6Q NQECNK\G VJG KOOWPQINQDWNKP +I DKPFKPI FQOCKPU QH VJG JWOCP O[GNQKF +I# (Eα TGEGRVQT (Eα4

Hele tekst

(1)IgA binds CD89 via EC-1. ,PPXQRJOREXOLQELQGLQJ 6LWHV RI +XPDQ )Fα5, &'

(2) DQG %RYLQH )Fγ5 $UH /RFDWHG LQ WKHLU 0HPEUDQHGLVWDO([WUDFHOOXODU'RPDLQV# *%TCKI/QTVQP

(3) )GTXCP<CPFDGTIGPT%GGUXCP-QQVGPT%JTKU,*QYCTFh,CP ),XCPFG9KPMGNgCPF2GT$TCPFV\CGI

(4)

(5) &GRCTVOGPVU. QH +OOWPQJKUVQEJGOKUVT[ CPF +OOWPQRCVJQNQI[ 6JG 0CVKQPCN *QURKVCN 7PKXGTUKV[ QH 1UNQ 1UNQ 0QTYC[ .GKFGP 7PKXGTUKV[ /GFKECN %GPVGT .GKFGP 6JG 0GVJGTNCPFU h&GRCTVOGPVU QH +OOWPQNQI[ CPF 2CVJQNQI[ 6JG +PUVKVWVG HQT #PKOCN *GCNVJ %QORVQP 7PKVGF -KPIFQO g&GRCTVOGPV QH +OOWPQNQI[ CPF /GFCTGZ 'WTQRG 7PKXGTUKV[*QURKVCN7VTGEJV7VTGEJV6JG0GVJGTNCPFU. T&GRCTVOGPV QH 0GRJTQNQI[. 6800$5<. 6Q NQECNK\G VJG KOOWPQINQDWNKP +I DKPFKPI FQOCKPU QH VJG JWOCP O[GNQKF +I# (Eα TGEGRVQT (Eα4+ %& CPF VJG DQXKPG (Eγ TGEGRVQT D(Eγ4 EJKOGTKE TGEGRVQTU YGTG IGPGTCVGF D[ GZEJCPIKPIEQORCTCDNGTGIKQPUDGVYGGP VJGUG VYQ RTQVGKPU (Eα4+ CPF D(Eγ4 CTG JKIJN[ JQOQNQIQWU CPF CTG OQTG ENQUGN[ TGNCVGF VQ GCEJ QVJGT VJCP VQ QVJGT JWOCP CPF DQXKPG (E4U 0GXGT VJGNGUU VJG[ CTG HWPEVKQPCNN[ FKUVKPEV KP VJCV (Eα4+ DKPFU JWOCP +I# J+I# CPFPQVDQXKPG+I) D+I) YJGTGCU D(Eγ4 DKPFU D+I) CPF PQV J+I# (Eα4+ CPF D(Eγ4 RQUUGUU GZVTCEGNNWNCT TGIKQPU EQPUKUVKPI QH VYQ +INKMG FQ OCKPU C OGODTCPGFKUVCN GZVTCEGNNWNCT FQOCKP '% C OGODTCPGRTQZKOCN '% FQOCKP '% C VTCPUOGODTCPG #. TGIKQP CPF C UJQTV E[VQRNCUOKE VCKN %JKOGTCU EQPUVTWEVGF D[ GZEJCPIKPI EQORNGVG FQOCKPU DGVYGGP VJGUG VYQ TGEGRVQTU YGTG VTCPUHGEVGF VQ %15 EGNNUCPFCUUC[GFHQTVJGKTCDKNKV[VQDKPF J+I# QT D+I)EQCVGF DGCFU 6JG TG UWNVU UJQYGF VJCV VJG +IDKPFKPI UKVG QP DQVJ (Eα4+ CPF D(Eγ4 KU NQECVGF YKVJKP'%5WRRQTVKPIVJKUQDUGTXCVKQP OQPQENQPCN CPVKDQFKGU VJCV DNQEMGF +I#DKPFKPI VQ (Eα4+ YGTG HQWPF VQ TGEQIPK\G GRKVQRGU NQECVGF KP VJKU FQ OCKP +P VGTOU QH (E4+I KPVGTCEVKQPU EJCTCEVGTK\GF VJWU HCT VJKU NQECVKQP KU WPKSWG CPF UWTRTKUKPI DGECWUG KV JCU DGGP UJQYP RTGXKQWUN[ VJCV NGWMQE[VG (Eγ4UCPF(Eε4+DKPF+IXKCUKVGURTKPEK RCNN[ NQECVGF KP VJGKT '% FQOCKPU. J Exp Med 189:1715, 1999. 33.

(6) Chapter 2. ,1752'8&7,21 +OOWPQINQDWNKP +I (E TGEGRVQTU (E4U GZRTGUUGF QP RJCIQE[VKE EGNNU RTQXKFG C ETWEKCN NKPM DGVYGGP VJG JWOQTCN CPF EGNNWNCT DTCPEJGU QH VJG KOOWPG U[UVGO .KICVKQP QH (E4 D[ CPVKIGPDQWPF +I NGCFU VQ EGNNWNCT CEVKXCVKQP CPF VTKIIGTKPI QH RQYGTHWNGHHGEVQTOGEJCPKUOU +P JWOCPU CPF QVJGT OCOOCNU +I# RTGFQOKPCVGU KP OWEQUCG CPF HWTVJGTOQTG EQORTKUGU C UWDUVCPVKCN RTQRQTVKQP QH VJG EKTEWNCVKPI +I RQQN #V OWEQUCN UWTHCEGU +I# RTQXKFGU C HKTUVNKPG RTQVGEVKXG HWPEVKQP VGTOGF KOOWPG GZENWUKQP YJGTGD[ KV KPJKDKVU OKETQDKCN EQNQPK\CVKQP QP GRKVJGNKCN EGNNU CPF RGPGVTCVKQPQHJCTOHWNCPVKIGPU+PCFFKVKQPVJGRTQVGEVKXGHWPEVKQPQH+I#DQVJKP OWEQUC CPF KP VJG EKTEWNCVKQP OC[DG TGKPHQTEGF D[ KPVGTCEVKQP QH +I#EQORNGZGF CPVKIGPUYKVJVJGO[GNQKF(Eα4+ %& (Eα4+KUGZRTGUUGFQPOQPQE[VGU OCETQRJCIGU RQN[OQTRJQPWENGCT ITCPWNQE[VGU CPF GQUKPQRJKNU CPF KVU ETQUUNKPMKPI VTKIIGTU C XCTKGV[ QH KOOWPQNQIKECN GHHGEVQT HWPEVKQPU KPENWFKPI RJCIQE[VQUKU CPVKDQF[FGRGPFGPV EGNNWNCT VQZKEKV[ CPF TGNGCUG QH KPHNCOOCVQT[ OGFKCVQTUCPFE[VQMKPGU #OQPI (E4U EJCTCEVGTK\GF WPVKN PQY (Eα4+ KU OQUV ENQUGN[ TGNCVGF VQ VJG DQXKPG(ETGEGRVQTHQT+I) D(Eγ4 GZRTGUUGFQPOQPQE[VGUCPFITCPWNQE[VGU +PHCEVVJGUGVYQ(E4UCTGOQTGENQUGN[TGNCVGFVQGCEJQVJGTVJCPVQCP[MPQYP JWOCP QT DQXKPG (E4U /QTG TGEGPVN[ KV JCU DGGP UJQYP VJCV (Eα4+ CPF D(Eγ4 CTG OGODGTU QH C PGY IGPG HCOKN[ VJCV CRRCTGPVN[ GXQNXGF HTQO C EQOOQP CPEGUVTCN IGPG 1VJGT JWOCP IGPGU DGNQPIKPI VQ VJKU HCOKN[ KPENWFG VJG PCVWTCN MKNNGT EGNN KPJKDKVQT[ TGEGRVQTU -+4U VJG +INKMG VTCPUETKRVU +.6U VJG NGWMQE[VGCPFOQPQE[VGOCETQRJCIG+INKMGTGEGRVQTU .+4U/+4U .#+4CPF */ 6JGUG IGPGU CTG NQECVGF ENQUG VQ VJG (Eα4+ IGPG YKVJKP VJG UQ ECNNGF NGWMQE[VG TGEGRVQT EQORNGZ QP EJTQOQUQOG S 5GXGTCN OWTKPG OGODGTU QH VJG UCOG IGPG HCOKN[ IR$ C UVTWEVWTCN JQOQNQIWG QH JWOCP -+4U CPF VJG RCKTGF +INKMG TGEGRVQTU # CPF $ 2+4# CPF 2+4$ JCXG CNUQDGGPFGUETKDGF (Eα4+ CPF D(Eγ4 CTG DQVJ VTCPUOGODTCPG IN[EQRTQVGKPU EQORQUGF QH VYQ GZVTCEGNNWNCT '% +INKMG FQOCKPU '% CPF '% C VTCPUOGODTCPG TGIKQP EQPVCKPKPI C EJCTIGF CTIKPKPG TGUKFWG CPF C UJQTV E[VQRNCUOKE VCKN FGXQKF QH UKIPCNKPI OQVKHU 5KIPCN VTCPUFWEVKQP XKC (Eα4+ KU OGFKCVGF XKC VJG (E4 γ EJCKPYJKEJCUUQEKCVGUYKVJ(Eα4+VJTQWIJVJGEJCTIGFCTIKPKPGTGUKFWGYKVJKPKVU VTCPUOGODTCPGFQOCKPDWVFQGUPQVCHHGEVKVUCHHKPKV[HQT+I# &GURKVGVJG JKIJNGXGNQHCOKPQCEKFKFGPVKV[ YKVJKPVJG'%CPFVTCPUOGODTCPGTGIKQPU QH (Eα4+ CPF D(Eγ4 VJGUG VYQ TGEGRVQTU CTG HWPEVKQPCNN[ SWKVG FKUVKPEV KP VJCV (Eα4+ DKPFU +I# DWV PQV D+I) YJGTGCU D(Eγ4 DKPFU D+I) DWV PQV +I# 34.

