【課題】 本発明は、抗 HLA-DRモノクローナル抗体を提供する。
Contents
1.2.
3. 0 HLA 1000000000000000000 DRO 1000000000000000000 HLA DRO O 100000000000
4. DR 10000000 JJ UJU U UU DDD 0 10000 1000 lU U GvHD LU U DOO LU
5. CD20 Ri t uxi 2 neu Ri 0000 B 10000000000000000 Ri tuxi 00 0000000000000 Ri tuxi 00 00 0000000000000 000000000 00 Ri na
6. 00 9 00000000000000000000000000000000000 100000 900000000100000000 HD4 281 0000 1 0 10000000000
7. 0 50u LO 0 96000000000000 4 30000000000 100 RPM 16400 000000 10 1000000000000 99000000000 0 100 00000 370 5 H 11 04 000000000000 37 eco con 000 160 20000000 itiu Su ka 100000000000000000 HLA DRO 00000000 2 430 ATCC HB 550 00000 10000 HLA DR 1000 0000000 HLA 100000 12430 ATCC H 55 101
8. D000000000 00000 0000000000 HLA DRO 0000000000000 0000 000 0000 HD80 HD10 O 20011 100 10 00 0 0 0 0 000 00 00 060 1100100 1006 0000000000000000 HD8 O O FERM 7773 0000000000 000 O O FERM 777 40000000000 HD40 00000 FERM 77711 0000000000 HD6 l FERM BP 77720 000 70000000000000000000000000 0 0 0 HLA 000000000000000000000000000 HLA 10 0 0 HLA 1000000 81000000000000 1000000000
9. 11000000000000000000000000000000000000 1000000000000000000000000000000000 100 100000000000000000000 100 zuka et al Proc Acad Sci USA 2000 97 7220 100 100000000000000000000 000 10 20 000401 000000000080 100000000000000000000000 100000000000000000000000 100030 5000000000000000000 0000000000000 0000000000 ELISA 0000000600 20 ELISAI 0000000000000 0000 960 00000 00 0 BSAU 0
10. z 000000 ELI 5 9 100000000000000000000 m gt
11. 1000 1000094900000 16 1000 20 000 90000000000 81994000 0 9 0000 HD8G2Ser O0 0000 0 HD8G1S er 000 25 0 0 0 D O 0 HD8G2Ser HD8G2Ser 1000 0 HD80 0 0 0 0O O0 O O HD8G1Ser O HD8G2Ser O0 0 0 O O 0 0 0 0 RD8G2Ser0 ADCCO 0 0 00000 000000000 17 HD40 HD6 HD 0000000 110000 10 00000000000 11000000 9 79000000000000 HRP conj ugated anti rabb t DAKA 00000 5 DAKO 1000 LAS 1000000000000 HLA DRO 00 5 4 ATCC TI B 215 HLA DRO K28 000000000 0 K28 F
12. Lark N5KG10 0 0 O 19610000000 N5KG4 Lark I gG4 00000000 19620 0000 5 1 9621 1000000 12000 HD4 19610 0 962 1000000000000 331100000 10 Hua et 1993 00000 00 0 00 0 0 0 5 15 N5KG2Ser 1 2000000000000 Hoeg 00000 1000000000 19610 O O 9610 HD4G1 100 331100 150 00000 1000000000000 HDA 15 15 2 amp ayy 7 5 N5KG1 Val Lark HD4GI N5KG2Ser IgG2Ser HD4G2Ser N5KG4 Lark HD4G4
13. lg 1 0000000000000000000 lg 100000 14000000 19 KCo5SD 0000000000000 000 019 1190000 00000000000000 14 5 zuka al Proc Natl Sci USA 2000 97 72211 gk 0 KCoSD OOOO000000000000000000 Fishwld Nat Biotechnol 19960 114 845100000 1000000000000000000000000000000000 10000000000 190 000 0000000000000 Ishida BC s 11th Anti body ng Abstract 2000100000000000 100000000000000 DR 000000000000 UU D DU
14. JU DU DU 1000110001110 DI O00000000000000 40 0 O DNAD 1000000000000000000000000000000000 0 0000000000000000000000000000 1000000000000000000000000000000000 50 16 2005 325133 2005 11 24
15. 0000000000 JU JU DU UU DDD DDD 9000 JII UPDP UP UPDP UID DDD 10000000000000000000 VO02 43478 000000 DOO UU DD DU UU lU DOO 1000000000000000000 D01000000000000000000000000000000000000007 JU UU UU U DDD DDD D DUH JU UJU UU U DDD DDD DD Ul 0000000000000000000 10 2005 325133 2005 11 24 site specific mtagenesi s Proc Sci USA 1984 Vol 81 56621 DDDDDDD0D0D00D00000000000UU0
