Induction and analysis of antigen-specific T cell responses in
melanoma patients and animal model
Bins, A.D.
Citation
Bins, A. D. (2007, March 15). Induction and analysis of antigen-specific T cell responses in
melanoma patients and animal model. Retrieved from https://hdl.handle.net/1887/11457
Version: Corrected Publisher’s Version
License: Licence agreement concerning inclusion of doctoral thesis in the
Institutional Repository of the University of Leiden
Downloaded from: https://hdl.handle.net/1887/11457
Note: To cite this publication please use the final published version (if applicable).
'HVLJQDQGXVHRIFRQGLWLRQDO0+&FODVV,
OLJDQGV
0LUHLOOH7RHEHV$GULDDQ%LQV0LULDP&RFFRULV%RULV5RGHQNR5DTXHO
*RPH]1HOOD-1LHXZNRRS:LOOHNHYDQGH.DVWHHOH*XXV)5LPPHO]ZDDQ
-RKQ%$*+DDQHQ+XLE2YDDDQG7RQ106FKXPDFKHU
'LYLVLRQ RI ,PPXQRORJ\ 7KH 1HWKHUODQGV &DQFHU ,QVWLWXWH 3OHVPDQODDQ
&;$PVWHUGDP7KH1HWKHUODQGV
'LYLVLRQRI&HOOXODU%LRFKHPLVWU\7KH1HWKHUODQGV&DQFHU,QVWLWXWH3OHVPDQODDQ
&;$PVWHUGDP7KH1HWKHUODQGV
'HSDUWPHQWRI9LURORJ\DQG:+21DWLRQDO,QIOXHQ]D&HQWHU(UDVPXV0HGLFDO
&HQWHU32%R['55RWWHUGDP7KH1HWKHUODQGV
7KHVHDXWKRUVFRQWULEXWHGHTXDOO\
&RUUHVSRQGHQFHVKRXOGEHDGGUHVVHGWR7106WVFKXPDFKHU#QNLQORU+2
KRYDD#QNLQO
$EVWUDFW
0DMRUKLVWRFRPSDWLELOLW\FRPSOH[0+&FODVV,PROHFXOHVDVVRFLDWHZLWKDODUJH
YDULHW\ RI SHSWLGH OLJDQGV GXULQJ ELRV\QWKHVLV DQG SUHVHQW WKHVH OLJDQGV RQ WKH
FHOO VXUIDFH IRU UHFRJQLWLRQ E\ F\WRWR[LF 7 FHOOV :H KDYH GHVLJQHG FRQGLWLRQDO
0+&OLJDQGVWKDWIRUPVWDEOHFRPSOH[HVZLWK0+&PROHFXOHVEXWWKDWGHJUDGH
RQFRPPDQGE\H[SRVXUHWRDGH¿QHGSKRWRVWLPXOXV³(PSW\0+&PROHFXOHV´
JHQHUDWHGLQWKLVPDQQHUFDQEHORDGHGZLWKDUUD\VRISHSWLGHOLJDQGVWRGHWHUPLQH
0+&ELQGLQJSURSHUWLHVDQGWRPRQLWRUDQWLJHQVSHFL¿F7FHOOUHVSRQVHVLQDKLJK
WKURXJKSXWPDQQHU7KHYDOXHRIWKLVDSSURDFKLVGRFXPHQWHGE\WKHLGHQWL¿FDWLRQ
RI F\WRWR[LF 7 FHOO HSLWRSHV ZLWKLQ WKH +1 LQÀXHQ]D $9LHWQDP
JHQRPH
,QWURGXFWLRQ
0+&FODVV,PROHFXOHVDUHKHWHURWULPHUVWKDWFRQVLVWRIDQLQYDULDQWOLJKWFKDLQ
D SRO\PRUSKLF KHDY\ FKDLQ DQG DQ HLJKW WR WHQ DPLQR DFLG SHSWLGH OLJDQG 7KH
SHSWLGH IRUPV DQ HVVHQWLDO VXEXQLW RI WKH 0+& FODVV , FRPSOH[ DV 0+& FODVV
,PROHFXOHVWKDWIDLOWRDVVRFLDWHZLWKSHSWLGHOLJDQGDUHXQVWDEOH$VVRFLDWLRQ
RI SHSWLGHV ZLWK 0+& FODVV , PROHFXOHV LV LQ ODUJH SDUW EDVHG RQ VKDSH DQG
HOHFWURVWDWLFFRPSOHPHQWDULW\EHWZHHQWZRDPLQRDFLGVLGHFKDLQVDWWKHDQFKRU
SRVLWLRQVRIWKHSHSWLGHDQG0+&DOOHOHVSHFL¿FSRFNHWV,QDGGLWLRQELQGLQJRI
SHSWLGHOLJDQGVGHSHQGVRQWKHLQWHUDFWLRQRIWKH0+&PROHFXOHZLWKWKHWHUPLQDO
Į1+DQGFDUER[\OJURXSVRIWKHSHSWLGH%HFDXVHWKHVHTXHQFHUHTXLUHPHQWV
IRUELQGLQJWR0+&DUHODUJHO\UHVWULFWHGWRWZRGRPLQDQWDQFKRUUHVLGXHVWKH
DYHUDJHSURWHLQFRQWDLQVVHYHUDOGR]HQSRWHQWLDO7FHOOHSLWRSHVDQGDVDQH[DPSOH
WKH DSSUR[LPDWHO\ RSHQ UHDGLQJ IUDPHV RI D PHPEHU RI WKH KHUSHVYLULGDH
IDPLO\ FRQWDLQ DQ HVWLPDWHG SRWHQWLDO7 FHOO HSLWRSHV 'H¿QLWLRQ RI WXPRU
DQGSDWKRJHQHQFRGHG0+&OLJDQGVDQGGHWHFWLRQRI7FHOOUHVSRQVHVVSHFL¿FIRU
VXFK OLJDQGV UHPDLQV D PDMRU FKDOOHQJH 7KH YLVXDOL]DWLRQ RI DQWLJHQVSHFL¿F 7
FHOOUHVSRQVHVZDV¿UVWPDGHSRVVLEOHZLWKWKHGHYHORSPHQWRIWHWUDPHULF0+&
UHDJHQWV E\$OWPDQ HW DO ,Q WKLV VWUDWHJ\ VROXEOH 0+& PRQRPHUV FRPSOH[HG
ZLWKDSHSWLGHRILQWHUHVWDUHELRWLQ\ODWHGDQGFRQYHUWHGWRWHWUDYDOHQWVWUXFWXUHV
E\ELQGLQJWRÀXRURFKURPHFRQMXJDWHGVWUHSWDYLGLQRUDYLGLQ7KHUHVXOWLQJ0+&
WHWUDPHUVKDYHEHFRPHHVVHQWLDOUHDJHQWVIRUWKHGHWHFWLRQRIDQWLJHQVSHFL¿F&' DQG &'7 FHOOV E\ ÀRZ F\WRPHWU\ 0RUH UHFHQW ZRUN LQGLFDWHV WKDW GH¿QLWLRQ
RIDQWLJHQVSHFL¿F7FHOOUHVSRQVHVE\0+&PLFURDUUD\EDVHGVWUDWHJLHVLVDOVR
IHDVLEOH)XUWKHUPRUH0+&EDVHGVHOHFWLRQRIDQWLJHQVSHFL¿F7FHOOVKDVEHHQ
SURSRVHGDVDVWUDWHJ\WRERRVWPHODQRPDVSHFL¿F7FHOOUHVSRQVHVLQPHODQRPD
SDWLHQWV DQG WR SURYLGH GH¿QHG PLQRU KLVWRFRPSDWLELOLW\ DQWLJHQVSHFL¿F DQG
YLUXVVSHFL¿F 7 FHOOV LQ UHFLSLHQWV RI DOORJHQHLF VWHP FHOO WUDQVSODQWV DQG RWKHU
LPPXQRFRPSURPLVHG LQGLYLGXDOV $W SUHVHQW WKH PDMRU OLPLWDWLRQ RI 0+&
WHWUDPHUEDVHGWHFKQRORJLHVLVIRUPHGE\WKHLQYROYHGQDWXUHRIWKHUHIROGLQJDQG
SXUL¿FDWLRQ VWHSV WKDW DUH UHTXLUHG IRU HYHU\ LQGLYLGXDO SHSWLGH0+& FRPSOH[
&RQVHTXHQWO\LWKDVQRWEHHQIHDVLEOHWRDSSO\0+&WHWUDPHUWHFKQRORJ\IRUKLJK
WKURXJKSXWDSSOLFDWLRQV7RDGGUHVVWKLVLVVXHZHKDYHH[SORUHGWKHSRVVLELOLW\RI
