Characterization and quantification of porcine circulating endothelial cells

Area of Science:

• Immunology
• Cell Biology
• Transplantation Science

Background:

• Endothelial damage is a key factor in transplantation and cardiovascular diseases.
• Circulating endothelial cells (CEC) in humans indicate disease severity.
• Limited antibodies for porcine endothelium hinder CEC research in pig models.

Purpose of the Study:

• To develop a method for detecting porcine CEC.
• To utilize anti-human antibodies for porcine endothelium epitope detection.

Main Methods:

• Assessed cross-species immunoreactivity of 21 anti-human antibodies against swine endothelial cells (SUVEC) and porcine kidney-derived endothelial cells (PKEC).
• Employed fluorescence-activated cell sorting (FACS) and confocal microscopy.
• Developed a FACS-based whole blood CEC quantification method using CD105 and CD146 staining after CD45 exclusion and osmotic erythrolysis.

Main Results:

• CD105 (MEM-229 clone) and CD146 (P1H12 clone) antibodies showed immunoreactivity with porcine endothelial cells.
• Baseline porcine CEC count was 673.1 ± 551.4 CEC/ml.
• Blood samples showed elevated CEC counts (1118 ± 661.4 CEC/ml) in a vascular damage context.

Conclusions:

• Anti-human CD146 and CD105 antibodies specifically label porcine endothelium.
• These antibodies enable identification and quantification of CEC in porcine whole blood via FACS.