(7) IgA binds CD89 via EC-1. 6JGTGHQTGVQOCRVJGNKICPFDKPFKPIFQOCKPUQHVJGUGVYQ(E4UYGWVKNK\GFVJGKT JKIJ FGITGG QH KFGPVKV[ CPF GZEJCPIGF JQOQNQIQWU TGIKQPU DGVYGGP VJGO $CUGF QPMPQYNGFIGQHKPVGTCEVKQPUDGVYGGPQVJGTVYQFQOCKP(E4U (Eγ4++(Eγ4+++CPF (Eε4+ YKVJVJGKTTGURGEVKXGNKICPFU +I)QT+I' YGGZRGEVGFVJG+IDKPFKPIUKVGU VQ DG NQECVGF YKVJKP VJG OGODTCPGRTQZKOCN '% FQOCKP 5WTRTKUKPIN[ JQYGXGTQWTTGUWNVUFGOQPUVTCVGFVJCVVJGNKICPFDKPFKPITGIKQPQHDQVJ(Eα4CPF D(Eγ4 KU NQECVGF KP VJGKT OGODTCPGFKUVCN '% FQOCKPU +P RCTV VJKU HKPFKPI RTQDCDN[ TGHNGEVU VJG GXQNWVKQPCT[ FGXGNQROGPV QH (Eα4+ CPF D(Eγ4 HTQO CP CPEGUVTCNIGPGFKUVKPEVHTQOVJGRWVCVKXG(Eγ4(Eε4RTGEWTUQT. 0$7(5,$/6$1'0(7+2'6 &HOOFXOWXUH %15EGNNUYGTGOCKPVCKPGFKP&/'/ $KQ9JKVVCMGT UWRRNGOGPVGFYKVJ (%5O/.INWVCOKPGCPFµIONIGPVCO[EKP .KHG6GEJPQNQIKGU 7- 6JG OWTKPG ++# $ EGNN NKPG VJCV EQGZRTGUUGU (Eα4+ CPF VJG (E4 γEJCKP JCU DGGP FGUETKDGF F'1$VDQGFRQVWUXFWLRQRIFKLPHULF)F5V E&0#UGPEQFKPIVJGEQORNGVG(Eα4+EQFKPITGIKQPCPFCOWVCPVE&0#GPEQFKPI C UQNWDNG HQTO QH (Eα4+ YGTG IKHVU HTQO &T % /CNKU\GYUMK +OOWPGZ %QTR 5GCVVNG 9# E&0# HQT D(Eγ4 JCU DGGP FGUETKDGF RTGXKQWUN[ %JKOGTKEE&0#UYGTGEQPUVTWEVGFD[QXGTNCRGZVGPUKQP2%4 #NVJQWIJVJG IGPQOKEUVTWEVWTGQHD(Eγ4KUWPMPQYPVJGJKIJJQOQNQI[VQ(Eα4+CNNQYGFWU VQ KPHGT KPVTQPGZQP DQWPFCTKGU DGECWUG COKPQ CEKF TGUKFWGU VJCV NKPM VJG (Eα4+ GZQPUCTGKFGPVKECNVQVJQUGCVEQORCTCDNGRQUKVKQPUKPD(Eγ4 (KIWTG 2TKOGTU YGTGVJWUFGUKIPGFVQCNNQYVJGHWUKQPQHGZQPUCVVJGUGTGUKFWGU6QEQPUVTWEVVJG. D'% (Eα4+OWVCPVRTKOGTUYGTGFGUKIPGFVQCNNQYVJGHWUKQPQHVJGHKTUV COKPQ CEKFU QH D(Eγ4 VQ (Eα4+ CV KUQNGWEKPG DQVJ (Eα4+ CPF D(Eγ4 JCXG KUQNGWEKPG TGUKFWGU CV VJKU RQUKVKQP YJKEJ NKGU CNOQUV GZCEVN[ KP VJG OKFFNG QH VJG '% FQOCKPU 6JG KPVGITKV[ QH CNN EJKOGTKE E&0#U YCU EQPHKTOGF D[ UGSWGPEG CPCN[UKU %JKOGTKE (E4 E&0#U YGTG ENQPGF KPVQ VJG R%&0# OCOOCNKCP GZRTGUUKQPXGEVQT +PXKVTQIGP6JG0GVJGTNCPFU RTKQTVQVTCPUHGEVKQP 7UDQVIHFWLRQV %15EGNNUYGTGVTCPUKGPVN[VTCPUHGEVGFYKVJµIEJKOGTKE(E4E&0#EQPUVTWEVU D[ OGCPU QH (WIGPG VTCPUHGEVKQP TGCIGPV $QGJTKPIGT /CPPJGKO )GTOCP[ CEEQTFKPI VQ VJG OCPWHCEVWTGTNU KPUVTWEVKQPU +P UQOG GZRGTKOGPVU µI R%/8. 35.