LGHQWLI\LQJFRQGLWLRQDO0+&OLJDQGVWKDWFDQEHXVHGWRJHQHUDWHSHSWLGHUHFHSWLYH
0+&FODVV,PROHFXOHVDWZLOO
5HVXOWV
'HVLJQRIFRQGLWLRQDO0+&FODVV,OLJDQGV
:H VHW RXW WR FUHDWH 0+& FODVV , OLJDQGV WKDW ZRXOG GLVLQWHJUDWH RQ FRPPDQG
ZKLOH ERXQG WR WKH 0+& FRPSOH[ $V D EXLOGLQJ EORFN IRU VXFK FRQGLWLRQDO
OLJDQGVZHV\QWKHVL]HGDÀXRUHQ\OPHWK\OR[\FDUERQ\O)PRFGHULYDWLYHRI
DPLQRQLWURSKHQ\OSURSLRQLF DFLG - 6XEVHTXHQWO\ ZH XVHG WKLV EXLOGLQJ
EORFNWRJHQHUDWHYDULDQWVRIWKHKXPDQOHXNRF\WHDQWLJHQ+/$$UHVWULFWHG
LQÀXHQ]D$PDWUL[0HSLWRSHVHTXHQFH*,/*)9)7/$QH[DPSOHRIVXFK
D YDULDQW LQ ZKLFK WKH7KU LV UHSODFHG E\ DPLQRQLWURSKHQ\OSURSLRQLF
DFLG UHVXOWLQJ LQ FRPSRXQG , *,/*)9)-/ LV JLYHQ LQ )LJ D &RQVLVWHQW
ZLWK SULRU OLWHUDWXUH WKLV FRPSRXQG GLVLQWHJUDWHV XSRQ H[SRVXUH WR 89 OLJKW
)LJDDQGIROORZLQJ89H[SRVXUHDPROHFXOHZLWKDPDVVRIEHFRPHV
DSSDUHQWFRUUHVSRQGLQJWRWKHPDVVRIWKHKHSWDPHULFSHSWLGHDFHWDPLGHIUDJPHQW
)LJDFRPSRXQG,,)LJE7KHVPDOOFDUER[\WHUPLQDOSHSWLGHIUDJPHQWWKDW
LVDOVRJHQHUDWHGGXULQJWKLVFOHDYDJHUHDFWLRQZDVQRWIXUWKHUVWXGLHG
7RHVWDEOLVKZKHWKHU89VHQVLWLYHOLJDQGVFDQEHXVHGWRJHQHUDWHSHSWLGH0+&
FRPSOH[HVZHXVHGWZR0YDULDQWVZLWKDPLQRQLWURSKHQ\OSURSLRQLF
DFLGLQFRUSRUDWHGDWSRVLWLRQIRXUFRPSRXQG,,,*,/-)9)7/6XSSO)LJRU
SRVLWLRQHLJKWFRPSRXQG,)LJDLQ0+&FODVV,UHIROGLQJUHDFWLRQV0+&
UHIROGLQJUHDFWLRQVZLWKHLWKHU*,/-)9)7/RU*,/*)9)-/SURGXFHGKLJK\LHOGV
RI+/$$FRPSOH[HV$VD¿UVWFUXGHWHVWRIWKHVHQVLWLYLW\RIWKHQRQQDWXUDO
0+&ERXQG OLJDQGV WRZDUGV 89PHGLDWHG FOHDYDJH ZH H[SRVHG +/$$
FRPSOH[HV FRQWDLQLQJ HLWKHU WKH XQPRGL¿HG LQÀXHQ]D $ 0 HSLWRSH RU WKH
*,/-)9)7/RU*,/*)9)-/YDULDQWWR89DQGDQDO\]HGWKHVHUHDFWLRQVE\JHO
¿OWUDWLRQFKURPDWRJUDSK\:KHUHDVWKHFRQYHQWLRQDO+/$$SHSWLGHFRPSOH[
LV QRW DIIHFWHG E\ H[SRVXUH WR QP OLJKW H[SRVXUH RI HLWKHU +/$$
*,/-)9)7/QRWVKRZQRU+/$$*,/*)9)-/6XSSO)LJDOHDGVWRD
VXEVWDQWLDOUHGXFWLRQLQ0+&UHFRYHU\)XUWKHUPRUHZKHQ+/$$FRPSOH[HV
DUH SUHSDUHG ZLWK DQ LQÀXHQ]D $ 0 GHULYDWLYH LQ ZKLFK WKH 89UHVLVWDQW
EXLOGLQJ EORFN DPLQRSKHQ\OSURSLRQLF DFLG LV LQFRUSRUDWHG WKH UHVXOWLQJ
0+& FRPSOH[HV DUH LQVHQVLWLYH WR 89 H[SRVXUH GHPRQVWUDWLQJ WKDW WKH 89
LQGXFHG GHFD\ RI +/$$*,/-)9)7/ DQG +/$$*,/*)9)-/ UHTXLUHV
WKHSUHVHQFHRIWKHQLWURSKHQ\OPRLHW\1RWDEO\WKHREVHUYHGORVVRIIROGHG0+&
XSRQ 89 H[SRVXUH RI WKH +/$$*,/*)9)-/ FRPSOH[ FDQ EH SUHYHQWHG
E\LQFOXVLRQRI+/$$ELQGLQJSHSWLGHVGXULQJWKHFOHDYDJHUHDFWLRQEXWQRW
E\ LQFOXVLRQ RI D FRQWURO +/$$ ELQGLQJ SHSWLGH 6XSSO )LJ E VXJJHVWLQJ
HI¿FLHQWSHSWLGHH[FKDQJH
:KLOH WKHVH GDWD GHPRQVWUDWH WKH 89VHQVLWLYLW\ RI +/$ FRPSOH[HV FRQWDLQLQJ
DPLQRQLWURSKHQ\OSURSLRQLF DFLGEDVHG SHSWLGH OLJDQGV LW LV GLI¿FXOW WR
HVWDEOLVKWKHHI¿FLHQF\RIWKLVFOHDYDJHUHDFWLRQE\JHO¿OWUDWLRQFKURPDWRJUDSK\
VHH /HJHQG WR 6XSSO )LJ D$V D PRUH VWULQJHQW WHVW IRU UHSODFHPHQW RI WKH
FRQGLWLRQDOOLJDQGE\WKHQHZO\DGGHGSHSWLGHZHH[SRVHG+/$$*,/*)9)-/
FRPSOH[HVWR89OLJKWLQWKHSUHVHQFHRIWKHF\WRPHJDORYLUXV&09SS HSLWRSH DQG IROORZLQJ WKLV UHDFWLRQ SXUL¿HG WKH SHSWLGH0+& FRPSOH[HV DQG
DQDO\]HGWKHVHFRPSOH[HVE\PDVVVSHFWURPHWU\7KHPDMRUSHSWLGHPDVVWKDWLV
YLVLEOHSULRUWR89H[SRVXUHFRUUHVSRQGVWRWKH1DLRQRI*,/*)9)-/SURYLGLQJ
IRUPDO SURRI WKDW WKLV OLJDQG IRUPV D VWDEOH FRPSOH[ ZLWK +/$$ )LJ
)ROORZLQJ89PHGLDWHGFOHDYDJHQRGHWHFWDEOHDPRXQWRI*,/*)9)-/UHPDLQV
DVVRFLDWHG ZLWK +/$$ )XUWKHUPRUH DOVR WKH OHYHOV RI FOHDYDJH SURGXFW ,,
FRPSOH[HG ZLWK +/$$ DUH EHORZ EDFNJURXQG VXJJHVWLQJ WKDW GLVVRFLDWLRQ
RI WKLV KHSWDPHULF SHSWLGH IUDJPHQW LV HVVHQWLDOO\ FRPSOHWH ,QVWHDG XSRQ 89
PHGLDWHG FOHDYDJH WKH VROH SHSWLGH PDVV YLVLEO\ DVVRFLDWHG ZLWK +/$$
FRUUHVSRQGVWRWKHPDVVRIWKH&09SSHSLWRSH)LJ&ROOHFWLYHO\WKHVH
H[SHULPHQWVLQGLFDWHWKDWFRQGLWLRQDO0+&OLJDQGVFDQEHGH¿QHGWKDWDUHUHOHDVHG
IURP0+&PROHFXOHVDWZLOODQGWKDW0+&PROHFXOHVJHQHUDWHGLQWKLVSURFHVV
FDQEHORDGHGZLWKHSLWRSHVRIFKRLFH7RH[WHQGWKHVHGDWDWRRWKHUFRQGLWLRQDO
+/$$ OLJDQGV ZH V\QWKHVL]HG D SHSWLGH WKDW LV SUHGLFWHG WR ELQG DYLGO\ WR
+/$$ ZLWK WKH 89VHQVLWLYH EXLOGLQJ EORFN - LQFRUSRUDWHG DW SRVLWLRQ
HLJKW,/$(79$-95HIROGLQJUHDFWLRQVZLWKWKLVFRQGLWLRQDOOLJDQGJDYHKLJK
\LHOGVRIIROGHG+/$$FRPSOH[HVDQGDQDORJRXVWRWKHGDWDREWDLQHGZLWK
+/$$*,/*)9)-/ FRPSOH[HV WKHVH FRPSOH[HV GLVLQWHJUDWH IROORZLQJ 89
H[SRVXUHGDWDQRWVKRZQ 0+&H[FKDQJHWHWUDPHUV
7RWHVWWKHSRWHQWLDOYDOXHRIWKLV0+&H[FKDQJHWHFKQLTXHIRUWKHYLVXDOL]DWLRQRI
