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To create the recombinant mouse interleukin-7 (IL7), CUSABIO uses genetic engineering techniques to insert the DNA fragment that encodes the 26-154aa of IL7 protein into a plasmid and then transform the recombinant plasmid into E.coli cells for protein expression. The E.coli serves as a factory for producing the IL7 protein using its cellular machinery. The recombinant mouse IL7 is extracted from the cell lysate and then subjected to affinity chromatography purification. Its purity reaches over 96% as assessed by SDS-PAGE. It contains endotoxin less than 1.0 EU/μg as measured by the LAL method. This mouse IL protein has also been validated to be biologically active in a cell proliferation assay. Its ED50 is less than 0.2 ng/ml using murine 2E8 cells, corresponding to a specific activity of >5.0x106 IU/mg.
IL7 is a crucial cytokine that plays a significant role in the development and maintenance of B and T cells in mice [1][2]. It has been identified as a proliferation factor for B-cell progenitors [3]. Studies have shown that IL7 is essential for the survival of memory T lymphocytes and for promoting the clonal expansion of activated lymphocytes [4]. Furthermore, IL7 has been found to enhance the development of functional human T cells and boost thymic human T cell development [5][6]. In mouse models, IL7 has been demonstrated to regulate the survival and generation of memory CD4 cells [7].
IL7 has also been associated with immune responses against cancer, as it can enhance vaccine-induced antitumor immunity and improve mouse survival in tumor-bearing models by increasing the survival and cytolytic activity of activated T cells [8]. Additionally, IL7 has been shown to induce the expansion and cytotoxic activity of cytotoxic T lymphocytes (CTL) in the presence of other cytokines like IL15.
References:
[1] D. Munfus, C. Haga, P. Burrows, & M. Cooper, A conserved gene family encodes transmembrane proteins with fibronectin, immunoglobulin and leucine-rich repeat domains (figler), BMC Biology, vol. 5, no. 1, 2007. https://doi.org/10.1186/1741-7007-5-36
[2] H. Uenishi, H. Hiraiwa, T. Sawazaki, S. Kiuchi, & H. Yasue, Genomic organization and assignment of the interleukin 7 gene (il7) to porcine chromosome 4q11→q13 by fish and by analysis of radiation hybrid panels, Cytogenetic and Genome Research, vol. 93, no. 1-2, p. 65-72, 2001. https://doi.org/10.1159/000056951
[3] E. Gerace, P. Pasquali, B. Oesch, M. Falduto, F. Mandanici, M. Fiasconaroet al., Stimulation of bovine whole-blood samples cultured in media supplemented with recombinant interleukin-7 (il-7) and il-12 extends the life span of the gamma interferon assay to detect mycobacterium bovis-infected cattle, Journal of Clinical Microbiology, vol. 54, no. 9, p. 2315-2320, 2016. https://doi.org/10.1128/jcm.00629-16
[4] E. Coppin, B. Sundarasetty, S. Rahmig, J. Blume, N. Verheyden, F. Bahlmannet al., Enhanced differentiation of functional human t cells in nsgw41 mice with tissue-specific expression of human interleukin-7, Leukemia, vol. 35, no. 12, p. 3561-3567, 2021. https://doi.org/10.1038/s41375-021-01259-5
[5] A. Lent, W. Dontje, M. Nagasawa, R. Siamari, A. Bakker, S. Pouwet al., Il-7 enhances thymic human t cell development in “human immune system” rag2−/−il-2rγc−/− mice without affecting peripheral t cell homeostasis, The Journal of Immunology, vol. 183, no. 12, p. 7645-7655, 2009. https://doi.org/10.4049/jimmunol.0902019
[6] R. Kondrack, J. Harbertson, J. Tan, M. McBreen, C. Surh, & L. Bradley, Interleukin 7 regulates the survival and generation of memory cd4 cells, The Journal of Experimental Medicine, vol. 198, no. 12, p. 1797-1806, 2003. https://doi.org/10.1084/jem.20030735
[7] H. Jeong, S. Lee, H. Seo, & B. Kim, Recombinantmycobacterium smegmatisdelivering a fusion protein of human macrophage migration inhibitory factor (mif) and il-7 exerts an anticancer effect by inducing an immune response against mif in a tumor-bearing mouse model, Journal for Immunotherapy of Cancer, vol. 9, no. 8, p. e003180, 2021. https://doi.org/10.1136/jitc-2021-003180
[8] A. Parmigiani, M. Pallin, H. Schmidtmayerova, M. Lichtenheld, & S. Pahwa, Interleukin-21 and cellular activation concurrently induce potent cytotoxic function and promote antiviral activity in human cd8 t cells, Human Immunology, vol. 72, no. 2, p. 115-123, 2011. https://doi.org/10.1016/j.humimm.2010.10.015
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