16. Mayt ansi Chari al Cancer Res 1992 0 52 127 Liu et al Proc Natl Acad Sci USA 1996 Vol 93 8681 1 00000000100 1000000 1100000000000000 1000000000 20 100000000 100000000000000000 0 30000000000 410000000 50000000000 100000006 7 100000000 8100000000000000000 100000000 HLA 1000000000000 1000000000000 10 0 0100 9000000000000 1000000000 008000000000 0000 Steven al 20000 T FactsBook Academic Press 0 HLA DR 1000 100000000000000 10000 HLA DRO 929 DR 0 0 10000000 pEF HLA DRa neo HLA DRB 10000000 HLA DRa 0 DNAD O 0 O DRBR 0000000000 0 0 DNAD BOS M 2 5 1 S 6 Nagata S Nucleic Acids R es 19901 18 53320
17. 19 79 2005 325133 2005 11 24
18. HLA DRI HLA DRI DRI 14161 G2 gG4 Fc gG1 gG3 Fc 0 D0000000 Janeway Jr Travers 1997 ol Third Edition Curre Phulishing 1 00000000000000000000
19. 33 2005 325133 2005 11 24 l HD8G2CHO HDSG1 HD8G1Ser 5 000000000000000 00000000 1000000000000 0 0 HD8G2 HD8G40 HD4G1 HD4G2Se HD4c40 000000000 0 540 000000000000000 HD8G1 HD8G2 HD8G2Ser HD8G4 HD4G1 HD4G2Se
20. HLA DRI DRB10 150110 0 0 1000000000 13 0 00002000 5 0 00000000000000000000000000000000 080 0 00000000000000000000000000 100000 HLA DRI DRB10 150110 0 0000000000000 13 0000 2000 o A E a 10000000000000000000000000000000 080 0 00000000000000000000000000 HLA DRI DRB10 150110 0 0 1000000000 13 00002000 100000000000000 HLA DRI 100000000000000 a u g o B HLA DRI DRB10 150110 0 0 1000000000 13 0 0000000 a 00000000000000000000000000000000 ED tiy T tn Bul 080 000000000000000 100000000000000000 HLA DRI DRB10 150110 0 0 1000000000 13 10000000000000000000000000000000 000000000000000 au HLA DRI DRB10 150110 0 0 1000000000 13 0 0000000 100 00000000000 00000000000 0000000000 HLA DRI 000000
21. 0 0 4005 10000 0 61 0000000000 16400 0 25 9 37 30 16400 0 0 0000 x 1057 1640 10 96 1000 100 30 1004 7000 0000 0 0 00 0000 0 0 96000000 10 37 co0000000000 Aner sham Phar mci a Bi ot ech 1 Ou Ci 0 10000000000 37 160 200 100 96 Harvester 5 10000 100 3 Printed 100 Bet Scint VWallacD 0000000 1205 JU Uu l HLA DRU 000000100000000000 HDI HD40 HD6 13 20010 100 1100 0 0000 0 00 0000 00 0 100 1101 1006 000000000000 00 HDS J FERM 77 20 730 HD10 FERM 77740 HD40 FERM 77711 FERM 77720 8 l L l 4105000 00000 JU JU lD l rnp Protein 0 8 00000 100000000000000
22. 6 000 0 0000080 10000 0 5 Anersham Pharmacia iotech 60094000000 4 00051 5 2 1006 100 0 0 0 0 0 0 0 0 0 0 0 0 0 0 00 0 1000 0 1000 1000 CD14 00 00 macso O0 00 MB CD140 10000100000 macso 0000 10 0000 014 5019 10 000000000000000000000 200 nt 100 FCSU 1000 0 O 40 1000 100000 7 0000000000 0000 l l 0000000 100010 000 0000000000000 10000000000000 human eukocyte anti gen HLA DRO 000000110 0000000000010 000 1000 H A 00000000 10000000000000 1000 10000000000 000001000 0 1000 O 1000 vitro 0000000000000
23. 100000 068 800000 00000000 MSVSFLI FLPVLGLPWsVLSQVQLQHSGPGLVKPSQTILSLTCAI SGDSVSSNSASVINW RQSPSRGLEW GRTYYRSKW NDYAVSVKSRI VI NPDTSKNQF 52 5 TVTVSSAS 100000 068 800000 5 0000000 10 20 30 50 28 2005 325133 2005 11 24
24. 100000 54 100000 9 0000000 METPAQLLFLLLLVWLPDTTGELVLTQSPGTLSLSPGERATLSCRASQSVSSRYLAW QQKPGQAPRLLI YGASSRATGI DRFSGSGSGTDFTLTI SRLEPEDF AVYYCQQYGSSPLTF GGGTKVEI
25. a DO oana oana naan 0O oaa 14 2005 325133 2005 11 24
26. 19830 80 2713 2717 Jonker et al J Autoi mun 19880 1 399 414 St evens HP et al Transplant 19900 22 1783 1784 100000000 0 10000000 1000000 100000 Billing amp Chatterjee 1983 Transplant Proc 15 649 650 Jonker t al Transplant 19911 23 264 2651 1000 100 O 0 10100 HLA DRO O O 000000 100 O HLA DRI Sheri AK et al Int J 20010 93 556 5650 D HLA DR 0 0000000 100 199 IgA IgE IgM 0 100 19900000000 100 10000000000000000000 HLA 00 11100 Mco 0 0 0 0 0 0 O HLA DRO HLA HLA DRO HLA DRI 0 DRO 100000 00000 000000000000 000000000 100 10000100000000000 0 20 Fab Fv 10000 Fv scFv Applications and Engi neeri of Mono Anti bodi es 1998 T J Internati onal Ltd