DQWLJHQVSHFL¿F7FHOOVZHSHUIRUPHG0+&H[FKDQJHUHDFWLRQVZLWKELRWLQ\ODWHG
+/$$*,/*)9)-/RU+/$$,/$(79$-9FRPSOH[HV6XEVHTXHQWO\
SK\FRHU\WKULQ 3(VWUHSWDYLGLQ ZDV DGGHG DQG WKH UHVXOWLQJ 0+& WHWUDPHUV
KHUHDIWHU UHIHUUHG WR DV 0+& H[FKDQJH WHWUDPHUV ZHUH XVHG WR GHWHFW DQWLJHQ
VSHFL¿F7FHOOVE\ÀRZF\WRPHWU\0+&H[FKDQJHWHWUDPHUVFRQWDLQLQJWKHKLJK
DI¿QLW\ $/ YDULDQW RI WKH 0HODQ$0$57 HSLWRSH VWDLQ D 0$57
VSHFL¿F &7/ FORQH DV HI¿FLHQWO\ DV FRQYHQWLRQDO 0+& FODVV , WHWUDPHUV )LJ
D/LNHZLVH0+&H[FKDQJHWHWUDPHUVFDQEHXVHGWRGHWHFWORZPDJQLWXGH7
FHOOUHVSRQVHVLQSHULSKHUDOEORRGVDPSOHV)LJE6XSSO)LJD89LQGXFHG
FOHDYDJH RI 0+&ERXQG OLJDQGV DOVR DOORZV WKH V\QWKHVLV DQG XVH RI 0+&
WHWUDPHUV WKDW FRQWDLQ WKH QDWXUDOO\ RFFXUULQJ 0$57 SHSWLGH WKDW ELQGV WR
0+&FODVV,ZLWKORZDI¿QLW\GXHWRWKHDEVHQFHRIDOHXFLQHPHWKLRQLQHUHVLGXHDW
WKH3DQFKRUSRVLWLRQ)LJD7KLVLQGLFDWHVWKDW89LQGXFHGSHSWLGHH[FKDQJH
LV VXI¿FLHQWO\ UREXVW WR VFUHHQ FROOHFWLRQV RI SXWDWLYH 7 FHOO HSLWRSHV ZLWKRXW D
SULRULNQRZOHGJHRI0+&ELQGLQJDI¿QLWLHV,PSRUWDQWO\0+&WHWUDPHUVSUHSDUHG
IURP +/$$*,/*)9)-/ FRPSOH[HV RU$.E*,/*)9)-/ FRPSOH[HV VHH
IXUWKHUWKDWKDYHQRWEHHQH[SRVHGWR89OLJKWDQGWKDWDUHWKHUHIRUHXQLIRUPO\
RFFXSLHG E\ WKH 89VHQVLWLYH 0 YDULDQW GR QRW VWDLQ SRO\FORQDO LQÀXHQ]D
$ 0VSHFL¿F 7 FHOOV 6XSSO )LJ E DQG F 7KLV LQGLFDWHV WKDW HLWKHU WKH
DOWHUDWLRQLQWKHSHSWLGHEDFNERQHGXHWRWKHLQWURGXFWLRQRIDQXQQDWXUDOȕDPLQR
DFLG RU WKH UHSODFHPHQW RI WKH WKUHRQLQH VLGH FKDLQ LV LQFRPSDWLEOH ZLWK7 FHOO
UHFRJQLWLRQ E\ 0±VSHFL¿F 7 FHOOV &RQVHTXHQWO\ HYHQ LQ VHWWLQJV ZKHUH
UHOHDVH RI FRQGLWLRQDO OLJDQG ZRXOG QRW EH RSWLPDO 0+& H[FKDQJH UHDJHQWV
DUHQRWH[SHFWHGWRGLVSOD\EDFNJURXQGUHDFWLYLW\GXHWRWKHSUHVHQFHRIUHVLGXDO
FRQGLWLRQDOOLJDQG,WLVQRWHGWKDWLIFRQGLWLRQDOOLJDQG0+&FRPSOH[HVIRURWKHU
DOOHOHVZRXOGGLVSOD\EDFNJURXQGUHDFWLYLW\LWVKRXOGEHVWUDLJKWIRUZDUGWRSUHYHQW
VXFK UHDFWLYLW\ E\ PRGL¿FDWLRQ RI 7 FHOO UHFHSWRUH[SRVHG VLGHFKDLQV 1RWDEO\
0+&WHWUDPHUVJHQHUDWHGE\89H[FKDQJHFRPSDUHIDYRUDEO\WR0+&,JGLPHUV
JHQHUDWHGE\SDVVLYHSHSWLGHH[FKDQJHERWKZLWKUHVSHFWWRVLJQDOLQWHQVLW\DQG
VLJQDOWRQRLVHUDWLRV6XSSO)LJDE
7RWHVWZKHWKHUFRQGLWLRQDOOLJDQGVPD\UHDGLO\EHLGHQWL¿HGIRURWKHU0+&FODVV
, DOOHOHV ZH V\QWKHVL]HG IRXU YDULDQWV RI WKH +'EUHVWULFWHG $61(10(70
LQÀXHQ]D $ 13HSLWRSH 7ZR RI WKRVH YDULDQWV ,9 DQG 9 6XSSO )LJ
IXO¿OOHG ERWK FULWHULD LQ WKDW +'E FRPSOH[HV FRXOG EH JHQHUDWHG ZLWK WKHVH
FRQGLWLRQDO OLJDQGV DQG WKDW WKHVH OLJDQGV FRXOG EH FOHDYHG LQ WKH 0+&ERXQG
VWDWH&RQVLVWHQWZLWKWKHGDWDREWDLQHGIRU+/$$+'EH[FKDQJHWHWUDPHUV
SUHSDUHG IURP 89VHQVLWLYH +'E$61(1-(70 FRPSOH[HV VWDLQ DQWLJHQ
VSHFL¿F7FHOOVZLWKKLJKVSHFL¿FLW\)LJF)XUWKHUPRUHDVLVWKHFDVHIRU+/$
$*,/*)9)-/ WHWUDPHUV +'E$61(1-(70 WHWUDPHUV WKDW DUH XQLIRUPO\
RFFXSLHGE\WKH89VHQVLWLYHYDULDQWRI13GRQRWVWDLQLQÀXHQ]D$13 VSHFL¿F7FHOOV)LJF
+LJKWKURXJKSXWVFUHHQLQJZLWK0+&H[FKDQJHUHDJHQWV
$V D ¿UVW WHVW RI WKH SRWHQWLDO RI 0+& H[FKDQJH IRU KLJKWKURXJKSXW HSLWRSH
PDSSLQJZHFORQHGDQGVHTXHQFHGWKHIRXUJHQHVHQFRGLQJWKHLPPXQRGRPLQDQW
SURWHLQVRI$9LHWQDPDQLQÀXHQ]D$+1VWUDLQLVRODWHGIURPDIDWDO
KXPDQFDVHLQ9LHWQDPDQGVFDQQHGWKHHQFRGHGSURWHLQVIRUSRWHQWLDO+/$$
ELQGLQJSHSWLGHV:LWKLQWKHKHPDJJOXWLQLQ+$QHXUDPLQLGDVH1$PDWUL[
0DQGQXFOHRSURWHLQ13JHQHSURGXFWVZHLGHQWL¿HGSRWHQWLDOHSLWRSHV
ZLWKDVFRUHIRUSUHGLFWHG0+&ELQGLQJRIDQGWKHVHSHSWLGHVZHUHSURGXFHG
E\ PLFURVFDOH V\QWKHVLV ,Q SDUDOOHO ZH XVHG WKH VDPH VHW RI JHQHV WR SUHSDUH
YHFWRUV IRU '1$ YDFFLQDWLRQ DQG JURXSV RI +/$$ WUDQVJHQLF PLFH ZHUH
YDFFLQDWHGE\'1$WDWWRR$WWKHSHDNRIWKHYDFFLQDWLRQLQGXFHG7FHOOUHVSRQVH
ZHJHQHUDWHGDFROOHFWLRQRI0+&H[FKDQJHWHWUDPHUVE\SHUIRUPLQJSDUDOOHO
89PHGLDWHGH[FKDQJHUHDFWLRQVRQ$.E*,/*)9)-/FRPSOH[HVLQPLFURWLWHU
IRUPDWDQGWKHUHVXOWLQJ0+&WHWUDPHUFROOHFWLRQZDVXVHGWRVFUHHQSHULSKHUDO
EORRGVDPSOHVRIYDFFLQDWHGPLFH7KLVDQDO\VLVGHPRQVWUDWHGWKHSUHVHQFHRIWZR
7FHOOHSLWRSHVZLWKLQWKHVHIRXUJHQHSURGXFWVRI$9LHWQDP6SHFL¿FDOO\
WKLVVFUHHQFRQ¿UPHGWKHLPPXQRJHQLFLW\RIWKHNQRZQLQÀXHQ]D$0HSLWRSH
WKDWLVFRQVHUYHGEHWZHHQWKHPDMRULW\RILQÀXHQ]D$VWUDLQV,QDGGLWLRQWKLVVFDQ
UHYHDOHGWKHSUHVHQFHRIDSUHYLRXVO\XQNQRZQ+/$$UHVWULFWHG7FHOOHSLWRSH
ORFDWHG LQ WKH $9LHWQDP QXFOHRSURWHLQ )LJ ,QWHUHVWLQJO\ LQ WKLV
