Correspondence: Claudio Sorio, MD, Ph.D., Department of Pathology, General Pathology Section, University of Verona, Strada Le Grazie 87134 Verona, Italy. Tel: +39-045-8027688; Fax: +39-045-8027127;
Email: claudio.sorio@univr.it
Please note that this article may not be used for commercial purposes. For further information please refer to the copyright
Protein Tyrosine Phosphatase Gamma
(PTP
!) is a Novel
Leukocyte Marker Highly Expressed by CD34
+Precursors
Andrea Mafficini1, Marzia Vezzalini1, Loris Zamai2,3,4, Laura Galeotti2,3
Gabriella Bergamini5, Marco Della Peruta1, Paola Melotti5 and Claudio Sorio1
1Department of Pathology, University of Verona, Italy.
2Institute of Histology and Laboratory Analysis, University of Urbino “Carlo Bo”, Italy.
3Flow Cytometry and Cytomorphology Center, University of Urbino “Carlo Bo”, Italy.
4INFN-Gran Sasso National Laboratory, SS17bis km 18+910, 67010 Assergi, L’Aquila, Italy.
5Cystic Fibrosis Center, Azienda Ospedaliera of Verona, Verona, Italy.
!bstra't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in vitro(%B-.'+&#'(&'<;3%<(7*(GH4M(.'<(IKJ(/"#:$,(8.34 $#"+9(=%($,%"%8#"%(-"#-#+%(!)!!(.+(.('%:(8;'3$&#'.55*("%/;5.$%<(5%;6#3*$%(0."6%"(:,#+%("#5%(&'('#"0.5(.'<(-.$,#5#/&3.5( 3#'$%B$(<%+%">%(8;"$,%"(&'>%+$&/.$&#'9( Key-ords:(P//(*#56(&00;'#/5#7;5&'(1G/Q2R(,;0.'(-"#$%&'($*"#+&'%(-,#+-,.$.+%(/.00.(1!)!!2R((;#"%+3%'3%(.3$&>.$%<( 3%55(+#"$&'/(1JAKI2R(,.%0.$#-#&%$&3(-"#/%'&$#"+9 !bbreviations:(G/QC(K,&36%'(%//(*#56(&00;'#/5#7;5&'+R(3,!)!!C(A'$&4!)!!(G/Q(.'$&7#<*R(!)!!C(,;0.'(-"#$%&'($*"#+&'%( -,#+-,.$.+%(/.00.9
Introduction
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in vitro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
Materials and Methods
Cells, puri cation and culture
),%(3%55(5&'%+(_[\S(1A'<%"++#'(%$(.59(UYZY2C(`H4\T( 1a.55./,%"( %$( .59( UYZY2C( )`!4U( 1)+;3,&*.( %$( .59( UYbT2(.'<(^YMZ(1I;'<+$"#0(.'<(@&5++#'C(UYZ\2( :%"%( -;"3,.+%<( 8"#0(A)KKC( EH4M( 1c,*.+,&6&( %$( .59( UYb\2( :.+( .( 6&'<( /&8$( 8"#0( -"#89( E9A9( K.++.$%55.9(K&"3;5.$&'/(,;0.'(0#'#3*$%+(1dZTe( -;"%(.+(.++%++%<(7*(%B-"%++&#'(#8(KLUN2C(-#5*4 0#"-,#';35%."(3%55+(1dY[e(-;"%(.+(.++%++%<(7*( KLU[(%B-"%++&#'2(.'<(5*0-,#3*$%+(1dY[e(-;"%( .+(.++%++%<(7*(0#"-,#5#/*(.'<($,%(5.36(#8(KLUN2( :%"%( -;"&8&%<( 7*( !%"3#55( 1!,."0.3&.( ^--+.5.C( I:%<%'2(/".<&%'$(3%'$"&8;/.$&#'(8"#0(5%;6#3*$%4 "&3,( 7;88*( 3#.$+( #7$.&'%<( 8"#0( ,;0.'( 75##<( #8( ,%.5$,*( <#'#"+C( .+( <%+3"&7%<( %5+%:,%"%( 1E;D&#( %$(.59(UYYN29(G00.$;"%(0#'#3*$%4<%"&>%<(,;0.'( <%'<"&$&3(3%55+(10#LK2(:%"%(#7$.&'%<(in vitro .+( -"%>&#;+5*(<%+3"&7%<(1I#DD.'&(%$(.59(UYYZ29(F"&%(*C( -;"&(%<(0#'#3*$%+(:%"%(3;5$;"%<(&'(?!EG(U\NT( 1S("#UT\( 3%55+W:%552( 3#'$.&'&'/( UTe( ,%.$4&'.3$&4
