Comprehensive Immunological Profiling in Acute Ischemic Stroke: Myeloid Cell Activation and Molecular Signatures in Blood and Thrombus during Mechanical Thrombectomy

Wirginia Krzyściak^1*Tadeusz Popiela²

• ¹Department of Medical Diagnostics, Faculty of Pharmacy, Medical College, Jagiellonian University, Krakow, Poland, Kraków, Poland
• ²Department of Diagnostic Imaging, University Hospital, 30-688 Kraków, Poland, Krakow, Poland

Background: Acute ischemic stroke (AIS) induces a complex local and systemic inflammatory response; however, most studies rely solely on peripheral blood, providing an incomplete view of immune activity at the occlusion site and within the thrombus. Objective: To characterize immune activation and transcriptomic signatures of myeloid cells across three compartments—arterial blood at the occlusion site, peripheral blood, and thrombus—and to evaluate their associations with radiological and clinical outcomes following mechanical thrombectomy. Methods: This prospective, single-center study will include AIS patients treated with mechanical thrombectomy. Matched arterial, peripheral, and thrombus samples will undergo spectral flow cytometry, cytokine profiling, cell-free DNA (cfDNA) quantification, microscopy, and RNA sequencing. Immune and molecular readouts will be correlated with clinical scores (NIHSS, mRS), imaging markers (e.g., hyperdense middle cerebral artery sign [HMCAS]), and procedural outcomes (TICI score, number of passes). Significance: Integrating local and systemic immune profiles with clinical and radiological parameters may identify biomarkers predictive of thrombectomy efficacy and functional recovery, thereby supporting precision-medicine approaches in AIS.