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Co-insertion of the gene encoding the human interleukin-6 (IL6) protein (30-212aa) with the N-terminal His-tag gene into a plasmid vector results in the formation of a recombinant plasmid, which is then introduced into E.coli cells. Positive E.coli cells are selected relying on their ability to survive in the presence of a specific antibiotic. The E.coli cells containing the recombinant plasmid are cultured under conditions conducive to the expression of the gene of interest. Following expression, the recombinant human IL6 protein is isolated and purified from the cell lysate through affinity purification. The resultant recombinant human IL6 protein is analyzed using denaturing SDS-PAGE, revealing a purity greater than 85%.
Interleukin-6 (IL6) is a versatile cytokine that plays numerous roles in the body's functions and diseases. It helps regulate genes for acute-phase proteins and other cytokines, affecting inflammation, blood cell formation, and immediate responses to injury or infection [1]. IL6 also manages the production of acute-phase proteins like albumin by liver cells, which is crucial for overall protein balance in the body [2]. Moreover, IL6 influences basic cell activities, like cell growth, communication, and movement, particularly in endothelial cells lining blood vessels [3]. In health issues, IL6 is linked to lung function decline and chronic lung conditions like COPD, hinting at its part in lung inflammation [4]. It's also associated with aggressive types of brain tumors, suggesting a role in cancer spread [5]. Additionally, IL6 has been highlighted in severe COVID-19 cases, indicating its involvement in widespread inflammation and immune responses [6]. IL6 acts through various pathways and connections with other proteins. For example, in aggressive brain tumors, it's linked with NFAT1-controlled signaling [5]. It also affects the body's response to antiproliferative actions of IFNα2 through a protein called IRF9, impacting immune responses [7]. Additionally, IL6 teams up with another protein, leukemia inhibitory factor (LIF), to fuel ovarian cancer growth, possibly showing overlap in their functions through a protein called STAT3 [8]. Furthermore, IL6 levels can serve as markers for disease progression in specific groups of patients with amyotrophic lateral sclerosis, indicating its potential as a measure of disease worsening [9].
References:
[1] S. Akira, H. Isshiki, T. Sugita, O. Takahashi, S. Kinoshita, Y. Nishioet al., "A nuclear factor for il-6 expression (nf-il6) is a member of a c/ebp family.", The Embo Journal, vol. 9, no. 6, p. 1897-1906, 1990. https://doi.org/10.1002/j.1460-2075.1990.tb08316.x
[2] J. Rossi, Z. Liu, M. Jourdan, & B. Klein, "Interleukin-6 as a therapeutic target", Clinical Cancer Research, vol. 21, no. 6, p. 1248-1257, 2015. https://doi.org/10.1158/1078-0432.ccr-14-2291
[3] L. Ljungberg, M. Zegeye, C. Kardeby, K. Fälker, D. Repsilber, & A. Sirsjö, "Global transcriptional profiling reveals novel autocrine functions of interleukin 6 in human vascular endothelial cells", Mediators of Inflammation, vol. 2020, p. 1-12, 2020. https://doi.org/10.1155/2020/4623107
[4] J. He, M. Foreman, K. Shumansky, X. Zhang, L. Akhabir, D. Sinet al., "Associations of il6 polymorphisms with lung function decline and copd", Thorax, vol. 64, no. 8, p. 698-704, 2009. https://doi.org/10.1136/thx.2008.111278
[5] Y. Jiang, S. Han, W. Cheng, Z. Wang, & A. Wu, "Nfat1-regulated il6 signalling contributes to aggressive phenotypes of glioma", Cell Communication and Signaling, vol. 15, no. 1, 2017. https://doi.org/10.1186/s12964-017-0210-1
[6] N. Aarskog, H. Aass, J. Holter, M. Rostrup, & A. Holten, "Interleukin-6 in critical coronavirus disease 2019, a driver of lung inflammation of systemic origin?", Critical Care Explorations, vol. 3, no. 10, p. e0542, 2021. https://doi.org/10.1097/cce.0000000000000542
[7] H. Erb, R. Langlechner, P. Moser, F. Handle, T. Casneuf, K. Verstraetenet al., "Il6 sensitizes prostate cancer to the antiproliferative effect of ifnα2 through irf9", Endocrine Related Cancer, vol. 20, no. 5, p. 677-689, 2013. https://doi.org/10.1530/erc-13-0222
[8] K. McLean, L. Tan, D. Bolland, L. Coffman, L. Peterson, M. Talpazet al., "Leukemia inhibitory factor functions in parallel with interleukin-6 to promote ovarian cancer growth", Oncogene, vol. 38, no. 9, p. 1576-1584, 2018. https://doi.org/10.1038/s41388-018-0523-6
[9] M. Wosiski‐Kuhn, J. Caress, M. Cartwright, G. Hawkins, & C. Milligan, "Interleukin 6 (il6) level is a biomarker for functional disease progression within il6r358ala variant groups in amyotrophic lateral sclerosis patients", Amyotrophic Lateral Sclerosis and Frontotemporal Degeneration, vol. 22, no. 3-4, p. 248-259, 2020. https://doi.org/10.1080/21678421.2020.1813310
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