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23 Immunohistopathological analysis of rheumatoid synovitis
Abstracts
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IMM1.JNOttlSTOPATItOL(IGI(:AL ANALYSIS ()F RtlE/!MA'I'CJlI) SYN()VITIS Xl Kyt)goku. M.D.. P h . D . K Murakami M.I)., I Itoh I).\:.M. and T Sawai M.I).. P h D l)epartment of Patholngy. "I'ohoku Ilniversity Sclmc)l of Medit:ine. Sendal. c.18(},]apan.
Rheumatrfid arthritis (RA), a systemic intra(:table disease, originates in the ioint arid terminates in the joint. It iS O[le (If the lllOSt vC)mlI)OT1 (:hrt)nic i)iflitlTilllatorv txlndititlllS ill ht.lFllal) beings, but the histol~athologi(: picture is not typical ¢)f chronic infianlmation. Synnvial tissue, the site c)f RA inflammation, consists of sm.eral cell types n[ u n k n o w n c~rigin, making the pathc)logic features of RA (luih~ urmsuiH. h) il i)t)rlllill slate, s?,'novial lining t:tlliSists (if A cells, xillm)s-sur{ac ed lysosome-ri~ h and Class lI imtigenpositive cell. and 13 cells, smooth surtaced, RER-rich CA(; producers which release various kinds (ff metalloprnteinilse, i n c l u d i n g collagenase Bath are vimentin and fikmme
1 Murakami K et al. (1988): Proc Sixth SEAPAL C~mgress. T~kvo 2. Kyogoku M 11~4881: Presidential Address in The 32nd (;emmd :\ssemblv at lapan Rheumatism ,'\sst)t:ialion, SmMai.
3. Kyo,qoku M ( l(.ll~): Proc t)t Intl S y m t m s i u m tm Tim Chemical Mediatt)rs in The Ski]) lnflammati(m. SmMai.
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TRANS-AC'I'IN(;-FA(:T(,~R(S) INVOI.k'H) IN TI IE ACTIVATI( )N C)t" t 1[ "M:kN IMMUN( )GI,()B[ H.IN IIEAVY-CtlAIN GENE ENIIANCER. Takeshi Watanabe Medical Institute at Bi(~regulation. Kyushu University. Fukuoka. ]apart.
Xlicrair)ieclion of proteins into living cells bv means of erythr(~cyn!-glmst fusion has made it feasi't)le to test the rum:film of cellular c m n t m n e n t s w h i c h participate in I?,'nH)hc~Q,'leactivatinn I()hara and D,atanabe, 1982, Eda et al., 1982. Watanabe et al.. 1.q8ti]. Using this approach, we demonstrated that nuclear prnteins obtained from B-lymllhoc?,te lineage, but not other types of cells, contained transactinR factors whi(:h spe~:ificall:..' i n d u c e d the transcription of a h u m a n heavy (:hain gene [1 li(; 1) integrated into I,-t:ell transformants (Maeda el al.. 1986). We have now use.d tlI'I,C, l)hc)spho(:ellulose ct,lumn, hepa[m Sel~harose. and I)NA affinity (:olumn c h r o m a t o g r a p h y Io resolve three types o1" nu(:lear proteins from illurine al)d hklIIlall myeloma cells. The first was finally eluted from an octanucleotide affinity column, was at)proximately 74 kDa in ,~ize. and recognized the sequence AT(;CAAAT in a gel relardatmn assay and h~otprinting N(~ heavv chain transcription resulted from microinjection of this octamer-binding protein purified from B-lymphoid cells into I.-cell transf()rmants. A second type of protein was highly enri(:hed by positive selection on an affinity column bearing the synthetic oligonucleotide TTTA(,CAAGCAAA (HE2 sequence). [t ',','us allproximately 96 kl)a in size. hmno~ehe(ms by SI)S-PAGt" analysis, an(I it rect)gnized the expected DNA seqtmn{:e from the hear?, (:ham e n h a n c e r segment ill gel retardation and footprinting assays This protein was sufficient fr)r induction ()f I]l(;l gen*t expressian in nut rni(:roinjectinn illclde]. The iII)IlltlI1Ogl()hUJill at:tamer (decza[ller) ill()tiJ has been d(![)l(ll)strilted t() be a fl)l)clio)la] (:is-at:ling (;OlllpOlllHlt (if prolnoters it))d e()hai1cers w h i c h mediates cell-type specific i!xpression of ilntl)un(lgltlbulill geiles. It hils be(?i) s h o w n that ther(~ is at [east (1)11}t)l'liquitous protein (NF-A I I and (me tissue-rf'stri(:ted [)rotei|) (NI"-A2) w h i c h re~:(Jg[lize this particular se(ltlenc:e I towe',er, w e havp [laD," delil(lrlstrated that remllval of o(:talilt!r-hinding proteins from a nut:h!ar extrac:t did not affect Sklbst?ql.)t![It yc:hain i)ldut il)g activity ill our in rive) assay s?,'sb.ml. It is still possible in our exI)t!ri[lle(lts that some [l(llll)(:tamer binding tlroteins m(Mitied and activated ubiquit(ms o(:tanler-binding I~rnteins alter nm:roinie(:tion, t lowever. it is clear that the el)riched octarl)er binding pr()teins from the N,'q-1 myelozna arl! [l(it thernsel,,'es sufficient for heavy (:hain Rene activati(m in the present systenl O u r lmrifled 9(i k[.}il [mJtein b o u n d to the sequem e TTTA(;(;AA(;CAAA in the h u m a n heavv chain
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IMM1.JNOttlSTOPATItOL(IGI(:AL ANALYSIS ()F RtlE/!MA'I'CJlI) SYN()VITIS Xl Kyt)goku. M.D.. P h . D . K Murakami M.I)., I Itoh I).\:.M. and T Sawai M.I).. P h D l)epartment of Patholngy. "I'ohoku Ilniversity Sclmc)l of Medit:ine. Sendal. c.18(},]apan.