>.$%<(JKIC(S0E(/5;$.0&'%(.'<(+;--5%0%'$%<(:&$,( [T( '/W05( "%3#07&'.'$( ,;0.'( aE4KIJ( .'<( ST( '/W05( "%3#07&'.'$( ,;0.'( GH4N( 1!%-"#$%3,C( ?#36*(`&55C(@f2(8#"([g\(<.*+9()#(&'<;3%(0.$;".4 $&#'C(&00.$;"%(0#LK(:%"%($"%.$%<(8#"(SN(,(:&$,( UTT('/W05(H!I(1P+3,%"&3,&.(3#5&(+%"#$*-%(TS\V(F\R( I&/0.C(I$9(H#;&+(Ec2(3%55+(:%"%(+;7]%3$%<($#((#:( 3*$#0%$"*(.'.5*+&+($#(.++%++($,%(.3$&>.$&#'W0.$;".4 $&#'( +$.$;+( .+( -"%>&#;+5*( <%+3"&7%<( 1H&++.'<"&'&( %$(.59(STT\29(J#"(KLMN3 3%55(+%5%3$&#'C(0#'#';35%."(
3%55+(:%"%(&+#5.$%<(8"#0(,%.5$,*(<#'#"+(7;88*(3#.$+( 7*(J&3#55W`&+$#-.h;%4UTZZ(1I&/0.4A5<"&3,C(E&5.'C(
G$.5*2( /".<&%'$( 3%'$"&8;/.$&#'C( "&'+%<( .'<( .<,%"4 %'3%4<%-5%$%<(8#"(#'%(,#;"(.+(<%+3"&7%<(&'(1i.0.&( %$(.59(STTT2(:&$,(0#<&(3.$&#'+9(A8$%"("%0#>.5(#8( .<,%"%'$(3%55+C(KLMNO(3%55+(&+#5.$&#'(:.+(.33#04
-5&+,%<(7*(&00;'#0./'%$&3.5(-#+&$&>%(+%5%3$&#'( ;+&'/(0./'%$&3(7%.<+(3#.$%<(:&$,(.'$&4KLMN(0A7( 1KLMN( &+#5.$&#'( 6&$( -5;+( X."EAKI( +*+$%0C( E&5$%'*&(F&#$%3,C(F#5#/'.C(G$.5*2C(.33#"<&'/($#($,%( 0.';8.3$;"%"j+(&'+$";3$&#'+R(3%55+(:%"%(+;7]%3$%<( $#((#:(3*$#0%$"*($#(.++%++(-;"&$*(.'<(!)!!(%B-"%+4 +&#'9( !;"&8&%<( KLMNO( 3%55+( 1[TCTTTW052( :%"%( 3;5$;"%<(8#"(M(<.*+(&'(U(05(k4X&>#(ST(1F&#=&$$.6%"C( =.56%"+>&55%C(EAC(^9I9A2(0%<&;0(+;--5%0%'$%<( :&$,(UTe(,;0.'(AF(+%";0(&'($,%(-"%+%'3%(#8(+$%0( 3%55(8.3$#"(1IKJC([T('/W052(.'< GH4M(1UT('/W0529( A55(3*$#6&'%+(:%"%(-;"3,.+%<(8"#0(!%-"#$%3,(PK( H$<9(1H#'<#'C(^9_929
Antibody production
A(3,&36%'(-#5*35#'.5(.'$&7#<*(:.+(".&+%<(./.&'+$( $,%(+%h;%'3%(Ki(@PL(P_P(_)J()_L(IL_(LH_( 1"%+&<;%+( lMYTgNTZ( #8( %B$".( 3%55;5."( !)!!( +%h;%'3%2R( -"#<;3%<( G/Q( :%"%( .8('&$*( -;"&(%<( ./.&'+$($,%(+.0%(+%h;%'3%(7*(A>%+(H.7+(1)&/."<C( c?C( ^9I9A29( !"%4&00;'%( 3,&36%'( G/Q( :%"%( 3#55%3$%<(.'<(-;"&(%<(8"#0($,%(+.0%(,%'(7%8#"%( $,%(&00;'&D.$&#'(-"#3%++9(F"&%(*C($,%(.'$&/%'(:.+( &']%3$%<(&'$#($,%(-%3$#".5(0;+35%(#8(#'%(A0%"&3.'4 5.*&'/(,%'9(),"%%(7##+$%"(&']%3$&#'+(3#'$.&'&'/([T( m/( #8( .'$&/%'( 0&B%<( :&$,( &'3#0-5%$%( J"%;'<j+( .<];>.'$(:%"%(/&>%'(.$(SC(N(.'<(\(:%%6+9(),%(%//+( :%"%(3#55%3$%<(<.&5*(.'<(+$#"%<(.$(N(nK(;'$&5($,%( .'$&7#<&%+( :%"%( %B$".3$%<9(),%( 3";<%( .'$&4!)!!( 3,&36%'(G/Q(13,!)!!2(:.+(8;"$,%"(-;"&(%<(;+&'/( .88&'&$*( 3#5;0'( 3,"#0.$#/".-,*( ./.&'+$( $,%( ST4.0&'#(.3&<(-%-$&<%9(),%('#'4+-%3&(3(-"#$%&'+( :%"%(:.+,%<(8"#0($,%(3#5;0'(:&$,(:.+,&'/(7;88%"( 1T9U(E()"&+g(`K5C(T9[(E(@.K5C(-`(b9T2C(;'$&5($,%( .7+#"7.'3%(.$(SbT('0(<%3"%.+%<($#(D%"#(.'<($,%( .'$&7#<*(:.+($,%'(%5;$%<(:&$,(.(<%+#"7&'/(./%'$( 1T9U(E(/5*3&'%C(-`(M9T2(PTP
! extra cellular domain-enriched
