Short Communication: Cloning, production and responses of the sMTL-13 protein (13 kDa lectin secretion) Mycobacterium tuberculosis against Interleukin 6

##plugins.themes.bootstrap3.article.main##

ROSANA AGUS
WA ODE SITI PURNAMASARI
RISA DENGEN PARURA
FERDINANDO SILANG
MUHAMMAD NASRUM MASSI

Abstract

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.

##plugins.themes.bootstrap3.article.details##

References
Briand L, Marcion G, Kriznik A, Heydel JM, Artur Y, Garrido C, Seigneuric R, Neiers. 2016. A self-inducible heterologous protein expression system in Escherichia coli. Scientific Reports 6: 33037. doi:10.1038/srep33037.
Bafica A, Morales S, Eto C, Souza N, Nogueira L, Riley L, Mansur D. 2015.
Biologics International Corp. 2018. IPTG Induction Theory. https://www.biologicscorp.com/blog/iptg-induction-protein-expression
Donovan RS, Robinson CW, Glick BR. 1996. Review: Optimizing inducer and culture conditions for expression of foreign proteins under the control of the lac promoter. Journal of Industrial Microbiology 16: 145-154. doi:10.1007/bf015
Gong W, Liang Y, Wu X. 2018. The current status, challenges, and future developments of new tuberculosis vaccines. Human vaccines & immunotherapeutics 14: 1697–1716. doi: 10.1080/21645515.2018.1458806.
Kolbe K, Veleti SK, Reiling N, Lindhorst TK. 2019. Lectins of Mycobacterium tuberculosis – rarely studied proteins Beilstein J Org Chem 15: 1–15. doi:10.3762/bjoc.15.1.
Liang Y, Zhang X, Xiao L, Bai X, Wang X, Yang Y, Zhang J, Song J, Liu Y, Li N, Wu X. 2016. Immunogenicity and Therapeutic Effects of pVAX1-rv1419 DNA from Mycobacterium tuberculosis. Current Gene Therapy 16: 249-255. doi:10.2174/1566523216666161102170123
Lopes FHA, Lívia Coelho de Assis, Roberto da Justa Pires Neto, Karine Paschoal Botelho, Kélvia Miranda Sá, Cristiane Cunha Frota, José Walter Correia, Max Victor Carioca Freitas, Serum levels of interleukin-6 in contacts of active pulmonary tuberculosis. Jornal Brasileiro de Patologia e Medicina Laboratorial 49(6) :410-414
Merck. Blue-White Screening & Protocols for Colony Selection. 2019. [viewed 18 November 2019]. Available from: https://www.sigmaaldrich.com/ID/en/technical-documents/technical-article/genomics/cloning-and-expression/blue-whitescreening.
Martinez AN, Mehra S, Kaushal D. 2013. Role of Interleukin 6 in Innate Immunity to Mycobacterium tuberculosis Infection. Journal of Infectious Diseases 207(8): 1253–1261.
Nogueira L, Cardoso FC, Mattos AM, Bordignon J, Figueiredo CP, Dahlstrom P, Frota CC, Santos CNDD, Chalhoub M, Cavada BS, Teixeira HC, Oliveira SC, Netto MB, Bafica A. 2010. Mycobacterium tuberculosis Rv1419 encodes a secreted 13 kDa lectin with immunological reactivity during human tuberculosis. Eur J Immunol 40:744–753. doi: 10.1002/eji.200939747.
Patra D, Srikalaivani R, Misra A, Singh DD, Selvaraj M, Vijayan M. 2010. Cloning, expression, purification, crystallization and preliminary X-ray studies of a secreted lectin (Rv1419) from Mycobacterium tuberculosis. Acta Cryst 66: 1662–1665. doi:10.1107/s1744309110042892.
Saunders BM, Anthony A. Frank, Ian M. Orme and Andrea M. Cooper, Interleukin-6 Induces Early Gamma Interferon Production in the Infected Lung but Is Not Required for Generation of Specific Immunity to Mycobacterium tuberculosis Infection. Infect Immun 68(6): 3322–3326.
Sundar S, Lokesh T, Shanmughavel P, Jeyakumar N. 2021. Discovering mycobacterial lectins as potential drug targets and vaccine candidates for tuberculosis treatment: a theoretical approach. Journal of Proteins and Proteomics 12: 93–104. doi:10.1007/s42485-021-00065-y
Sambrook J, Fritsch EF, Maniatis T.1989. Molecular Cloning A Laboratory Manual 2 edn. Cold Spring Harbor Laboratory Press. USA.
Vaccine Knowledge Project. 2019. BCG Vaccine (TB vaccine). Oxford Vaccine Group. https://vk.ovg.ox.ac.uk/vk/
WHO. 2019. Global Tuberculosis Report Executive Summary 2019. World Health Organization publishers. USA.
Wilkie MEM, McShane H. 2015. TB vaccine development: where are we and why is it so difficult?. Chest clinic 70: 299–301. doi: 10.1136/thoraxjnl-2014-205202