Autoantigen cell activation for rapid diagnosis of different autoimmune disorders: Comparison of proliferation assays with a static cytometer (CellScan)

Boris Gilburd, Ora Shovman, Giesle Zandman-Goddard, Yael Schiffenbauer, Ela Trubniykov, Mia Severin, Yehuda Shoenfeld*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Objective. We analyzed the cellular response to different autoantigens: nucleosomes; lysophosphatidylcholine, Mycobacterial heat shock protein 65 kD, and human β2 glycoprotein 1 in patients with systemic lupus erythematosus and patients with stable and unstable angina pectoris comparing the CellScan system and routine proliferation assay. Patients and methods. One hundred thirty-eight patients with different autoimmune diseases and 85 healthy control were studied. The patient's group included: Twenty-seven patients with systemic lupus erythematosus; 74-with unstable angina and 37-with stable angina. Activation of peripheral blood mononuclear cells induced by different antigenic stimulus was measured by fluorescence polarization changes using the CellScan or routinely used proliferation assay. Results. The incubation with nucleosomes led to significant proliferation of peripheral blood mononuclear cells from patients with systemic lupus erythematosus (p<0.01) in comparison to patients with atherosclerosis and healthy subjects (8567±786; 3965±987 and 3687±456 cpm respectively). The nucleosomes were found to induce a higher percentage of peripheral blood mononuclear cells stimulation in systemic lupus erythematosus patients as was measured by CellScan (5.8±0.7%). The nucleosomal antigen had no effect on the stimulation of peripheral blood mononuclear cells from healthy subjects and patients with atherosclerosis (1.65±0.28 and 1.9±0.23% respectively). A strong correlation was found between the CellScan system and proliferation assay (r=0.76). Stimulation of peripheral blood mononuclear cells from patients with unstable and stable angina pectoris with 10 μM lysophosphatidylcholine resulted in pronounced cell proliferation (4752±924; 3758±743 cpm) compared to healthy subjects (1722±616 cpm; p<0.01). The 30 min incubation of peripheral blood mononuclear cells with lysophosphatidylcholine was also effective in inducing significant activation (4.7±0.89; 3.42±0.85%) in patient's groups compared to healthy subjects (1.23±0.23%; p<0.01), measured by the CellScan. Human β2 glycoprotein 1 significantly increased proliferation of peripheral blood mononuclear cells in patients with unstable and stable angina (5678±756 and 3789±954 cpm respectively) in comparison to healthy subjects (1132±345 cpm; p<0.01). The same stimulation (3.87±0.34; 2.34±0.78 and 1.27±0.32% p<0.01) of peripheral blood mononuclear cells was observed by the CellScan. Conclusion. We have presented an alternative approach to direct measuring of peripheral blood mononuclear cell activation at the single-cell level by monitoring changes in fluorescence polarization using the CellScan system. This method of detection of cell reactivity was compared to a routinely used proliferation assay. We conclude that the fluorescence polarization measurements can serve as a powerful tool to assess the response of cells to diverse autoantigens involved in ongoing pathologic processes of autoimmunity.

Original languageEnglish
Pages (from-to)316-323
Number of pages8
JournalClinical Application of Immunology
Volume3
Issue number1
StatePublished - Mar 2004
Externally publishedYes

Keywords

  • Atherosclerosis
  • Autoantigens
  • Autoimmunity
  • Cell immunity
  • Fluorescence polarization
  • Lupus
  • Nucleosomes

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