supernatants production
SYMJ(3%55+(1G'>&$"#/%'C(E&5.'C(G$.5*2(:%"%($".'+4 8%3$%<(:&$,(.(3L@A(%'3#<&'/(8#"($,%(%B$".(3%55;5."( <#0.&'(#8(!)!!(1-!)!B2C(3#'$.&'&'/($,%(%-&$#-%( #8(&'$%"%+$C(35#'%<(&'(-?KWKEX(>%3$#"(1G'>&$"#/%'C( E&5.'C( G$.5*29( )".'+8%3$&#'( :.+( .3,&%>%<( ;+&'/( H&-#8%3$.0&'%STTT)E( 1G'>&$"#/%'C( E&5.'C( G$.5*2C(
.33#"<&'/($#($,%(0.';8.3$;"%"j+(#-$&0&D%<(-"#$#3#5( 8#"( SYMJ( 3%55+9( !#+&$&>%( 3%55+( :%"%( +%5%3$%<( 7*( -5.$&'/($".'+8%3$%<(+.0-5%+(.$(.7#;$(UT[(3%55+W0H(
&'( ?!EG( U\NTC( UTe( JFIC( Ue( ^5$"./5;$.0&'%C( T9Y( 0/W0H( aNUb( 1G'>&$"#/%'C( E&5.'C( G$.5*2( .'<( &'3;7.$&'/($,%0(&'(.(,;0&<&(%<(.$0#+-,%"%(:&$,( [e(KcS(.$(MZ(nK9(PB,.;+$(0%<&;0(:.+(,.">%+$%<( %>%"*(Nb(,#;"+(.'<(3%'$"&8;/%<([(0&'(.$(M[T("(/( $#( %5&0&'.$%( "%+&<;.5( 3%55+9(A8$%"( <%3.'$&'/C( $,%( +;-%"'.$.'$(:.+(8;"$,%"(3%'$"&8;/%<(8#"(UT(0&'(.$( MTTT("(/($#(.>#&<(3%55;5."(<%7"&+9(),%(35."&(%<( -##5+(:%"%(+$#"%<(.$(gST(nK(;'$&5('%%<%<9(
Western blot analysis
Ph;.5(.0#;'$(#8(+%";048"%%(+-%'$(0%<&;0(:%"%( 0&B%<($#(T9M(>#5;0%+(#8(+.0-5%(7;88%"(1U\T(0E( )"&+C( STe( a5*3%"#5C( [e($40%"3.-$#%$,.'#5C( Ne(ILIC(T9TUe(7"#0#-,%'#5(75;%2(.'<(5#.<%<(#'( .(UTe(-#5*.3"*5.0&<%C(T9Ue(ILI(/%59(a%5+(:%"%( ";'(8#"(U(,#;"(.$(STTXC(UN(0A(&'(.(E&'&(!"#$%.'(M( A--.".$;+( 1F&#".<C( E&5.'C( G$.5*2( :&$,( T9Me)"&+C( U9N[e(a5*3&'%C(T9Ue(ILI(";''&'/(7;88%"C(-`(b9Y9( A8$%"(ILI4!AaPC(/%5+(:%"%(&'3;7.$%<(8#"(ST(0&'( &'( $".'+8%"( 7;88%"( 1ILI( ";''&'/( 7;88%"C( STe( 0%$,.'#52( .'<( $,%'( %5%3$"#475#$$%<( #'( `*7#'<( '&$"#3%55;5#+%(1aP(`%.5$,3."%C(E&5.'C(G$.5*2(8#"(U( ,#;"(.$(UTTXC(UST(0A(&'(.(E&'&()".'+4F5#$(P5%34 $"#-,#"%$&3()".'+8%"( K%55( 1F&#".<C( E&5.'C( G$.5*2R( $,%( ,#0#/%'%#;+( $".'+8%"( #8( $,%( +.0-5%+( #'( 0%07".'%+(:.+(3,%36%<(7*(!#'3%.;(+$.&'&'/9(J#"( ;+%(:&$,(3,!)!!C(0%07".'%+(:%"%(+.$;".$%<(7*( &'3;7.$&#'( 8#"( U( ,#;"( &'( UTe( F5#36`%'( GGo( 1A>%+H.7+C( )&/."<( c"%/#'C( ^9I9A29(A8$%"( $,"%%( :.+,%+(&'(!FIC(T9T[e($:%%'p(ST($,%(0%07".'%+(
:%"%(&'3;7.$%<(#>%"'&/,$(:&$,(3,!)!!(1Um/W052( <&5;$%<(&'(!FIC(T9Ue($:%%'p(ST9(E%07".'%+(:%"%(
$,%'( :.+,%<( $,"%%( $&0%+( :&$,( !FIC( T9T[e( $:%%'p(ST( .'<( &'3;7.$%<( 8#"( U( ,#;"( :&$,( /#.$(
.'$&43,&36%'( `?!( 1A>%+H.7+C( )&/."<( c"%/#'C( ^9I9AR( <&5;$&#'( UVU[CTTT29(A8$%"( $,"%%( 8;"$,%"( :.+,%+C(0%07".'%+(:%"%(.++.*%<(:&$,(PKH(1aP( `%.5$,3."%C(E&5.'C(G$.5*29(
RNA extraction and northern blot
UT(%/(#8($#$.5(?@A(:.+(%B$".3$%<(8"#0(3%55(5&'%+( ;+&'/( $,%( )?GD#5p( "%./%'$( 1G'>&$"#/%'C( E&5.'C(
G$.5*2(.33#"<&'/($#($,%(0.';8.3$;"%"(&'+$";3$&#'+( .'<( %5%3$"#-,#"%+%<( #'( .( Ue( 8#"0.5<%,*<%4 ./."#+%(/%5C(75#$$%<(#'$#(.(`*7#'<(@O(0%07".'%( .'<( ,*7"&<&D%<( $#(MS!45.7%5%<( 3L@A( -"#7%+(
