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• W1958429596 abstract "Neutrophil granulocytes in the peripheral blood represent the central cell population in innate immunity. Expression of the Fc receptor CD64 on their surface has been shown to correlate with complications in sepsis 1-3, infectious diseases 4, 5, and even in solid organ transplanted patients 6. CD64, also called FcγRI (Fc γ receptor I), is a class of plasma membrane receptors on human myeloid cells. It contains three extracellular immunoglobulin-like domains that represent binding sites for the Fc portion of IgG 3. Expression level of CD64 on neutrophil granulocytes can be detected easily. It is upregulated in inflammatory and septic complications; upregulation over an index of 1.5 as calculated against bead standard is of diagnostic value for patients with inflammatory response 2. In health, CD64 is constitutively presented by monocytes, macrophages, eosinophils, and neutrophil granulocytes. With <1,000 sites per cell, the neutrophil granulocyte CD64 expression is negligible in a state of health and is at the level of detection by flow cytometry 2. CD64 expression by monocytes has no diagnostic impact. On neutrophil granulocytes it becomes upregulated by various interleukins and depends on the intensity of the stimulation 2, 3, 7. It is mainly induced by bacterial infection via CD14 and toll-like receptors 1-3, 8-13. In situations of increased inflammatory response like sepsis, SIRS (systemic inflammatory response syndrome), local infections, or tissue injuries, the cell surface expression of CD64 by neutrophil granulocytes elevates within 4–6 h after stimulation, and the amount of detectable CD64 mRNA increases and can be measured by Northern Blot analysis already within 1–3 h 2. CD64 expression by neutrophil granulocytes also varies by the type of infection. Gram-negative bacteria induce higher expression compared to Gram-positive bacteria 2. This appears to directly correlate with enhanced bactericidal and antifungal activity 2. The antibody-dependent crosslinking of expressed CD64 14 contributes to cellular cytotoxicity, phagocytosis, and the clearance of immune complexes 3. Compared to other hematological indices, expression level of CD64 by neutrophil granulocytes shows a biphasic response to lipopolysaccharide (LPS) administration in humans. A correlation between pro-inflammatory cytokine serum levels, but not of the anti-inflammatory IL-10, and CD64 expression was demonstrated, illustrating that CD64 expression by neutrophil granulocytes is a measure of innate immunity 15. For clinical application, a CE approved in vitro diagnostic test kit is available (Leuko64, Trillium diagnostics, Brewer, Maine). This test guarantees a fair internal quality control 16. Within our course in clinical flow cytometry, we decided to use this test system. After collecting blood samples (50 µl EDTA-anticoagulated blood), the measurement of CD64 expression on neutrophil granulocytes was performed by quantitative flow cytometry on a Gallios (Beckman-Coulter, Krefeld, Germany) by using the Leuko64™ kit, a whole-blood lysed no-wash method containing internal calibration beads for quantification. The proprietary CD64 software was used for data analysis. This kit reports leukocyte expression of CD64 and CD163 as an index (Fig. 1) using fluorescein-labeled calibration beads 2. Expression of CD64 by neutrophil granulocytes is calculated by comparing median of CD64 expression to FITC-labeled beads as internal standard. [Color figure can be viewed at wileyonlinelibrary.com] Validation of this test kit reveals good sensitivity (90.5%) and specificity (96.3%) as well as high positive and negative predictive values for sepsis 2. In our hands, the interassay imprecision is below 15%, and calibration beads allow day-to-day comparisons of quantitative values. Given that the neutrophil is the major cellular component of acute response to infection, the detection of molecular changes of neutrophil granulocytes' activation could provide a rapid and sensitive indicator of a systemic inflammatory response 1, 2, 9-11, 14. Diagnostic use of a quantitative flow cytometric assay for expression strength of CD64 by neutrophil granulocytes as an in vitro indicator of neutrophil granulocytes' activation seems to be a valuable marker as an additional utility in the evaluation of patients with suspected acute inflammation or infection 1, 3. CD64 expression by neutrophil granulocytes has also been shown to be a useful parameter for diagnostic evaluation of suspect postoperative complications in transplanted patients 6; the high diagnostic value for infectious complications shown in other studies is not influenced by immunosuppression in this population. The use of CD64 expression by neutrophil granulocytes as an infection marker also in the postoperative course of transplanted patients should allow early determination of further treatment already within 24 h without waiting for the definitive microbiologic culture results 9. Besides the fast performance of the CD64-kit, an anticipated benefit of measuring CD64 expression by neutrophil granulocytes is that it will make the need for performing subjective and laborious manual microscopic differential counts to determine band counts and immature-to-total myeloid ratios superfluous 2, in particular if it is integrated into routine laboratory procedure. In comparison to various other parameters including C-reactive protein (CRP), CD64 expression by neutrophil granulocytes has a very good performance as summarized in Ref. 6 and it is shown there for transplanted patients in detail. Regarding procalcitonin, at least in preterm children, CD64 is even better than procalcitonin (PCT) 17. Various studies show superior diagnostic performance in sepsis as compared to other inflammatory parameters 2, 7. Nevertheless, it must be noted that the test is implemented as a diagnostic test in only very few laboratories. Although the potential diagnostic value of CD64 has been known for some time, most laboratories analyze CD64 expression by neutrophil granulocytes for clinical research only. A major barrier could be the nature of the indication (infection/sepsis), which requires a fast turn-around time, which is usually not possible if the test is not yet available on automated instruments or if the necessary specialized 24 × 7-laboratory is not available on-site. Ulrich Sack* Institute of Clinical Immunology Medical Faculty, Universität Leipzig Leipzig, Germany" @default.
• W1958429596 created "2016-06-24" @default.
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• W1958429596 date "2015-04-17" @default.
• W1958429596 modified "2023-09-26" @default.
• W1958429596 title "CD64 expression by neutrophil granulocytes" @default.
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• W1958429596 doi "https://doi.org/10.1002/cyto.b.21216" @default.
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