(8) Chapter 2. )(2 YCU EQVTCPUHGEVGF VQIGVJGT YKVJ VJG (E4 EQPUVTWEVU %GNNU YGTG KPEWDCVGF CV u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s EGNN UQTVGT. $GEVQP&KEMKPUQP CPFOKZGFYKVJ+IEQCVGF&[PCDGCFUKPCHKPCNXQNWOGQHµN RGT YGNN KP 8DQVVQOGF OKETQVKVGT RNCVGU #HVGT C OKP KPEWDCVKQP CV TQQO VGORGTCVWTGVJGRNCVGYCUURWPCVIHQTOKPCPFKPEWDCVGFHQTCPCFFKVKQPCN OKPCVTQQOVGORGTCVWTG%GNNUCPFDGCFUYGTGTGUWURGPFGFCPFGZCOKPGFHQT VJGRTGUGPEGQHTQUGVVGUWUKPICEQODKPCVKQPQHNKIJVCPFHNWQTGUEGPVOKETQUEQR[ KPC0KMQP'ENKRUG'OKETQUEQRG4QUGVVGUYGTGFGHKPGFCU)(2 EGNNUDKPFKPI HQWTQTOQTG+IEQCVGFDGCFUCPFCVNGCUV)(2 %15EGNNUYGTGEQWPVGFHQT GCEJ FGVGTOKPCVKQP (QT DNQEMKPI UVWFKGU EGNNU YGTG KPEWDCVGF YKVJ GKVJGT O#D /[ µNEWNVWTGUWRGTPCVCPV QT%%) µNCUEKVGUHNWKFFKNWVGF HQT OKPCVTQQOVGORGTCVWTGDGHQTGVJGCFFKVKQPQH+IEQCVGFDGCFU 3URGXFWLRQDQGSXULILFDWLRQRIUHFRPELQDQWVROXEOH&' #E&0#VJCVGPEQFGUCUQNWDNGHQTOQH(Eα4+YCUGZRTGUUGFKP%JKPGUGJCOUVGT QXCT[EGNNUD[OGCPUQHVJGR''GZRTGUUKQPU[UVGO .QP\C7- CPFVJGRTQVGKP YCU KUQNCVGF HTQO VJG EWNVWTG UWRGTPCVCPV D[ CHHKPKV[ EJTQOCVQITCRJ[ YKVJ 5GRJCTQUGDQWPFJWOCP+I# XCP<CPFDGTIGP)CPF%XCP-QQVGPWPRWDNKUJGF FCVC 0RQRFORQDODQWLERGLHV 6JGRTGXKQWUN[FGUETKDGF(Eα4+O#DNU/[ OWTKPG+I/ ###CPF # CNNOWTKPG+I) YGTGWUGFKPVJKUUVWF[ /[CPF#YGTGIKHVU HTQO&T.K5JGP &CTVOQWVJ/GFKECN5EJQQN.GDCPQP0* CPF&T/CZ%QQRGT. 7PKXGTUKV[ QH #NCDCOC $KTOKPIJCO #. TGURGEVKXGN[ # YCU UWRRNKGF D[ /GFCTGZ 'WTQRG 6JG 0GVJGTNCPFU CPF # CPF # YGTG RWTEJCUGF HTQO 36.

(9) IgA binds CD89 via EC-1. +OOWPQVGEJ (TCPEG CPF 4GUGCTEJ &KCIPQUVKEU +PE TGURGEVKXGN[ 6JG D(Eγ4 O#D %%) OWTKPG +I/ YCU IGPGTCVGF D[ KOOWPK\KPI OKEG YKVJ D(Eγ4 RTQVGKP RWTKHKGF HTQO ECVVNG NGWMQE[VGU CPF VJG URGEKHKEKV[ YCU EQPHKTOGF D[ UVCKPKPI%15EGNNUVTCPUHGEVGFYKVJE&0#GPEQFKPIVJGD(Eγ4QTDQXKPG(Eγ4++. *QYCTF %, WPRWDNKUJGF FCVC 6Q QDVCKP PGY (Eα4+ O#DU HGOCNG $#.$% OKEG YGTG KOOWPK\GF YKVJ RWTKHKGF UQNWDNG (Eα4+ 5RNGPQE[VGU KUQNCVGF HTQO KOOWPK\GFCPKOCNUYGTGHWUGFYKVJO[GNQOCEGNNU 52 KPVJGRTGUGPEGQH RQN[GVJ[NGPGIN[EQN6JGEGNNUWURGPUKQP YCU FKNWVGF KP +/&/ UWRRNGOGPVGF YKVJ (%5 J[RQZCPVJKPG µOQN COKPQRVGTKP µ/ VJ[OKFKPG µ / RION +. 7NKVGT RGPKEKNNKP CPF µION UVTGRVQO[EKP %GNNU RTQFWEKPICPVKDQFKGUVQ(Eα4+CPVKDQFKGUYGTGUWDENQPGFD[NKOKVKPIFKNWVKQP(KXG ENQPGURTQFWEKPIO#DVQ(Eα4+YGTGGZRCPFGFCPFVJGURGEKHKEKV[YCUFGVGTOKPGF D[(#%5sCPCN[UKU UGGDGNQY 6QFGHKPGVJGECRCEKV[QHVJGUGPGYO#DVQKPJKDKV DKPFKPIQH+I#VQ(Eα4+DNQEMKPIUVWFKGUYGTGRGTHQTOGFCUHQNNQYU(Eα4+O#DU QT EQPVTQN O#DUQH VJG UCOG KUQV[RG YGTG FKNWVGF KP (#%5 DWHHGT CPF KPEWDCVGF VQIGVJGT YKVJ (Eα4+VTCPUHGEVGF ++# EGNNU HQT OKPWVGU CV u% 2WTKHKGF JWOCPUGTWO+I#YJKEJJCFRTGXKQWUN[DGGPJGCVCIITGICVGFHQTJQWTCVu%. C+I# YCU VJGP CFFGF HQT J CV u% %GNNU YGTG YCUJGF CPF DQWPF +I# YCU FGVGEVGFD[KPEWDCVKQPYKVJCIQCVCPVKJWOCP+I#( CD 2'RQN[ENQPCNCPVKDQF[ EQPLWICVG 5QWVJGTP $KQVGEJPQNQI[ CUUQEKCVGU +PE WUKPI (#%5 s CPCN[UKU +UQV[RG EQPVTQN CPVKDQFKGU HQT OWTKPG +I) YGTG RWTEJCUGF HTQO $GEVQP &KEMKPUQP CPF VJQUG HQT OWTKPG +I/ YGTG RTQXKFGF D[ &T 4QDGTV $WTPU 5EQVVKUJ #ITKEWNVWTCN 5EKGPEG#IGPE['FKPDWTIJ7- )$&6 DQDO\VLV %GNNU Z YGTGYCUJGFVYKEGYKVJ(#%5DWHHGT 2$5$5#C\KFG CPFKPEWDCVGFYKVJGKVJGT(Eα4+O#D OWTKPG +I/ QT+I) EWNVWTG UWRGTPCVCPV QT VJG CRRTQRTKCVG KUQV[RG EQPVTQN UWRGTPCVCPV HQT J CV u% %GNNU YGTG VJGP YCUJGFVYKEGYKVJ(#%5DWHHGTCPFKPEWDCVGFHQTJYKVJGKVJGTIQCVCPVKOQWUG. )#/ +I/(+6%EQPLWICVG HKPCNFKNWVKQP QTC)#/+I)2'EQPLWICVG. HKPCN FKNWVKQP DQVJ HTQO 5QWVJGTP $KQVGEJPQNQI[ #UUQEKCVGU +PE +P GZRGTKOGPVUYJGTG)(2 EGNNUYGTGCPCN[\GFHQTEJKOGTKE(E4GZRTGUUKQPC)#/ +I) 6TKEQNQT UGEQPFCT[ TGCIGPV HKPCN FKNWVKQP %CNVCI .CDU YCU WUGF #HVGT YCUJKPI VYKEG YKVJ (#%5 DWHHGT EGNNU YGTG HKZGF KP 2$5DWHHGTGF . YVXQN RCTCHQTOCNFGJ[FG CV u% CPF CPCN[\GF QP (#%5ECPs &CVC CESWKUKVKQP YCU EQPFWEVGF YKVJ .[UKU ++ UQHVYCTG $GEVQP &KEMKPUQP CPF FCVC CPCN[UKU YCU RGTHQTOGFWUKPI9KP/&+UQHVYCTG CXCKNCDNGHTQO6JG5ETKRRU4GUGCTEJ+PUVKVWVG .C,QNNC%# . 37.