-"%-."%<(7*(".'<#0(-"&0&'/(6&$(1?%.<*($#4/#(L@A(
5.7%5&'/( F%.<+C(A0%"+,.0( !,."0.3&.( F&#$%3,C( ^--+.5.C(I:%<%'2(;+&'/(&4MS!(<K)!(1GK@C(K#+$.(
E%+.C(KA29(),%(,*7"&<&D.$&#'(:.+(-%"8#"0%<(.+( <%+3"&7%<( 1I.07"##6( %$( .59( UYbY29( !)!!C(!)!'C( KLUNb(.'<(.3$&'(3L@A(:%"%(;+%<(.+(-"#7%+9(!)!!# 3L@A(:.+(.0-5&(%<(7*(!K?(8"#0(!)!!(-K?pM9U( -5.+0&<(1G'>&$"#/%'2(;+&'/($,%(8#55#:&'/(-"&0%"+V( 8#":."<([j(Ka)(KAK(KAa()K)(KK)(MjC("%>%"+%( [j(aAA(aAa(aKA(aaA(aAa(Mj9(!)!'(3L@A( -"#7%(:.+(#7$.&'%<(7*("%>%"+%($".'+3"&-$&#'(#8($#$.5( ?@A(8"#0(,;0.'(.+$"#3*$#0.($&++;%(.'<(.0-5&(%<( ;+&'/($,%(8#55#:&'/(-"&0%"+V(8#":."<([j(K)A(aK)( aAa(aaa())a(aAA()K(MjC("%>%"+%([j(a)a(KK)( a))(K))(KKA(AK)(KK(Mj9(KLUNb(3L@A(-"#7%( :.+(#7$.&'%<(7*("%>%"+%($".'+3"&-$&#'(#8($#$.5(?@A( 8"#0( ,;0.'( -%"&-,%".5( 75##<( /".';5#3*$%+( .'<( .0-5&(%<(;+&'/($,%(8#55#:&'/(-"&0%"+V(8#":."<([j( )aK(KAK(AKA(Aaa(AKK(MjC("%>%"+%([j()aA( )))( aK)( KKK( KAK( Mj9( ),%( &<%'$&$*( #8( 7#$,( -"#7%+(:.+(3#'("0%<(7*(+%h;%'3&'/9(
Flow cytometry of puri ed cells
and whole blood
!;"&8&%<( 3%55+( .'<( :,#5%( 75##<( +.0-5%+( :%"%( +$.&'%<( :&$,( 0#'#35#'.5( .'$&4KLMN( !P( 135#'%( AKUM\C( E&5$%'*&( F&#$%3C( F#5#/'.C( G$.5*2C( .'$&4 KLUN( !PC( .'$&4KLU[( JG)KC( .'$&4KLM( !PC( .'$&4 KLUY( !PC( .'$&4KLU3( 7&#$&'C( .'$&4KLMTM( 7&#$&'C( .'$&4KLU.( JG)KC( .'$&4KLbT( !PC( .'$&4KLb\( !PC( .'$&4KLbM(JG)K(1F&#H%/%'<C(I.'(L&%/#C(KA2(.'<( $,%(.'$&4!)!!(3,&36%'(G/Q(%+$.75&+,%<(&'(#;"(5.79( F&#$&'*5.$%<(.'$&7#<&%+(:%"%(<%$%3$%<(:&$,(+$"%-4 $.>&<&'(!PWK*[(1F&#H%/%'<C(I.'(L&%/#C(KA29(A+( .'(&+#$*-&3(3#'$"#5C(-"%4&00;'%(G/Q(.'<(0.$3,%<( G/aU4!P( #"( G/aS.4!P( 1F&#H%/%'<C I.'( L&%/#C( KA2( :%"%( ;+%<9( F"&%(*C( -;"&(%<( 3%55+( :%"%( ("+$( 75#36%<(:&$,(UTe(>W>(,;0.'(+%";0(1G'>&$"#/%'C( E&5.'C(G$.5*2(8#"(U[(0&';$%+(.$(?)C($,%'(&'3;7.$%<( :&$,(3,!)!!(1S(%/WUT\(3%55+2(8#"(U(,#;"(.$(?)9(A8$%"(
#'%(:.+,(:&$,(+$.&'&'/(7;88%"(1[e(JFI(&'(!FIC( T9Ue( @.@M2C( 3%55+( :%"%( +$.&'%<( :&$,( /#.$( .'$&4 3,&36%'(A5%B.Nbb(1G'>&$"#/%'C(E&5.'C(G$.5*2(8#"(MT( 0&'(.$(?)(.'<W#"(:&$,(0#'#35#'.5(.'$&7#<&%+(8#"( U[(0&'(.$(?)9(=,%"%(7&#$&'*5.$%<(.'$&7#<&%+(:%"%( &'35;<%<C(.(8;"$,%"(:.+,(:.+('%3%++."*C(8#55#:%<( 7*(.'(&'3;7.$&#'(:&$,(+$"%-$.>&<&'(!PWK*[(8#"(U[( 0&'( .$( ?)9( !%"&-,%".5( 75##<( +.0-5%+( :%"%( ("+$( &'3;7.$%<(:&$,(S(%/(3,!)!!(#"(-"%4&00;'%(G/Q( 8#"(U(,#;"(.$(?)C($,%'(+;7]%3$%<($#(%"*$,"#3*$%+( 5*+&+( 7*( .<<&$&#'( #8( ST( >#5;0%+( #8( %"*$,"#3*$%+( 5*+&+( +#5;$&#'( 1T9bb\e( @`NK5C( T9Ue( _`KcMC(