+/$$WUDQVJHQLFPRXVHPRGHOWKHLPPXQRJHQLFLW\RIWKLVHSLWRSH13 LVVXEVWDQWLDOO\KLJKHUWKDQWKDWRIWKHFODVVLFDO0HSLWRSH0RQHRXW
RI ¿YH UHVSRQGLQJ PLFH DIWHU SULPDU\ YDFFLQDWLRQ WKUHH RXW RI ¿YH UHVSRQGLQJ
PLFHDIWHUVHFRQGDU\YDFFLQDWLRQ13¿YHRXWRI¿YHUHVSRQGLQJPLFHDIWHU
SULPDU\YDFFLQDWLRQ7KLVQRYHO7FHOOHSLWRSHLVVKDUHGEHWZHHQ+1VWUDLQVRI
WKHSDVW\HDUVEXWLVGLVWLQFWLQROGHULQÀXHQ]D$VWUDLQV
'LVFXVVLRQ
+HUH ZH KDYH GHVFULEHG FRQGLWLRQDO 0+& OLJDQGV WKDW FDQ GLVLQWHJUDWH LQ WKH
0+& ERXQG VWDWH XQGHU FRQGLWLRQV WKDW GR QRW DIIHFW WKH LQWHJULW\ RI WKH 0+&
PROHFXOH WKHUHE\ SHUPLWWLQJ WKH UHORDGLQJ RI DVVHPEOHG 0+& PROHFXOHV ZLWK
HSLWRSHV RI FKRLFH ,PSRUWDQWO\ WKH SHSWLGH OLJDQGV WKDW DUH ERXQG WR WKH 0+&
LQWKHVHUHDFWLRQVDUHXQPRGL¿HGQRULVWKH0+&EDFNERQHDOWHUHG,QOLQHZLWK
WKLV0+&FRPSOH[HVJHQHUDWHGE\WKLVH[FKDQJHWHFKQRORJ\GLVSOD\WKHSUHGLFWHG
ELQGLQJVSHFL¿FLW\LQDOOFDVHVWHVWHG7KLVVWUDWHJ\DQGUHODWHGFKHPLFDOFOHDYDJH
VWUDWHJLHV VKRXOG EH RI VXEVWDQWLDO XVH LQ WKH KLJKWKURXJKSXW LGHQWL¿FDWLRQ RI
ERWK0+&OLJDQGVDQGF\WRWR[LF7FHOOUHVSRQVHV,QWKLVVWUDWHJ\DVLQJOHEDWFK
RI 89VHQVLWLYH 0+& FRPSOH[ LV SUHSDUHG E\ WKH FODVVLFDO LQ YLWUR 0+& FODVV
, UHIROGLQJ DQG SXUL¿FDWLRQ SURWRFROV DQG WKLV 89VHQVLWLYH 0+& FRPSOH[ LV
VXEVHTXHQWO\ XVHG WR JHQHUDWH ODUJH DUUD\V RI GHVLUHG S0+& FRPSOH[HV LQ K
H[FKDQJHUHDFWLRQV
0+&,J GLPHUV SXUL¿HG IURP HXNDU\RWLF FHOOV KDYH SUHYLRXVO\ EHHQ XVHG
WR JHQHUDWH SHSWLGH0+& UHDJHQWV E\ SHUIRUPLQJ H[FKDQJH UHDFWLRQV ZLWK
H[RJHQRXVO\DGGHGSHSWLGH:KLOHWKHRYHUDOOJRDORIWKLVWHFKQRORJ\LVVLPLODU
WRWKDWRI89LQGXFHGSHSWLGHH[FKDQJHWKHWHFKQRORJLHVGLIIHUDWHVVHQWLDOSRLQWV
6SHFL¿FDOO\ZKLOHWKH0+&,JGLPHUWHFKQRORJ\GHSHQGVRQWKHVORZUHOHDVHRID
SRRORIXQNQRZQHQGRJHQRXVSHSWLGHVDQGLVIDFLOLWDWHGE\FRQGLWLRQVHJORZRU
KLJKS+WKDWDOVRGHVWDELOL]HWKH0+&PROHFXOH89LQGXFHGSHSWLGHH[FKDQJHLV
EDVHGRQWKHUHOHDVHRIDVLQJOHOLJDQGE\H[SRVXUHWRDGH¿QHGWULJJHUWKDWGRHV
QRWDIIHFWWKHLQWHJULW\RIWKH0+&PROHFXOH)XUWKHUPRUHWKHFDSDFLW\RI0+&,J
EDVHGUHDJHQWVWRUHYHDODQWLJHQVSHFL¿F7FHOOUHVSRQVHVLVOLPLWHGDVFRPSDUHGWR
0+&H[FKDQJHWHWUDPHUV6XSSO)LJ
7KH REVHUYDWLRQ WKDW OLJDQGV WKDW GLVLQWHJUDWH RQ FRPPDQG FRXOG UHDGLO\ EH
LGHQWL¿HGIRUERWKWHVWHG0+&DOOHOHVVXJJHVWVWKDWLWZLOOEHVWUDLJKWIRUZDUGWR
LGHQWLI\ QLWURSKHQ\OEDVHG FRQGLWLRQDO OLJDQGV IRU RWKHU 0+& FODVV , DOOHOHV
&RQGLWLRQDO OLJDQGV FDQ EH GHVLJQHG E\ UHSODFHPHQW RI DPLQR DFLGV LQ HLWKHU D
NQRZQSHSWLGHOLJDQGRULQDSUHGLFWHGKLJKDI¿QLW\OLJDQGEDVHGRQWKHSHSWLGH
ELQGLQJPRWLIIRUWKLVDOOHOHDQGIRU+/$$ERWKDSSURDFKHVSURYHGVXFFHVVIXO
)RU 0+& DOOHOHV IRU ZKLFK VWUXFWXUDO LQIRUPDWLRQ LV DYDLODEOH WKH ZDWHU
DFFHVVLELOLW\ RI VLGHFKDLQV PD\ DOVR EH XVHG DV D FULWHULRQ WR VHOHFW SRVLWLRQV DW
ZKLFKWKH89VHQVLWLYHEXLOGLQJEORFNFDQEHLQFRUSRUDWHG
9DULRXV W\SHV RI IXQFWLRQDO DVVD\V IRU DQWLJHQVSHFL¿F 7 FHOO GHWHFWLRQ VXFK DV
LQWUDFHOOXODU F\WRNLQH VWDLQLQJ DQG F\WRNLQH FDSWXUH KDYH EHHQ GHYHORSHG LQ WKH
SDVWIHZ\HDUVDQGWKHVHDVVD\VPD\EHXWLOL]HGIRUKLJKWKURXJKSXWDQDO\VLVRI7
FHOOVWKDWGLVSOD\DJLYHQHIIHFWRUIXQFWLRQ0+&H[FKDQJHPXOWLPHUWHFKQRORJ\
VKRXOG FRPSOHPHQW WKHVH WHFKQRORJLHV E\ DOORZLQJ KLJKWKURXJKSXW DQDO\VLV
RI 7 FHOO UHVSRQVHV LUUHVSHFWLYH RI WKH FDSDFLW\ RI 7 FHOOV WR SURGXFH D JLYHQ
F\WRNLQH$OVRWKHFRPELQDWLRQRIWKHWZRWHFKQRORJLHVLQZKLFKKLJKFRPSOH[LW\
0+&PXOWLPHUDUUD\VDUHXVHGWRSUREH7FHOOUHDFWLYLW\E\PRQLWRULQJF\WRNLQH
SURGXFWLRQ PD\ EH SDUWLFXODUO\ XVHIXO 2QFRPPDQG FOHDYDJH RI 0+& OLJDQGV
DOVR VHHPV DWWUDFWLYH IRU WKH SURGXFWLRQ RI VHWV RI FOLQLFDO JUDGH 0+& UHDJHQWV
IRUDGRSWLYH7FHOOWKHUDS\6SHFL¿FDOO\WKHJHQHUDWLRQRIDVLQJOHODUJHEDWFKRI
FOLQLFDOJUDGH0+&PROHFXOHVFRPSOH[HGZLWKFRQGLWLRQDOOLJDQGVKRXOGDOORZWKH
VWUDLJKWIRUZDUGDVVHPEO\RIDQ0+&UHDJHQWGHVLUHGIRUFOLQLFDOXVHE\SHUIRUPLQJ
0+&H[FKDQJHUHDFWLRQVZLWKWKHUHOHYDQWSHSWLGHOLJDQGV6XFKKLJKJUDGH0+&
UHDJHQWVVKRXOGEHSDUWLFXODUO\DWWUDFWLYHIRUWKHLVRODWLRQRIPHODQRPDVSHFL¿F7
FHOOVDQGIRUWKHLVRODWLRQRIGH¿QHGPLQRUKLVWRFRPSDWLELOLW\DQWLJHQVSHFL¿F7
FHOOV)LQDOO\LQDGGLWLRQWRWKHXVHRIFRQGLWLRQDO0+&FODVV,OLJDQGVIRUKLJK
WKURXJKSXWGLDJQRVWLFVFUHHQLQJDQGIRUDGRSWLYH7FHOOWKHUDS\ZHVSHFXODWHWKDW
FOHDYDJHRIOLJDQGVERXQGWR0+&PROHFXOHVLQWKHFU\VWDOOLQHVWDWHPLJKWEH
XVHGWRREWDLQWKHHOXVLYHVWUXFWXUHRIHPSW\0+&FODVV,
0HWKRGV
3HSWLGH V\QWKHVLV DQG SUHSDUDWLRQ RI UHFRPELQDQW 0+& :H REWDLQHG WKH
