Human granzyme-B enzyme spot detection The particle enzyme-B enzyme spot detection kit provided by Sanquin applies the latest technology and has high sensitivity. Only one positive cell can detect the presence of particle enzyme-B. Strong specificity and good repeatability. Provide new and powerful detection tools for anti-virus and anti-tumor research. Comparison of enzyme point technology and enzyme labeling technology 1. The obvious feature of enzyme point technology is the increased sensitivity 2. The enzyme spot technology only requires 1 positive cell / well, and the sensitivity reaches 1: 1,000,000 for positive accurate analysis 3. Enzyme labeling technology requires at least 400 cells / well to reach a meaningful detection level 4. T cells that can produce cytokines can be analyzed at the single cell level Introduction to Granzyme-B Granzymes are exo-serine proteases derived from cytoplasmic granules released by cytotoxic T lymphocytes (CTLs) and natural killer cells (NK). These particles contain proenzyme granules and other proteases, including perforated proteins. Because CTL cells bind to target cells (through CTL receptor and antigen peptide-MHC class I molecular complex on the surface of target cells), the granulase enters the target cells through the exocytosis process of the particles, and the perforated protein aggregates on the cell membrane to enter the target The cells form small pores across the membrane, perforating the target cell membrane. Perforin releases granzyme by perforating the granzyme-endosomal membrane. Granzyme B activates the intracellular caspases reaction and finally causes the death of target cells. Granzyme A can also induce apoptosis of target cells. In many diseases, the percentage of granulase B-positive CTL can be determined by flow cytometry and immune cells. Some granzymes can also "leak" into peripheral blood and other body fluids. Granzymes can also be measured in healthy people. These soluble granzymes can be measured by ELISA. The PeliSPOT Human Granzyme B kit is used to repetitively and quantitatively determine the frequency of granzyme B secreted and released by human CTL cells and NK cells. method The cells were incubated overnight in pre-coated microplates with pre-coated high affinity anti-granase-B monoclonal antibodies. The secreted granzyme-B binds to the pre-coated anti-granase-B antibody. Wash to remove unbound components, and then add biotin-labeled anti-granase-B antibody. With the addition of streptavidin-HRP polymer and substrate, blue / purple spots can be seen where secreted cytokines are present. Granzyme-B clinical significance Viral infections EBV, HIV, CMV, hepatitis virus and dengue virus infections can enhance the CTL response of patients and increase the content of soluble particulate enzymes in the body. Lymphoma and sarcoma. The concentration of granzyme B in lymphoma patients such as Hodgkin's disease increases. Rheumatoid arthritis Soluble granzyme B is significantly higher in the synovial fluid of patients with rheumatoid arthritis than in primary systemic osteoarthropathy. Transplantation Granzyme is involved in acute rejection after kidney transplantation. Infiltrating lymphocytes strongly express plasma soluble granzymes, and elevated plasma soluble granzyme levels in kidney transplant patients suggest systemic viral infections, such as CMV infection
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The enzyme point (ELISPOT) technology is a new type of measurement method developed in recent years and applied to antiviral immunity and antitumor immunity.
