Recombinant human IL-3 protein

Summary

• Highly pure human protein (UniProt number: P08700)
• >98%, by SDS-PAGE quantitative densitometry
• Source: Expressed in E. coli
• 15.2 kDa monomer
• Animal origin-free (AOF) and carrier protein-free
• Manufactured in Cambridge, UK
• Lyophilized from acetonitrile/TFA
• Resuspend in 10 mM HCl, prepare single-use aliquots, add carrier protein if desired and store frozen at -20°C or -80°C

Featured applications

• Generation of iPSC-derived myeloid progenitors
• Maintenance of peripheral blood cells
• Proliferation of hematopoietic stem cells
• Differentiation of peripheral blood cells to myeloid lineages
• Differentiation of myeloid cells into antigen-presenting cells/monocytes
• Differentiation of myeloid progenitors into erythrocytes and megakaryocytes

Bioactivity

Purity

Qkine IL-3 is as biologically active as the comparable alternative supplier protein

Stimulation of proliferation of TF-1 cells with Qkine IL-3 (Qk090, green) and alternative supplier IL-3 (Supplier B, black). Cells were treated in triplicate with a serial dilution of IL-3 for 72 hours and proliferation measured using the CellTiter-Glo (Promega) luminescence assay.

Protein Background

Interleukin 3 (IL-3) also known as B-cell stimulatory factor 2/interferon beta 2, is a pleiotropic cytokine with a crucial role in hematopoiesis and multi-lineage growth factor [1–4]. It promotes the proliferation, differentiation, and survival of hematopoietic stem cells and myeloid progenitors [3–6]. IL-3 induces the development of mast cells, basophils, monocytes, erythrocytes, and megakaryocytes and mediates the release of mediators from these cells. It is also involved in the recruitment of myeloid cells to inflammation sites and regulating T-cell function [2].

IL-3 is a monomer glycoprotein composed of 140 amino acids with a molecular weight of approximately 15.7 kDa [7]. The primary structure of IL-3 includes four alpha helices and a bundle of beta sheets stabilized by disulfide bridges [8]. It is produced by various cells including activated T cells, B cells, macrophages, monocytes, mast cells, vascular endothelial cells, and fibroblasts [5]. IL-3 binds with high affinity to the IL-3 receptor (IL-3R) of the hematopoietic receptor superfamily expressed on hemopoietic progenitor cells, monocytes, and B lymphocytes. Ligand stimulation of IL-3R induces tyrosine phosphorylation of effector proteins which subsequently trigger a cascade of intracellular responses such as the PI3k/Akt, Ras/ERK, and MAPK pathways [9,10]. IL-3R is heterodimeric and is composed of two subunits: an alpha subunit (IL-3Rα) and a beta subunit (βc) [11]. IL-3Rα is specific to IL-3, while βc is shared with other cytokine receptors, including those for IL-5 and granulocyte-macrophage colony-stimulating factor (GM-CSF) [5,10,12].

IL-3 is used in cell culture for the maintenance of peripheral blood cells13 and to induce the expansion and differentiation of hematopoietic stem cells [14]. IL-3 acts synergistically with IL-6 to enhance the proliferation of hemopoietic progenitors1. IL-3 is also used as an early regulator for the differentiation of induced pluripotent stem cells into myeloid and more specific lineages using other lineage instructive cytokines [14,15]. For example, antigen-presenting cells can be derived with the addition of GM-CSF and IL-4 for dendritic cells or M-CSF for macrophages [15]. Finally, IL-3 is also used as a growth factor for the differentiation of erythrocytes with IL-12 and megakaryocytes with IL-6 [16,17].

IL-3 has been studied for conditions which could benefit from the production of early myeloid progenitors and hematopoietic recovery [2]. These include bone marrow reconstitutions, aplastic anemias, and bone marrow disorders such as acute myeloid leukemia and myelodysplastic syndromes [2,18]. Research into the use of IL-3 in immunotherapy and cancer treatment is ongoing, as it may enhance the anti-tumor immune response. For example, the IL-3 and IL-3R axes could be potential therapeutic targets to treat acute myeloid leukemia [10,18,19]. Finally, the potency of IL-3 in amplifying acute inflammation makes it a potential therapeutic target in sepsis [20].

