Based on our advanced Cytokine Production Measurement Service and powerful technology, Creative Biolabs provides a complete range portfolio of assays for Interleukin (IL) production services to meet our customers' different purposes. Our highly skilled scientists are ready to get started on your projects.

Introduction of Interleukins

Interleukins (ILs), one kind of the most widely studied cytokines, are expressed and secreted by leukocytes and many other body cells. They serve as key signaling molecules (as shown in Fig.1) for the communication between innate and adaptive immune cells as well as non-immune cells and tissues. In consideration of their critical role in cancer development, and progression, the properties of interleukins are urgently needed to be exploited to promote the effectiveness of Engineered Cancer Cells in self-destruction immunotherapy.

Fig.1 Interleukins in carcinogenesis. (Briukhovetska; et al., 2021)

• Functions of ILs

ILs consist of a large group of more than 50 secreted proteins and elicit complex interactions between cells and tissues by binding to high-affinity receptors in cell surfaces. They promote the development and differentiation of T and B lymphocytes, and hematopoietic cells. The primary function of ILs is to modulate proliferation, activation, differentiation, cell maturation, migration, and adhesion during inflammatory and immune responses. Due to their anti-tumor effects, ILs, such as IL-2, could act as potential therapeutic agents applicable to cancer immunotherapy.

However, accumulated knowledge of cancer immune evasion mechanisms has revealed a notion that some ILs, such as IL-4, IL-9, IL-10, and IL-18, may help nurture an environment favoring cancer growth, metastasis, invasion, and immune escape. As Fig.2 shows, the IL-12 family interleukins maintain a balance between activating immune responses and promoting tumor growth in tumorigenesis. These ILs with regulatory immune functions are produced by either host immune cells or tumor cells themselves.

Fig.2 The role of IL-12 family cytokines in tumorigenesis. (Mirlekar & Pylayeva-Gupta, 2021)

IL Production Measurement Services at Creative Biolabs

The production of ILs could be measured by methods including immunohistochemistry (IHC), western blot (WB), real-time quantitative PCR (QPCR), and the methods flow cytometry (FC), enzyme-linked immunoabsorbent assay (ELISA) and immunofluorescence (IF) as we described in the Interferon (IFN) Production Measurement.

• IL Production Measurement by IHC

IHC is a visualized immunochemical technique that permits the detection and localization of Interleukins in the normal control tissue and benign or malignant cancer tissues. For the immunohistochemical staining, paraffin sections will be prepared and cut with a rotary microtome, and then de-waxed through gradients ethanol into water. After antigen retrieval, sections will be washed and blocked to eliminate the nonspecific binding. With the antibody incubated overnight, the sections will be washed and then incubated with a secondary antibody before counterstaining with hematoxylin for imaging. The lab of Creative Biolabs owns all the equipment needed for the whole process of immunohistochemical staining. Our experienced experts will do IHC for IL production measurement you needed.

• IL Production Measurement by WB

WB could be used for the quantification of intracellular localized ILs. In this technique, an equal amount of protein mixtures will be separated by gel electrophoresis based on molecular weight. The gels will be then transferred to a PVDF membrane by equipment Trans-Blot to produce a band for each protein. After being blocked, the membrane will then be incubated with labeled antibodies specific to the protein of interest. The membrane results will be visualized with the chemiluminescence imaging system after incubation of the ECL mix in the dark room. The lab of Creative Biolabs is equipped with electrophoresis apparatus and automated imaging systems for WB. Our experienced experts will do ELISA for the ILs production measurement you needed.

• IL Production Measurement by QPCR

QPCR assay for the analysis of human ILs' gene expression in infected or stimulated cells has been developed for immunological investigations. QPCR could help us understand the origin and functions of IL-producing and assess the immune system and identify ideal disease biomarkers. As for the QPCR procedure, samples will be collected to extract RNA for the following reverse transcription. QPCR will run with specific primers we designed for interested genes related to the ILs. The lab of Creative Biolabs possesses real-time qPCR equipment to achieve the optimization and validation of the experiment parameters to have the qPCR results. QPCR is a routine project for our experienced experts, and they will do this measurement of IL production with veritable results.

With the in-depth study of interleukins, Creative Biolabs offers advanced custom analytic service through the combined use of our software and multiplex analyzers, including FC, ELISA, IF, IHC, WB, and QPCR assay. If you are interested in this, please feel free to contact us directly.

References

1. Briukhovetska, D.; et al. Interleukins in cancer: from biology to therapy. Nat Rev Cancer. 2021, 21(8): 481-499.
2. Mirlekar, B.; Pylayeva-Gupta, Y. IL-12 family cytokines in cancer and immunotherapy. Cancers. 2021, 13: 167.

For Research Use Only | Not For Clinical Use