AI Article Synopsis
- The study aimed to create an ELISA assay to measure the release of a specific propeptide (DSGISPR) from activated granulocytes, which relates to type 1-phospholipase A2 (1-proPLA2).
- Researchers developed a sensitive assay using antibodies that target the propeptide, allowing for precise measurement of its release during granulocyte activation, while confirming the presence of its inactive precursor.
- The findings revealed that DSGISPR and its precursor were specifically present in human granulocytes, with their release triggered by TNF alpha or GM-CSF, highlighting potential implications for understanding acute lung injury.
Article Abstract
Objective: To establish a ELISA assay to measure release of type 1-phospholipase A2 propeptide from activated granulocytes. Human type 1-prophospholipase A2 (1-proPLA2) is biosynthesized and stored as inactive zymogen. Activation involves tryptic-like cleavage at the N-terminus, with equimolar release of the heptapeptide DSGISPR.
Methods: Using antibodies directed to the carboxyterminus of synthetic DSGISPR we developed a sensitive solid-phase ELISA specific for the released propeptide that accurately reports the activation of 1-proPLA2. The presence of the 1-proPLA2 precursor itself can be determined by trypsinization of the sample and subsequent assay for free DSGISPR.
Results: Using this ELISA, we demonstrated the presence of immunoreactive DSGISPR and its 14 kDa 1-proPLA2-like precursor in human granulocytes, but their absence in human macrophages and lymphocytes. Stimulation of cultured granulocytes with 1 pM of TNF alpha or GM-CSF caused rapid release of DSGISPR and precursor into the surrounding medium. The immunoreactive signal coeluted with standard synthetic DSGISPR on G50 Sephadex chromatography.
Conclusion: Release of DSGISPR immunoreactivity appears to be a specific consequence of granulocyte activation of potential relevance to the clinical pathophysiology of conditions like acute lung injury.
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| http://dx.doi.org/10.1016/0009-9120(94)00068-7 | DOI Listing |
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Activation of human granulocyte type 1-prophospholipase A2.
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View Article and Find Full Text PDFType 1-prophospholipase A2 propeptide immunoreactivity is released from activated granulocytes.
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Article Synopsis
- The study aimed to create an ELISA assay to measure the release of a specific propeptide (DSGISPR) from activated granulocytes, which relates to type 1-phospholipase A2 (1-proPLA2).
- Researchers developed a sensitive assay using antibodies that target the propeptide, allowing for precise measurement of its release during granulocyte activation, while confirming the presence of its inactive precursor.
- The findings revealed that DSGISPR and its precursor were specifically present in human granulocytes, with their release triggered by TNF alpha or GM-CSF, highlighting potential implications for understanding acute lung injury.
Type 1 prophospholipase A2 propeptide in acute lung injury.
Lancet
November 1994
Department of Surgery, St George's Hospital Medical School, London, UK.
Neutrophil sequestration and activation in the pulmonary vasculature and interstitium are important in acute lung injury. Phospholipase A2 plays an important part in the production of potent inflammatory mediators in this syndrome. We used our ELISA for type 1 prophospholipase A2 activation peptides, which have the aminoacid sequence Asp-Ser-Gly-Ile-Ser-Pro-Arg (DSGISPR), to show that DSGISPR concentrations in plasma and urine are a sensitive and specific marker of acute lung injury in patients admitted to intensive care.
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