T9TT\e(PL)A2(.'<(&'3;7.$&#'(8#"(UT(0&'(.$(?)( :&$,(/%'$5%(./&$.$&#'9(A8$%"(.<<&$&#'(#8((T9[(>#5;0%+( #8(+$.&'&'/(7;88%"(.'<(-%55%$&'/(#8(3%55+C(.(8;"$,%"( :.+,( :&$,( +$.&'&'/( 7;88%"( :.+( -%"8#"0%<9( K%55+( :%"%($,%'(+$.&'%<(:&$,(/#.$(.'$&43,&36%'(A5%B.Nbb( 8#"(MT(0&'(.$(?)(.'<W#"(:&$,(0#'#35#'.5(.'$&7#<&%+( .+(.7#>%(<%+3"&7%<9(J5#:(3*$#0%$"*(:.+(-%"8#"0%<( #'(.(F%3$#'(L&36&'+#'(JAKI3.'p((#:(3*$#0%$%"9( A'.5*+&+(#8((#:(3*$#0%$"*(<.$.(:.+(-%"8#"0%<( :&$,( JKI( PB-"%++(XM( +#8$:."%( 1L%( @#>#( I#8$4 :."%2(.'<(/%#0%$"&3(0%.'((;#"%+3%'3%(&'$%'+&$*( 1EJG2(".$&#(7%$:%%'(3,!)!!(.'<(&+#$*-%(3#'$"#5( .'$&7#<&%+(,.+(7%%'(;+%<($#(%>.5;.$%(!)!!(<%'+&$*( #8(%B-"%++&#'9(
Results
PTP
! expression in selected cell lines
and antibody validation
),%(3.-.7&5&$*($#("%3#/'&D%(!)!!(7*($,%(3,&36%'( .'$&7#<*( :.+( %>.5;.$%<( 7*( :%+$%"'( 75#$( #'( $,%( "%3#07&'.'$( %B$".( 3%55;5."( <#0.&'9( =%( 5#.<%<( 3#'<&$&#'%<(0%<&.(8"#0(SYMJ(3%55+($".'+8%3$%<(:&$,( %&$,%"( .'( %0-$*( >%3$#"( #"( :&$,( .( 3L@A( 8#"( $,%( %B-"%++&#'(#8((!)!!(%B$".(3%55;5."(<#0.&'($";'3.$%<( .$($,%(5%>%5(#8($,%(-;$.$&>%($".'+0%07".'%("%/&#'9( ),%(.'$&7#<*(+-%3&(3.55*("%3#/'&D%($,%(-"%<&3$%<( UST(_L(7.'<(&'($,%(/%5(5.'%(5#.<%<(:&$,($,%(+;-%"4 '.$.'$( 3#'$.&'&'/( $,%( "%3#07&'.'$( %B$".( 3%55;5."( <#0.&'C(7%&'/($,%(+;-%"'.$.'$(<%"&>%<(8"#0(0#36( $".'+8%3$%<(3%55+('%/.$&>%9(A(+&0&5."C(+5&/,$(7.364 /"#;'<(:.+(<%$%3$.75%(&'(7#$,($,%(5.'%+(1J&/9(UA29( )#(>.5&<.$%($,%(;+%(#8($,%(.'$&7#<*(8#"((#:(3*$#04 %$"*(:%(("+$(.'.5*D%<(!)!!(0?@A(%B-"%++&#'(#'( .(-.'%5(#8(3%55(5&'%+(7*('#"$,%"'(75#$(1J&/9(UF2(.'<( +%5%3$%<( $:#( 3%55;5."( 0#<%5+V( _[\S( 3#0-5%$%5*( 5.36(!)!!(%B-"%++&#'(.'<(:.+($,%'($".'+8%3$%<(:&$,( 7#$,(0#36(.'<(!)!!(3L@A(3#'$.&'&'/(-5.+0&<+( 1_[\S(!U29(^YMZ(:.+(;+%<(.+(.(-#+&$&>%(3#'$"#5(8#"( '.$&>%(-"#$%&'9(A+(%B-%3$%<C(3,!)!!(+-%3&(3.55*( 7&'<+( $#( ^YMZ( .'<( _[\S(!U( :,&5%( _[\S( :%"%( '%/.$&>%(1J&/9(UK29(
PTP
! is expressed in peripheral
blood leukocytes
=%( $,%'( 3,%36%<( !)!!(%B-"%++&#'(&'(-%"&-,%".5( 75##<(5%;6#3*$%+9(c;"(-"%>&#;+(:#"6(+,#:%<($,.$( -;"&(%<(0#'#3*$%+(%B-"%++(!)!!(0?@AC(5*0-,#4 3*$%+(+,#:(.(>%"*(5#:(%B-"%++&#'($,.$(:%(&'$%"4 -"%$%<(.+("%+&<;.5(1[e2(0#'#3*$%+(3#'$.0&'.$&#'( #8( $,%( -"%-.".$&#'C( :,&5%( -#5*0#"-,#';35%.$%<( 3%55+( 1!E@2( :%"%( '%/.$&>%( 1H&++.'<"&'&( %$( .59( STT\29(J5#:(3*$#0%$"*(1J&/9(S2(3#'("0%<(?@A( <.$.(8#"(0#'#3*$%+9(`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g2( .'<( -5.+0.3*$#&<( 1KLMTMO2( 3%55+( 3#'("0%<($,%("%+;5$+(-"%>&#;+5*(#7$.&'%<(7*(?@A( .'.5*+&+(1H&++.'<"&'&(%$(.59(STT\2C(:&$,(-5.+0.3*4 $#&<(LK(+,#:&'/(.(+5&/,$5*(+$"#'/%"(+$.&'&'/($,.'( 0*%5#&<(LK9PTP
! is expressed in
monocyte-derived dendritic cells (moDC)
and further induced by maturation
