Elucidating the immunological landscape of acute myocardial infarction using an integrated single-cell enzyme and antibody quantification (iSEAQ) assay

Abstract Funding Acknowledgements Type of funding sources: Public grant(s) – National budget only. Main source(s): Research Foundation Background Characterizing enzyme secretion with surface antigen expression circulating leukocytes provides deep immunological insights that may benefit diagnostics and therapeutics acute myocardial infarction (AMI). Current methods are limited by laborious sample preparation, lengthy assay time analytical throughput. Purpose We aimed to develop a point-of-care platform for real-time, granular immune profiling AMI during index hospitalization. Methods developed system 40-min termed integrated single-cell quantification (iSEAQ), consisting an microfluidic device streamlined process recover isolate single from whole blood, customized 5-channel high-speed droplet fluorescence cytometer sensitive key protein activity resembling live leukocyte in-vivo (Figure 1). Using panel 2 secreted enzymes (granzyme B (GzB) neutrophil elastase (NE)) 3 antigens (cluster differentiation (CD) 66b, CD3 CD31), we tracked the longitudinal shift functionality 9 patients admitted ST-segment elevation (STEMI) (median age 52 years, 100% males) at timepoints hospitalization: 1) on admission prior coronary revascularization, 2) within 24 hours 3) pre-discharge. The data were imported using python script pre-processed, before piping into Numpy, Pandas, Scipy UMAP library preparation classification. Seaborn Matplotlib used all visualization plots. Data was analyzed student two-tailed t-tests, significance level <5%. Results Leveraging its ability dissect phenotype class, iSEAQ unraveled distinct contributions CD31, NE GzB peripheral granulocytes, T lymphocytes other cells 2). majority response is expressed in CD66b+ granulocytes showing increased NE+ CD31 denoted Class 0 Figure 2. Compared samples healthy donors STEMI admission, CD31+ expressions significantly lower pre-discharge (p<0.05). also distinguished anterior inferior STEMI, as evident elevated NE+CD31-secretion (365±70 vs 83±35 fg, p < 0.001), preliminary promise risk stratify. Conclusion provided comprehensive atlas responses hospital admission. With open design commercial biosensors, potentially powerful tool taking snapshots many diseases.