Materials Provided

• Pre-coated microtiter plate(s)
• Plate Sealers
• Wash Solution Concentrate
• Human IFN-Alpha Standard, 10,000 pg/ml
• Dilution Buffer
• Antibody Concentrate
• HRP Conjugate Concentrate
• Concentrate Diluent
• TMB Substrate
• Stop Solution

Additional Materials Required (Not Provided) 

• Microplate reader capable of reading an OD at a wavelength of 450 nm
• Variable volume microtiter pipettes
• Adjustable multichannel pipette (50-300 μl)
• Reagent reservoirs
• Wash bottle or plate washing system
• Distilled or deionized water
• Serological pipettes (1, 5, 10 or 25 ml)
• Disposable pipette tips (polypropylene)
• Plate shaker

Tips, Tools and Troubleshooting:

• Human IFN-α ELISA Subtype Specificities Comparison Chart

Background

Interferons (IFNs) are a family of mammalian cytokines initially characterized by their ability to inhibit viral infection. They are synthesized and secreted by most cell types in response to pathogens. In addition to their antiviral properties, IFNs have also been shown to exhibit anti-proliferative, immunomodulatory, and many other activities.

In humans, IFN-α consists of a group of proteins that are greater than 85% homologous by amino acid sequence. Numerous individual human IFN-α subtypes have been identified; many display different properties. It remains unclear why there are multiple IFN-α subtypes. A variety of studies suggested they possess overlapping but also unique sets of biological activities.

34 Citations

1. Hansen, A. M. et al.,  (2022), "H3K9 dimethylation safeguards cancer cells against activation of the interferon pathway", Sci. Adv. 8:eabf8267, PMID: 3530284, (link)


2. Leylek, Rebecca, et al. (2019). Integrated Cross-Species Analysis Identifies a Conserved Transitional Dendritic Cell Population. Cell Reports, 39 pgs. PMID: 31825848. (link)


3. Osterlund, Pamela, et al. (2019). Asian and African lineage Zika viruses show differential replication and innate immune responses in human dendritic cells and macrophages. Scientific Reports, 15 pgs. PMID: 31673117. (link)


4. Kontny, et al. (2018). Subgroups of Sjögren's syndrome patients categorised by serological profiles: clinical and immunological characteristics. Reumatologia, 8 pgs. PMID: 30647480. (link)


5. Speer, Esther, et al. (2018). Pentoxifylline, Dexamethasone, and Azithromycin Demonstrate Distinct Age-Dependent and Synergistic Inhibition of TLR- and Inflammasome-Mediated Cytokine Production in Human Newborn and Adult Blood In Vitro. PLOS One, 29 pgs. PMID: 29715306. (link)


6. Riggs, Jeffrey, et al. (2018). Characterisation of Anifrolumab, a Fully Human Anti-Interferon Receptor Antagonist Antibody for the Treatment of Systemic Lupus Erythematosus. Lupus Science & Medicine, 12 pgs. PMID: 29644082. (link)


7. Bressy, Christian, et al. (2018). Cell Cycle Arrest in G2/M Phase Enhances Replication of Interferon-Sensitive Cytoplasmic RNA Viruses via Inhibition of Antiviral Gene Expression. JVI, 55 pgs. PMID: no PMID. (link)


8. Zecevic, Lamija, et al. (2017). Potential Immune Biomarkers in Diagnosis and Clinical Management for Systemic Lupus Erythematosus. Journal of Medical Biochemistry, 9 pgs. PMID: 30581353. (link)


9. Smith, Nikaia, et al. (2017). Natural Amines Inhibit Activation of Human Plasmacytoid Dendritic Cells Through CXCR4 Engagement. Nature Communications, 13 pgs. PMID: 28181493. (link)


10. Oon, Shereen, et al. (2016). A Cytotoxic Anti-IL-3R-alpha Antibody Targets Key Cells and Cytokines Implicated in Systemic Lupus Erythematosus. JCI Insight, 15 pgs. PMID: 27699260. (link)


11. Dowell, Nicholas, et al. (2016). Acute Changes in Striatal Microstructure Predict the Development of Interferon-Alpha Induced Fatigue. Biological Psychiatry, 9 pgs. PMID: 26169252. (link)


12. Huang, Cheng, et al. (2015). Highly Pathogenic New World and Old World Human Arenaviruses Induce Distinct Interferon Responses in Human Cells. JVI, 10 pgs. PMID: 25926656. (link)


13. Bonaparte, Sarah, et al. (2015). HPV-Positive Cervical Cancer Cells Show Generally Reduced Levels of IFN-Alpha. Belmont University, 11 pgs. PMID: no PMID. (link)


14. Huang, Yanyan, et al. (2014). Hcmv-miR-UL112 Attenuates NK Cell Activity by Inhibition Type I Interferon Secretion. Immunology Letters, 30 pgs. PMID: 25530545. (link)


15. Lederle, Alexandre, et al. (2014). Neutralizing Antibodies Inhibit HIV-1 Infection of Plasmacytoid Dendritic Cells by an Fc-gamma-Rlla Independent Mechanism and Do Not Diminish Cytokines Production. Scientific Reports, 10 pgs. PMID: 25132382. (link)


16. Vogelsang, Petra, et al. (2014). Altered Phenotype and Stat1 Expression in Toll-Like Receptor 7/8 Stimulated Monocyte-Derived Dendritic Cells from Patients with Primary Sjoergen's Syndrome. Arthritis Research & Therapy, 22 pgs. PMID: 25113744. (link)


17. Maeyama, Jun-ichi, et al. (2014). A Palindromic CpG-Containing Phosphodiester Oligodeoxynucleotide as a Mucosal Adjuvant Stimulates Plasmacytoid Dendritic Cell-Mediated T_H1 Immunity. PLOS One, 10 pgs. PMID: 24586411. (link)