89VHQVLWLYH EXLOGLQJ EORFN IRU SHSWLGH V\QWKHVLV 1ÀXRUHQ\OPHWK\OR[\FDU ERQ\O DPLQRQLWURSKHQ\OSURSLRQLF DFLG E\ SURWHFWLRQ RI DPLQR
QLWURSKHQ\OSURSLRQLF DFLG /DQFDVWHU ZLWK ÀXRUHQ\OPHWK\O FKORURIRUPDWH
6LJPD$OGULFK LQ GLR[DQH DTXHRXV 1D&2 YY DFFRUGLQJ WR D
SXEOLVKHG SURFHGXUH :H V\QWKHVL]HG QDWXUDOO\ RFFXUULQJ SHSWLGHV DQG 89
VHQVLWLYHSHSWLGHYDULDQWVE\VWDQGDUG)PRFV\QWKHVLV:HSHUIRUPHG0+&FODVV,
UHIROGLQJUHDFWLRQVDVGHVFULEHGDQGZHSXUL¿HGUHIROGHG0+&FODVV,PROHFXOHV
E\ JHO¿OWUDWLRQ FKURPDWRJUDSK\ RQ D 3KHQRPHQH[ %LRVHS 6(& 6 FROXPQ
3KHQRPHQH[LQP07ULV+&OS+P01D&O3XUL¿HG0+&FODVV,
FRPSOH[HVDUHVWRUHGDW&LQP07ULV+&OS+P01D&ODQG
JO\FHURO
0+& H[FKDQJH UHDFWLRQV7R SURGXFH VLQJOH RU VPDOO VHWV RI 0+& UHDJHQWV E\
0+& H[FKDQJH ZH H[SRVHG ELRWLQ\ODWHG +/$$ *,/*)9)-/ +/$$
,/$(79$-9 RU +'E$61(1-(70 FRPSOH[HV 0 LQ P0 7ULV+&O
S+P01D&OP0GLWKLRWKUHLWRO'77WR89QP89ODPS
&$0$* LQ WKH SUHVHQFH RI 0 RI WKH LQGLFDWHG SHSWLGHV IRU K RQ LFH
)ROORZLQJH[FKDQJHVDPSOHVZHUHVSXQDWJIRUPLQ3(VWUHSWDYLGLQ
ZDVDGGHGDQGZHXVHGWKHUHVXOWLQJ0+&H[FKDQJHWHWUDPHUVIRU7FHOOVWDLQLQJ
ZLWKRXWIXUWKHUSXUL¿FDWLRQ
7RJHQHUDWHWKHFROOHFWLRQRI+1$.EWHWUDPHUVZHSUHGLFWHGSRWHQWLDOSHSWLGH
HSLWRSHV ZLWKLQ IRXU JHQH VHJPHQWV RI LQÀXHQ]D $9LHWQDP XVLQJ WKH
6<)3(,7+,SUHGLFWLRQSURJUDPDQGDOOSHSWLGHVZLWKD6<)3(,7+,VFRUH
ZHUH SURGXFHG E\ PLFURVFDOH QPROH V\QWKHVLV -37 3HSWLGH 7HFKQRORJLHV
*PE+2IWKHSRWHQWLDOHSLWRSHVWHUPLQDWHGLQHLWKHUDYDOLQHOHXFLQHRU
LVROHXFLQHUHVLGXHDQGWKHVHZHUHV\QWKHVL]HGZLWKWKHQDWXUDOO\RFFXUULQJFDUER[\
WHUPLQDO DPLQR DFLG 7KH UHPDLQLQJ SHSWLGHV WHUPLQDWLQJ LQ YDULRXV QRQ
DOLSKDWLFDPLQRDFLGVZHUHDOOV\QWKHVL]HGZLWKDFDUER[\WHUPLQDOLVROHXFLQHWR
IDFLOLWDWHSHSWLGHSURGXFWLRQ:HSUHSDUHG0+&H[FKDQJHWHWUDPHUVE\SHUIRUPLQJ
SDUDOOHO VPDOO VFDOH H[FKDQJH UHDFWLRQV RQ ELRWLQ\ODWHG$.E*,/*)9)-/ FODVV
, FRPSOH[HV 0 ZLWK WKH FDQGLGDWH LQÀXHQ]D $9LHWQDP
HSLWRSHVDQGDVHWRIFRQWUROSHSWLGHVDOOSHSWLGHVDW0E\H[SRVXUHWR89
QP89ODPS&$0$*IRUKRQLFHLQP07ULV+&OS+P0
1D&OP0'77LQZHOOWLVVXHFXOWXUH9ERWWRPSRO\VW\UHQHSODWHV181&
6XEVHTXHQWO\VDPSOHVZHUHVSXQDWJIRUPLQ3(VWUHSWDYLGLQJPO
¿QDOFRQFHQWUDWLRQZDVDGGHGDQGZHXVHGWKHUHVXOWLQJ0+&H[FKDQJHWHWUDPHUV
IRU7FHOOVWDLQLQJZLWKRXWIXUWKHUSXUL¿FDWLRQ
0LFHDQGYDFFLQDWLRQV
:H REWDLQHG &%/ PLFH DQG PLFH WUDQVJHQLF IRU WKH +/$$.E IXVLRQ
JHQH IURP WKH DQLPDO GHSDUWPHQW RI WKH 1HWKHUODQGV &DQFHU ,QVWLWXWH $OO
DQLPDOH[SHULPHQWVZHUHFDUULHGRXWLQDFFRUGDQFHZLWKLQVWLWXWLRQDODQGQDWLRQDO
JXLGHOLQHV DQG ZHUH DSSURYHG E\ WKH ([SHULPHQWDO $QLPDO &RPPLWWHH RI WKH
1HWKHUODQGV&DQFHU,QVWLWXWH'(&
)RU OLYH LQÀXHQ]D$ LQIHFWLRQV ZH LQWUDQDVDOO\ DGPLQLVWHUHG O RI +(3(6
EXIIHUHG VDOLQH VROXWLRQ /LIH 7HFKQRORJLHV FRQWDLQLQJ KHPDJJOXWLQDWLQJ
XQLWV LQÀXHQ]D $+.[ YLUXV WR DQHVWKHWL]HG PLFH )RU YDFFLQDWLRQ RI +/$
$ WUDQVJHQLF PLFH ZLWK +1 JHQH VHJPHQWV ZH REWDLQHG WKH LQGLFDWHG JHQH
VHJPHQWV IURP LQÀXHQ]D $9LHWQDP LVRODWHG IURP D KXPDQ FDVH LQ
9LHWQDPDQGFORQHGWKHVHLQWRS9$;*URXSVRIIRXUWRVL[PLFHZHUHYDFFLQDWHG
E\ '1$ WDWWRR RQ GD\ DQG ZLWK J RI S9$; H[SUHVVLQJ HLWKHU WKH
LQÀXHQ]D$9LHWQDP0+$1$RU13JHQHXQGHUFRQWURORIWKH&09
SURPRWHU
&HOOV DQG )ORZ F\WRPHWU\ )RU DQDO\VLV RI 0+& PXOWLPHU ELQGLQJ DQG 7 FHOO
UHVSRQVHV LQ PRXVH VDPSOHV ZH REWDLQHG SHULSKHUDO EORRG DQG HU\WKURF\WHV
ZHUH UHPRYHG E\ LQFXEDWLRQ LQ HU\WKURF\WH O\VLV EXIIHU P0 1+&O
P0.+&2P0('7$S+RQLFH&HOOVZHUHVWDLQHGZLWKDQWLERG\WR
&'Į%'%LRVFLHQFHVDQGWKHLQGLFDWHG0+&WHWUDPHUVIRU±PLQDWURRP
WHPSHUDWXUH
)RUDQDO\VLVRI0+&PXOWLPHUELQGLQJDQG7FHOOUHVSRQVHVLQKXPDQVDPSOHV
ZH REWDLQHG SHULSKHUDO EORRG PRQRQXFOHDU FHOOV RI KHDOWK\ YROXQWHHUV E\ )LFROO
JUDGLHQW VHSDUDWLRQ 7KH 0$57 VSHFL¿F &7/ FORQH ZDV REWDLQHG E\
UHSHWLWLYHVWLPXODWLRQDQGFORQLQJRIWXPRULQ¿OWUDWLQJO\PSKRF\WHVRIDPHODQRPD
SDWLHQW&HOOVZHUHVWDLQHGZLWKWKHLQGLFDWHG0+&WHWUDPHUVIRUPLQDW&
6XEVHTXHQWO\&'VSHFL¿FDQWLERG\%'%LRVFLHQFHVZDVDGGHGDQGFHOOVZHUH
LQFXEDWHGIRU±PLQDWURRPWHPSHUDWXUH'DWDDFTXLVLWLRQDQGDQDO\VLVZDV
FDUULHGRXWRQD)DFV&DOLEXU%HFWRQ'LFNLQVRQXVLQJ&HOO4XHVWVRIWZDUH
$FNQRZOHGJPHQWV
:HZRXOGOLNHWRWKDQN9&HUXQGROR:HDWKHUDOO,QVWLWXWHRI0ROHFXODU0HGLFLQH
2[IRUG8.IRUWKH$.EH[SUHVVLRQFRQVWUXFWDQG)/HPRQQLHU,QVWLWXW3DVWHXU
3DULV)UDQFHIRU+/$$WUDQVJHQLFPLFH:HZRXOGOLNHWRWKDQN$3IDXWK