27. 10 HLA DR 0 00000000000000000000000 1191 0 1000190 000000000000000000000000 1000 01190 0 1000m 1 oana DO 2 000000000000000000 10000000000000000000000000000000
28. 100000000000000000 10000 10000 5 10000 HLA 10000 0 00 100000 0000 10000 1000000000000000000000000 1000000000000000000000000 1000000000000000000000000 1000000000000000000000000 1000000000000000000000000 Wight et al 19911 Bio Technology 9 830 834
29. TGGAGT CTGGGG GAGGCTTGGTACAGCCTGGGGGGTCCCTGAGACTCTCCTGTGCAGCCTCTGGATTCACCTTTAGCAGCTATGCCATGACC TGGGTCCGCCAGGCTCCAGGGAAGGGGCTGGAGTGGGTCTCAGGTATTAGTGGTGGTGGTGATAGCACATACTACGCAGA CTCCGTGAAGGGCCGGTTCACCATCTCCAGAGACAATTCCAAGAACACGCTGTATCTGCAAATGAACAGCCTGAGAGCCG AGGACACGGCCGTATATTACTGTGCGAGAGATCATGGTTCGGGGAGTTATTATCCCTACTGGTTTGACTACTGGGGCCAG GGAACCCTGGTCACCGTCTCCTCAGCTAGC 100000 04 00000 aa 100000 MEFGLSWFLVAI LKGVQCEVQLLESGGCLVQPGGSLRLSCAASGFTFSSYAMTVWRQAPGKGLEVWSGI SGGGDSTYYA DSVKGRFTI SRDNSKNTLYLQVNSLRAEDTAVYYCARDHGSGSYYPYW DYWGQGTLVTVSSA 100000 10 20 30 50 27 2005 325133 2005 11 24 lt D4 000000 040000000
30. 100000 08 000000000000000 MDMRVPAQLLGLLLLW PGARCAI QLTQSPSSLSASVGDRVTI TCRASQGI 55 YDASSLESGV PSRFSGSGSGTDFTLTI 551 5 0000 4960 10 49700000000 10000000000000000000 000 1000 1520000000 00000000 153
31. HD40 HD6 HD80 HD100 hi 99 HDS HDAD O HD10 2430 000000 L243 100 I 6 10 DRI 10 0 0 0 O HLA DRO O L929 00 0000000 300000 1000000 107 0 0 19290 00 O 19291 0000 1 100000000000000 5 Staining 0 0000000000 60 C 10000300000 l I l 50 100000000000000 30000000000 6009 000000 1004 1 0 000 5 000000000 3 5 2 20 m Q x 5 0 30 1000000 300 0 5 1
32. 50 In 50u 000000000000000 DMM H 00000 DMEM 10 DMEM 20 10000 DEM 0000000000000 100 FCS DMEM DMEM 37 C 1060 0 m SP2 CRL 1581 DMEM 5 1100000000000000000000000000000000 50 w 0000 1500 00000000 mnnn 000 0 37 DMEM 1mp 2 mt DMEM 000 100 FCSD 11 60 10 9 0000 7100000 HAT DMEM HT 10 CS IL 6 DMEM 96 HLA ELI SAD 000000000000000000000000004005000 Cell 54 1000000 HLA 000000 10000000 000000 4 00000000000 19 290 0000000 0 Daudi ATCC 2130 0000000 10000000 2000 1000000000000 5 10000000000000000 19 50u 9 2000000000000 Fcsg 0 PBS 0000 00000000 100 0 20 100 0 5M 0 0 100 1100000000 100000 45017 0000 570 00000000000000014 20 ARVO 5 000000000000000000 0000000 10 20 000 100 5 0 00000000000 961 19620 0 963 9640 2000 50u L The Bi ndi ng Site 000 10000000060 H PBSD 0 0 1
33. 32 2005 325133 2005 11 24 HD40 000000000000 0 130 0000 610 00 730000 10000000 3500 0 0 EMBL EBI 16 1100000 1210000 000 LAS20000 28 HD6 000
34. 6 2005 325133 2005 11 24 1000000000000000 19610 19620 0 19640 3310 0 0 0 0 0 0 19610 1 15 19620 1 gG2Ser 1965400000000 196 11 19 15 O O 19610 O 19620 0 0 o 000 00000 HD80 HD100 HLA DRI 0000000 1000000 D U JUD JU U 10 00000 0000000000 U 10000000000000000000 HLA DRO B DRB1 15011 U U U 0 0 0 00 0 0 00 00 00 00 00 00 00 00 0000 13 21100000000000000 20 29 227000 1991 00 13 A E J lI l 0 0000000 DRO DRB1 1501 1100000000000 000 13 000000000000000000000 100 1000000000000 0 HLA 1000000000 DRO DRB1 150111 000000000000000000 1000000 100 100 30 JUD JJ UJU l 100 0 HLA DRO 0 000 000 JU U SCI D Raj i 1000 1000 000 JUD JD HJ U 000 20900000 0 1u 9 9 0 5 g kg 50u
35. 00000000000000000 100000 10000 12430 000000000 SCI 0 100 00000000000000 100000 ATCC ccL 860 0 0 10 0 100000 570000000 50 00000000 Hugk 100000 900 1000 FCS RPM 1640 0 0 O 80 200 8 100 H H 16400 0 00000 10 0000000000 50 9600 0000000000000 40 3000 0 100 Fcsg 0 H 16400 000000 x 10 0000000000 10000000000000000 99 17 2005 325133 2005 11 24 3 37 C000C00000000000000 10000 1 0u 0 0 0 C0000 161 20000000000000000 1000 1000000000000000000000000000000 0 0 12430 ATCC HB 550 0 0 0 0 000 O FERM