Background references

1. Ikebuchi, K. et al. Interleukin 6 enhancement of interleukin 3-dependent proliferation of multipotential hemopoietic progenitors. Proc. Natl. Acad. Sci. 84, 9035–9039 (1987).
2. Ihle, J. N. Interleukin-3 and hematopoiesis. Chem. Immunol. 51, 65–106 (1992).
3. Schrader, J. W. Interleukin-3. in Growth Factors and Cytokines in Health and Disease (eds. Leroith, D. & Bondy, C.) vol. 2 49–84 (JAI, 1997).
4. Pixley, F. J. & Stanley, E. R. Chapter 319 – Cytokines and Cytokine Receptors Regulating Cell Survival, Proliferation, and Differentiation in Hematopoiesis. in Handbook of Cell Signaling (Second Edition) (eds. Bradshaw, R. A. & Dennis, E. A.) 2733–2742 (Academic Press, 2010). doi:10.1016/B978-0-12-374145-5.00319-3.
5. Broughton, S. E. et al. The GM–CSF/IL-3/IL-5 cytokine receptor family: from ligand recognition to initiation of signaling. Immunol. Rev. 250, 277–302 (2012).
6. Kölle, J. et al. Targeted deletion of Interleukin-3 results in asthma exacerbations. iScience 25, 104440 (2022).
7. Clark-Lewis, I. et al. Automated Chemical Synthesis of a Protein Growth Factor for Hemopoietic Cells, Interleukin-3. Science 231, 134–139 (1986).
8. Lindemann, A. & Mertelsmann, R. Interleukin-3: Structure and Function. Cancer Invest. 11, 609–623 (1993).
9. Peso, L. del, González-Garcı́a, M., Page, C., Herrera, R. & Nuñez, G. Interleukin-3-Induced Phosphorylation of BAD Through the Protein Kinase Akt. Science 278, 687–689 (1997).
10. Testa, U. et al. Interleukin-3 receptor in acute leukemia. Leukemia 18, 219–226 (2004).
11. Miyajima, A., Mui, A., Ogorochi, T. & Sakamaki, K. Receptors for granulocyte-macrophage colony-stimulating factor, interleukin-3, and interleukin-5. Blood 82, 1960–1974 (1993).
12. Hodi, F. S. & Soiffer, R. J. Interleukins. in Encyclopedia of Cancer (Second Edition) (ed. Bertino, J. R.) 523–535 (Academic Press, 2002). doi:10.1016/B0-12-227555-1/00110-6.
13. Staerk, J. et al. Reprogramming of peripheral blood cells to induced pluripotent stem cells. Cell Stem Cell 7, 20–24 (2010).
14. Ackermann, M. et al. A 3D iPSC-differentiation model identifies interleukin-3 as a regulator of early human hematopoietic specification. Haematologica 106, 1354–1367 (2020).
15. Ackermann, M., Dragon, A. C. & Lachmann, N. The Immune-Modulatory Properties of iPSC-Derived Antigen-Presenting Cells. Transfus. Med. Hemotherapy 47, 444–453 (2020).
16. Ploemacher, R. E., van, S. P., Boudewijn, A. & Neben, S. Interleukin-12 enhances interleukin-3 dependent multilineage hematopoietic colony formation stimulated by interleukin-11 or steel factor. Leukemia 7, 1374–1380 (1993).
17. Majka, M. B.-K., Jacek Kijowski, Ryan Reca, Janina Ratajczak, Mariusz Z. Ratajczak, Marcin. In vitro expansion of human megakaryocytes as a tool for studying megakaryocytic development and function. Platelets 12, 325–332 (2001).
18. Munoz, L. et al. Interleukin-3 receptor alpha chain (CD123) is widely expressed in hematologic malignancies. Haematologica 86, 1261–1269 (2001).
19. Jordan, C. T. et al. The interleukin-3 receptor alpha chain is a unique marker for human acute myelogenous leukemia stem cells. Leukemia 14, 1777–1784 (2000).
20. Weber, G. F. et al. Interleukin-3 amplifies acute inflammation and is a potential therapeutic target in sepsis. Science 347, 1260–1265 (2015).