(10) Chapter 2. 5(68/76 (&RI&'DQGE)Fγ5PHGLDWHOLJDQGELQGLQJ 6Q OCR VJG NKICPFDKPFKPI FQOCKPU QH (Eα4+ CPF D(Eγ4 YG IGPGTCVGF HKXG EJKOGTKE TGEGRVQTU CU HQNNQYU J'%D(Eγ4 EQPUKUVKPI QH VJG VYQ '% FQOCKPU QH (Eα4+HWUGFVQVJGVTCPUOGODTCPGE[VQRNCUOKE 6/% FQOCKPQHD(Eγ4D'% (Eα4+ VJG VYQ DQXKPG '% FQOCKPU HWUGF VQ 6/% FQOCKP QH (Eα4+ J'% D(Eγ4 VJG '% FQOCKP QH (Eα4+ HWUGF VQ '%6/% TGIKQP QH D(Eγ4 D'% (Eα4+ VJG '% FQOCKP QH D(Eγ4 LQKPGF VQ '%6/% TGIKQP QH (Eα4+ CPF D'% (Eα4+ VJG HKTUV COKPQ CEKFU QH D(Eγ4 HWUGF CV KUQNGWEKPG VQ (Eα4+ (KIWTG Figure 1. Alignment of amino acid sequences of CD89 29 Fcα αRI and bFcγγ2R (Resp. bFcγ2R 29 X54154-Z37506). S1 exons are shown from the CD89 AIREAYLTQLMIIKNSTYREIGRRLKFWNETDPEFVIDHMDANKAGRYQC 79 methionine initiation cobFcγ2R GPPNTKSFS*NKEGD**PWN*HPS*EP*DKA--N*F*SNVREQQ****H* 77 dons. Because the gene EC2 structure of bFcγ2R is unCD89 QYRIGHYRFRYSDTLELVVTG------LYGKPFLSADRGLVLMPGENISL 123 known, intron-exon bFCγ2R SHF**VNWSEP*EP*D*L*A*EEPAGR*RDR*S**VRPSPSVA****VT* 127 boundaries are shown only for FcαRI. Amino acids CD89 TCSSAHIPFDRFSLAKEGE----LSLP-QHQSGEHPANFSLGPVDLNVSG 167 that link two FcαRI exons bFcγ2R L*Q*-GNRT*T*L*S***AAHRP*R*RS*D*D*WYQ*E***S**TSAHG* 176 are underlined, and the TM/C exon designation is shown CD89 IYRCYGWYNRSPYLWSFPSNALELVVTDSIHQDYTTQNLIRMAVAGLVLV 217 above the sequence. Note bFcγ2R T****RSLSTN***LSQ**EP*A*L*A*-----**M******GL*AS**L 221 that exon-linking amino acids are conserved beCD89 ALLAILVENWHSHTALNKEASADVAEPSWSQQMCQPGLTFARTPSVCK 265 tween FcαRI and bFcγ2R. bFcγ2R L*GIL*CQAR*D*GGA-R**ARS 243 The first 19 amino acids of both sequences are considered to represent NH2terminal signal peptides, which are removed before cell surface expression. Thus, glutamine (Q) 22 is proposed to be the first amino acid of both mature proteins and is therefore designated +1. Isoleucine (I) is also underlined. Residues of bFcγ2R identical to those of FcαRI are designated (*), and gaps that have been inserted to line up the sequences are designated (-), (also see reference 7). S1 S2 +1 EC1 MDPKQTTLLCLVLCLGQRIQAQEGDFPMPFISAKSSPVIPLDGSVKIQCQ *A*TLPA****G*SV*L*T*V*A*T**K*I*W*EP*S*V**GS**T*L**. 6QIGVJGT YKVJ YKNFV[RG (Eα4+ CPF D(Eγ4 E&0#U KPFKXKFWCN EJKOGTKE (E4 EQPUVTWEVU YGTG VTCPUHGEVGF VQ %15 EGNNU CPF VJGKT EGNN UWTHCEG GZRTGUUKQP YCU CUUGUUGF D[ (#%5s CPCN[UKU CPF D[ C URGEKHKE DKPFKPI CUUC[ WUKPI +IEQCVGF DGCFU. UGG /CVGTKCNU CPF /GVJQFU +PKVKCN GZRGTKOGPVU TGXGCNGF VJG VTCPUHGEVKQP GHHKEKGPE[QHKPFKXKFWCNEQPUVTWEVUVQDGSWKVGXCTKCDNG6JGOQUVGHHKEKGPVEQPUVTWEV FKTGEVGFGZRTGUUKQPQH(Eα4+QPVJGUWTHCEGQH`QH%15EGNNUYJGTGCUVJG NGCUVGHHKEKGPV D'%(Eα4+ YCUGZRTGUUGFD[QPN[QHVJGVTCPUHGEVCPVU (KIWTG # 6JGTGHQTG YG FGXGNQRGF C OGVJQF VQ UGNGEVKXGN[ GPTKEJ HQT VTCPUKGPVN[ VTCPUHGEVGF EGNNU D[ EQVTCPUHGEVKPI C RNCUOKF VJCV FKTGEVGF GZRTGUUKQP QH )(2 38.

(11) A. IgA binds CD89 via EC-1. XKUWCNK\GFD[ITGGPHNWQTGUEGPEG VQIGVJGTYKVJVJGEJKOGTKE(E4EQPUVTWEVU/QUV )(2 %15 EGNNU EQVTCPUHGEVGF YKVJ (Eα4+ YGTG TGEQIPK\GF D[ O#D # . (KIWTG$ CPFHQTOGFTQUGVVGUYKVJJ+I#EQCVGFDGCFU (KIWTG% B. A 1%. 0.2%. 62%. FSC-height. FcαRI. Control. 30%. 3%. A62. FcαRI. bEC1-FcαRI. C. Green fluorescent protein (GFP). 120. Figure 2. Expression of chimeric FcR by COS-1 cells. (A) COS-1 cells were transfected with the indicated constructs 2 d before har80 vesting and FACS® analysis. Cells were stained with either FcαRI EC2-specific mAb 60 A62 (mouse IgG1) (bottom panels) or an appropriate isotype control (top panels), followed 40 by a GAM IgG1 Tricolor reagent. Numbers in the top right corners of the plots refer to the 20 percentage of positive cells. (B) Enrichment of FcαRI expression in COS-1 cells cotransfected 0 with GFP. Cells transfected with both FcαRI Total cells GFP + cells GFP + cells and GFP were stained with FcαRI EC2-specific counted forming rosettes mAb A62 followed by GAM IgG1 Tricolor reagent and analyzed by FACS®. More than 60% of the GFP+ cells also express FcαRI. (C) Rosette formation by COS-1 cells cotransfected with both FcαRI and GFP and exposed to hIgA-coated beads. Rosettes were quantified as specified in Materials and Methods. Black bar, total number of cells counted; white bar, total number of GFP+ cells assessed by fluorescent microscopy; gray bar, number of GFP+ cells forming rosettes. Results shown are representative of three separate experiments. Cell number. 100. $[ VJKU RTQEGFWTG YG QDVCKPGF CP CRRTQZKOCVGN[ VYQHQNF GPTKEJOGPV QH EJKOGTKE (E4GZRTGUUKPI EGNNU YJKEJKP VJG ECUG QH (Eα4+ TGUWNVGF KP QH EGNNU DGKPI TGCEVKXGYKVJ(Eα4+O#DCPFHWTVJGTCDNGVQHQTOTQUGVVGUYKVJJ+I#EQCVGFDGCFU. (KIWTG $ CPF % +V UJQWNF DG PQVGF VJCV )(2 %15 CNOQUV PGXGT HQTOGF TQUGVVGU OQUV NKMGN[ DGECWUG VTCPUHGEVCPVU GZRTGUUKPI (Eα4+ CNQPG CEEQWPVGF HQT QPN[ ` QH VJG VQVCN EGNNU (KIWTG $ 9G HWTVJGTOQTG FGOQPUVTCVGF VJG. 39.

