0 19 Membrane-associated membrane-associated JJ 20 31 lymphotoxin lymphotoxin NN 32 34 on on IN 35 42 natural natural JJ 43 49 killer killer NN 50 55 cells cell NNS 56 65 activates activate VBZ 66 77 endothelial endothelial JJ 78 83 cells cell NNS 84 87 via via IN 88 90 an an DT 91 110 NF-kappaB-dependent nf-kappab-dependent JJ 111 118 pathway pathway NN 118 119 . . . 121 131 BACKGROUND background NN 131 132 : : : 133 143 Inhibition Inhibition NNP 144 146 of of IN 147 157 complement complement NN 158 160 in in IN 161 166 small small JJ 167 173 animal animal NN 174 180 models model NNS 181 183 of of IN 184 203 xenotransplantation xenotransplantation NN 204 207 has have VBZ 208 220 demonstrated demonstrate VBN 221 226 graft graft NN 227 239 infiltration infiltration NN 240 244 with with IN 245 252 natural natural JJ 253 259 killer killer NN 260 261 ( ( ( 261 263 NK NK NNP 263 264 ) ) ) 265 270 cells cell NNS 271 274 and and CC 275 284 monocytes monocyte NNS 285 295 associated associate VBN 296 300 with with IN 301 312 endothelial endothelial JJ 313 317 cell cell NN 318 319 ( ( ( 319 321 EC EC NNP 321 322 ) ) ) 323 333 activation activation NN 333 334 . . . 335 337 We we PRP 338 342 have have VBP 343 353 previously previously RB 354 366 demonstrated demonstrate VBN 367 371 that that IN 372 377 human human JJ 378 380 NK nk NN 381 386 cells cell NNS 387 395 activate activate VBP 396 403 porcine porcine JJ 404 406 EC EC NNP 407 409 in in FW 410 415 vitro vitro FW 415 416 , , , 417 422 which which WDT 423 430 results result VBZ 431 433 in in IN 434 442 adhesion adhesion NN 443 451 molecule molecule NN 452 462 expression expression NN 463 466 and and CC 467 475 cytokine cytokine NN 476 485 secretion secretion NN 485 486 . . . 487 489 In in IN 490 494 this this DT 495 500 study study NN 500 501 , , , 502 504 we we PRP 505 509 used use VBD 510 513 the the DT 514 516 NK NK NNP 517 521 cell cell NN 522 526 line line NN 527 531 NK92 nk92 NN 532 534 to to TO 535 541 define define VB 542 545 the the DT 546 555 molecular molecular JJ 556 559 and and CC 560 568 cellular cellular JJ 569 574 basis basis NN 575 577 of of IN 578 580 NK NK NNP 581 594 cell-mediated cell-mediated JJ 595 597 EC EC NNP 598 608 activation activation NN 608 609 . . . 610 617 METHODS methods NNS 617 618 : : : 619 621 EC EC NNP 622 626 were be VBD 627 638 transfected transfecte VBN 639 643 with with IN 644 650 either either CC 651 659 reporter reporter NN 660 670 constructs construct NNS 671 681 containing contain VBG 682 685 the the DT 686 696 luciferase luciferase NN 697 701 gene gene NN 702 708 driven drive VBN 709 715 either either CC 716 718 by by IN 719 729 E-selectin E-selectin NNP 730 732 or or CC 733 744 interleukin interleukin NN 745 746 ( ( ( 746 751 IL)-8 il)-8 NN 752 761 promoters promoter NNS 762 764 or or CC 765 766 a a DT 767 776 synthetic synthetic JJ 777 796 NF-kappaB-dependent nf-kappab-dependent JJ 797 805 promoter promoter NN 805 806 . . . 807 809 In in IN 810 818 addition addition NN 818 819 , , , 820 821 a a DT 822 839 dominant-negative dominant-negative JJ 840 846 mutant mutant JJ 847 852 tumor tumor NN 853 861 necrosis necrosis NN 862 868 factor factor NN 869 877 receptor receptor NN 878 879 I i CD 880 881 ( ( ( 881 886 TNFRI TNFRI NNP 886 887 ) ) ) 888 898 expression expression NN 899 905 vector vector NN 906 909 was be VBD 910 924 co-transfected co-transfecte VBN 925 927 in in IN 928 938 inhibition inhibition NN 939 946 studies study NNS 946 947 . . . 948 959 Forty-eight forty-eight CD 960 965 hours hour NNS 966 971 after after IN 972 984 transfection transfection NN 984 985 , , , 986 988 EC EC NNP 989 993 were be VBD 994 1004 stimulated stimulate VBN 1005 1009 with with IN 1010 1012 NK NK NNP 1013 1018 cells cell NNS 1019 1021 or or CC 1022 1024 NK NK NNP 1025 1029 cell cell NN 1030 1038 membrane membrane NN 1039 1047 extracts extract NNS 1048 1051 for for IN 1052 1053 7 7 CD 1054 1056 hr hr NN 1057 1060 and and CC 1061 1071 activation activation NN 1072 1075 was be VBD 1076 1084 measured measure VBN 1085 1087 by by IN 1088 1089 a a DT 1090 1100 luciferase luciferase NN 1101 1106 assay assay NN 1106 1107 . . . 