36. Hua et al 1993 9000 1000 10000000000 100000 1000 10000000000 100000 1000 0 100000 Sequences of protein i munol cal interest Publication No 91 3242 3311 00000 0 0 00 ADC 0 cocog 000000000 0 1000000000 0 ADC 1000000000 19620 00 19640 000000000000000000 G1 Igo2 0 0 19615 9 25 11 2005 325133 2005 11 24 19610 15 19620 1 gG2Ser 964 gG1 1962 19610 19615 100000 10 0 0 100000000000000000 J W Godi Mnvcl Anti bodi principles and practice 1993 Academ 55 0 0 0 100000000000000000 0 0 Applications Engi of Anti bodies 1998 Ltd M L 556 ard An
37. 14 12 10000000 138 0 00 0 0 00 00 00 000 0 000 0000 0 00000 1000000000000000000 CHO 1000 ATCC CRL 90960 Katakura Y et al Cytotechnol 31 103 109 1999 29 ATCC CRL 112681 000000000 5 100000 1000000000 0000 i 00 D D O 350 0 5004 FO D 0 D D x 10 CH 0 DO 0 9 culture plate0 00000000000 Pl us 100 10000 100 1000 110000 400000000000000000000000000 000000 80000000000 1 e
38. 10 20 30 50 30 2005 325133 2005 11 24 Anti body 1000000000000 HD4 enent Dependent Cytotoxicity I I O 1000000000000 hl gG 0000000 00000000000 oo0o0g0o000 cr y 0000000 Rajig ATCC ccL 8600 1 Fetal Calf Serum 5 0 0 0 0 0 50 LO 3 7 Cr gt OO DH D D D 10000 100 37900 0 00 1000 1301000000000 Cr5l Dn D 1000 20000 00000000 600000 0000000 200000
39. HLA DRO 0 L9290 0000000000 1929 JJ UJU DU U UD DD U lU HLA 11 HLA 80 0000000000 9 00000000000000000000 B 17 1 CR sci 0000000000 100000000 10u Raj ATCC CCL 86 i 10000000000000000 1000000000 1000000000 100 0000000 DR 100000000 25 2005 325133 2005 11 24
40. B 17 1 O 000000000000000000000000 00000000000 10 000000000000000000 ATCC 86 0 5 10 0000000000 i 0 10000000000 ID000o0 10000ig000000 0000000 1000 HD8G1Ser HD8G2Ser0 0 0 0 0 0 100 99000000000 0000000 100000600 9000000000000000 u 9 head0 1000 1000000000 go Hoc 0 u 1000 31 2005 325133 2005 11 24
41. 50 20 2005 325133 2005 11 24 neo HLA pEF neo HLA DRB O O Li pofect AM 100000000 19291 can Culture Collection U DD 0 000000000000000000 75cm 0 0 0 37 1000000 2400000000 64180000 100000000000000000000065 O0 0 0 0 O DRO O0 O0 000000000000000000 10000000000 1 0 0000000000000 1000 5440 00000000 39480 1000000000000000000000000000000000 100000000 Mtteucci MD and 19810 J Che 3185 391 0 0000000000000000000000000 100000000000000000000000000 20
42. 10 20 30 50 23 2005 325133 2005 11 24
43. 100000 20000 500000000000 0000000 21 100000900000000000000000000000000 21 10000 1520000000000000 00000000000000 5 341 00A ATG 5 0000 4200 0000 20 9007 220 0 LY Ph 100 2300000000 000 0000000000000000000000 00000000 23 129000000000 No 7547 5 to 3 EARS hh 3 GTG GCC GGT GGC GCC TG tnCHNhe gat ggg ccc ttg gtg cta gag gag acg g GCT TGT GGG GC 26 100000 19610 1000 N5KG1 Val O 19640 000 O Lark 0 O 0 O I DEC 50 US patent 60013580 0 0 O 0000000000 NSKGl Va 50 29 2005 325133 2005 11 24
44. 0000000 00 HD8G1Ser HD8G2Ser HD8G1 0 1u 10 0 00 HD8G1Ser HD8G2Ser HD8G1 0 1 Ou 9 HLA DRB HLA r HLA DRB HLA DRI 000 HD6 HLA DRB HLA DR 0 0000 0 0 HS HLA DRB 100 HL 20 90 o N i j i 9 1 i i 2 i i i l 4 T 8 5 L q 5 a bb X 5 A JS z 4 52 m T gt j 1 gt gt gt N gt CO lt H N x N gt UAR 2005 325133 2005 11 24 34 8S1980H W37 1
45. 60 0 5 GGT CTT 3 0000000000 989 6 SMART cDNA amplification 91 3000 5 GTG GGT GGC 3 100000000 98 200 0 0 000 0000 n kit 0000000000000 hh 40 5 GGT GG 3 10000000 1000000000 HD4 0000 tnHD4Sal 5 ata tgt 3 000000000 HD tnHD8Sal 5 ata cTGG 3 PP PE PT 0000 U 0000000 tnCHNhe 5 gat ggg ccc ttg gtg c ta gct acg 9 3 0 0O O 98 O O 60 301 72 30 Sal 10 10000000000000 N5KG 1 Lark I DEC Phar maceuti US patent 6001358 000000 direct 0 10000000000000000 SMART hk 20 5 GIT GCT TGT GGG 3 98 300
