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The innate immune system, also known as the non-specific immune system, is an important subsystem of the overall immune system that comprises the cells and mechanisms that defend the host from infection by other organisms. The cells of the innate system recognize and respond to pathogens in a generic way, differing from the adaptive immune system which provides a pathogen- specific response. In vertebrates, the innate immune system acts by several different mechanisms. It recruits immune cells to sites of infection, through the production of cytokines. It activates the complement cascade to target and kill bacterial pathogens. It uses specialised white blood cells to identify and remove pathogens. It also activates the adaptive immune system through antigen presentation. The physical barrier of the epithelium also forms part of the innate immune system.

Tissue resident immune cells identify pathogens and in response secrete cytokines that cause leukocytes to migrate from the circulation into inflamed tissue. There, they phagocytose the pathogen, attack the pathogen by releasing enzymes, and secrete other cytokines that maintain the inflammatory response.

“Cytokine” is a broad catch-all term used by researchers to describe a broad family of structurally diverse signalling molecules of varying functions that are best known as immune-modulators.

Cytokine functions are mediated by binding specific receptors and their activities include regulating cell activation, haematopoiesis, apoptosis, cell migration, and cell proliferation. In this capacity, they are involved in virtually all aspects of both innate and adaptive immune responses. Cytokines can be further divided into families, often based on structure or their primary function. 

Cytokine Detection

Secreted cytokines in the circulation are detected using classical immunoassays such as the enzyme-linked immunosorbent assay (ELISA). Through our partnership with Bio-techne, In Vitro offers the Quantikine ELISA, the recognised gold standard in immunoassay.

Quantikine ELISAs are exceptionally sensitive and accurate, allowing detection of picogram amounts of cytokines in serum, plasma, cell culture supernatants and other sample types from human, mouse and other sample types. Multiple simultaneous detection of cytokines is possible by using a Luminex assay.

This immuno-assay performs multiple ELISAs simultaneously, allowing researchers to profile more than one cytokine at a time. Luminex assays from R&D Systems save on valuable time and sample and are becoming more widespread The use of a Luminex assays allows researchers to study the context in which a particular cytokine is expressed as other related, dependant or inhibitory cytokines can be profiled at the same time. R&D Systems also offer the Proteome Profiler, a membrane based dot blot assay that allows for quick and easy screening of cytokine expression in a wide range of sample types.

luminex assays1

quantikinekits

proteomeprofiler

Blocking Cytokines

Once expressed by immune cells at the site of infection, cytokines play diverse roles in activating and repressing different functional outcomes. For example, interleukin 18 (IL-18) is an immuno-modulatory cytokine that is produced by macrophages and stimulates other cells of the immune system, natural killer (NK) cells and some T-cells to produce Interferon gamma, itself an important activator of macrophages. However, the secretion of Interferon gamma can be neutralised by using an IL-18 blocking antibody to deplete the available IL-18. R&D Systems offers over 860 validated antibodies for blocking and neutralising different cytokines. All R&D Systems antibodies come with an application guarantee, offering risk-free experiments.

R&D Systems offers over 860 validated antibodies for blocking and neutralising different cytokines. All R&D Systems antibodies come with an application guarantee, offering risk-free experiments. 

anticytokine antibodies

Functional Outcomes of cytokine secretion

Classically, cell invasion, such as neutrophil extravasation, has been studied by the use of Boyden chambers and transwell membranes, however these end-point assays are laborious, time-consuming and provide data for only a single time point. In contrast, ACEA’s cell invasion and migration plate (CIM-Plate®; used with the xCELLigence® RTCA DP system) contains electronically integrated Boyden chambers that provide, in real-time and without the use of labels, quantitative kinetic data for migration/invasion with minimal hands-on time by the researcher. As cells move from the upper chamber towards chemoattractant in the lower chamber they pass through a membrane containing pores and then adhere to gold impedance microelectrodes. The resultant change in impedance signal perfectly correlates with the number of cells attached to these electrodes, enabling facile collection of highly reproducible data over time ranges spanning from minutes to days.

Register for an xCelligence DP

The xCELLigence® RTCA DP instrument uses noninvasive electrical impedance monitoring to quantify cell proliferation, morphology change, and attachment quality in a label-free, real-time manner.

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17.01.2017