DQG)YDQ'LHSHQIRUÀRZF\WRPHWU\DVVLVWDQFH,%ORQNIRUKHOSLQJHQHUDWLRQ
RI +1 '1$ YDFFLQHV DQG + +LONPDQQ IRU SHSWLGH V\QWKHVLV7KLV ZRUN ZDV
IXQGHGE\1:23LRQHHU76DQG9,',+2JUDQWV
,
,,
P]
P]
72)06(6
H
72)06(6
H
>,,+@
72)06(6
>,1D@
)LJXUH 3KRWRFOHDYDJH VWUDWHJ\ D 7RS 6WUXFWXUH RI , WKH SKRWRFOHDYDEOH
DQDORJRIWKHLQÀXHQ]D$0HSLWRSHREWDLQHGDVDQDSSUR[LPDWHRQHWRRQH
PL[WXUHRIGLDVWHUHRLVRPHUV6LQJOHDPLQRDFLGVHTXHQFH*,/*)9)-/%RWWRP
6WUXFWXUH RI ,, WKH KHSWDPHULF SHSWLGH IUDJPHQW JHQHUDWHG XSRQ 89LQGXFHG
GHJUDGDWLRQ RI ,$VWHULVNV LQ , DQG ,, LQGLFDWH WKH SUHIHUUHG FOHDYDJH VLWH E
/LTXLGFKURPDWRJUDSK\PDVVVSHFWURPHWU\/&06RI,SULRUWRDQGIROORZLQJ
H[SRVXUHWRQPOLJKW&$0$*IRUPLQLQWKHSUHVHQFHRIP0'77
([SHFWHGPDVVHVVLQJOHSURWRQDWHG,VLQJOHSURWRQDWHG,,
72)06(6
P]
>&09SS1D@
)LJXUH 89PHGLDWHG SHSWLGH H[FKDQJH 0DVV VSHFWURPHWU\ RI +/$$
DVVRFLDWHGSHSWLGHRI+/$$*,/*)9)-/FRPSOH[HVSULRUWROHIWSDQHODQG
IROORZLQJULJKWSDQHO89H[SRVXUHLQWKHSUHVHQFHRI&09SSSHSWLGH
2EVHUYHGPDVVHVDQGDVVLJQPHQWVDUHLQGLFDWHG([SHFWHGPDVVHV1DLRQRI,
1DLRQRI&09SS1RWHWKHDEVHQFHRIGHWHFWDEOH,1D IROORZLQJ89LQGXFHSHSWLGHH[FKDQJHDUURZ0+/$$*,/*)9)-/
LQP07ULV+&OS+P01D&ODQGP0'77ZDVH[SRVHGWR
QPOLJKWIRUPLQRQLFHLQWKHSUHVHQFHRI0&09SSSHSWLGH
6XEVHTXHQWO\ +/$$SHSWLGH FRPSOH[HV ZHUH SXUL¿HG E\ JHO¿OWUDWLRQ
FKURPDWRJUDSK\ RQ D %LRVHS 6(& 6 FROXPQ LQ P0 1+2$F S+
DQGGLUHFWO\XVHGIRUDQDO\VLVRQD:DWHUV/&7(6,PDVVVSHFWURPHWHUE\GLUHFW
LQIXVLRQ XQGHU RSWLPL]HG FRQGLWLRQV 9 FRQH YROWDJH & GHVROYDWLRQ
WHPSHUDWXUH1RWHWKDWWKHXVHRIKLJKFRQHYROWDJHKLJKWHPSHUDWXUHUHVXOWVLQ
WKHGHWHFWLRQRISUHGRPLQDQWO\VRGLDWHGVSHFLHV
$ %
$,890$57:73(
$,890$57$/3(
$0$57$/3(
$,89+<3(
$03(
$,8903(
$,89SS3(
$SS3(
$,89+<3(
)LJXUH 7 FHOO VWDLQLQJ ZLWK 0+& H[FKDQJH WHWUDPHUV D )ORZ F\WRPHWULF
DQDO\VLV RI 0+& WHWUDPHU VWDLQLQJ RI D 0$57VSHFL¿F &7/ FORQH ZLWK
FODVVLFDO0+&WHWUDPHUVFRQWDLQLQJWKH0$57$/HSLWRSHWRSOHIWSDQHORU
0+&H[FKDQJHWHWUDPHUVFRQWDLQLQJWKH0$57$/HSLWRSHWRSULJKWSDQHO
D FRQWURO SHSWLGH +< ERWWRP OHIW SDQHO RU WKH QDWXUDOO\ RFFXUULQJ ORZ
DI¿QLW\ 0$57 HSLWRSH ERWWRP ULJKW SDQHO E )ORZ F\WRPHWULF DQDO\VLV
RI SHULSKHUDO EORRG PRQRQXFOHDU FHOOV IURP DQ +/$$ SRVLWLYH LQGLYLGXDO
VWDLQHGZLWKFODVVLFDO0+&WHWUDPHUVFRQWDLQLQJWKH&09SSHSLWRSHWRS
OHIWSDQHORULQÀXHQ]D$0HSLWRSHWRSULJKWSDQHORUZLWK0+&H[FKDQJH
WHWUDPHUV FRQWDLQLQJ D FRQWURO SHSWLGH +< ERWWRP OHIW SDQHO WKH &09
SSHSLWRSHERWWRPPLGGOHSDQHORUWKHLQÀXHQ]D$0HSLWRSHERWWRP
ULJKWSDQHO
$
3HSWLGH 89
VWUHSSH
%
'E,989133(
'E,989133( 'E,98960&<3( 'E,93(
&RQWURO
$+.[
LQIHFWHG
F)ORZF\WRPHWULFDQDO\VLVRI0+&WHWUDPHUVWDLQLQJRISHULSKHUDOEORRGFHOOV
RID&%/PRXVHWRSSDQHORUD&%/PRXVHHLJKWGD\VSRVWLQWUDQDVDO
LQIHFWLRQZLWKLQÀXHQ]D$+.[HQFRGLQJWKH$3513$61(10(70
HSLWRSHERWWRPSDQHOV$QDO\VLVZDVSHUIRUPHGZLWK+'EH[FKDQJHWHWUDPHUV
FRQWDLQLQJWKH$3513HSLWRSHOHIWSDQHOVDFRQWUROSHSWLGH60&<
ERWWRPPLGGOHSDQHORU+'EWHWUDPHUVSUHSDUHGIURPELRWLQ\ODWHG+'E
$61(1-(70PRQRPHUVWKDWKDGQRWXQGHUJRQHH[FKDQJHUHDFWLRQVERWWRPULJKW
SDQHO
)LJXUH+LJKWKURXJKSXWVFUHHQRI+17FHOOHSLWRSHVD)ORZF\WRPHWULF
DQDO\VLVRIPRXVHSHULSKHUDOEORRGPRQRQXFOHDUFHOOVVWDLQHGZLWK$.E13 H[FKDQJHWHWUDPHUVRIHLWKHUDQRQYDFFLQDWHG+/$$WUDQVJHQLFPRXVHOHIW RURIDQ+/$$WUDQVJHQLFPRXVHYDFFLQDWHGDWGD\VZLWKJRI$
9LHWQDP13HQFRGLQJ'1$
$.E,89133( $.E,89133(
&
$
E7RS7FHOOUHVSRQVHVRILQGLYLGXDOPLFHFORVHGVTXDUHVDQGDYHUDJH7FHOO
UHVSRQVHV VWULSHV RI QRQYDFFLQDWHG PLFH ± DQG PLFH YDFFLQDWHG ZLWK WKH
LQÀXHQ]D$9LHWQDP0OHIWDQG13ULJKWJHQHVDQDO\]HGZLWK
WKHLQGLFDWHG$.EH[FKDQJHWHWUDPHUVDWGD\SRVWSULPDU\YDFFLQDWLRQ$WWKH
SHDN RI WKH YDFFLQDWLRQLQGXFHG 7 FHOO UHVSRQVH SHULSKHUDO EORRG ZDV GUDZQ
%ORRG RI PLFH LQ HDFK RI WKH IRXU JURXSV ZDV SRROHG DQG DQDO\]HG E\ 0+&
WHWUDPHUVWDLQLQJ0+&WHWUDPHUVWKDWVFRUHGSRVLWLYHZHUHXVHGIRUUHDQDO\VLVRI
LQGLYLGXDOPLFH%RWWRP7FHOOUHVSRQVHVRILQGLYLGXDOPLFHFORVHGVTXDUHVDQG
DYHUDJH7FHOOUHVSRQVHVVWULSHVRIQRQYDFFLQDWHGPLFH±DQGPLFHYDFFLQDWHG
ZLWK WKH LQÀXHQ]D $9LHWQDP 0 OHIW DQG 13 ULJKW JHQHV
DQDO\]HG ZLWK WKH LQGLFDWHG$.E H[FKDQJH WHWUDPHUV DW GD\ SRVW VHFRQGDU\
YDFFLQDWLRQ7KHLGHQWLW\RIWKH13HSLWRSHZDVFRQ¿UPHGE\V\QWKHVLVRIWKLV
VHTXHQFHRQDSUHSDUDWLYHVFDOHIROORZHGE\+3/&SXUL¿FDWLRQDQGVFUHHQLQJRI
YDFFLQDWHGPLFHE\0+&H[FKDQJHWHWUDPHUVWDLQLQJDQGLQWUDFHOOXODULQWHUIHURQȖ
