A putative role for platelet-derived PPARγ in vascular homeostasis demonstrated by anti-PPARγ induction of bleeding, thrombocytopenia and compensatory megakaryocytopoiesis.
Widely known for its role in adipogenesis and energy metabolism, PPARγ also plays a role in platelet function. To further understand functions of platelet-derived PPARγ, we produced rabbit polyclonal (PoAbs) and mouse monoclonal (MoAbs) antibodies against PPARγ 14mer/19mer peptide-immunogens. Unexpectedly, our work produced two key findings. First, MoAbs but not PoAbs produced against PPARγ peptide-immunogens displayed antigenic crossreactivity with highly conserved PPARα and PPARβ/δ. Similarly, Santa Cruz PoAb sc-7196 was monospecific for PPARγ while MoAb sc-7273 crossreacted with PPARα and PPARβ/δ. Second, immunized rabbits and mice exhibited unusual pathology including cachexia, excessive bleeding, and low platelet counts leading to thrombocytopenia. Spleens from immunized mice were fatty, hemorrhagic and friable. Although passive administration of anti-PPARγ PoAbs failed to induce experimental thrombocytopenia, megakaryocytopoiesis was induced 4-8-fold in mouse spleens. Similarly, marrow megakaryocytopoiesis was enhanced 1.8-4-fold in immunized rabbits. These peptide-immunogens are 100% conserved in human, rabbit and mouse; thus, immune-mediated platelet destruction via crossreactivity with platelet-derived PPARγ likely caused bleeding, thrombocytopenia, and compensatory megakaryocytopoiesis. Such overt pathology would cause significant problems for large-scale production of anti-PPARγ PoAbs. Furthermore, a major pitfall associated with MoAb production against closely related molecules is that monoclonicity does not guarantee monospecificity, an issue worth further scientific scrutiny.
Simpson-Haidaris
Patricia J
PJ
Department of Medicine, University of Rochester School of Medicine and Dentistry, 601 Elmwood Ave, Rochester, NY 14642, USA. pj simpsonhaidaris@urmc.rochester.edu
Seweryniak
Kathryn E
KE
Spinelli
Sherry L
SL
Garcia-Bates
Tatiana M
TM
Murant
Thomas I
TI
Pollock
Stephen J
SJ
Sime
Patricia J
PJ
Phipps
Richard P
RP
D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000378:metabolism*; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000378:metabolism; D002100:Cachexia; D018604:Epitope Mapping; D005260:Female; D006651:Histocytochemistry; D006706:Homeostasis / Q000502:physiology*; D006801:Humans; D007422:Intestines / Q000473:pathology; D008099:Liver / Q000473:pathology; D051379:Mice; D008807:Mice, Inbred BALB C; D010053:Ovary / Q000473:pathology; D047495:PPAR gamma / Q000037:antagonists & inhibitors / Q000737:chemistry / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D010455:Peptides / Q000737:chemistry / Q000276:immunology* / Q000378:metabolism; D020033:Protein Isoforms; D011817:Rabbits; D013154:Spleen / Q000473:pathology; D013921:Thrombocytopenia / Q000473:pathology / Q000503:physiopathology; D034061:Thrombopoiesis
D000906:Antibodies; D000911:Antibodies, Monoclonal; D047495:PPAR gamma; D010455:Peptides; D020033:Protein Isoforms