I ns t i t ut e of DNA Medi ci ne
Depar t ment of Mol ecul ar I mmunol ogy
Saburo Saito,Associate Professor and Director Daitaro Kurosaka,Associate Professor Yuji Ohno,Assistant Professor
Gener al Summar y
Our r e s e a r c h ha s f oc us e d on a na l y s i s of t he ba s i c i mmune s y s t e m t o pr ot e c t us f r om di s e a s e s a nd of i mmune di s or de r s ,s uc h a s hy pe r s e ns i t i v i t y di s e a s e s a nd a ut oi mmune di s e a s e s .
Res ear ch Act i v i t i es
I nt e r l e uki n 3 1 i nc r e as e s s e r um I g E l e v e l s i n mi c e
I nt e r l e uki n( I L) - 3 1 t r a ns g e ni c mi c e t ha t we ha v e e s t a bl i s he d s how hi g he r s e r um I g E l e v e l s t ha n do t he i r nont r a ns g e ni c l i t t e r ma t e s . To c onf i r m t he a s s oc i a t i on of I L- 3 1 wi t h s e r um I g E l e v e l s obs e r v e d i n t he t r a ns g e ni c mi c e ,r e c ombi na nt( r )mur i ne I L- 3 1 wa s a dmi ni s t e r e d t o nor ma l C5 7 BL / 6 a nd BALB / c mi c e a nd t o I L- 4 r e c e pt or α - de f i c i e nt knoc k- out mi c e . Se r um I g E l e v e l s of mi c e we r e me a s ur e d wi t h a 2 - s i t e c a pt ur e a s s a y . To i nv e s t i g a t e t he pr oduc t i on of I L- 4 ,I L- 5 ,I L- 1 3 ,a nd i nt e r f e r onγ ,s pl e e n c e l l s f r om mi c e we r e c ul t ur e d f or 5 da y s ,a f t e r whi c h c ul t ur e s upe r na t a nt s we r e ha r v e s t e d a nd t e s t e d f or c y t oki ne a c t i v i t y by me a ns of s a ndwi c h e nz y me - l i nke d i mmunos or be nt a s s a y . Se r um I g E l e v e l s of C5 7 BL / 6 a nd BALB / c mi c e t ha t r e c e i v e d i nj e c t i ons of r I L- 3 1 we r e s i g ni f i c a nt l y hi g he r t ha n t hos e of mi c e t r e a t e d wi t h phos pha t e - buf f e r e d s a l i ne( PBS) . Howe v e r ,no s i g ni f i c a nt i nc r e a s e i n s e r um I g E l e v e l s wa s obs e r v e d i n I L- 4 r e c e pt or α - de f i c i e nt knoc k- out mi c e t ha t r e c e i v e d i nj e c t i ons of I L- 3 1 . Fur t he r mor e ,pr oduc t i on of I L- 1 3 a nd I L- 5 wa s s i g ni f i c a nt l y hi g he r i n t he r I L- 3 1 - t r e a t e d C5 7 BL / 6 mi c e t ha n i n PBS- t r e a t e d c ont r ol mi c e . I n c ont r a s t ,no s i g ni f i c a nt di f f e r e nc e i n i nt e r f e r on γpr oduc - t i on a c t i v i t y wa s f ound be t we e n r I L- 3 1 - t r e a t e d mi c e a nd PBS- t r e a t e d mi c e ,whe r e a s t he c onc e nt r a t i on of I L- 4 wa s be l ow t he de t e c t a bl e l e v e l i n bot h g r oups . The s e r e s ul t s s ug g e s t t ha t I L- 3 1 i s i nv ol v e d i n t he e nha nc e me nt of s e r um I g E l e v e l s v i a i nduc t i on of I L- 1 3 e x pr e s s i on.