(12) Chapter 2. % inhibition of rosette formation. URGEKHKEKV[ QH QWT DKPFKPI CUUC[ D[ WUKPI DNQEMKPI O#DU URGEKHKE HQT (Eα4+ QT D(Eγ4 VQ KPJKDKV TQUGVVG HQTOCVKQP (KIWTG 6JG HKPFKPI VJCV VJG KPJKDKVKQP QDVCKPGF YKVJ /[ YCU QPN[ RCTVKCN ` YCU OQUV NKMGN[ GZRNCKPGF D[ VJG WUG QH EWNVWTG UWRGTPCVCPV QVJGT VJCP C JKIJGT EQPEGPVTCVKQP QH 100 RWTKHKGF CPVKDQF[ YJKEJ YCU PQV CXCKNCDNG 80 60 40 20. FcR transfected Coating Ig. +C CG2 4 +c on IgM trol. Blocking Ab. +M y4 3 +c on IgM trol. 0. FcαRI. bFcγ2R. hIgA. bIgG2. Figure 3. Specificity of the bead rosetting assays. COS-1 cells were cotransfected with the indicated FcR construct together with pCMV-GFP. GFP+ cells were purified as described in Materials and Methods and incubated with the relevant murine IgM-blocking mAb or an irrelevant murine IgM control mAb for 30 min before addition of Igcoated beads. Results are shown, as percentages of inhibition of rosette formation when compared with transfectants that were not incubated with mAb before rosetting analysis. Results shown are representative of two experiments.. (Eα4+FQGUPQVDKPFD+I)CPFD(Eγ4FQGUPQVDKPFJ+I# CEEQTFKPIN[YG PGKVJGT QDUGTXGF TQUGVVGU YJGP J+I#EQCVGF DGCFU YGTG OKZGF YKVJ D(Eγ4 VTCPUHGEVCPVU PQT YJGP D+I)EQCVGF DGCFU YGTG OKZGF YKVJ (Eα4+ VTCPUHGEVCPVU. (KIWTG $KPFKPI UVWFKGU YKVJ %15EGNNU GPTKEJGF HQT (E4 GZRTGUUKQP CU FGUETKDGF CDQXG UJQYGF PQV WPGZRGEVGFN[ VJCV YKNFV[RG (Eα4+ CPF D(Eγ4 VTCPUHGEVCPVU RTQFWEGF VJG JKIJGUV NGXGNU QH TQUGVVG HQTOCVKQP YKVJ J+I# CPF D+I)EQCVGF DGCFU TGURGEVKXGN[ (KIWTG 6TCPUHGEVCPVU GZRTGUUKPI EJKOGTCU EQFKPIHQTVJGGPVKTG'%RQTVKQPUQHVJGTGEGRVQTU J'%D(Eγ4CPFD'%(Eα4+ DQWPFVJGKTTGURGEVKXG+IEQCVGFDGCFUGHHKEKGPVN[CNVJQWIJCVCUNKIJVN[NQYGTNGXGN VJCP VJGKT YKNFV[RG EQWPVGTRCTVU (KIWTG 6JG HCEV VJCV VJG J'%D(Eγ4 EJKOGTCTGVCKPGF+I#DKPFKPIECRCEKV[FGOQPUVTCVGFVJCVVJGDKPFKPIUKVGQH(Eα4+ NKGU YKVJKP VJG OGODTCPGFKUVCN '% FQOCKP (WTVJGTOQTG DGECWUG VJKU EJKOGTC FKFPQVHQTOTQUGVVGUYKVJD+I)EQCVGFDGCFUQWTHKPFKPIHWTVJGTUWIIGUVGFVJCV VJG DKPFKPI UKVG HQT D+I) KP D(Eγ4 YCU PQV NQECVGF YKVJKP VJG '% FQOCKP QH VJKUTGEGRVQT%QPXGTUGN[VJGD'%(Eα4+EJKOGTCFKFHQTOTQUGVVGUYKVJD+I) EQCVGFDGCFUDWVPQVYKVJDGCFUEQCVGFYKVJJ+I# #NVQIGVJGT VJGUG TGUWNVU UJQYGF VJCV KP EQOOQP YKVJ (Eα4+ VJG NKICPF DKPFKPIUKVGQHD(Eγ4CRRGCTGFVQNKGYKVJKPVJG'%FQOCKP+VUJQWNFDGPQVGF JQYGXGT VJCV VJG NGXGN QH DKPFKPI QDVCKPGF YKVJ VJG VYQ '% EJKOGTCU J'%. 40.

(13) IgA binds CD89 via EC-1. D(Eγ4 CPF D'%(Eα4+ YCU TGFWEGF YJGP EQORCTGF YKVJ VJG YKNFV[RG TGEGRVQTUCPFVJG'%EJKOGTCU J'%D(Eγ4CPFD'%(Eα4+ (KIWTG (WTVJGTOQTG VQ DGVVGT NQECNK\G VJG +IDKPFKPI UKVGU QH VJGUG VYQ TGEGRVQTU C HWTVJGT EJKOGTC YCU EQPUVTWEVGF KP YJKEJ VJG HKTUV COKPQ CEKFU QH VJG '% FQOCKPYGTGHTQOD(Eγ4YJKNGVJGTGOCKPKPI'% COKPQCEKFU CPFVJGTGUV QH VJG TGEGRVQT YGTG HTQO (Eα4+ =D'% (Eα4+? #NVJQWIJ VJKU EJKOGTC YCU GZRTGUUGF CV VJG EGNN UWTHCEG CPF EQWNF DG TGEQIPK\GF D[ VJG OCLQTKV[ QH (Eα4+ O#DKVDQWPFPGKVJGTJ+I#PQTD+I)EQCVGFDGCFU. cDNA. Structure. % rosette formation 0. FcαRI. 20. 40. 60. 80. 20. 40. 60. 80. S EC1 EC2 TM/C. hEC-bFcγ2R hEC1-bFcγ2R bFcγ2R bEC-FcαRI bEC1-FcαRI bEC1(1-50)-FcαRI hIgA-coated beads. bIgG2-coated beads. Figure 4. Rosette formation by FcR/GFP cotransfected COS-1 cells. Schematic representation of wild type or chimeric FcRs. Unshaded regions are derived from FcαRI, shaded regions from bFcγ2R. S, signal peptide; EC1, extracellular domain 1; EC2, extracellular domain 2; TM/C, transmembrane - cytoplasmic tail. GFP+ transfectans were purified from COS-1 cells cotransfected with GFP and FcR constructs as described. Ig-binding to FcRs coexpressed with GFP was assessed by rosetting with either hIgA- or bIgG2-coated beads. More than 200 cells were counted for each determination, and the number of cells binding four or more Ig-coated beads is expressed as percentage rosette formation. Results are representative of three separate experiments.. (&VSHFLILFDQWLERGLHVEORFN&'ELQGLQJRI,J$ 6Q EQPHKTO VJCV VJG +I#DKPFKPIUKVG QH (Eα4+ NKGU YKVJKP VJG '% FQOCKP YG OCRRGFVJGURGEKHKEGRKVQRGUHQTCPWODGTQHDNQEMKPICPFPQPDNQEMKPIO#DU1H. 41.

(14) Chapter 2. VJGRTGXKQWUN[FGUETKDGF(Eα4+O#DUQPN[/[ OWTKPG+I/ YCUCDNGVQDNQEM VJG DKPFKPI QH J+I# VQ (Eα4+ (QWT QVJGTU # # # CPF # CNN OWTKPG +I) FKF PQV KPJKDKV DKPFKPI 9G CNUQ KPENWFGF C PWODGT QH PGY (Eα4+O#DU '&)*CPF&CNNOWTKPG+I) TCKUGFCICKPUV C UQNWDNG HQTO QH (Eα4+ 6JGUG O#DU YGTG UJQYP VQ DG URGEKHKE HQT (Eα4+ D[ TGCEVKQPYKVJ++#EGNNUGZRTGUUKPIVJKUTGEGRVQT (KIWTG# A A77. 100 101. 2E6. 102 103 104 100 101. 2D11. 102 103 104 100 101. 7G4. 102 103 104 100 101. 102 103 104 100 101. 2H8. 102 103 104 100 101. 7D7. 102 103 104. B. 100 101. C. 2E6 + aIgA. 102 103 104 100 101. 2D11 + aIgA. 102 103 104 100 101. 7G4 + aIgA. 102 103 104 100 101. 102 103 104 100 101. 2H8 + aIgA. 102 103 104 100 101. 7D7 + aIgA. 102 103 104. 7D7 CC -G2 4. aIgA. 2H8. 2D1 1 7G4. 2E6. A77. A59 A62. My4 3 A3. Figure 5. FACS® analysis cDNA of newly produced mAbs to Fcα αRI compared to + + + + + + + + + + FcαRI mAb A77 of similar specificity. A. FcαRI-expressing + + + + + + + + + + hEC-bFcγ2R murine B cells (IIA1.6) were + + + + + incubated with mAbs as inhEC1-bFcγ2R dicated (white peaks or with - + bFcγ2R concentration and isotype– matched control mAbs - + bEC-FcαRI (hatched peaks), followed by a GAM IgG1-PE con- + + + + bEC1-FcαRI jugate. B. FcαRI-expressing + + + + + +/ - + - +/ - + bEC1(1-50)-FcαRI IIA1.6 cells were incubated without (left panel) or with mAbs as indicated, followed by heat-aggregated hIgA (aIgA). After 1 h at 4°C, cells were washed and bound hIgA was detected by incubation with goat anti-human IgA F(ab)2-PE conjugate (white peaks). Cells incubated with only the secondary reagent served as negative controls (hatched peaks). C. Reactivity of chimeras (left) with a panel of FcαRI and bFcγ2R mAb (top) as measured by FACS® analysis. Binding was graded as follows: +, strong binding; +/-, weak binding; -, no binding.. 6JG[ YGTG PGZV CUUC[GF HQT CDKNKV[ VQ DNQEM DKPFKPI QH JGCVCIITGICVGF J+I# VQ (Eα4+O#DU'&)CPF*RTQFWEGFUWEJKPJKDKVKQPYJKNG&FKF 42.