1108 1115 RESULTS results NNS 1115 1116 : : : 1117 1127 Co-culture Co-culture NNP 1128 1130 of of IN 1131 1133 NK NK NNP 1134 1139 cells cell NNS 1140 1144 with with IN 1145 1156 transfected transfecte VBN 1157 1159 EC EC NNP 1160 1168 enhanced enhance VBD 1169 1179 E-selectin E-selectin NNP 1179 1180 , , , 1181 1185 IL-8 il-8 NN 1185 1186 , , , 1187 1190 and and CC 1191 1210 NF-kappaB-dependent nf-kappab-dependent JJ 1211 1219 promoter promoter NN 1220 1228 activity activity NN 1228 1229 . . . 1230 1232 NK NK NNP 1233 1237 cell cell NN 1238 1246 membrane membrane NN 1247 1255 extracts extract NNS 1256 1264 retained retain VBD 1265 1268 the the DT 1269 1277 capacity capacity NN 1278 1280 to to TO 1281 1289 activate activate VB 1290 1292 EC EC NNP 1293 1296 and and CC 1297 1304 induced induce VBD 1305 1312 nuclear nuclear JJ 1313 1326 translocation translocation NN 1327 1329 of of IN 1330 1339 NF-kappaB NF-kappaB NNP 1340 1341 ( ( ( 1341 1344 p50 p50 NN 1345 1348 and and CC 1349 1352 p65 p65 NN 1352 1353 ) ) ) 1353 1354 . . . 1355 1362 Western Western NNP 1363 1371 blotting blotting NN 1372 1374 of of IN 1375 1377 NK NK NNP 1378 1382 cell cell NN 1383 1386 and and CC 1387 1395 membrane membrane NN 1396 1404 extracts extract NNS 1405 1413 detected detect VBD 1414 1417 the the DT 1418 1426 presence presence NN 1427 1429 of of IN 1430 1447 Lymphotoxin-alpha Lymphotoxin-alpha NNP 1448 1449 ( ( ( 1449 1456 LTalpha LTalpha NNP 1456 1457 ) ) ) 1458 1461 but but CC 1462 1465 not not RB 1466 1471 tumor tumor NN 1472 1480 necrosis necrosis NN 1481 1493 factor-alpha factor-alpha NN 1493 1494 . . . 1495 1506 Furthermore furthermore RB 1506 1507 , , , 1508 1515 LTalpha LTalpha NNP 1516 1519 was be VBD 1520 1528 secreted secrete VBN 1529 1531 in in IN 1532 1537 NK:EC NK:EC NNP 1538 1549 co-cultures co-culture NNS 1549 1550 . . . 1551 1566 Co-transfection co-transfection NN 1567 1571 with with IN 1572 1589 dominant-negative dominant-negative JJ 1590 1596 mutant mutant JJ 1597 1602 TNFRI TNFRI NNP 1603 1612 inhibited inhibit VBD 1613 1615 EC EC NNP 1616 1626 activation activation NN 1627 1629 by by IN 1630 1632 NK NK NNP 1633 1637 cell cell NN 1638 1646 membrane membrane NN 1647 1655 extracts extract NNS 1656 1659 and and CC 1660 1662 by by IN 1663 1665 NK NK NNP 1666 1671 cells cell NNS 1672 1674 by by IN 1675 1677 80 80 CD 1677 1678 % % NN 1679 1682 and and CC 1683 1685 47 47 CD 1685 1686 % % NN 1686 1687 , , , 1688 1700 respectively respectively RB 1700 1701 . . . 1702 1705 The the DT 1706 1710 same same JJ 1711 1718 pattern pattern NN 1719 1721 of of IN 1722 1732 inhibition inhibition NN 1733 1736 was be VBD 1737 1745 observed observe VBN 1746 1751 using use VBG 1752 1762 anti-human anti-human JJ 1763 1765 LT lt NN 1766 1770 sera serum NNS 1770 1771 . . . 1772 1783 CONCLUSIONS conclusions NNS 1783 1784 : : : 1785 1790 Human human JJ 1791 1793 NK NK NNP 1794 1798 cell cell NN 1799 1813 membrane-bound membrane-bound JJ 1814 1816 LT lt NN 1817 1824 signals signal NNS 1825 1831 across across IN 1832 1839 species specie NNS 1840 1843 via via IN 1844 1849 TNFRI TNFRI NNP 1849 1850 , , , 1851 1858 leading lead VBG 1859 1861 to to TO 1862 1871 NF-kappaB NF-kappaB NNP 1872 1879 nuclear nuclear JJ 1880 1893 translocation translocation NN 1894 1897 and and CC 1898 1911 transcription transcription NN 1912 1914 of of IN 1915 1925 E-selectin E-selectin NNP 1926 1929 and and CC 1930 1934 IL-8 IL-8 NNP 1934 1935 , , , 1936 1941 which which WDT 1942 1949 results result VBZ 1950 1952 in in IN 1953 1955 EC EC NNP 1956 1966 activation activation NN 1966 1967 . . . 1968 1971 The the DT 1972 1983 discrepancy discrepancy NN 1984 1986 in in IN 1987 1990 the the DT 1991 1997 degree degree NN 1998 2000 of of IN 2001 2011 inhibition inhibition NN 2012 2014 by by IN 2015 2023 membrane membrane NN 2024 2032 extracts extract NNS 2033 2036 and and CC 2037 2039 NK nk NN 2040 2045 cells cell NNS 2046 2050 with with IN 2051 2057 mutant mutant JJ 2058 2063 TNFRI TNFRI NNP 2064 2072 suggests suggest VBZ 2073 2077 that that IN 2078 2088 additional additional JJ 2089 2097 pathways pathway NNS 2098 2101 are be VBP 2102 2110 utilized utilize VBN 2111 2113 by by IN 2114 2116 NK NK NNP 2117 2122 cells cell NNS 2123 2125 to to TO 2126 2134 activate activate VB 2135 2137 EC EC NNP 2137 2138 . . .