VWDLQLQJGDWDQRWVKRZQ
$.E,8913
±
$.E,890
±
±
±
9DFFLQDWLRQ 0+&WHWUDPHU &'FHOOV0+&WHWUDPHU &'FHOOV
%
&
,,,
$0
$,
7LPHPLQ
$,89
$089
6XSSOHPHQWDU\ )LJXUH &RQGLWLRQDO OLJDQGV IRU +/$$ DQG +'E 7RS
6WUXFWXUH RI ,,, WKH SKRWRFOHDYDEOH DQDORJ RI WKH LQÀXHQ]D $ 0 HSLWRSH
LQ ZKLFK *O\ KDV EHHQ UHSODFHG E\ SKRWRODELOH DPLQRQLWURSKHQ\O
SURSLRQLF DFLG VLQJOH OHWWHU VHTXHQFH *,/-)9)7/ 0LGGOH 6WUXFWXUH RI ,9
WKH SKRWRFOHDYDEOH DQDORJ RI WKH LQÀXHQ]D$35 13HSLWRSH LQ ZKLFK
0HWKDVEHHQUHSODFHGE\DPLQRQLWURSKHQ\OSURSLRQLFDFLGVLQJOHOHWWHU
VHTXHQFH$61(1-(70 %RWWRP 6WUXFWXUH RI9 WKH SKRWRFOHDYDEOH DQDORJ RI
WKHLQÀXHQ]D$3513LQZKLFK*OXKDVEHHQUHSODFHGE\DPLQR
QLWURSKHQ\OSURSLRQLFDFLGVLQJOHOHWWHUVHTXHQFH$61(10-70
6XSSOHPHQWDU\)LJXUH&RQGLWLRQDOSHSWLGH0+&FRPSOH[HVDUHVHQVLWLYHWR
89 D *HO¿OWUDWLRQ FKURPDWRJUDSK\ RI +/$$ FRPSOH[HV RFFXSLHG ZLWK
WKHSDUHQWDO0HSLWRSHWRSRU+/$$FRPSOH[HVRFFXSLHGZLWKWKH89
VHQVLWLYH FRPSRXQG *,/*)9)-/ ERWWRP ZLWK JUHHQ RU ZLWKRXW UHG SULRU
H[SRVXUHWR89IRUKRQLFHLQP07ULV+&OS+P01D&ODQG
P0'77
$
,9
9
,QIOXHQ]D$0VSHFLILF7FHOOV &09SSVSHFLILF7FHOOV
0+&WHWUDPHUFRQFHQWUDWLRQJPO 0+&WHWUDPHUFRQFHQWUDWLRQJPO 0+&H[FKDQJHWHWUDPHUV 0+&WHWUDPHUV
E*HO¿OWUDWLRQFKURPDWRJUDSK\RI+/$$FRPSOH[HVRFFXSLHGZLWKWKH89
VHQVLWLYHSHSWLGH*,/*)9)-/H[SRVHGWR89IRUKRQLFHLQP07ULV+&O
S+P01D&ODQGP0'77ZLWKRXWSHSWLGHUHGRULQWKHSUHVHQFH
RI0RIWKH+/$$ELQGLQJSHSWLGHV0$57$/JUHHQRU+<
EOXH RU WKH +/$$ELQGLQJ SHSWLGH JS EODFN 1RWH )UHH 0+&
FODVV,KHDY\FKDLQVKDYHDSURSHQVLW\WRDJJUHJDWHDQGWKLVDJJUHJDWLRQRIIUHH
0+& FODVV , KHDY\ FKDLQV UHVXOWV LQ D ORVV RI PDWHULDO HOXWLQJ ZLWK D UHWHQWLRQ
WLPH RI PLQ 7KLV DQDO\VLV XQGHUHVWLPDWHV WKH H[WHQW RI 89LQGXFHG
0+&GHVWUXFWLRQDVDJJUHJDWLRQRI0+&FODVV,KHDY\FKDLQVLVLQFRPSOHWHDQG
VHSDUDWLRQRI0+&FODVV,FRPSOH[HVDQGIUHH0+&FODVV,KHDY\FKDLQVE\JHO
¿OWUDWLRQFKURPDWRJUDSK\LVLQHI¿FLHQW
6XSSOHPHQWDU\)LJXUH6HQVLWLYLW\DQGVSHFL¿FLW\RI0+&H[FKDQJHWHWUDPHUV
D3HULSKHUDOEORRGPRQRQXFOHDUFHOOVRIDQ+/$$SRVLWLYHLQGLYLGXDOZHUH
VWDLQHG ZLWK WKH LQGLFDWHG FRQFHQWUDWLRQV RI FRQYHQWLRQDO +/$$ WHWUDPHUV
RU ZLWK +/$$ H[FKDQJH WHWUDPHUV FRQWDLQLQJ HLWKHU WKH LQÀXHQ]D$ 0 HSLWRSHRUWKH&09SSHSLWRSH0HDQÀXRUHVFHQFHLQWHQVLW\RIWKH0+&
WHWUDPHU FHOOV ZDV GHWHUPLQHG DQG LV SORWWHG DV D IXQFWLRQ RI 0+& WHWUDPHU
FRQFHQWUDWLRQ
7LPHPLQ
$,89 0$57$/$ELQGHU
$,89
$,89 +<$ELQGHU
$,89 JS$ELQGHU
%
$
$,8903(
$,3(
$.E,3(
$.E,8903(
E)ORZF\WRPHWULFDQDO\VLVRISHULSKHUDOEORRGPRQRQXFOHDUFHOOVRIDQ+/$
$ SRVLWLYH LQGLYLGXDO ZLWK +/$$ WHWUDPHUV FRQWDLQLQJ WKH 89VHQVLWLYH
SHSWLGH*,/*)9)-/OHIWSDQHORUWKHSDUHQWDO0HSLWRSHULJKWSDQHOF
)ORZF\WRPHWULFDQDO\VLVRISHULSKHUDOEORRGPRQRQXFOHDUFHOOVRIDQ+/$$
WUDQVJHQLFPRXVHYDFFLQDWHGZLWKWKHLQÀXHQ]D$9LHWQDP0HQFRGLQJ
H[SUHVVLRQSODVPLGZLWK+/$$.EWHWUDPHUVFRQWDLQLQJWKH89VHQVLWLYHSHSWLGH
*,/*)9)-/OHIWSDQHORUWKHSDUHQWDO0HSLWRSHULJKWSDQHO
%
'LPHU;
+/$$,J
89H[FKDQJH +/$$7HWUDPHUV
FRQWURO &09 FRQWURO &09
6XSSOHPHQWDU\ )LJXUH 'HWHFWLRQ RI DQWLJHQ VSHFL¿F 7 FHOOV ZLWK 0+&,J
GLPHUVDQG0+&H[FKDQJHWHWUDPHUVD)ORZF\WRPHWULFDQDO\VLVRISHULSKHUDO
EORRG PRQRQXFOHDU FHOOV RI IRXU &09SRVLWLYH +/$$ SRVLWLYH LQGLYLGXDOV
ZLWK 0+&,J 'LPHU; UHDJHQWV OHIW SUHSDUHG ZLWK D FRQWURO SHSWLGH +<
RUWKH&09SSSHSWLGHRUZLWK+/$$H[FKDQJHWHWUDPHUVULJKW SUHSDUHGZLWKDFRQWUROSHSWLGH+<RUWKH&09SSSHSWLGH
$
E3ORWVRIWKHEDFNJURXQGDQGVSHFL¿F
VWDLQLQJREWDLQHGZLWK0+&,JUHDJHQWV
DQG 0+& H[FKDQJH WHWUDPHUV LQ IRXU
GRQRUV 1RWH WKDW WKH LQWHQVLW\ RI
VWDLQLQJ RI &09 SS VSHFL¿F 7
FHOOVLVKLJKHUZLWK+/$$H[FKDQJH
WHWUDPHUV WKDQ ZLWK 0+&,J GLPHUV
DQG WKDW WKH EDFNJURXQG REVHUYHG ZLWK
FRQWURO0+&PXOWLPHUVLVVXEVWDQWLDOO\
ORZHU WKDQ WKH EDFNJURXQG REVHUYHG
ZLWK 0+&,J GLPHUV 0+&,J GLPHUV
IRU +/$$ %' %LRVFLHQFHV ZHUH
ORDGHG ZLWK WKH +<RU &09
SS SHSWLGH SHU PDQXIDFWXUHU¶V
SURWRFRO ,Q EULHI 'LPHU; UHDJHQWV
ZHUHSUHSDUHGRYHUQLJKWDW&XVLQJ
D IROG PRODU H[FHVV RI SHSWLGH
RYHU 0+&,J SURWRFRO SHSWLGH
FRQFHQWUDWLRQ RI 0 &HOOV ZHUH
VWDLQHGZLWKJRI'LPHU;UHDJHQW
IRUKDW&IROORZHGE\VWDLQLQJZLWK
3(ODEHOHGDQWLPRXVH,J*FORQH$
%' %LRVFLHQFHV SOXV ),7&ODEHOHG
&'VSHFL¿F DQWLERG\ FORQH *
%' %LRVFLHQFHV IRU PLQ DW URRP
WHPSHUDWXUHSURWRFRO
FRQWURO &09 FRQWURO &09
FRQWURO &09 FRQWURO &09
FRQWURO &09 FRQWURO &09