A+(0#LK(%B-"%++%<(,&/,(!)!!(5%>%5+(:%(:&+,%<( $#(3#'("0($,%(%B-"%++&#'(.'<(;-40#<;5.$&#'(#8( $,%(-"#$%&'(;-#'(&'<;3$&#'(#8(LK(0.$;".$&#'(7*( H!I9(K%55+(:%"%(#7$.&'%<(8"#0(0#'#3*$%+(3;5$;"%<( &'(-"%+%'3%(#8(GH4N(.'<(aE4KIJ(.'<(.'.5*D%<(.$( <.*(\(#8(3;5$;"%C(.8$%"(SN(,(#8(&'3;7.$&#'(:&$,(#"( :&$,#;$( H!I9( G'( $,&+( 3.+%C( (#:( 3*$#0%$"*( :&$,( 3,!)!!(8;55*(3#'("0%<(-"%>&#;+(<.$.C(+,#:&'/( $,.$(0#LK(%B-"%++($,%(-,#+-,.$.+%(<;"&'/(<&88%"4 %'$&.$&#'( 1/%#0%$"&3( EJG( ".$&#( q( N9U2( .'<( $,.$( $,%( %B-"%++&#'( &+( 8;"$,%"( %',.'3%<( ;-#'( H!I4 &'<;3%<(0.$;".$&#'(1/%#0%$"&3(EJG(".$&#(q(SS9Z2( 1J&/(MA29
PTP
! is strongly expressed in puri ed
CD34
+peripheral blood haematopoietic
progenitors and down-modulated
during in vitro expansion
=%( <%0#'+$".$%<( $,.$( !)!!(%B-"%++&#'(3.;+%(.( 75#36( &'( ,.%0#-#&%$&3( <&88%"%'$&.$&#'( :&$,( .'( .33;0;5.$&#'(#8(3_G)WKLMNO(-"%3;"+#"+(&'(0;"&'%(
PI( 3%55+( 1I#"&#( %$( .59( UYYZ2( .'<C( 0#"%( "%3%'$5*C( <%+3"&7%<(,&/,(!)!!(%B-"%++&#'(&'(%'<#$,%5&.5(3%55+( 1X%DD.5&'&( %$( .59( STTZ29( =%( $,%"%8#"%( :&+,%<( $#( %B.0&'%( $,%( -"%+%'3%( #8( !)!!(#'(KLMNO( 3%55+C(
3#'+&<%"%<(-"%3;"+#"+(8#"(7#$,(,.%0#-#&%$&3(.'<( %'<#$,%5&.5(3%55+(1I;<.(%$(.59(STTT29(=%(-;"&(%<(
KLMNO(3%55+(8"#0(5%;6#3*$%4"&3,(7;88*(3#.$+(.'<(
.'.5*D%<( $,%( %B-"%++&#'( #8( $,&+( -,#+-,.$.+%9( J"%+,5*(&+#5.$%<(KLMNO(3%55+(+,#:%<($,%(,&/,%+$(
5%>%5(#8(%B-"%++&#'(<%$%3$%<(+#(8."(1/%#0%$"&3(EJG( ".$&#(q(Y\9Z(r(Y9Z[(ILC('(q(N29(A8$%"(.($,"%%(<.*+( $"%.$0%'$( :&$,( IKJ( .'<( GH4M( ,.%0.$#-#&%$&3( /"#:$,(8.3$#"+C(6'#:'($#(&'<;3%(-"#5&8%".$&#'(.'<( <&88%"%'$&.$&#'( .5#'/( %"*$,"#40*%5#&<( 5&'%./%( 1i.0.&(%$(.59(STTT2C($,%(5%>%5+(#8(!)!!(%B-"%++&#'( <%3"%.+%+(1/%#0%$"&3(EJG(".$&#(q(N9\Y(r(U9[T(ILC( '(q(N2(1J&/9(MF29(
Discussion
),%(<.$.(,%"%(-"%+%'$%<(."%(.&0%<($#(3#0-5%0%'$( $,#+%( .5"%.<*( %B-#+%<( &'( #;"( -"%>&#;+( +$;<&%+( 1I#"&#(%$(.59(UYYZC(H&++.'<"&'&(%$(.59(STT\2("%/."<&'/( $,%( %B-"%++&#'( #8( !)!!( &'( $,%( ,.%0.$#-#&%$&3( +*+$%09(),%(3,!)!!(.'$&7#<*("%3%'$5*(<%>%5#-%<( 7*( #;"( /"#;-( .55#:%<( ;+( $#( 0#>%( 8"#0( 0?@A( %B-"%++&#'(<.$.($#(+;"8.3%(5.7%5&'/(#8(3%55+(.'<((#:( 3*$#0%$"*(.'.5*+&+9(=&$,(8%:(%B3%-$&#'+C($,%(<.$.( 3#'("0%<(.$($,%(-"#$%&'(5%>%5(:,.$(,.<(7%%'(+%%'( .$($,%(?@A(5%>%5V(!)!!(&+(-"%8%"%'$&.55*(%B-"%++%<( 7*(0#'#3*$%+( .'<( <%'<"&$&3( 3%55+9(=&$,($,%( +#5%( %B3%-$&#'( #8( !E@+( $,%( 0?@A( 5%>%5+( .--%."( $#( 3#""%5.$%(".$,%"(:%55(:&$,(-"#$%&'(5%>%5+C(&'<&3.$&'/( .($".'+5.$&#'.5(3#'$"#5(#8($,%(%B-"%++&#'(5%>%5+(#8( $,&+(-,#+-,.$.+%9(E#"%#>%"C(&$+(%B-"%++&#'(5%>%5+( ."%(+;7+$.'$&.55*(0#<;5.$%<(:,%'(3%55+(."%(&'<;3%<( $#(-"#5&8%".$%(#"(<&88%"%'$&.$%9(=%(.5+#(<%0#'+$".$%( $,.$(!)!!(&+(%B-"%++%<(7*(F(3%55+($,.$("%-"%+%'$(.( 35.++(#8(5*0-,#3*$%+(:&$,($,%(3.-.7&5&$*($#(.3$(.+( .'$&/%'4-"%+%'$&'/( 3%55+( $#/%$,%"( :&$,( 35.++&3( 0*%5#&<(A!K+9(KLMNO(3%55+C(-"%3;"+#"+(8#"(7#$,(