46. 24 2005 325133 2005 11 24 96 HLA DR 10 16400 10 5 HLA 8 9 RPM 10 FCSD 0 9 0000000 100 000 HLA DRO 37 otech 1 Ou Ci 10 37 CO 96 SKATRONG Printed Fi Betap 5 peil 1205BETAPLATE IgGI 1990000 12430 ATCC HB 55 2000 40 10 5 50u LO 960 100 L Aner sham Phar 160 20
47. ssuwuwant ust wi TS a Sannoo E os AUIWA c ROSA rw Se wm a se he es ee ES my voavoyoswwa muososswsava 0 oswwavoason shwwayaduon3l awaa soms aso S LAvsdyaongsasy 3owosadansaa lt lt S330NAJAYONAY BONAdANg duAY 1 SaD AYAY 180000 160000 140000 120000 100000 80000 60000 40000 20000 HD6 1
48. 1000000000000000000000000000000000 7110000000 10 Dg000000000000000000000000000000 0 1000000000000000000000000000000000 DDD000000000000000000000000000000 H 20 DDD00000000000000000000000000000 0 100000000000 1 00000 HLA DRO 00000000000000000000000 DR 000 BI 0 JJ j B u u i ul DNA al 0 0 0 O 30 0008000000 CDNAU 0000000000 F neo HLA DRa pEF neo DNAD O HLA DRR DNAD 0 5 HLA 1000000000 5 3 1000000 0 5 c 3 0000000 0 5 CCGGAATTCCTGGTCCTG 3 0 0 0 0000 0 0 5 3 10000000 0 0 40
49. 5 0 e 0 38 E 09 0 9814 5 9 8 7 198dH 1 30 Hgy T 120 1070 52 F 1080H 14 31 O 466 0 AAEH CDC HD8G4 9 025 HD4G4 116 gt 2 HD4G1 1 0 1 0 01 1 0 pg head Raji HERE hg mL e HD8GISer amp 8 25 HD8GI higG 35 0 1hg head RE JP 2005 325133 A 2005 11 24 HD4 30 HD8G1Ser A HD8G2Ser higG
50. 10 20 30 50 2005 325133 2005 11 24 3 N lt gt A El C s O
51. SMART amplification hk 6 5 GGT 98 6 8 purification O0000000000000 hk 60000000000000000000000 tnHD4Bg 5 ata tag atc TGC TCA GAA 3 0000000 HPIOOO0D0000000tn HD8Bgl 5 ata tag atc tGG TCA 000000 O0000000000000000000000 tnCkBsi 5 aag gat ggt gta cgt ttg at 3 PCRO O 980 60 C30 0 72 cDNA Bgl11 1 Lark 000000 direct sequence 00 00 0000000000000 0 DUUUUUUUUUUUUUUD 54 100000 5 0000000
52. 0 6 3 100000000000 1MTris 0 pH 9 00 0000 0 pH 7 22 00 0000000000000000 0 0 00 100000 0 0 O Spectrum Laborator ie 00000 PBS 0 22u m0 0000000000 30 JUD JU U lI 000 0 0 0 0 00 0 0000 0000 00 0 2801 000000 00 w m 01400000000 100 DR 1000000000000000000 HLA DRI 10 0 O 10 9 IL 60 10 Fetal Calf 5 5 SI eRDF a a E A H 9 1000000000000000000 01nM 2 5x 10 mM D D 0 00 0 0 D Low 196 40 10 0 24 00000000000 0000000 00000000 9 00000 00000000000000000000000000 1000 1000000000000000000000000000000000 0000000000000000000000000000in vitrol O0000000000000000000in vitroj00000000 1000000000000000000000000000000 50
53. DO0000000000000 HLA DRI neo HLA DRa HLA DRB O 929 000 00 000000 1000 DAKO 00 00 0 O 00000 DRO 0000000000000000 2929 0000000000 2000000000000 12 2005 325133 2005 11 24
54. g g ee 1 gt gt rT E m m c g E45 lt gt go I o 5 23 AN en C lt O i O 1 O m m m ono c lt lt lt lt lt lt l l E T 4 2005 325133 2005 11 24
55. L 37 C0 00 C o0 0004000000 O00 20000 0000000100 1000 000000 96 1000000 200 Ln 1000 37 c0 00 2 1 50u L 10 00 96 0 96 000000000000 55 00000000000 TopSeal A 96 30 0 30 8 gG1Ser HD8I gG2Ser HD8I HD4c40 0000 0 O HD8I 9610 HD8I gG2 HD4G1 00 00000000000 00 gG1 gG1Ser O 0000000000000 1 5
56. ODD000000000000000000000000000000 0 1000 199000000000000000000000000000 40 0000000000 HLA DRI N 1 2100000000000000000000000000000000 gt g IjUUUUUUUUUUD 10 5 Q ant Human IL 60 10000000 35019 21 2005 325133 2005 11 24