(15) IgA binds CD89 via EC-1. PQV (KIWTG$ #EEQTFKPIN[YGRTGUWOGFVJCVVJGDNQEMKPIO#DUYQWNFRTQXG VQDG'%URGEKHKEYJGTGCUVJGPQPDNQEMKPIQPGUYQWNFDG'%URGEKHKE6QVGUV VJKUJ[RQVJGUKUYGUETGGPGFVJGTGCEVKXKV[QHCNNO#DUCICKPUVVJGRCPGNQHEJKOGTKE (E4 GZRTGUUGF KP %15 EGNNU D[ (#%5s CPCN[UKU +PFGGF CNN O#DU ECRCDNG QH DNQEMKPIVJGDKPFKPIQHJGCVCIITGICVGFJ+I#VQ(Eα4+ (KIWTG$ OCRRGFVQVJG '%FQOCKP (KIWTG% #NUQCNNPQPDNQEMKPIO#DUYGTGFKTGEVGFCICKPUVVJG'%FQOCKPGZEGRVHQT O#D # VJCV CRRCTGPVN[ TGEQIPK\GF CP GRKVQRG FGRGPFKPI QP RCTVU QH DQVJ FQOCKPU 7PHQTVWPCVGN[ DGECWUG QPN[ QPG O#D CICKPUV D(Eγ4 YCU CXCKNCDNG C UKOKNCT FGVCKNGF UVWF[ EQWNF PQV DG RGTHQTOGF HQT VJKU TGEGRVQT 9G UJQYGF VJCV VJGO#D%%)QPN[TGEQIPK\GFYKNFV[RG D(Eγ4 CPF VJG D'%(Eα4+ EJKOGTC. (KIWTG% 6JWUNKMGO#D#KVKUOQUVNKMGN[FKTGEVGFCICKPUVCEQPHQTOCVKQPCN GRKVQRGFGRGPFKPIQPDQVJ'%CPF'%. ',6&866,21 $[OGCPUQHCRCPGNQHEJKOGTKE(E4UYGKFGPVKHKGFEQPENWUKXGN[HQTVJGHKTUVVKOG VJGNKICPFDKPFKPIUKVGUQH(Eα4+CPFD(Eγ45WTRTKUKPIN[VJGUGUKVGUYGTGHQWPF VQDGNQECVGFKPVJG'%FQOCKPU(Eα4+CPFD(Eγ4CTGJKIJN[JQOQNQIQWUDQVJ CV VJG RTQVGKP CPF PWENGQVKFG NGXGN CPF KFGPVKV[ TGURGEVKXGN[ DWV UJQY OWEJ NGUU JQOQNQI[ YKVJ QVJGT JWOCP CPF DQXKPG (E4U 6JKU UWIIGUVU VJCV (Eα4+ CPF D(Eγ4 GXQNXGF HTQO C EQOOQP CPEGUVTCN IGPG CNUQ UJCTGF D[ -+4+.6/+4.+4.#+4*/2+4CPFIR$IGPGU CPFPQVUJCTGFD[ QVJGT JWOCP QT DQXKPG (E4U /CRRKPI QH VJG D(Eγ4 IGPG VQ DQXKPG EJTQOQUQOGYJKEJEQTTGURQPFUVQJWOCPEJTQOQUQOGHWTVJGTUWRRQTVU VJKUPQVKQP 1WT HKPFKPI VJCV VJG +IDKPFKPI UKVGU QH DQVJ (Eα4+ CPF D(Eγ4 CTG NQECVGF YKVJKPVJGKT'%FQOCKPUYCUDCUGFQPVJGHCEVVJCVTQUGVVKPIYKVJ+IEQCVGFDGCFU YCUQPN[UGGPHQTVJGEQTTGURQPFKPI(Eα4+QTD(Eγ4'%EJKOGTC+PDQVJECUGU JQYGXGTCTGFWEVKQPKPDKPFKPICEVKXKV[YCUUGGPEQORCTGFYKVJVJCVQDVCKPGFHQT VJGYKNFV[RGTGEGRVQTUCPFHQTVJGEQORCTCDNG'%EJKOGTCU (KIWTG #NVJQWIJ VJGUG FKHHGTGPEGU KP RCTV OC[ DG CVVTKDWVGF VQ VJG GZRTGUUKQP NGXGNU QH KPFKXKFWCN EQPUVTWEVU GURGEKCNN[HQTVJGD'%(Eα4+EJKOGTCUGGCDQXG KVKUCNUQRQUUKDNG VJCVVJG'%FQOCKPUCPFOGODTCPGRTQZKOCNTGIKQPUQHVJGTGEGRVQTUEQPVTKDWVG GKVJGTFKTGEVN[ D[HQTOKPIPUGEQPFCT[QEQPVCEVUKFGU QTKPFKTGEVN[ D[RTGUGTXKPI VJTGGFKOGPUKQPCNUVTWEVWTG VQVJGCHHKPKV[ CPF UVCDKNKV[QH VJG NKICPF KPVGTCEVKQPU 6JKUYQWNFDGCPCNQIQWUVQVJGCEVKXKV[QHQVJGTVYQFQOCKP(E4UPCOGN[(Eγ4++ (Eγ4+++CPF(Eε4+KPYJKEJVJGNKICPFDKPFKPIUKVGUCTGNQECVGFKPVJGOGODTCPG. 43.