FRQWURO &09 FRQWURO &09
'LPHU;
+/$$,J
89H[FKDQJH +/$$
7HWUDPHUV
0+& H[FKDQJH WHWUDPHUV ZLWK WKH +<RU &09 SS SHSWLGH ZHUH
SUHSDUHG E\ SHUIRUPLQJ K 89H[FKDQJH UHDFWLRQV RQ +/$$*,/*)9)-/
PRQRPHUVLQWKHSUHVHQFHRI0RIWKHLQGLFDWHGSHSWLGHRQLFH6XEVHTXHQWO\
VDPSOHV ZHUH VSXQ DW J IRU PLQ 3(VWUHSWDYLGLQ JPO ¿QDO
FRQFHQWUDWLRQZDVDGGHGDQGWKHUHVXOWLQJ0+&H[FKDQJHWHWUDPHUVZHUHXVHG
IRU7FHOOVWDLQLQJZLWKRXWIXUWKHUSXUL¿FDWLRQ
%
/MXQJJUHQ+*HWDO(PSW\0+&FODVV,PROHFXOHVFRPHRXWLQWKHFROG
1DWXUH
6FKXPDFKHU71 HW DO 'LUHFW ELQGLQJ RI SHSWLGH WR HPSW\ 0+& FODVV ,
PROHFXOHVRQLQWDFWFHOOVDQGLQYLWUR&HOO
)UHPRQW'+0DWVXPXUD06WXUD($3HWHUVRQ3$ :LOVRQ,$
&U\VWDOVWUXFWXUHVRIWZRYLUDOSHSWLGHVLQFRPSOH[ZLWKPXULQH0+&FODVV,+
.E6FLHQFH
6LOYHU0/*XR+&6WURPLQJHU-/ :LOH\'&$WRPLFVWUXFWXUHRI
DKXPDQ0+&PROHFXOHSUHVHQWLQJDQLQÀXHQ]DYLUXVSHSWLGH1DWXUH
%RXYLHU 0 :LOH\ '& ,PSRUWDQFH RI SHSWLGH DPLQR DQG FDUER[\O
WHUPLQLWRWKHVWDELOLW\RI0+&FODVV,PROHFXOHV6FLHQFH
6FKXPDFKHU71HWDO3HSWLGHVHOHFWLRQE\0+&FODVV,PROHFXOHV1DWXUH
)DON . 5RW]VFKNH 2 6WHYDQRYLF 6 -XQJ * 5DPPHQVHH +*
$OOHOHVSHFL¿FPRWLIVUHYHDOHGE\VHTXHQFLQJRIVHOISHSWLGHVHOXWHGIURP0+&
PROHFXOHV1DWXUH
$OWPDQ-'HWDO3KHQRW\SLFDQDO\VLVRIDQWLJHQVSHFL¿F7O\PSKRF\WHV
6FLHQFH
6RHQ<&KHQ'6.UDIW'/'DYLV00 %URZQ32'HWHFWLRQDQG
FKDUDFWHUL]DWLRQ RI FHOOXODU LPPXQH UHVSRQVHV XVLQJ SHSWLGH0+& PLFURDUUD\V
3/R6%LRO(
6WRQH -' 'HPNRZLF] :( -U 6WHUQ /- +/$UHVWULFWHG HSLWRSH
LGHQWL¿FDWLRQDQGGHWHFWLRQRIIXQFWLRQDO7FHOOUHVSRQVHVE\XVLQJ0+&SHSWLGH
DQGFRVWLPXODWRU\PLFURDUUD\V3URF1DWO$FDG6FL86$
<HH&6DYDJH3$/HH33'DYLV00 *UHHQEHUJ3',VRODWLRQ
RI KLJK DYLGLW\ PHODQRPDUHDFWLYH &7/ IURP KHWHURJHQHRXV SRSXODWLRQV XVLQJ
SHSWLGH0+&WHWUDPHUV-,PPXQRO
)DONHQEXUJ -+ YDQ GH &RUSXW / 0DULMW (: :LOOHP]H 5 0LQRU
KLVWRFRPSDWLELOLW\DQWLJHQVLQKXPDQVWHPFHOOWUDQVSODQWDWLRQ([S+HPDWRO
0RVV3 5LFNLQVRQ$&HOOXODULPPXQRWKHUDS\IRUYLUDOLQIHFWLRQDIWHU
+6&WUDQVSODQWDWLRQ1DW5HY,PPXQRO
3HJJV .6 HW DO $GRSWLYH FHOOXODU WKHUDS\ IRU HDUO\ F\WRPHJDORYLUXV
LQIHFWLRQDIWHUDOORJHQHLFVWHPFHOOWUDQVSODQWDWLRQZLWKYLUXVVSHFL¿F7FHOOOLQHV
/DQFHW
5RRQH\&0HWDO8VHRIJHQHPRGL¿HGYLUXVVSHFL¿F7O\PSKRF\WHVWR
FRQWURO(SVWHLQ%DUUYLUXVUHODWHGO\PSKRSUROLIHUDWLRQ/DQFHW
:DOWHU ($ HW DO 5HFRQVWLWXWLRQ RI FHOOXODU LPPXQLW\ DJDLQVW
F\WRPHJDORYLUXV LQ UHFLSLHQWV RI DOORJHQHLF ERQH PDUURZ E\ WUDQVIHU RI 7FHOO
FORQHVIURPWKHGRQRU1(QJO-0HG
&REEROG 0 HW DO$GRSWLYH WUDQVIHU RI F\WRPHJDORYLUXVVSHFL¿F &7/ WR
VWHPFHOOWUDQVSODQWSDWLHQWVDIWHUVHOHFWLRQE\+/$SHSWLGHWHWUDPHUV-([S0HG
%RVTXHV&- ,PSHULDOL%3KRWRO\WLFFRQWURORISHSWLGHVHOIDVVHPEO\-
$P&KHP6RF
:LHEROGW 5 HW DO 6\QWKHVLV DQG FKDUDFWHUL]DWLRQ RI SKRWRODELOH R
QLWUREHQ]\OGHULYDWLYHVRIXUHD-2UJ&KHP
*DUERF]L '1 +XQJ '7 :LOH\ '& +/$$SHSWLGH FRPSOH[HV
UHIROGLQJ DQG FU\VWDOOL]DWLRQ RI PROHFXOHV H[SUHVVHG LQ (VFKHULFKLD FROL DQG
FRPSOH[HGZLWKVLQJOHDQWLJHQLFSHSWLGHV3URF1DWO$FDG6FL86$
6XO\RN *$ HW DO 6ROLGSKDVH V\QWKHVLV RI D QRQSHSWLGH 5*' PLPHWLF
OLEUDU\QHZVHOHFWLYHDOSKDYEHWDLQWHJULQDQWDJRQLVWV-0HG&KHP
5DPPHQVHH +* %DFKPDQQ - (PPHULFK 11 %DFKRU 2$ 6WHYDQRYLF 6 6<)3(,7+, GDWDEDVH IRU 0+& OLJDQGV DQG SHSWLGH PRWLIV
,PPXQRJHQHWLFVDFFHVVYLDZZZV\ISHLWKLGH
9DOPRUL ' HW DO (QKDQFHG JHQHUDWLRQ RI VSHFL¿F WXPRUUHDFWLYH &7/
LQ YLWUR E\ VHOHFWHG 0HODQ$0$57 LPPXQRGRPLQDQW SHSWLGH DQDORJXHV -
,PPXQRO
*UHWHQ7)HWDO'LUHFWYLVXDOL]DWLRQRIDQWLJHQVSHFL¿F7FHOOV+7/9
7D[VSHFL¿F&'7FHOOVDUHDFWLYDWHGLQSHULSKHUDOEORRGDQGDFFXPXODWH
LQFHUHEURVSLQDOÀXLGIURP+$0763SDWLHQWV3URF1DWO$FDG6FL86$
2HONH0 6FKQHFN-3+/$,JEDVHGDUWL¿FLDODQWLJHQSUHVHQWLQJFHOOV
VHWWLQJWKHWHUPVRIHQJDJHPHQW&OLQ,PPXQRO
)LUDW+HWDO+FODVV,NQRFNRXW+/$$WUDQVJHQLFPLFHDYHUVDWLOH
DQLPDOPRGHOIRUSUHFOLQLFDOHYDOXDWLRQRIDQWLWXPRULPPXQRWKHUDSHXWLFVWUDWHJLHV
(XU-,PPXQRO
%LQV$HWDO$UDSLGDQGSRWHQW'1$YDFFLQDWLRQVWUDWHJ\GH¿QHGE\LQ
YLYRPRQLWRULQJRIDQWLJHQH[SUHVVLRQ1DW0HG
&KRL(03DOPRZVNL0&KHQ- &HUXQGROR97KHXVHRIFKLPHULF
$.EWHWUDPHUVWRPRQLWRU+/$$LPPXQHUHVSRQVHVLQ+/$$WUDQVJHQLF
PLFH-,PPXQRO0HWKRGV
'XGOH\ 0( HW DO &DQFHU UHJUHVVLRQ DQG DXWRLPPXQLW\ LQ SDWLHQWV DIWHU
FORQDOUHSRSXODWLRQZLWKDQWLWXPRUO\PSKRF\WHV6FLHQFH
6FKOLFKWLQJ , HW DO 7LPHUHVROYHG ;UD\ FU\VWDOORJUDSKLF VWXG\ RI WKH
FRQIRUPDWLRQDO FKDQJH LQ +D5DV S SURWHLQ RQ *73 K\GURO\VLV 1DWXUH