Recognition of apoptotic cells by viable cells is specific, ubiquitous, and species independent: analysis using photonic crystal biosensors.
Apoptotic recognition is innate and linked to a profound immune regulation (innate apoptotic immunity [IAI]) involving anti-inflammatory and immunosuppressive responses. Many of the molecular and mechanistic details of this response remain elusive. Although immune outcomes can be quantified readily, the initial specific recognition events have been difficult to assess. We developed a sensitive, real-time method to detect the recognition of apoptotic cells by viable adherent responder cells, using a photonic crystal biosensor approach. The method relies on characteristic spectral shifts resulting from the specific recognition and dose-dependent interaction of adherent responder cells with nonadherent apoptotic targets. Of note, the biosensor provides a readout of early recognition-specific events in responder cells that occur distal to the biosensor surface. We find that innate apoptotic cell recognition occurs in a strikingly species-independent manner, consistent with our previous work and inferences drawn from indirect assays. Our studies indicate obligate cytoskeletal involvement, although apoptotic cell phagocytosis is not involved. Because it is a direct, objective, and quantitative readout of recognition exclusively, this biosensor approach affords a methodology with which to dissect the early recognition events associated with IAI and immunosuppression.
Department of Microbiology and Immunology, University of Illinois College of Medicine, Chicago, IL 60612.
Goutham
G
Pattabiraman
Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801.
Erich A
EA
Lidstone
Department of Microbiology and Immunology, University of Illinois College of Medicine, Chicago, IL 60612.
Karol
K
Palasiewicz
Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801Department of Electrical and Computer Engineering, University of Illinois at Urbana-Champaign, Urbana, IL 61801Cancer Center, University of Illinois College of Medicine, Chicago, IL 60612 duck@uic.edu bcunning@illinois.edu.
Brian T
BT
Cunningham
Department of Microbiology and Immunology, University of Illinois College of Medicine, Chicago, IL 60612Cancer Center, University of Illinois College of Medicine, Chicago, IL 60612Department of Bioengineering, University of Illinois College of Medicine, Chicago, IL 60612 duck@uic.edu bcunning@illinois.edu.
David S
DS
Ucker
D000818:Animals; D017209:Apoptosis; D015374:Biosensing Techniques / Q000379:methods*; D002448:Cell Adhesion; D002450:Cell Communication; D002470:Cell Survival; D003460:Crystallization; D003599:Cytoskeleton / Q000378:metabolism; D006801:Humans; D019169:Jurkat Cells; D051379:Mice; D010587:Phagocytosis; D017785:Photons; D013045:Species Specificity