Rheumatrfid arthritis (RA), a systemic intra(:table disease, originates in the ioint arid terminates in the joint. It iS O[le (If the lllOSt vC)mlI)OT1 (:hrt)nic i)iflitlTilllatorv txlndititlllS ill ht.lFllal) beings, but the histol~athologi(: picture is not typical ¢)f chronic infianlmation. Synnvial tissue, the site c)f RA inflammation, consists of sm.eral cell types n[ u n k n o w n c~rigin, making the pathc)logic features of RA (luih~ urmsuiH. h) il i)t)rlllill slate, s?,'novial lining t:tlliSists (if A cells, xillm)s-sur{ac ed lysosome-ri~ h and Class lI imtigenpositive cell. and 13 cells, smooth surtaced, RER-rich CA(; producers which release various kinds (ff metalloprnteinilse, i n c l u d i n g collagenase Bath are vimentin and fikmme
1 Murakami K et al. (1988): Proc Sixth SEAPAL C~mgress. T~kvo 2. Kyogoku M 11~4881: Presidential Address in The 32nd (;emmd :\ssemblv at lapan Rheumatism ,'\sst)t:ialion, SmMai.
3. Kyo,qoku M ( l(.ll~): Proc t)t Intl S y m t m s i u m tm Tim Chemical Mediatt)rs in The Ski]) lnflammati(m. SmMai.
24
TRANS-AC'I'IN(;-FA(:T(,~R(S) INVOI.k'H) IN TI IE ACTIVATI( )N C)t" t 1[ "M:kN IMMUN( )GI,()B[ H.IN IIEAVY-CtlAIN GENE ENIIANCER. Takeshi Watanabe Medical Institute at Bi(~regulation. Kyushu University. Fukuoka. ]apart.
Xlicrair)ieclion of proteins into living cells bv means of erythr(~cyn!-glmst fusion has made it feasi't)le to test the rum:film of cellular c m n t m n e n t s w h i c h participate in I?,'nH)hc~Q,'leactivatinn I()hara and D,atanabe, 1982, Eda et al., 1982. Watanabe et al.. 1.q8ti]. Using this approach, we demonstrated that nuclear prnteins obtained from B-lymllhoc?,te lineage, but not other types of cells, contained transactinR factors whi(:h spe~:ificall:..' i n d u c e d the transcription of a h u m a n heavy (:hain gene [1 li(; 1) integrated into I,-t:ell transformants (Maeda el al.. 1986). We have now use.d tlI'I,C, l)hc)spho(:ellulose ct,lumn, hepa[m Sel~harose. and I)NA affinity (:olumn c h r o m a t o g r a p h y Io resolve three types o1" nu(:lear proteins from illurine al)d hklIIlall myeloma cells. The first was finally eluted from an octanucleotide affinity column, was at)proximately 74 kDa in ,~ize. and recognized the sequence AT(;CAAAT in a gel relardatmn assay and h~otprinting N(~ heavv chain transcription resulted from microinjection of this octamer-binding protein purified from B-lymphoid cells into I.-cell transf()rmants. A second type of protein was highly enri(:hed by positive selection on an affinity column bearing the synthetic oligonucleotide TTTA(,CAAGCAAA (HE2 sequence). [t ',','us allproximately 96 kl)a in size. hmno~ehe(ms by SI)S-PAGt" analysis, an(I it rect)gnized the expected DNA seqtmn{:e from the hear?, (:ham e n h a n c e r segment ill gel retardation and footprinting assays This protein was sufficient fr)r induction ()f I]l(;l gen*t expressian in nut rni(:roinjectinn illclde]. The iII)IlltlI1Ogl()hUJill at:tamer (decza[ller) ill()tiJ has been d(![)l(ll)strilted t() be a fl)l)clio)la] (:is-at:ling (;OlllpOlllHlt (if prolnoters it))d e()hai1cers w h i c h mediates cell-type specific i!xpression of ilntl)un(lgltlbulill geiles. It hils be(?i) s h o w n that ther(~ is at [east (1)11}t)l'liquitous protein (NF-A I I and (me tissue-rf'stri(:ted [)rotei|) (NI"-A2) w h i c h re~:(Jg[lize this particular se(ltlenc:e I towe',er, w e havp [laD," delil(lrlstrated that remllval of o(:talilt!r-hinding proteins from a nut:h!ar extrac:t did not affect Sklbst?ql.)t![It yc:hain i)ldut il)g activity ill our in rive) assay s?,'sb.ml. It is still possible in our exI)t!ri[lle(lts that some [l(llll)(:tamer binding tlroteins m(Mitied and activated ubiquit(ms o(:tanler-binding I~rnteins alter nm:roinie(:tion, t lowever. it is clear that the el)riched octarl)er binding pr()teins from the N,'q-1 myelozna arl! [l(it thernsel,,'es sufficient for heavy (:hain Rene activati(m in the present systenl O u r lmrifled 9(i k[.}il [mJtein b o u n d to the sequem e TTTA(;(;AA(;CAAA in the h u m a n heavv chain