57. sovsasswsvan G stvsasuwwani ssuvuusnuyasi ES wwsunastaSqa IUS messnssranoo Em SZ scranoouws SAHID NOS Fosaa amA 5 JWnmubdlwdo omprshmovya3 TDlSsAmowa309 Svazu ABIDIN TANASI DNAN aaalS9dh39 MNSOdAIOSA Aarsodhd56AS 1Sodhd9aASOAT dkdasnsoATN MoSAsommNHH swsanmINHHDn ASGATINHHOTdO ATnRHHOndou NHHOTGO LXSaAAN HTeolxsdwuA ILODI DiysdNuAoldDy HSAH DNU SALDANA ALIOR 3 SDASSOAL4SSAA A88OA12SSAA9A MDAL4S3AASANH AL3SSAA9A8882 SahAOANHMOAL 3nAOANHaoALGA AQANHaDALGnvy ANHYOAIOAYYYY HaoAtayvavoa DA10AYVSVO3Y 10AyYuYO3 YY vayoanoybs imayoswwiavo osNwwsSyadaon yor LAWS AGGIE unsaon Au SSSONA3A8014
58. 12430 000 O00000000000000000000000 HLA DRO 100 1000 000000000000 0000000 0 12430 ATCC 550 0000 12430 0000 0 12430 000000000000 DR 100000 100 100000000 10000 2 2005 325133 2005 11 24
59. 45 5 0 L L E 0 RRA HHH 4 WNAQKDLLEQRRA 47 WN 5 L EQRRA 36 HD8 JP 2005 325133 A 2005 11 24 xspvsasuyuwaN E seysasyvawaAl VEIEN Id ISYANA swumanidsins EE lt masuwssranoo AsdHaADoLAa SdH3A0O1AA39S gt wwaosadnial IVAN SYAWAL Od AMIANO NATLOILMODNO ATLOILMOONDIT 104LMOONONDL 4MODNONOLSA MOONONDLSAND snonolsAwowm E onolsnmayvyaa 19LSAWDYY330 BYIN IMYA ooNidwanats wbABISGdhdD MA315DdAdSSA ABIS93A29SASD 1S94A39SAS3A7 S3A398ASOATIN AdoSASOnTINHH SSAS3ATINHHO1 nmTINRHDndDJl NHHD10D12SdAA HO38DIXSAAA1A ASJALE AJAMaDANSDALI AdDAUSOALIS3 OANYDALIS3AA SBOA13S3AASANH 5 3SBAASANHSOAL AASANHHOALOA ADANHHOALanwY ANHNOALOAwYary 1 AvysvoawQyDS YuvOaYONOSNM IOXDSNNW3SVQd yDSNwwayadaon SNNw
60. cDNA 50 26 2005 325133 2005 11 24
61. 10000 10000 Ouchterlony O SA 1000000000000000 1000000000000000 00 2 00 00000 1 4 002800 00000000 0 1 0000000000000 0000000 000 000 100000000000000000000 1000000000000000000000 10 20 30 50 15 2005 325133 2005 11 24 10000000000000000000000000000 10000000000000000000000000000 5 10000000000000000000000000000 100 1000000000000000000000000 0000000000000000000000000000000 0000000000000000000000000000000 100 100000 BODY PRODUCTI ON ESSEN 10 AL QUES 1997 W LEY P She
62. 19 JP 2005 325133 A 2005 11 24 4 2005 325133 2005 3251334 43 17 11 24 2005 11 24 51 AG1K 39 395 A61K 39 395 T 4BO24 A61P 35 00 A61P 55 00 4CO85 A61P 37 00 A61P 37 00 4HO45 AG1P 37 08 A61P 37 08 GO1N 33 53 1 33 53 12 OL 6385 21 2005 202038 P2005 202038 71 000253503 22 17 7 11 2005 7 11 62 2004 337448 2004 337448 1 O 1 DAE 4 8 100091096 14 10 159 2002 10 15 31 BARRERES 2001 317054 2001 317054 74 100096183 32 13 10 15 2001 10 15 Et AR 33 AE JP 74 100118773 GAREA 100111741 Et 72 3 lt 54 PH L A DR 5 0000 1000 0 0 0 DRO 1000000000000 0 0 0000000000000000000000
63. 77711 0000 0000 0 0 FERM 77731 000000000000000 FERM 7774 0 0000000000000 1000 10000000000000000000000 2430 1000 000000000000000000000000 100 FCS RPM 16400 0 0 0 0 0 80 200 9 50u LO 100 FCSH H H RPM 16400 000000 190 00 0 D D 000 504 960 00000000000000 C 100 FCS 0 0 RPM 1640 10 100 99 100 L 37 C0 0 0 1 1000000000 37 c0 0 0 160 20 D 100 DRO 243 ATCC HB 55 0000 HD60 RM BP 7772 000000000000000000000000
64. LU U DN 5 0000 790 0000000 50000 5040 00 9 100000000000000 1000 1420000000 5 10 0000 UUUUUD lU 100000 19 10000000000 200000000 Elin 0000000000000000000 UUUUUD 20 100000 1420000000000 0000000000000 5 35 00 00050 4188 0000000400000 1000 00 128 10 00 2000 0000000000000 00210 000000000E 0000000000000000000 10000000 21 128 O HD8D 00000000000000000 JUUUUUUUUUUUUUD 068 800000 9 0000000
65. U LU U Raji 0 5000000000 0000 5ng kg 000 000 0000 HD4 lU U HD8 1000 1000 JU UU DU UU DD DD 0 JJ UU UU DU DU UU Hl HD4 1000 10000 UU DD UD DD Ul 0 0 HD 00000000 JJ UJU DU DD UD U lU HLA DRO l 1100000 JU UJU UU UU DDD UU UU JU DU D Ul HLA DRO DODD0UUUUUUUUUUUUUD HLA DRO 000000000000 10000000000000000 10000000 1000 1000000 10000000 10 20 30 50 9 2005 325133 2005 11 24 0000000000000000000000000000000000 0 000000000000000 00000000010 00000 100000000000000000000 Rosenbaum JT et J Exp 19810 154 1694 1702 et al Proc Natl Acad Sci USA
