Primary porcine endothelial cells express membrane-bound B7-2 (CD86) and a soluble factor that co-stimulate cyclosporin A-resistant and CD28-dependent human T cell proliferation.
Increasing evidence suggests that endothelial cells can directly activate syngeneic, allogeneic and xenogeneic T cells. In this study we demonstrate that unstimulated, paraformaldehyde-fixed primary porcine aortic endothelial cells (PAEC) and microvascular endothelial cells (PMVEC) can provide co-stimulation for human T cell IL-2 secretion and proliferation. EC-mediated co-stimulation has both cyclosporin A (CsA)-sensitive and CsA-resistant components. The CsA-resistant component is completely suppressed either by blocking with anti-CD28 F(ab) fragments or CTLA-4-Ig. Northern blot analysis of unstimulated PAEC and PMVEC with porcine-specific probes reveals constitutive expression of B7-2 mRNA while B7-1 message was not detected. hCTLA-4-Ig and anti-B7-2 mAb immunoprecipitates a single 79 kDa PMVEC surface protein. Surprisingly, PMVEC conditioned media also has soluble co-stimulatory activity that is blocked by anti-CD28 F(ab) fragments or anti-B7-2 mAb. These findings demonstrate that primary unstimulated porcine EC can co-stimulate CsA-resistant human T cell proliferation through binding of membrane bound, constitutively expressed EC B7-2 (CD86) to human T cell CD28, providing one of the first demonstrations of functional B7-2 on cells outside the immune system. In addition, PMVEC secrete or shed a soluble factor that mediates CD28-dependent human T cell proliferation, demonstrating the existence of soluble mediators of CD28 activation.
Davis
T A
TA
Immune Cell Biology Program, Naval Medical Research Institute, Bethesda, MD 20889-5067, USA.
Craighead
N
N
Williams
A J
AJ
Scadron
A
A
June
C H
CH
Lee
K P
KP
D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis*; D001011:Aorta; D051940:B7-2 Antigen; D018106:CD28 Antigens / Q000494:pharmacology*; D002478:Cells, Cultured; D017077:Culture Media, Conditioned; D016572:Cyclosporine / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D004730:Endothelium, Vascular / Q000166:cytology / Q000276:immunology* / Q000378:metabolism*; D006801:Humans; D008213:Lymphocyte Activation / Q000187:drug effects*; D008562:Membrane Glycoproteins / Q000096:biosynthesis*; D015398:Signal Transduction / Q000276:immunology; D012995:Solubility; D013552:Swine; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology*
D015703:Antigens, CD; D051940:B7-2 Antigen; D018106:CD28 Antigens; C497964:CD86 protein, human; D017077:Culture Media, Conditioned; D008562:Membrane Glycoproteins; D016572:Cyclosporine