Detection of expressed IL-32 in human stomach cancer using ELISA and immunostaining.

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  • Additional Information
    • Source:
      Publisher: Korean Society for Microbiology and Biotechnology Country of Publication: Korea (South) NLM ID: 9431852 Publication Model: Print Cited Medium: Print ISSN: 1017-7825 (Print) Linking ISSN: 10177825 NLM ISO Abbreviation: J Microbiol Biotechnol Subsets: MEDLINE
    • Publication Information:
      Publication: 2002- : Seoul : Korean Society for Microbiology and Biotechnology
      Original Publication: Seoul : Korean Society for Applied Microbiology, 1991-2001.
    • Subject Terms:
      Enzyme-Linked Immunosorbent Assay/*methods ; Immunohistochemistry/*methods ; Interleukins/*metabolism ; Stomach Neoplasms/*metabolism; Antibodies, Monoclonal/biosynthesis ; Humans ; Immunoglobulin G/biosynthesis ; Interleukins/blood ; Interleukins/immunology ; K562 Cells ; Recombinant Proteins/metabolism ; Sensitivity and Specificity
    • Abstract:
      Interleukin (IL)-32 is a recently identified proinflammatory cytokine that is one of the IL-18 inducible genes, and plays an important role in autoimmune and inflammatory diseases. We produced antibodies against IL-32 and studied the expression of IL-32 in human stomach cancer. We detected IL-32 secreted from K-562 cells that werw stably transfected with IL-32 and in the sera of stomach cancer patients, by a sandwich ELISA using a monoclonal antibody KU32-52 and a polyclonal antibody. In order to optimize a sandwich immunoassay, recombinant IL-32alpha was added, followed by the addition of a biotinylated KU32-52 into microtiter plate wells precoated with a goat anti-IL-32 antibody. The bound biotinylated KU32-52 was probed with a streptavidin conjugated to HRP. This sandwich ELISA was highly specific and had a minimal detection limit of 80 pg/ml (mean+/-SD of zero calibrator) and measuring up to 3,000 pg/ml. This ELISA showed no cross-reaction with other cytokines such as hIL-1alpha, hIL-1beta, hIL-2, hIL-6, hIL-8, hIL-10, hIL-18, and hTNF-alpha. Intra-assay coefficients of variation were 18.5% to 4.6% (n=10), and inter-assay coefficients were 23% to 9% (n=10). The average IL-32 level in the sera of 16 stomach cancer patients (189 pg/ml) was higher than that of 12 healthy control men (109 pg/ml). Our results indicate that serum IL-32 level can be detected by using an established ELISA, and that this immunoassay and mAb KU32-09 specific for immunohistochemistry can be used in the detection of expressed and secreted IL- 32 in stomach cancer patients.
    • Accession Number:
      0 (Antibodies, Monoclonal)
      0 (IL32 protein, human)
      0 (Immunoglobulin G)
      0 (Interleukins)
      0 (Recombinant Proteins)
    • Publication Date:
      Date Created: 20081015 Date Completed: 20090203 Latest Revision: 20220330
    • Publication Date:
      20240628
    • Accession Number:
      18852519