(16) Chapter 2. RTQZKOCN '% FQOCKPU YJGTGCU UVTWEVWTGU YKVJKP VJG '% FQOCKPU EQPVTKDWVG VQ VJGDKPFKPIRTQEGUU +V UJQWNF CNUQ DG PQVGF VJCV VJG TGIKQP QH J+I# KPVGTCEVKPI YKVJ (Eα4+ JCU TGEGPVN[DGGPOCRRGFVQVJG%α%αDQWPFCT[ 6JKUKUKPEQPVTCUVVQVJG TGIKQPQHJWOCP+I)TGURQPUKDNGHQTKPVGTCEVKQPYKVJ(Eγ4UYJKEJKURTQRQUGFVQ NKGOWEJENQUGTVQVJGJKPIG 6JGTGHQTGKPVGTOUQHVJGGXQNWVKQPQH+I(E4 KPVGTCEVKQPU KV UJQWNF DG KPVGTGUVKPI VQ OCR VJG TGIKQP QH D+I) VJCV DKPFU VQ D(Eγ4 $GECWUG TGEQODKPCPV D+I) KU CXCKNCDNG GZRGTKOGPVU ECP TGCFKN[ DG FGUKIPGF VQ FGVGTOKPG YJGVJGT VJKU TGIKQP NKGU ENQUG VQ VJG JKPIG TGIKQP CU KP JWOCP+I)QTKPCRQUKVKQPCPCNQIQWUVQVJCVQHJ+I# 6QUWDUVCPVKCVGQWTQDUGTXCVKQPVJCV+I#DKPFUVQVJG'%FQOCKPQH(Eα4+ YGOCRRGFVJGGRKVQRGUQHCRCPGNQHDNQEMKPICPFPQPDNQEMKPICPVK(Eα4+O#DU VJCVDQWPFGSWCNN[VQVJG'%RCTVUQHYKNFV[RG(Eα4+CPFVJGJ'%(Eγ4EJKOGTC 6JGPQPDNQEMKPIO#DU ###CPF& YGTGUJQYPVQTGCEVYKVJVJG OGODTCPGRTQZKOCN'%FQOCKPDGECWUGVJG[DQWPFQPN[VQYKNFV[RG(Eα4+CPF VJG J'%D(Eγ4 D'%(Eα4+ CPF D'% (Eα4+ EJKOGTCU 6JG QPN[ GZEGRVKQP YCU VJG PQPDNQEMKPI O#D # VJCV DQWPF QPN[ VQ YKNFV[RG (Eα4+ CPF VJG J'% D(Eγ4 CPF D'% (Eα4+ EJKOGTCU UWIIGUVKPI VJCV KVU GRKVQRG KU EQPHQTOCVKQPCN CPF FGRGPFU QP TGIKQPU QH DQVJ '% CPF '% UKOKNCT VQ VJG D(Eγ4O#D%%)UGG4GUWNVUUGEVKQP +PEQPVTCUVCNNDNQEMKPIO#DU /[ '&)CPF* YGTGUJQYPVQTGCEVYKVJVJG'%FQOCKPQH(Eα4+ DGECWUG VJGKT DKPFKPI CEVKXKV[ YCU TGVCKPGF YKVJ VJG J'%D(Eγ4 EJKOGTC 6JG GRKVQRGU TGEQIPK\GF D[ /[ CPF & YGTG HWTVJGT NQECNK\GF VQ VJG TGIKQP QH '% FKTGEVN[ CFLCEGPV VQ '% DGECWUG VJG[ YGTG UJQYP VQ DKPF VJG D'% (Eα4+ EJKOGTC ' CPF * QP VJG QVJGT JCPF DQWPF QPN[ YGCMN[ VQ VJKU EJKOGTC YJGTGCU ) FKF PQV DKPF CV CNN 5KOKNCT OCRRKPI UVWFKGU YKVJ DNQEMKPI O#DU JCXG RTGXKQWUN[ DGGP WUGF VQ NQECNK\G VJG +I)DKPFKPI UKVGU QH (Eγ4++ CPF (Eγ4+++VQVJGKT'%FQOCKPU # PWODGT QH (Eα4+ O40#U JCXG DGGP KUQNCVGF CPF UJQYP VQ GPEQFG URNKEG XCTKCPVUQHVJGTGEGRVQT 1PGUWEJTGRQTVFGUETKDGFEGNNUWTHCEGGZRTGUUKQP QHCP(Eα4+XCTKCPVVJCVNCEMGFVJGEQORNGVG'%FQOCKPCPFUWIIGUVGFVJG'% FQOCKPVQDGKPXQNXGFKPJ+I#DKPFKPI *QYGXGTKPEQPVTCUVVQQWTTGUWNVU O#D /[ YCU RTQRQUGF VQ TGCEV YKVJ '% CPF O#D # YKVJ '% 9G DGNKGXG VJCV QDUGTXCVKQP VQ DG URWTKQWU GKVJGT FWG VQ KPEQTTGEV EGNN UWTHCEG GZRTGUUKQPQH VJG URNKEG XCTKCPV CPFQT CDGTTCPV TGEGRVQT UVTWEVWTG ECWUGF D[ NCEM QH VJG EQORNGVG '% FQOCKP 6JKU RQUUKDKNKV[ YCU UWRRQTVGF D[ QWT CVVGORVU VQ GZRTGUU XCTKQWU (Eα4+ URNKEG XCTKCPVU KP %15 EGNNU YKVJ PQ UWEEGUU #FFKVKQPCNN[EJKOGTCUEQPUVTWEVGFDGVYGGP(Eα4+CPF(Eγ4++YGTGPQVGZRTGUUGF. 44.

(17) IgA binds CD89 via EC-1. GHHKEKGPVN[ /QTVQP *% CPF ,), XCP FG 9KPMGN WPRWDNKUJGF QDUGTXCVKQPU RQUUKDN[TGHNGEVKPICFGITGGQHUVTWEVWTCNKPEQORCVKDKNKV[DGVYGGPVJGUGVYQ(E4U+P HCEV QWT WPUWEEGUUHWN GZRGTKGPEG YKVJ VJQUG CRRTQCEJGU NGF WU VQ EQPUVTWEV EJKOGTCU DGVYGGP (Eα4+ CPF D(Eγ4 CU TGRQTVGF JGTG DGECWUG VJGKT NGXGNU QH JQOQNQI[ CPFJGPEGRTGUWOCDN[VJGKTQXGTCNNUVTWEVWTG CTGOQTGUKOKNCTVJCPHQT QVJGT (E4U 6JWU UYCRRKPI QH JKIJN[ JQOQNQIQWU TGIKQPU UJQWNF JCXG OKPKOCN CHHGEV QP VJG QXGTCNN UVTWEVWTCN KPVGITKV[ QH VJG TGUWNVCPV EJKOGTCU 6JGTGHQTG YG HGGNVJCVVJGRTGUGPVCRRTQCEJKUOQTGRJ[UKQNQIKECNVJCPRTGXKQWUCVVGORVUVQVJKU GPF 6JG UWTRTKUKPI FKHHGTGPEG UGGP DGVYGGP VJG NKICPFDKPFKPI UKVGU QH (Eα4+ CPF D(Eγ4 XGTUWU VJQUG QH QVJGT NGWMQE[VG (Eγ4U CPF (Eε4+ OC[ JCXG KPVGTGUVKPI KORNKECVKQPU KP VGTOU QH +I KPVGTCEVKQPU #U OGPVKQPGF CDQXG VJKU FKURCTKV[ EQWNF UKORN[TGHNGEVVJGRTQRQUGFGXQNWVKQPQH(Eα4+CPFD(Eγ4HTQOCPCPEGUVTCNIGPG FKUVKPEVHTQOVJCVIKXKPITKUGVQQVJGT(Eγ4UCPF(Eε4+6JKUPQVKQPKUUWRRQTVGFD[ VJG QDUGTXCVKQP VJCV TGUKFWGU YKVJKP VJG OGODTCPGFKUVCN FQOCKP QH VYQ -+4 RTQVGKPUFGVGTOKPGVJGKTCDKNKV[VQDKPFVQVJGKTTGURGEVKXGNKICPFUVJGVYQITQWRU QH *.#% CNNQV[RGU /QTGQXGT FWG VQ VJGKT JKIJ NGXGNU QH JQOQNQI[ VJG VJTGGFKOGPUKQPCNUVTWEVWTGQH(Eα4+CPFD(Eγ4OC[OQTGENQUGN[TGUGODNGVJCV QH VJG -+4 RTQVGKPU VJCP VJCV QH OQTG FKUVCPVN[ TGNCVGF (E4U +PFGGF OQTG FGVCKNGF OWVCVKQPCN CPCN[UKU FKTGEVGF D[ OQFGNKPI UVWFKGU WUKPI VJG TGEGPVN[ RWDNKUJGFVJTGGFKOGPUKQPCNUVTWEVWTGQHVJGR-+4CUCVGORNCVGHQTVJGRTQVGKP DCEMDQPGU QH (Eα4+ CPF D(Eγ4 CTG EWTTGPVN[ WPFGTYC[ KP QWT NCDQTCVQT[ VQ HWTVJGTNQECNK\GVJG+IDKPFKPIUKVGUYKVJKPVJGUGVYQTGEGRVQTU #P CNVGTPCVKXG GXQNWVKQPCT[ GZRNCPCVKQP RQUUKDN[ CRRNKECDNG CV NGCUV HQT (Eα4+ OKIJV DG VJCV KVU NKICPFDKPFKPI UKVG FGXGNQRGF VQ GPUWTG KPVGTCEVKQP YKVJ CNN OQNGEWNCT HQTOU QH +I# OQPQOGTKE +I# FKOGTKE +I# KPENWFKPI ,EJCKP CPF UGETGVQT[ +I# KPENWFKPI ,EJCKP CPF UGETGVQT[ EQORQPGPV (Eα4+ KU TGRQTVGF VQ DKPF CNN VJGUG NKICPF XCTKCPVU 6JGTGHQTG DGECWUG VJG UKVG QH KPVGTCEVKQP YKVJ(Eα4+CVVJG%α%αDQWPFCT[CRRGCTUVQDGCEEGUUKDNGVQVJGTGEGRVQTKP CNNVJGUGHQTOUQH+I#(Eα4+EQWNFJCXGGXQNXGFVQCEEQORNKUJVJKUKPVGTCEVKQPXKC KVU '% FQOCKP VQ CXQKF RQVGPVKCN RTQDNGOU QH UVGTKE JKPFTCPEG QH C OQTG OGODTCPGRTQZKOCNDKPFKPIUKVGKPTGNCVKQPVQNCTIG+I#RQN[OGTU +PEQPENWUKQPYGJCXGUJQYPVJCVVJGENQUGN[TGNCVGF(Eα4+CPFD(Eγ4DKPF VJGKT NKICPFU XKC UKVGU NQECVGF KP VJGKT OGODTCPGRTQZKOCN '% FQOCKPU 6JG FKHHGTGPEG KP VJG +IDKPFKPI UKVGU QH VJGUG VYQ TGEGRVQTU XGTUWU QVJGT NGWMQE[VG (Eγ4UCPF(Eε4+OC[TGHNGEVVJGRTQRQUGFFKXGTIGPVGXQNWVKQPCT[RCVJYC[HTQOC FKUVKPEV IGPGVKE RTGEWTUQT QT CV NGCUV KP VJG ECUG QH (Eα4+ C URGEKHKE CFCRVCVKQP HQTGHHKEKGPVKPVGTCEVKQPYKVJNCTIGOQNGEWNCTHQTOUQH+I#. 45.

