Suppression of human anti-porcine natural killer cell xenogeneic responses by combinations of monoclonal antibodies specific to CD2 and NKG2D and extracellular signal-regulated kinase kinase inhibitor.
Natural killer (NK) cells can destroy xenogeneic tissues by antibody-dependent cell cytotoxicity (ADCC) and direct lysis. Unlike ADCC, activating interactions between human NK receptors and their cognate ligands in pigs are not fully elucidated. We set up this study to identify human NK activating receptors recognizing porcine cells isolated from distinct organs, e.g., aorta, cornea and liver, and to provide a molecular basis for effective immunosuppressive regimens. Among the array of NK receptors tested, NKp46, 2B4, CD49d, CD48, CD2 and NKG2D, only CD2 and NKG2D were shown to be involved in both cytotoxicity and cytokine (interferon-gamma and tumour necrosis factor-alpha) production against porcine targets. Simultaneous blocking of CD2 and NKG2D by combining its monoclonal antibodies further suppressed xenogeneic NK responses. Moreover, addition of a suboptimal dose of PD98059, an extracellular signal-regulated kinase (ERK) kinase inhibitor, to those cells maximally reduced NK cytotoxicity, suggesting that ERK plays an important role in NK-mediated xenoreactivity. These impairments in NK cells were tightly associated with defective intracellular calcium mobilization and the subsequent degranulation process. Therefore, our data demonstrate a distinct role of CD2 and NKG2D on human NK cells in recognizing porcine grafts and further provide a potentially efficacious combinational regimen using anti-CD2 and anti-NKG2D monoclonal antibodies with PD98059 in a pig-to-human transplantation model.
Kim
Tae-Jin
TJ
Global Research Lab, Department of Biochemistry and Division of Brain Korea 21 Program for Biomedical Science, Korea University College of Medicine, Seoul, Korea.
Kim
Nayoung
N
Kim
Eun-Ok
EO
Choi
Jong-Rip
JR
Bluestone
Jeffrey A
JA
Lee
Kyung-Mi
KM
D000818:Animals; D000910:Antibodies, Heterophile / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D018801:CD2 Antigens / Q000276:immunology*; D002118:Calcium / Q000378:metabolism; D002478:Cells, Cultured; D003602:Cytotoxicity, Immunologic; D048049:Extracellular Signal-Regulated MAP Kinases / Q000037:antagonists & inhibitors*; D006801:Humans; D007371:Interferon-gamma / Q000096:biosynthesis / Q000276:immunology; D007694:Killer Cells, Natural / Q000276:immunology* / Q000378:metabolism; D055655:NK Cell Lectin-Like Receptor Subfamily K / Q000276:immunology*; D009930:Organic Chemicals / Q000494:pharmacology*; D013552:Swine; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000276:immunology
D000910:Antibodies, Heterophile; D000911:Antibodies, Monoclonal; D018801:CD2 Antigens; D055655:NK Cell Lectin-Like Receptor Subfamily K; D009930:Organic Chemicals; C466090:PD 98058; D014409:Tumor Necrosis Factor-alpha; D007371:Interferon-gamma; D048049:Extracellular Signal-Regulated MAP Kinases; D002118:Calcium
Nat Immunol. 2008 May;9(5):495-502
18425106
Mol Immunol. 2005 Apr;42(6):665-71
15781110
Transplantation. 2007 Feb 15;83(3):323-32
17297407
Mol Immunol. 2008 Jul;45(12):3354-62
18533264
Transpl Immunol. 1996 Dec;4(4):293-9
8972559
Science. 1995 Apr 21;268(5209):405-8
7716543
Biochem Biophys Res Commun. 1988 May 31;153(1):46-50
3288207
Science. 2002 Feb 8;295(5557):1089-92
11778012
Nat Rev Immunol. 2007 Jul;7(7):519-31
17571072
J Immunol. 2005 Mar 1;174(5):2653-60
15728472
Nat Med. 2006 Mar;12(3):301-3
16491083
Nat Biotechnol. 2002 Mar;20(3):251-5
11875425
Am J Transplant. 2008 Oct;8(10):1972-81
18828764
Nat Med. 2009 Jul;15(7):746-9
19584865
Cell Cycle. 2009 Apr 15;8(8):1168-75
19282669
J Immunol. 1997 Oct 1;159(7):3655-61
9317166
Nat Immunol. 2000 Nov;1(5):419-25
11062502
Exp Mol Med. 2009 Mar 31;41(3):161-70
19293635
Transplant Proc. 2006 Dec;38(10):3312-4
17175258
Transplantation. 1998 Mar 15;65(5):727-34
9521210
Blood. 2006 Feb 1;107(3):994-1002
16204312
Am J Transplant. 2005 Sep;5(9):2085-93
16095487
J Exp Med. 1998 Oct 5;188(7):1267-75
9763606
Transplantation. 2002 Mar 15;73(5):789-96
11907429
Lab Anim Care. 1968 Feb;18(1):104-9
4230573
J Immunol. 2005 Oct 15;175(8):5463-70
16210654
Transplantation. 2009 Jan 15;87(1):35-43
19136889
Nature. 1986 Feb 20-26;319(6055):675-8
3951539
Blood. 2000 Sep 15;96(6):2181-90
10979964
J Immunol Methods. 2004 Nov;294(1-2):15-22
15604012
J Immunol. 1997 Apr 1;158(7):3148-54
9120268
Annu Rev Med. 2002;53:133-47
11818467
J Immunol. 2005 Jul 1;175(1):213-8
15972651
Mol Immunol. 2005 Jun;42(10):1205-14
15829309
J Exp Med. 1998 Jun 15;187(12):2065-72
9625766
Xenotransplantation. 2007 Mar;14(2):126-34
17381687
Nat Med. 2006 Mar;12(3):304-6
16501570
Am J Pathol. 1997 Jul;151(1):111-20
9212737
J Immunol. 1998 Dec 15;161(12):6648-56
9862693
Xenotransplantation. 2005 May;12(3):181-8
15807768
Biochem Biophys Res Commun. 2003 Oct 3;309(4):992-8
13679072
Mol Immunol. 2005 Feb;42(4):489-94
15607804
J Immunol. 2009 Jun 1;182(11):6933-42
19454690
J Clin Oncol. 2004 Nov 15;22(22):4456-62
15483017
Exp Hematol. 2005 Oct;33(10):1160-71
16219538
Xenotransplantation. 2009 Jan-Feb;16(1):19-26
19243557
Nat Immunol. 2008 May;9(5):503-10
18425107
Transpl Int. 2005 May;17(12):787-94
15711982
Cell Biol Int. 2005 Aug;29(8):638-46
15950500