred in the number of macrophages, Th1, and Tregs in the combination group compared to the Mtb infection group. The number of Th2 and Th17 cells in the combination group was compared with the control but not significantly. TNF-α cytokine levels increased in the combination group compared to Mtb infection, while in IL-4, we found between all groups, there was no significant difference. Bacterial colonies on culture in the Lowenstein-Jensen medium were only seen in positive controls. Conclusion: Our study concluded that administration of a combination between Ag38kDa recombinant antibody and rifampicin could inhibit granuloma formation and enhance immune response.

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Section: Overexpression Of Ag38kdamentioning

confidence: 99%

Immune Response on the Administration of Recombinant Protein Antibodies Ag-38 Kda Mycobacterium Tuberculosis and Rifampicin Ex-Vivo.

Raras

Fahrinda²,

Yuliati³

et al. 2022

AJID

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“…4,5 Antigen 38 kDa M.tb merupakan zat yang berkemampuan untuk pengembangan vaksin TB karena memiliki epitop khas sel TCD4+, sel T CD8+ (CTL) dan epitop untuk sel B. [6][7][8] Penelitian lain tentang protein 38 kDa M.tb menyebutkan bahwa zat ini membangkitkan respons sel T CD4+ dan T CD8+ di tikus, tetapi di manusia hanya T CD8+. 9 Pada penelitian oleh Kusumaningsih dkk tahun 2009 ditemukan peningkatan jumlah sel T CD4+, T CD8+ dan IFN-γ + di jaringan paru dan usus mencit yang diberi vaksin oral dengan protein ini.…”

Section: Pendahuluanunclassified

Protein Rekombinan 38 Kda Mycobakterium Tuberkulosis Dapat Mengimbas Pembuatan Interleukin-2 Dan Interferon-Î³ Limfosit T Di Kultur Sel Mononuklear Darah Tepi

Arthamin¹,

Wahono²,

Primardianti³

et al. 2018

Indonesian J. Clin. Pathol. Med. Lab.

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Tuberculosis (TB) is caused by Mycobacterium tuberculosis (M.tb) and is one of the significant mortality causes WHO (2012). Theprimary immune response in TB pathogenesis is Cell Mediated Immunity (CMI), roled by T lymphocytes. Interleukin-2 (IL-2) is a growthfactor for T lymphocytes. Gamma Interferon is the key cytokine in M.tb infection control, synthezised by T lymphocytes. An effectivevaccination strategy is achieved by giving vaccine which is able to stimulate T lymphocytes in synthezising cytokines. The 38 kDa M.tbprotein is potential in the vaccine development program, because it has specific epitopes for T lymphocytes. The aim of this study was toknow how to determine that the 38 kDa recombinant protein of M.tb Malang strain could induce cellular immune response by IL-2 andIFN-γ synthezised by T lymphocytes. The study was carried out by an experimental in vitro study on PBMC from healthy endemic subjects,those having TB contact, and the TB patients themselves. PBMC from subjects was cultured with 38 kDa recombinant protein of M.tbMalang strain, with PPD and without any protein. The analysis of IL-2 and IFN-γ used flowcytometry. The result showed that the highestpercentage of IL-2 was found in the culture with 38 kDa recombinant protein of M.tb Malang strain, in healthy endemic (p=0.000)and in those who had TB contact (p=0.000). the highest percentage of IFN-γ was found in the culture with 38 kDa recombinant proteinof M.tb Malang strain, in healthy endemic (p=0.007) and those who had TB contact (p = 0.105). The 38 kDa recombinant proteinof M.tb Malang strain was able to induce IL-2 and IFN-γ synthezised by TCD3+ lymphocytes from healthy endemic subjects and thosewho had TB contact.

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“…Production of Ag38-rec was conducted according to a previous study with slight modifications 11 . The pab gene coding for Ag38 was amplified via a PCR method using chromosomal DNA from M. tuberculosis which was isolated from a severe pulmonary TB patient in Malang, Indonesia.…”

Section: Purification Of Ag38-recmentioning

confidence: 99%

Evaluation of immunologic response of salivary sIg-A in pediatric tuberculosis patients to antigen Ag38-rec of Mycobacterium tuberculosis Indonesian strain

2015

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We studied the immune response of salivary secretory Objective immunoglobulin A (sIg-A) from a pediatric tuberculosis (TB) group (scoring of 6) and non-TB group (scoring <6) against recombinant Ag38 (Ag38-rec) from Indonesian strain. Mycobacterium tuberculosis: Seventy-eight children were divided into three Materials and Methods groups; those with TB (n=26), those with suspected TB (n=26), and healthy children (n=26), their saliva was collected, and salivary sIg-A was challenged with purified Ag38-rec using the dot blot method. A change of color from white to dark blue indicated a positive reaction.: The immune response of sIg-A of children with TB and those with Results suspected TB to Ag38-rec was not significantly different. In the TB group, Ag38-rec showed a higher sensitivity than protein purified derivative (PPD) (70.8% vs. 62.5%), but a lower specificity (26.9% vs. 34.62%). However, within both groups (scoring of 6) as well as non-TB group (scoring <6) Ag38-rec was able to identify children with a positive TST (tuberculin skin test) better than PPD.: The antigen Ag38-rec could not distinguish between children with Conclusion TB scores of 6 and <6.1. However, it demonstrated the potential of Ag38-rec for use in screening for TB infection among children with suspect TB (scores <6).

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Cloning and expression of pab gene of M. tuberculosis isolated from pulmonary TB patient in E.coli DH5α

Cited by 10 publications

References 6 publications

Immune Response on the Administration of Recombinant Protein Antibodies Ag-38 Kda Mycobacterium Tuberculosis and Rifampicin Ex-Vivo.

Immune Response on the Administration of Recombinant Protein Antibodies Ag-38 Kda Mycobacterium Tuberculosis and Rifampicin Ex-Vivo.

Protein Rekombinan 38 Kda Mycobakterium Tuberkulosis Dapat Mengimbas Pembuatan Interleukin-2 Dan Interferon-Î³ Limfosit T Di Kultur Sel Mononuklear Darah Tepi

Evaluation of immunologic response of salivary sIg-A in pediatric tuberculosis patients to antigen Ag38-rec of Mycobacterium tuberculosis Indonesian strain

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