,.%0#-#&%$&3( .'<( %'<#$,%5&.5( 3%55+( 1I;<.( %$( .59( STTT2C( %B-"%++%<( !)!!( &'<&3.$&'/( $,.$(
Figure 1A.) Western blot analysis of conditioned media with chPTP! antibody: Nctr = medium from 293 cells transfected with empty vector, PTP! = medium from 293 cells transfected with PTP! extra cellular domain cDNA, producing and secreting a soluble 120 KD protein.
B) Northern blot analysis of haemopoietic cell lines for PTP!; PTP' is the other member of subtype V receptor type tyrosine phosphatase and is used as a negative control; $-actin is showed for total RNA estimation. As a positive control for PTP' hybridization we utilized human brain RNA on the same blot (data not shown). C) FACS analysis of K562, K562 transfected with PTP! full-length cDNA (K562 !12 and U937 cell lines for PTP! using chPTP! antibody. Isotype control staining is showed in gray.
;'<&88%"%'$&.$%<(-"#/%'&$#"+(."%(3,.".3$%"&D%<(7*( %B-"%++&#'(#8($,&+(-,#+-,.$.+%9(),%"%(0&/,$(7%($,%( -#++&7&5&$*($,.$(!)!!(&+("%h;&"%<(8#"($,%(0.&'$%4 '.'3%(#8($,%(;'<&88%"%'$&.$%<(-,%'#$*-%(&'(,.%0#4 -#&%$&3(-"%3;"+#"+9(),&+(&+(+;//%+$%<(7*(-"%>&#;+( <.$.(&'($,%(0;"&'%(,.%0#-#&%$&3(+*+$%0(:,%"%(&$+( #>%"%B-"%++&#'( &',&7&$%<( %"*$,"#40*%5#&<( <&88%"4 %'$&.$&#'( :&$,( $,%( .33;0;5.$&#'( #8( 3_G)WKLMNO(
-"%3;"+#"+( 1I#"&#( %$( .59( UYYZ2( .'<( 7*( $,%( <#:'( 0#<;5.$&#'(8#55#:&'/(in vitro(3;5$;"%(:&$,(/"#:$,( 8.3$#"+(,%"%(#7+%">%<9(),&+(5.$$%"(('<&'/C(&'(./"%%4 0%'$(:&$,(&$+(-"#-#+%<($;0#"(+;--"%++#"(8;'3$&#'C( :#;5<( +;//%+$( .'( &'>%"+%( 3#""%5.$&#'( 7%$:%%'( ,.%0.$#-#&%$&3( -"#5&8%".$&#'W<&88%"%'$&.$&#'( .'<( !)!!( %B-"%++&#'( #'( ,.%0.$#-#&%$&3( -"#/%'&$#"( 3%55+9(c8(3#;"+%(&$(0;+$(7%(3#'+&<%"%<($,.$($,%(,&/,( %B-"%++&#'( #7+%">%<( #'( 8"%+,5*( -;"&(%<( KLMNO( 3%55+(0&/,$(.5+#(7%(<;%($#($,%(-;"&(3.$&#'(-"#3%4 <;"%9( c8( '#$%( &+( $,%( #7+%">.$&#'( $,.$( .55( $,%( .'.-5.+$&3(5."/%(3%55(5*0-,#0.+(.'<(?%%<4I$%"'4 7%"/(3%55+(&'(`#</6&'j+(<&+%.+%(+$.&'%<(-#+&$&>%(8#"(
!)!!( %B-"%++&#'( :,&5%( #'5*( UNe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c;"( 8&'<&'/+(+;//%+$(.(<&+$&'3$("#5%(#8(!)!!(%B-"%+4 +&#'( &'( $,%( ,.%0#-#&%$&3( +*+$%0C( :,%"%( &$+( %B-"%++&#'(&+("%<;3%<(8#55#:&'/($,%(&'&$&.5(+$%-+( #8(0.$;".$&#'(.5#'/($,%(%"*$,"#40*%5#&<(+%"&%+C( .'<(%'<#$,%5&.5(3%55+(:,%"%(&$(&+(8#;'<(+$"#'/5*( %B-"%++%<(&'(0.$;"%(3%55+9 A5$#/%$,%"(#;"(<.$.(>.5&<.$%($,%(;+%(#8(.('#>%5( .'$&7#<*(8#"($,%(<%$%3$&#'(#8(!)!!9(),%("%+;5$+(.5+#(
Figure 2. FACS analysis of peripheral blood PMN (CD15+; CD14–), T lymphocytes (CD3+), B lymphocytes (CD19+), monocytes (CD14+),