66. 00 L D DAKO 0 0 00 200 0 5M 1004 1 00000000000000000 450 5701 000000000000000000 1420 00000000 100000000000000000000000000600 10 20 30 50 22 2005 325133 2005 11 24 1 000 0 0000000 00000001991 000000000000000 0000000000060 100000 1000 7 7 7752
67. 00000 T Q H g kg DDD DD JJ DU DDD DDD DDD DDD 0010 10000000000 9000000000000 000000000000000 HLA DR HLA DRO DRB1 15011 1990 0000000000 00 290 0 0 2270 0 0 199 13 J 000000000000 DOO DOO DODO 18 2005 325133 2005 11 24 100 10 HLA DR 71 HLA DR 3500
68. 00000 O HD100 O O0 O 2 HSAD OO0O00000 Rajji I 6000000000000 10000 2090 0 10 00 9 000000000 50 10 H g kg Hoeg JU U UU U 1000000000000000 99100000000 HD8 0 12 0 HD40 HD8 100 10000 10000 HD80 10 9 6 0 10 Fetal Bovine Serum SIGMAU J 0 0 O0 20 SMART cDNA amplification 1000000000000000000000000000 90 total 151 strand cDNAU 00000 Ist strand cDNA 0 0 O0 Total 9 0 5 CDS J H L SMART oligo L 100000 70 0 0 000 0000 30 Buffer H L DIT H L DNTP mx H L Superscript II H L 42 1 500000000000 100 Tricine Buffer 0 0000 72 15 st 40 0 O Takara 0 2 000 0 cDNA uL 10xZ Taq Buffer dNTP mx L 2 u L 10 10 10000 1000 10000000000000 504 1000 0 UMPO SMART
69. 000000 0000 00 100000 000000000000 50000 5000 1000 H 100 0 O 20000 20000 30000 0 3000 0 500 00 0 500 0 0 0 650 660 690 72 000 000 000 0000000 00 000000000000 65 66 69 720 0 00000000000 0 0 HLA 100000000 000000 0 0 Pan HLA DR E Innnnnnn HLA 100 100000000 000000000 ARH77 ATCC CRL 1621 Daudi ATCC CCL 213 HS 54 ATCC CRL 1484 M 9 ATCC CCL 159 CAR ATCC CRL 8083 CCL 86 CRL 1596 L ATCC 2261 SKV 4 ATCC TI 215 DRB1 150110 0000000 0000 ATCC CCL 1 10000 Ul 00000 00000 l ull 10000 500 0 150 0 UUUUUD HLA DRI RPM 8226 ATCC CCL 155 I m gt g
70. 000000000000 0000000000000 0000000000 100000 HDI 0000 10000 G 100000000000 HD8 7773 HD1o0 00 0 FERM BP 77741 20011 100 1110000 0 10001001 10000000000000 HD4 7771 20 1000000 1101 ERM 77720 0 O 20011 100 110000000000 100000 0 0 10000000100100 1006000 100000000000000000 100000000000000000 19610 19620 19630 0 0 0 1964 620 1963 19640 l 10000 gG1 19620 1963 404 0 1961 464 100000000000000 U 0 Sequences of pro ological interest Publication 91 3242 331
71. 0000000000000 00000000000 000000 30 3100000000000 100000 1000000 0 100000000000000 100000 1000000 0 100000000000000 100000 18 0 0000 P3X63Ag8U 10 P3 U10 D E et al Current Topics in M crobiology and m unol ogy 81 1 7 1978 P3 NSI 1 Ag4 10 NS 1 Kohler G et al European J munol ogy 6 511 519 19760 O Sp2 14 SP 20 Shul et al Nat ure 276 269 270 1978 P3X63Ag8 6530 653 Kearney J F et al J mu 123 1548 15500 19790 6 X63 K and Harris W Nature 256 495 497 19750 1000000000000 40 000000000 8 0000 1000 1002 0 0000000000000 1000000000000 5 RPM 16401 0 0 8 0 0000 10000001000000 10000000 5 5 Modified 5 u Dul becco Modified Eag 0 0 0 D M 000000 10030 400000000 0000 100 D M O 0000000000000 2 1070000000000 10000 100000 1400000 100000000000000000000000000000000000000 50 13 2005 325133 2005 11 24
72. 243 1243 10000000 100 5 1640 8u 50 100 FcCSH RPM 16400 x 100 nt 100 100000 50 0 96 4 lt 0 30000000000 100 50 0 H RPM 16400 100 00000000000000000000000000600 00 99 1000 L 37 50 3 Hg 1 Ou Ci 0 37 0 160 20 100000000000000000 HLA DRO 0 L2430 ATCC HB 55 100000 10000000000 FERM 77730 0 HD10 FERM 77740 00000000 77710 HD6 FERM 77720 00000000000000000000 961 33100000000 HD80 O0 H 0 0 HD8 615 0 0 0 0 0 0 0 331 Se 2 962 EUD 0 33100000000 0 000 0 0 000 54625 0 1000 10000000000 00000000 1000000000000000000 rHsi 0000 FERM 77730 0 HD100 00 0 O FERM 77744 00000000 0000 FERM 77710 6 0 0 FERM 77720 00000000000000000000 HLA DPO 00000000000000000000000000000000600 00000000000000000000000000000000000000 8 2005 325133 2005 11 24
