BIODIVERSITAS ISSN: 1412-033X Volume 23, Number 12, December 2022 E-ISSN: 2085-4722 Pages: 6208-6212 DOI: 10.13057/biodiv/d231215 Short Communication: Cloning, production and responses of the sMTL-13 protein (13 kDa lectin secretion) Mycobacterium tuberculosis against Interleukin 6 ROSANA AGUS 1, , WA ODE SITI PURNAMASARI 1 , RISA DENGEN PARURA 1 , FERDINANDO SILANG 1 , MUHAMMAD NASRUM MASSI 2 1 Department of Biology, Faculty of Mathematics and Natural Sciences, Universitas Hasanuddin. Jl. Perintis Kemerdekaan Km.10, Tamalanrea, Makassar 90245, South Sulawesi, Indonesia. Tel./Fax.: +62-8124136912,  email: rosanagus65@gmail.com 2 Department of Microbiology, Faculty of Medical, Universitas Hasanuddin. Jl. Perintis Kemerdekaan Km.10, Tamalanrea, Makassar 90245, South Sulawesi, Indonesia Manuscript received: 1 September 2022. Revision accepted: 4 December 2022. Abstract. Agus R, Purnamasari WOS, Parura RD, Silang F, Massi MN. 2022. Short Communication: Cloning, production and responses of the sMTL-13 protein (13 kDa lectin secretion) Mycobacterium tuberculosis against Interleukin 6. Biodiversitas 23: 6208- 6212. The Bacillus Calmette-Guérin (BCG) vaccine protected effect against meningitis and disseminated Tuberculosis TB in children. However, it does not prevent primary infection and reactivation of latent pulmonary infection. Therefore, an effective vaccine is needed to prevent tuberculosis. One of the potentials is the sMTL-13 protein (13 kDa lectin secretion), which causes an increase in interleukin 6 (IL-6) as a proinflammatory cytokine and shows high titers in the serum of TB patients. This study aimed to clone, express and evaluate the sMTL-13 recombinant protein. The research steps were ligation of Rv1419 to pQE-30Xa, transformation to Escherichia coli BL-21, and production of recombinant protein with IPTG induction. Furthermore, immunoreactivity tests with the serum of TB patients and healthy serum using ELISA. The results showed that the PCR product Rv1419 showed a band of 474 bp and white colonies as a recombinant clone of pQE-30Xa-Rv1419. The production of sMTL-13 recombinant protein has been successfully carried out and produces 13 kDa protein. The results of the immunoreactivity test showed that the serum level of IL-6 in patients with active TB was 84.22 pg/mL and the serum level of IL-6 in healthy people was 63.74 pg/mL. Keywords: BCG, clone, expression, lectin, sMTL-13 INTRODUCTION Tuberculosis is caused by Mycobacterium tuberculosis (Mtb). These bacteria usually attack the lungs but can attack other body parts such as the kidneys, spine, and brain (CDC 2020). In 2020, 30 countries accounted for 86% of new TB cases. More than 95% of cases and deaths occur in developing countries (WHO 2021). The Bacillus Calmette-Guérin BCG is the most widely used vaccine in the world but its effectiveness varies, between 0 and 80% (Heemskerk et al. 2015). BCG can prevent TB, but its effectiveness varies in high-burden countries and is not effective for preventing of pulmonary TB in adults (WHO. 2019; Wilkie and McShane. 2015). Recent data show that 154 countries have BCG vaccination policies for the entire population. There are 53 countries reporting BCG vaccination coverage of at least 95% and in 2019 there were 31 countries with a decrease in BCG coverage of 5% or more (WHO 2021). Some of the side effects caused by the BCG vaccine include hardness at the injection site, an ulcer arising from blisters at the injection site and an abscess at the injection site (Vaccine Knowledge Project. 2019). Although more than 20 vaccines are currently in clinical trials, it is very important to choose a TB vaccine candidate that is effective and safe against TB infection, especially in the presence of MDR TB and HIV-TB coinfection (WHO 2019). Therefore, developing new vaccines for TB prevention and control is very important. Surface and secreted mycobacterial proteins play a major role in the infection process by mediating macrophage-bacterial interactions and cell death. One of the potential M. tuberculosis proteins is the 13-kDa lectin (sMTL-13) encoded by the Rv1419. Bioinformatics analysis showed that sMTL-13 belongs to the ricin-type β- trefoil family of proteins containing a Sec-type signal peptide present in Mtb complex species. The sMTL-13 causes an increase in IFN-γ production by PBMCs from active tuberculosis patients (Nogueira et al. 2010). sMTL- 13 has an R-type lectin on the surface of Mtb cells and functions in interaction with macrophages. R-type lectins have been detected in plants, animals, and bacteria (Kolbe et al. 2019). In addition, lectins can induce the production of proinflammatory cytokines by APCs and cause the proliferation of CD4+ T lymphocyte cells indicating that sMTL-13 has immunogenic activity (Souza 2012). Interleukin-6 (IL-6) is a cytokine with high serum levels in active pulmonary tuberculosis (TB). IL-6 screening in patients in contact with TB is useful for monitoring the infectious process and for observing the risk of disease progression (Lopes et al. 2013). Interactions between macrophages infected with M. tuberculosis and cell T lymphocytes will cause the release of various