myeloid (CD1c+/CD19–) and plasmacytoid (CD303+) DC using chPTP! antibody. Isotype control is shown in gray, one representative
&'<&3.$%( $,.$( !)!!( ,.+( $,%( 8%.$;"%+( #8( .( '#>%5( 5%;6#3*$%(0."6%"(:,#+%(%B-"%++&#'(&+(0#<;5.$%<( <;"&'/(<&88%"%'$&.$&#'(.'<(0.$;".$&#'(#8(+-%3&(3( 5%;6#3*$%(+;7+%$+9(A5$,#;/,(0#"%(:#"6(&+('%%<%<( &'(#"<%"($#(%5;3&<.$%($,%(7#/&3.5(8;'3$&#'(#8($,&+( -,#+-,.$.+%C($,%(%>.5;.$&#'(#8(&$+(%B-"%++&#'(0&/,$( "%-"%+%'$(.(;+%8;5($##5(8#"($,%(3,.".3$%"&D.$&#'(#8( ,.%0.$#-#&%$&3C( %'<#$,%5&.5( .'<( +$%0( 3%55+( +;7+%$+9(
Acknowledgements
),&+( +$;<*( :.+( +;--#"$%<( 7*( /".'$+( 8"#0( K#'+#"D&#(I$;<&(^'&>%"+&$."&C(X%"#'.C(G$.5*(1K9I92C( J#'<.D&#'%( K."&>%"#'.C( F.'<#( STTM( 1K9I92C( G$.5&.'( K*+$&3( J&7"#+&+( ?%+%."3,( J#;'<.$&#'( 1a".'$+(JJKlTMWSTTSR(JJKlUTWSTT[(7*(G+$&$;$&( I3#5.+$&3&(X%"#'%+&($#(!9E9(.'<(K9I92R(K#0&$.$#( <&(X&3%'D.(<%55jA++#3&.D&#'%(X%'%$.(-%"(5.(5#$$.( 3#'$"#(5.(J&7"#+&(K&+$&3.(.'<(AD&%'<.(c+-%<.5&%".( <&(X%"#'.V(H%//%([NbWYM(J&'.'D&.0%'$#(?&3%"3.( J&7"#+&(K&+$&3.(STTN(1!9E92C(@.$&#'.5(K#('(STT[( 1H9i92( #8( $,%( G$.5&.'( E&'&+$"*( #8( I3&%'$&(3( .'<( )%3,'#5#/*( ?%+%."3,C( .'<( G@J@( 8;'<+( 8#"( K?Gc?AL(-"#]%3$(1H9i92C(G$.5*9References
A'<%"+%'C(f9@9C(E#"$%'+%'C(c9`9C(!%$%"+C(a9`9C(L".6%C(!9a9C(G>%"+%'C(H9J9C( c5+%'C(c9`9C(f.'+%'C(!9a9C(A'<%"+%'C(`9I9C()#'6+C(@9_9(.'<(E#55%"C(@9!9( STTU9(I$";3$;".5(.'<(%>#5;$&#'."*("%5.$&#'+,&-+(.0#'/(-"#$%&'($*"#4 +&'%(-,#+-,.$.+%(<#0.&'+9(Mol. Cell. Biol.C(SUVZUUZgM\9A'<%"++#'C(H9K9C(@&5++#'C(_9(.'<(a.,07%"/C(K9a9(UYZY9(_[\Ss.(,;0.'( %"*$,"#5%;6%0&3(3%55(5&'%9(Int. J. Cancer,(SMVUNMgZ9 F."'%.C(a9C(I&5>%''#&'%'C(c9C(I,..'.'C(F9C(`#'%//%"C(A9E9C(K.'#55C(!9L9C( Lj%;+$.3,&#C(!9C(E#"+%C(F9C(H%>*C(f9F9C(H.8#"/&.C(I9C(`;%7'%"C(_9(%$(.59( UYYM9(G<%'$&(3.$&#'(#8(.(3."7#'&3(.',*<".+%45&6%(<#0.&'(&'($,%(%B4 $".3%55;5."("%/&#'(#8(?!)!(/.00.(<%('%+(.('%:(+;78.0&5*(#8("%3%-4 $#"($*"#+&'%(-,#+-,.$.+%+9(Mol. Cell. Biol.,(UMVUNYZg[T\9
Figure 3. A) FACS analysis of immature (upper) or LPS-matured (lower) monocyte-derived DC using chPTP! antibody; isotype control is shown in gray. B) double staining of puri ed peripheral blood precursors (upper) at the time of separation and after three days of culture in the presence of SCF and IL-3 (lower). In red: cells treated with anti-CD34 and chPTP!#; in black: cells treated with anti-CD34 and chicken isotype IgY. One representative experiment of four is shown. Each experiment involved puri cation, differentiation (for DC) and staining of cells from individual donors.
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