(18) Chapter 2. 5()(5(1&(6 4CXGVEJ,8CPF,2-KPGV(ETG EGRVQTU#PPW4GX+OOWPQN. UQOGQHYJKEJDKPFVQ/*%ENCUU+OQNGEWNGU, +OOWPQN. &CGTQP / (E TGEGRVQT DKQNQI[ #PPW4GX+OOWPQN. #TO , 2 % 0YCPMYQ CPF - ( #WUVGP /QNGEWNCT KFGPVKHKECVKQP QH C PQXGN HCOKN[ QH JWOCP +I UWRGTHCOKN[ OGODGTU VJCV RQUUGUU KOOWPQTGEGRVQT V[TQUKPGDCUGF KP JKDKVKQP OQVKHU CPF JQOQNQI[ VQ VJG OQWUG IR$ KPJKDKVQT[ TGEGRVQT , +OOWPQN . /QTVQP*%/XCP'IOQPFCPF,), XCPFG9KPMGN5VTWEVWTGCPFHWPEVKQPQH JWOCP+I#(ETGEGRVQTU (ECNRJC4 %TKV4GX +OOWPQN 5JGP.4GEGRVQTUHQT+I#QPRJCIQ E[VKEEGNNU+OOWPQN4GU /QPVGKTQ 4 % * -WDCICYC CPF / & %QQRGT%GNNWNCTFKUVTKDWVKQPTGIWNCVKQP CPFDKQEJGOKECNPCVWTGQHCP(ECNRJCTGEGRVQT KPJWOCPU,'ZR/GF /QPVGKTQ 4 % 4 9 *QUVQHHGT / & %QQRGT , 4 $QPPGT ) . )CTVNCPF CPF * -WDCICYC&GHKPKVKQPQHKOOWPQINQDWNKP # TGEGRVQTU QP GQUKPQRJKNU CPF VJGKT GPJCPEGF GZRTGUUKQP KP CNNGTIKE KPFKXKFWCNU , %NKP +PXGUV <JCPI),4;QWPI%#6TGICUMGU2 5QRR CPF % , *QYCTF +FGPVKHKECVKQP QHCPQXGNENCUUQHOCOOCNKCP(EICOOCTGEGR VQT,+OOWPQN &WRQPV$#5GNXCMWOCTCPF75VGHHGPU 6JG MKNNGT EGNN KPJKDKVQT[ TGEGRVQT IGPQ OKE TGIKQP QP JWOCP EJTQOQUQOG S 6KUUWG#PVKIGPU 5COCTKFKU,CPF/%QNQPPC%NQP KPIQHPQXGNKOOWPQINQDWNKPUWRGTHCOKN[TGEGR VQTUGZRTGUUGFQPJWOCPO[GNQKFCPFN[ORJQKF EGNNU UVTWEVWTCN GXKFGPEG HQT PGY UVKOWNCVQT[ CPF KPJKDKVQT[ RCVJYC[U 'WT , +OOWPQN %QUOCP & 0 (CPIGT . $QTIGU / -WDKP 9 %JKP . 2GVGTUQP CPF / . *UW #PQXGNKOOWPQINQDWNKPUWRGTHCOKN[TG EGRVQT HQT EGNNWNCT CPF XKTCN /*% ENCUU + OQNG EWNGU+OOWPKV[ $QTIGU . / . *UW 0 (CPIGT / -WDKPCPF&%QUOCP#HCOKN[QHJW OCP N[ORJQKF CPF O[GNQKF +INKMG TGEGRVQTU. 46. 9CIVOCPP 0 5 4QLQ ' 'KEJNGT * /QJTGPYGKUGT CPF ' 1 .QPI # PGY JWOCPIGPGEQORNGZGPEQFKPIVJGMKNNGTEGNNKP JKDKVQT[ TGEGRVQTU CPF TGNCVGF OQPQ E[VGOCETQRJCIGTGEGRVQTU%WTT$KQN -TGOGT', 8 -CNCV\KU ' $CMGT & ( %CNNGP ) 4 5WVJGTNCPF CPF % 4 /CNKU\GYUMK 6JG IGPG HQT VJG JWOCP +I#(ETGEGRVQTOCRUVQS*WO)GPGV 9KNUQP / / 6QTMCT CPF , 6TQYUFCNG 6JG NGWMQE[VG TGEGRVQT EQORNGZ QP JW OCP EJTQOQUQOG S +OOWPQNQI[ #DUVTCEV #TO , 2 / ( )WTKUJ & 5 4G[PQNFU * % 5EQVV % 5 )CTVPGT - ( #WUVGP CPF *4 -CV\ /QNGEWNCT ENQPKPI QH IR C EGNNUWTHCEG CPVKIGP VJCV KU RTGHGTGPVKCNN[ GZ RTGUUGFD[OQWUGOCUVEGNNRTQIGPKVQTUCPFKUC PGY OGODGT QH VJG KOOWPQINQDWNKP UWRGTHC OKN[,$KQN%JGO 9CPI . . + - /GJVC 2 # .G$NCPE CPF 9 / ;QMQ[COC /QWUG PCVWTCN MKNNGT EGNNU GZRTGUU IR$ C UVTWEVWTCN JQOQ NQIWG QH JWOCP MKNNGT KPJKDKVQT[ TGEGRVQTU , +OOWPQN -WDCICYC * 2 & $WTTQYU CPF / & %QQRGT # PQXGN RCKT QH KOOWPQINQDW NKPNKMGTGEGRVQTUGZRTGUUGFD[$EGNNUCPFO[G NQKF EGNNU =UGG EQOOGPVU? 2TQE 0CVN #ECF 5EK 75# /CNKU\GYUMK % 4 % , /CTEJ / # 5EJQGPDQTP 5 )KORGN CPF . 5JGP 'ZRTGUUKQP ENQPKPI QH C JWOCP (E TGEGRVQT HQT +I#,'ZR/GF.

(19) IgA binds CD89 via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

(20) Chapter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

(21)

6Q NQECNK\G VJG KOOWPQINQDWNKP +I  DKPFKPI FQOCKPU QH VJG JWOCP O[GNQKF +I# (E&alpha; TGEGRVQT (E&alpha;4

6Q NQECNK\G VJG KOOWPQINQDWNKP +I DKPFKPI FQOCKPU QH VJG JWOCP O[GNQKF +I# (Eα TGEGRVQT (Eα4