ELISA applied in research

Cytokines and chemokines 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, but not Th2 cells, primarily secrete IFN-γ; while Th2, but not Th1 cells, 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, namely T regulatory cells (Treg) and Th17 cells. 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-17AF, IL-21, IL-22, IL-26, GM-CSF, TNF-α and CCL20 (MIP-3α).

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 within virus-infected 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 where it causes apoptosis by various pathways.  

Signaling proteins can best be detected and measured using ELISA techniques. U-CyTech has developed various high-quality ELISA kits for the detection of cytokines, chemokines and granzymes for human, monkey (including macaques and marmoset), mouse and rat models. These assays are widely applied in different research fields, including cancer research (1, 2), vaccine development (3, 4), infectious diseases (5, 6, 7), autoimmune diseases (8, 9), organ transplantation (10, 11) and parasitology (12, 13). 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. 

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

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

1. Pouw N, Treffers-Westerlaken E, Kraan J, Wittink F, Ten Hagen T, Verweij J and Debets R
    Combination of IL-21 and IL-15 enhances tumour-specific cytotoxicity and cytokine production of TCR-transduced
    primary T cells.
    Cancer Immunol Immunother 59:921-31 (2010).
    U-CyTech products used in this study:
    Mouse IFN-γ ELISA kit
    Mouse IL-10 ELISA kit

2. Skov L, Allen MH, Bang B, Francis D, Barker JN and Baadsgaard O
    Basal cell carcinoma is associated with high TNF-alpha release but nor with TNF-alpha polymorphism at position--308.
    Exp Dermatol 12:772-6 (2003).
    U-CyTech products used in this study:
    Human TNF-α ELISA kit

3. Bai B, Hu Q, Hu H, Zhou P, Shi Z, Meng J, Lu B, Huang Y, Mao P and Wang H
    Virus-like particles of SARS-like coronavirus formed by membrane proteins from different origins demonstrate
    stimulating activity in human dendritic cells.
    PLoS One 3:e2685 (2008).
    U-CyTech products used in this study:
    Human IL-6 ELISA kit
    Human IL-10 ELISA kit
    Human TNF-α ELISA kit
4. Zhang W, Ahmad G, Torben W, Noor Z, Le L, Damian RT, Wolf RF, White GL, Chavez-Suarez M, Podesta
     RB, Kennedy RC, and Siddiqui

     Sm-p80-based DNA vaccine provides baboons with levels of protection against Schistosoma mansoni infection
     comparable to those achieved by the irradiated cercarial vaccine.
     J Infect Dis 201:1105-12 (2010).
     U-CyTech products used in this study:
     Monkey IFN-γ ELISA kit
     Monkey IL–2 ELISA kit
     Monkey IL-4 ELISA kit
     Monkey IL-10 ELISA kit
     Monkey species: Papio anubis

5. Chang WL, Baumgarth N, Yu D and Barry PA
     Human cytomegalovirus-encoded interleukin-10 homolog inhibits maturation of dendritic cells and alters their
     functionality. J Virol 78:8720-31 (2004).
     U-CyTech products used in this study:
     Human IL-6 ELISA kit
     Human IL-10 ELISA kit
     Human IL-12p70 ELISA kit
     Human TNF-α ELISA kit

6. Duan R, Remeijer L, van Dun JM, Osterhaus AD, and Verjans GM
     Granulocyte macrophage colony-stimulating factor expression in human herpetic stromal keratitis: implications for the
     role of neutrophils in HSK.
     Invest Ophthalmol Vis Sci 48:277-84 (2007).
     U-CyTech products used in this study:
     Human GM-CSF ELISA

7. De Swart RL, Kuiken T, Timmerman HH, van Amerongen G, Van Den Hoogen BG, Vos HW, Neijens HJ,       
     Andeweg AC and
Osterhaus AD

     Immunization of macaques with formalin-inactivated respiratory syncytial virus (RSV) induces interleukin-13-associated
     hypersensitivity to subsequent RSV infection.
     J Virol 76:11561-9 (2002).
     U-CyTech products used in this study:
     Monkey IFN-γ ELISA kit
     Monkey IL-2 ELISA kit
     Monkey IL-4 ELISA kit
     Monkey IL-5 ELISA kit
     Monkey IL-10 ELISA kit
     Monkey IL-13 ELISA kit
     Monkey species: Macaca fascicularis

8. Bakker JM, Kavelaars A, Kamphuis PJ, Cobelens PM, van Vugt HH, van Bel F and Heijnen CJ
    Neonatal dexamethasone treatment increases susceptibility to experimental autoimmune disease in adult rats.
    J Immunol 165:5932-7 (2000).
    U-CyTech products used in this study:
    Rat TNF-α ELISA kit

9. Jagessar SA, Kap YS, Heijmans N, van Driel N, van Straalen L, Bajramovic JJ, Brok HP, Blezer EL, Bauer 
    J, Laman JD and t Hart BA

    Induction of progressive demyelinating autoimmune encephalomyelitis in common marmoset monkeys using   
    MOG34-56 peptide in incomplete freund adjuvant.
    J Neuropathol Exp Neurol 69:372-85 (2010).
    U-CyTech products used in this study:
    Human IL-1β ELISA kit
    Human IL-6 ELISA kit
    Human TNF-α ELISA kit
    Monkey IFN-γ ELISA kit
    Monkey TNF-α ELISA kit
    Monkey species: Callithrix jacchus

10. Demirkiran A, Bosma BM, Kok A, Baan CC, Metselaar HJ, Ijzermans JN, Tilanus HW, Kwekkeboom 
     J, and van der Laan LJ

     Allosuppressive donor CD4+CD25+ regulatory T cells detach from the graft and circulate in
     recipients after liver transplantation. J Immunol 178:6066-72 (2007).
     U-CyTech products used in this study:
     Human IFN-γ ELISA kit

11. Tha-In T, Metselaar HJ, Tilanus HW, Boor PP, Mancham S, Kuipers EJ, de Man RA and
     Kwekkeboom J

     Superior immunomodulatory effects of intravenous immunoglobulins on human T-cells and dendritic cells: comparison 
     to calcineurin inhibitors.
     Transplantation 81:1725-34 (2006).
     U-CyTech products used in this study:
     Human IFN-γ ELISA kit
     Human IL-4 ELISA kit

12. Janssen R, Van Wengen A, Verhard E, De Boer T, Zomerdijk T, Ottenhoff TH and Van Dissel JT
     Divergent role for TNF-alpha in IFN-gamma-induced killing of Toxoplasma gondii and Salmonella typhimurium
     contributes to selective susceptibility of patients with partial IFN-gamma receptor1 deficiency.
     J Immunol 169:3900-7 (2002).
     U-CyTech products used in this study:
     Human IFN-γ ELISA kit

13. Ozwara H, Langermans JA, Kocken CH, van der Wel A, van der Meide PH, Vervenne RA, Mwenda JM, 
      and Thomas AW

      Transfected Plasmodium knowlesi produces bioactive host gamma interferon: a new perspective for modulating
      immune responses to malaria parasites.
      Infect Immun 71:4375-81 (2003).
      U-CyTech products used in this study:
      Monkey IFN-γ ELISA kit
      Monkey species: Macaca mulatta