De c r e as e d numb e r s o f CD4 ＋ and CD8 ＋ c e l l s c o nt ai ni ng s i g nal - j o i nt T- c e l l r e c e p t o r e xc i s i o n c i r c l e s i n p at i e nt s wi t h s y s t e mi c l up us e r y t he mat o s us
Pa t i e nt s wi t h s y s t e mi c l upus e r y t he ma t os us( SLE)ha v e de c r e a s e d numbe r s of pe r i ph- e r a l bl ood T c e l l s c ont a i ni ng s i g na l - j oi nt T c e l l r e c e pt or e x c i s i on c i r c l e s( Sj TRECs ) , whi c h a r e c ons i de r e d a n i ndi c a t or of t hy mi c out put . The obj e c t i v e of t hi s s t udy wa s t o i nv e s t i g a t e t he me c ha ni s m of t he de c r e a s e i n s uc h T c e l l s . Pe r i phe r a l bl ood T c e l l s f r om pa t i e nt s wi t h SLE we r e c l a s s i f i e d a s CD4 ＋ or CD8 ＋ c e l l s . Le v e l s of Sj TREC we r e me a s ur e d wi t h t he r e a l - t i me pol y me r a s e c ha i n r e a c t i on. Te l ome r a s e a c t i v i t y wa s de t e r mi ne d wi t h t he t e l ome r i c r e pe a t a mpl i f i c a t i on pr ot oc ol a s s a y . The numbe r s of
169 Research Activities 2007 The Jikei University School of Medicine
CD4 ＋ a nd CD8 ＋ c e l l s c ont a i ni ng Sj TREC i n t he pe r i phe r a l bl ood we r e l owe r i n pa t i e nt s wi t h SLE t ha n i n c ont r ol s ubj e c t s . A c or r e l a t i on wa s f ound be t we e n t he numbe r of Sj TREC- pos i t i v e CD4 ＋ c e l l s a nd t he numbe r of CD8 ＋ c e l l s . The Sj TREC l e v e l i s i nf l ue nc e d by a n i nc r e a s e i n c e l l di v i s i on. To e x a mi ne t hi s i nc r e a s e , t e l ome r a s e a c t i v i t y ,a s a n i ndi c a t or of c e l l di v i s i on,wa s me a s ur e d s i mul t a ne ous l y ; howe v e r ,t he r e wa s no c or r e l a t i on be t we e n t he Sj TREC l e v e l a nd t e l ome r a s e a c t i v i t y . The s e r e s ul t s s ug g e s t t ha t t hy mi c out put i s de c r e a s e d i n pa t i e nt s wi t h SLE.
Te nas c i n- C i s r e q ui r e d f o r p r o l i f e r at i o n o f as t r o c y t e s i n p r i mar y c ul t ur e
As t r oc y t e s i n pr i ma r y c ul t ur e c a n be c l a s s i f i e d mor phol og i c a l l y i nt o 2 t y pe s :f i br ous a s t r oc y t e s a nd pr ot opl a s mi c a s t r oc y t e s . To e x a mi ne t he r ol e of t e na s c i n- C i n a n i n- v i t r o a s t r oc y t e c ul t ur e ,pr i ma r y c ul t ur e s of a s t r oc y t e s pr e pa r e d f r om t he br a i ns of wi l d- t y pe a nd of t e na s c i n- C- de f i c i e nt e mbr y oni c mi c e we r e a na l y z e d. I n pr i ma r y c ul t ur e of a s t r oc y t e s f r om t e na s c i n- C- de f i c i e nt mi c e ,f i br ous a s t r oc y t e s di d not a ppe a r ,a nd a s t r oc y t e s di d not be c ome t i l e - s ha pe d whe n t he y c a me i n c ont a c t wi t h e a c h ot he r . The r a t e of 5 - br omo- 2 ′ - de ox y ur i di ne i nc or por a t i on i n a c e l l pr ol i f e r a t i on a s s a y wa s muc h l owe r f or a s t r oc y t e s f r om t e na s c i n- C ‑ de f i c i e nt mi c e t ha n f or a s t r oc y t e s f r om wi l d- t y pe mi c e . The s e r e s ul t s s ug g e s t t ha t t e na s c i n- C i s a n e s s e nt i a l mol e c ul e f or ma i nt a i ni ng t he pr ol i f e r a t i on a nd pr ope r mor phol og y of a s t r oc y t e s i n pr i ma r y c ul t ur e .
Publications
Kurosaka D,Yasuda J,Ikeshima-Kataoka H, Ozawa Y,Yoshida K,Yasuda C,Kingetsu I,Saito S,Yamada A.Decreased numbers of signal- joint t cell receptor excision circle-containing CD4＋and CD8＋cells in systemic lupus eryth- ematosus patients.Mod Rheumatol 2007;17:
296‑300.
Kurosaka D,Yoshida K,Yasuda J,Yasuda C, Noda K,Furuya K,Ukichi T,Kingetsu I,Joh K,
Yamaguchi N,Saito S,Yamada A.The effect of endostatin evaluated in an experimental animal model of collagen-induced arthritis.Scand J Rheumatol.2007;36:434‑41.
Ikeshima-Kataoka H, Saito S, Yuasa S.
Tenascin-C is required for proliferation of astrocytes in primary culture.In Vivo 2007;21: 629‑33.
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Research Activities 2007 The Jikei University School of Medicine
