ELISA applied in research

Cytokines are a group of signaling proteins critically involved in various physiological processes such as immune regulation, cell differentiation, cell proliferation and chemotaxis. They are produced by a variety of different cell types in many vertebrate species and are active at very low concentrations mostly in the picogram to femtogram range. Cytokines are usually produced transiently and locally, acting in a paracrine or autocrine manner and interact with high affinity cell surface receptors specific for each cytokine or cytokine group. Chemokines are a family of small cytokines (with four conserved cysteine residues), which can mediate chemoattraction between cells. Some chemokines are homeostatic in nature and are constitutively produced and secreted, while others are considered inflammatory and are only produced by cells during infection or a pro-inflammatory stimulus.

Currently, it is well known that the type of an antigen-specific immune response largely depends on the selection or preferential activation of defined CD4+ T cell subsets (i.e. Th1, Th2 and Th17). Activation of these subsets is characterized by the secretion of distinct patterns of cytokines. Th1 cells primarily secrete IFN-γ and IL-2, while Th2 cells mainly produce IL-4, IL-5, IL-9 and IL-13. Other cytokines, such as TNF-α, are produced by both Th subsets. More recently, two novel subsets were found to differentiate from naïve CD4+ T cells: T regulatory cells (Treg), Th17  and T follicular T helper cells (Tfh). Treg cells are characterized by the production of TGF-β, IL-10 and IL-35, while Th17 cells can produce IL-17A, IL-17F, IL-17A/IL-17F, IL-21, IL-22, IL-26, GM-CSF, TNF-α and CCL20 (MIP-3α) and Tfh cells mainly produce IL-21.

T helper cell lineages

Figure: The different T helper cell lineages, their master regulators and characteristic cytokines. Figure source:  O'Shea JJ and Paul WE (2010). Mechanisms Underlying Lineage Commitment and Plasticity of Helper CD4+ T Cells. Science 327:1098-102.

Additionally, different critical cytokines are required for the differentiation of various cell types. The production of IL-12 and IL-10, produced by antigen presenting cells (APC) such as macrophages and dendritic cells, critically contributes to the preferential expansion of Th1- or Th2-type of cells. For instance, early production of IL-12 is considered essential for the development of Th1 cells. On the other hand, the absence or low concentrations of IL-12 and IFN-γ in the early phase of an immune response and concomitant production of IL-4 by cells of the mast cell/basophil lineage or T cells themselves is known to favor the development of Th2 cells. The differentiation of Treg and Th17 cells is interlinked through TGF-β, indicating potential plasticity between these two types of cells. The ultimate T cell subset is determined by the concentration of TGF-β along with the concentrations of IL-6 and IL-23.

Another important group of effector proteins involved in the immune system are perforin and granzymes (serine proteases). Granzymes and perforin are released by cytotoxic CD8+ T cells and natural killer cells to induce apoptosis in their target cells. For example, granzyme B and perforin seem to enter target cells in a multimeric complex enclosed in vesicles. Perforin allows granzyme B to pass the vesicle membrane into the cell to cause apoptosis by various pathways.  

Signaling proteins can best be detected and measured using ELISA techniques. U-CyTech has developed multiple high-quality ELISA kits for the detection of cytokines and granzymes for human, monkey (including macaques and marmoset), mouse and rat models. These assays are widely applied in different research fields, including cancer research1,2, vaccine development3,4, infectious diseases5,6,7 and autoimmune diseases8. Hundreds of peer-reviewed publications describe the successful use of U-CyTech’s ELISA systems and/or apply our high affinity antibodies in other immunoassays. Please find these references in our Reference database.

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Examples of studies in different research areas using one or more of our ELISA products:

Click on the authors for the abstract of the below mentioned acticles.

  1. Dong L et al. (2013). Human prostate stem cell antigen and HSP70 fusion protein vaccine inhibits prostate stem cell antigen-expressing tumor growth in mice. Cancer Biother. Radiopharm. 28: 391-7.
  2. Mirandola L et al. (2015). Novel antigens in non-small cell lung cancer: SP17, AKAP4, and PTTG1 are potential immunotherapeutic targets. Oncotarget. 6: 2812-26.
  3. Desallais L et al. (2016). Immunization against an IL-6 peptide induces anti-IL-6 antibodies and modulates the Delayed-Type Hypersensitivity reaction in cynomolgus monkeys. Sci. Rep. 6: 19549.
  4. Hussein J et al. (2018). A phase I, open-label trial on the safety and immunogenicity of the adjuvanted tuberculosis subunit vaccine H1/IC31® in people living in a TB-endemic area. Trails. 19: 24.
  5. Nelson M and Loveday M (2015). Exploring the innate immunological response of an alternative nonhuman primate model of infectious disease; the common marmoset. J. Immunol. Res. 2014: 913632.
  6. Mouser EE et al. (2016). Brugia malayi Antigen (BmA) Inhibits HIV-1 Trans-Infection but Neither BmA nor ES-62 Alter HIV-1 Infectivity of DC Induced CD4+ Th-Cells. PloS One. 11: e0146527
  7. Hoepel W et al. (2021). High titers and low fucosylation of early human anti-SARS-CoV-2 IgG promote inflammation by alveolar macrophages. Sci. Transl. Med. Online ahead of print.
  8. Singh S et al. (2015). Anti-inflammatory and antiarthritic activity of UNIM-301 (a polyherbal unani formulation) in Wistar rats. Pharmacognosy Res. 7: 188-92