73. 3yQda9131 AAAavaddon3ln 2 ad3DAOSQJHAS OS038ASS3DNA3 CsMAS33ONAdA AsaaDNAJABOT S33DNAdAHOnd 3ONA3A8O148AN NASABOT38A931 JAYOTANAYILON YTA ONA HUMIL INSHO Ab3l9NadH23 aloNadHo3ayd 9NddHD3aXdOA DUO de ll 300000 250000 200000 150000 100000 50000 0 37 2005 325133 2005 11 24 10000 2005325133000001 38 2005 325133 2005 11 24 000000000 16 28 1000 15 00 000 1509 1000 1628 72 10 10000000000 0000000000000000 00 48024 01 11 2 15 BM6 BAO 02 EAM HA 40085 14 1 023 0561 5 90 BAO 2 D50 6 22 28 5 51 FA2 5
74. 8 NAJANOTJUAY3L AUSA ONEAN BALONA MIDNA 19834423540 2 A aaydownabdet ydDwndHdaJOD 2 8 5 61 62 63 64 65 66 67 68 69 70 71 72 73 HD4 HD6 HD8 24 WN 5 0 F LEDRRA 25 5 0 DE EF ERR RA 6 SQ KD KD EFL 27 Ww NS 0 LE 28 WN 5 0 4 RA 2 WN 50 38 5 0 31 WNS ED 32 WN 5 Q KRA 33 YN 5 Q AR 34 K L EQRRA 35 WNS 0 1 36 WNS OKDLL EQKRG 37 WN 5 L EDRREIA 38 5 Q K 39 W N 5 Q K DI L E DE RA 4 o WN SQ KD 41 WN 5 PD LEE RA 2 0 1 0 43 WNS OKDL R RSG 48 WNSQADLLE QO RRA 1 49 WN 5 0 EK A L 50 5 51 W N SQ EQ RA A 44 5 L L QR RA
75. ERM BP 45771 19941 20 22 110000 00000000000000000 DRO 4 200 100 95 25 1 5hr0 0 0 0 15072 0 hr 0 1000000 HD40 HD6 00 0 O HLA DRB HLA DRO DRB1 15011 1990 00000000000 100 20000 0 2271 199 100 100000100 13 940 0 0 Reineke 2001 Epitope napping wit h synthetic peptides prepared SPOT synthesis Antibody Spri nge Manual Eds Dubel 433 4599 1000000000 Lu nager TM nger i 00000000000000000000000 61 73 17 29 63 75 65 77 B 61 73 0 570 0 69 590 0 71 0000000000000 0 HD80 61 73 63 75 65 77 HLA Dessen et Al 1997 7 473 481 61 73
76. b 00 000000000000 1000000000000000000 10000000000000011000 00000000 81 10 0 000000000000 i t uxi nab 1000000000000 0000 0 0000000000000 000 0000000000 101000 7150 00000000000 Il M Th Th 100000000 5 MLaughlin P al J Clin Oncol 1998 16 2825 2833 Coiffier et Blood 1998 92 1927 81 Babbin et al Nature 19850 317 359 361 et al cs 19800 11 617 625 JT et al J Exp 19811 154 1694 1702 et al Proc Natl Acad Sci USAQ 19831 80 2713 2717 5 2005 325133 2005 11 24 Jonker et al J Autoi mun 19880 1 399 414 Stevens et al Transplant 19900 22 1783 1784 Billing 6 Chatterjee 5 19830 Transplant Proc 15 649 6 Jonker et al Transplant 19911 23 264 265 DOO00000000000000000000000000U0U0000
77. g kg 9 body 50u g kg 0 1u g b 40 0 SH g kg 0 0 00 6 SCI D 10u 0 ATCC 86 x 10 0 100000 10 5000000 50 9 0 0 O 5 9 00000 10000 5 0000000000000000000000 10 FCSH RPM 1640 1000 O00 00 50 100 RPM 1640 100 0 000000 000001000 50 7 2005 325133 2005 11 24
78. neo HLA DRa neo HLA DRB 0 O O HLA DRa 94 15 0 55 300 0 72 600 x 300 100 EcoRI Not l 1000000000000000000 pEF neo 0 0 pEF BOS M zushi S 6 Nagata S Nucleic Acids Res 19900 18 53320 neonyci n 10000000000000000000000000 neo HLA DRa neo HLA DRB 0 pEF neo HLA DRa 0 O 765bp cDNA 000 0 O O pEF neo 8016 0 cDNAU 100000000000000 29000000000 1000 97000 000000000000000000000000 10000 HLA
79. pherd Dean Anti bodies 20 00 OXFORD VERSI PRESS J W Godi ng Anti bodi es pri and pr acti ce 1993 PRESS 1000 100 1000 100 100 1000 100 FR1 FR20 4 Sequences of proteins of i interest Publication 91 32 20 42 HLA 0 1243 0 000 0 0 0 0 0 0 0000 HLA 12430 ATCC HB 55 243 243 0 DNAD 900 000000000000 95000000000000000 97 30 30 10 25 J Sanbr ook Molecular Clonin A Laboratory Mnnual Second Edition Cold Spring Harbor Laboratory Press 1989 90011000
80. ti body of Cancer 1998 Marcel Dekker
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