18. Stone, Amy, et al. (2013). Hepatitis C Virus Pathogen Associated with Molecular Pattern (PAMP) Triggers Production of Lambda-Interferons by Human Plasmacytoid Dendritic Cells. PLOS One, 13 pgs. PMID: 23637605. (link)


19. Chen, Qian, et al. (2013). Interleukin-27 is a potent inhibitor of cis HIV-1 replication in monocyte-derived dendritic cells via a type I interferon-independent pathway. PLOS One, 10 pgs. PMID: 23527130. (link)


20. Tian, Ren-Rong, et al. (2012). IFN-lambda Inhibits HIV-1 Integration and Post-Transcriptional Events In Vitro, but there is Only Limited In Vivo Repression of Viral Production. Antiviral Research. PMID: 22584351. (link)


21. Drannik, Anna, et al. (2012). Trappin-2/Elafin Modulate Innate Immune Responses of Human Endometrial Epithelial Cells to Poly I:C. PLOS One, 16 pgs. PMID: 22545145. (link)


22. Zepp, Jarod A, et al. (2011). Protection from RNA and DNA viruses by IL-32. Journal of Immunology, 10 pgs. PMID: 21346229. (link)


23. Tomescu, et al. (2010). Increased Plasmacytoid Dendritic Cell Maturation and Natural Killer Cell Activation in HIV-1 Exposed, Uninfected Intravenous Drug Users. AIDS, 9 pgs. PMID: 20647906. (link)


24. Xu, Donghseng, et al. (2010). Viral transformation for production of personalized type I interferons. Biotechnology Journal. PMID: 20518060. (link)


25. Xu, Yongfen, et al. (2009). HBsAg inhibits TLR9-mediated activation and IFN-alpha production in plasmacytoid dendritic cells. Molecular Immunology, 7 pgs. PMID: (link)


26. Scagnolari, Carolina, et al. (2009). Gene Expression of Nucleic Acid-Sensing Pattern Recognition Receptors in Children Hospitalized for Respiratory Syncytial Virus-Associated Acute Bronchiolitis. Clinical and Vaccine Immunology, 8 pgs. PMID: 19386802. (link)


27. Mansson, Anne, et al. (2009). Role of atopic status in Toll-like receptor (TLR)7- and TLR9-mediated activation of human eosinophils. JLB, 9 pgs. PMID: 19129482. (link)


28. Nold, Marcel F, et al (2008). Endogenous IL-32 controls cytokine and HIV-1 production. Journal of Immunology, 10 pgs. PMID: 18566422. (link)


29. Fabricius, Dorit, et al. (2006). Human plasmacytoid dendritic cell function: inhibition of IFN-alpha secretion and modulation of immune phenotype by vasoactive intestinal peptide. Journal of Immunology, 9 pgs. PMID: 17056516. (link)


30. Gorski, Kevin, et al. (2006). Distinct indirect pathways govern human NK-cell activation by TLR-7 and TLR-8 agonists. International Immunology, 12 pgs. PMID: 16728430. (link)


31. Abel, Kristina, et al. (2005). Deoxycytidyl-Deoxyguanosine Oligonucleotide Classes A, B, and C Induce Distinct Cytokine Gene Expression Patterns in Rhesus Monkey Peripheral Blood Mononuclear Cells and Distinct Alpha Interferon Responses in TLR9-Expressing Rhesus Monkey Plasmacytoid Dendritic Cells. Clinical and Diagnostic Laboratory Immunology, 16 pgs. PMID: 15879022. (link)


32. Zhang, Weidong, et al. (2005). Inhibition of respiratory syncytial virus infection with intranasal siRNA nanoparticles targeting the viral NS1 gene. Nature Medicine, 7 pgs. PMID: 15619625. https://www.ncbi.nlm.nih.gov/pubmed/15619625 (link)


33. Coates, Toby, et al. (2003). Dendritic cell subsets in blood and lymphoid tissue of rhesus monkeys and their mobilization with Flt3 ligand. Blood Journal, 9 pgs. PMID: (link)


34. Kerkmann, Miren, et al. (2003). Activation with CpG-A and CpG-B oligonucleotides reveals two distinct regulatory pathways of type I IFN synthesis in human plasmacytoid dendritic cells. Journal of Immunology, 11 pgs. PMID: 12707322. (link)

References

1. Staehelin, T., Stähli, C., Hobbs, D.S., and Pestka, S. (1981) “A Rapid Quantitative Assay of High Sensitivity for Human Leukocyte Interferon with Monoclonal Antibodies,” in Methods in Enzymology, Vol. 79 (S. Pestka, ed.), Academic Press, New York, 589-595.


2. Kelder, B., Rashidbaigi, A., and Pestka, S. (1986) “A Sandwich Radioimmunoassay for Human IFNa” in Methods in Enzymology, Vol. 119 (S. Pestka, ed.), Academic Press, New York, 582-587.


3. Human IFN-α international reference standard provided by the NIH, reference no. Gxa01-901-535. Pestka, S. (1986) “Interferon Standards and General Abbreviations,” in Methods in Enzymology, Vol. 119 (S. Pestka, ed.), Academic Press, New York, 14-23.


4. Rubinstein, M., Levy, W.P., Moschera, J.A., Lai, C.-Y., Hershberg, R.D., Bartlett, R.T., and Pestka, S. (1981) “Human Leukocyte Interferon: Isolation and Characterization of Several Molecular Forms,” Arch. Biochem. Biophys. 210, 307-318.


5. Hobbs, D.S. and Pestka, S. (1982) “Purification and Characterization of Interferons from a Continuous Myeloblastic Cell Line,” J. Biol. Chem. 257, 4071-4076.