clinexpimmunol00093-0140
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Clin. exp. Immunol. (1988) 7 4 454-458 MHC Class I a n d Class I-like gene product expression b y malignant T cells: relationships between CD1a, HLA-ABC a n f2-microglobulin R . A . JONES, C . S . SCOTT, F . E . KATZ J . CHILDt Department o f Haematology, Cookridge Hospital, Leeds. *Department of Haematology/Oncology, Great Ormond S t . Hospital, a n d tDepartment o f Haematology, Leeds General Infirmary, Leeds. (Acceptedfor publication 2 9 July 1988) SUMMARY Beta2-microglobulin (f12m) forms t h e invariant light chain of t h e MHC-encoded HLA-ABC a n d t h e non-MHC-encoded C D I molecules. While HLA-ABC (MHC Class I molecules ar e virtually ubiquitious i n tissue distribution, CD 1 determinants by contrast a r e more restricted. We have assessed, b y indirect immunoenzymeassay, th e relative membrane densities o f these molecules on malignant thymic a n d post-thymic T cells. It was found that t h e T cells o f mature post-thymic proliferations expressed significantly more f2m-associated protein, predominantly HLA-ABC i n nature, than thymic-ALL blasts. This parallels t h e situation found i n normal peripheral T cells a n d thymocytes. I n contrast t o post-thymic T cells, thymic-ALL blasts showed considerable case t o case variation with respect t o non-HLA-associated f2m and, o f particular interest, n o t a ll of this excess 132m could b e accounted f or b y C D 1 a . W e therefore conclude that other f32m-containing molecules may be expressed on thymic-ALL blasts a n d possibly also on post-thymic leukaemic T cells. I n addition, i t was found that T cells from CD4+ cases o f post-thymic proliferations expressed more 2m-associated determinants than other T cells, whether o f either normal o r malignant origin, and that certain post-thymic malignancies express significantly increased levels o f f2m-associated protein relative t o normal peripheral T-cells. This i s i n direct contrast t o th e situation seen i n many solid malignancies. Keywords T cell leukaemia C D l I32m HLA-ABC INTRODUCTION T h e 12 k D protein beta2-microglobulin (f32m) constitutes t h e light chain of several structurally related glycoproteins. Th e most widespread o f these are t h e MHC-encoded 4 5 k D HLA- AB C (MHC Class I molecules (Cresswell e t al., 1974; Peterson, Rask Lindblom, 1974). Three other J32m-containing HLA-like glycoproteins of 43-49 k D form t h e first cluster o f differentia- tion (CDla, CDlb a nd CDlc) a n d a r e n o t MHC-encoded (McMichael Gotch, 1987; Milsten e t al., 1987). Functionally, HLA-ABC molecules ar e involved i n MHC- restricted T cell recognition events (Zinkernagel Doherty, 1979; McMichael, 1980) a nd a r e expressed o n t h e surface o f most nucleated cells (Daar e t a l , 1984; Natali e t al., 1984). C D1 molecules, b y contrast, have restricted tissue distribution a nd t h a s been suggested that they m a y be homologues of murine T I and or Q a antigens, defining functionally discrete lymphocyte Correspondence: R . A . Jones, Department o f Haematology, Cook- ridge Hospital, Leeds LS16 6QB, UK . populations (Flaherty, 1981; Knowles, 1984). Th e CDla anti- g e n i s expressed o n thymocytes i n reciprocal quantity t o HLA- A B C (Jones, Child, 1988), a n d is also expressed o n dermal dendrocytes a n d t h e blasts from a proportion o f cases o f thymic-acute lymphoblastic leukaemia (ALL) (Boumsell et al., 1986), a n d m a y function a s a receptor f o r extracellular / 2m (Kefford e t al., 1984). I t i s well documented that malignant cells i n some solid tumours exhibit defective expression o f f2m-associated determi- nants compared with their normal counterparts (Fleming e t al., 1981; Turbitt Mackie, 1981; Natali e t al., 1984), b u t little is known about t h e situation i n lymphoid malignancies. I n a previous report, w e detailed th e expression o f 12m, HLA-ABC a n d CD1 a antigens b y normal thymic a n d peripheral blood T cell subpopulations (Jones, Scott Child, 1988) a s part o f a wider investigation into th e expression a n d secretion o f f 2 m b y lymphoid tissue. W e have n o w extended this investigation t o include t h e T cell malignancies, i n which 32m, HLA-ABC and CD a antigen expression w as examined, a s assessed b y indirect immunoenzymeassay, b y t h e lymphoid cells from 3 9 cases o f T cell malignancy. 4 5 4
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Transcript of clinexpimmunol00093-0140
8/13/2019 clinexpimmunol00093-0140
http://slidepdf.com/reader/full/clinexpimmunol00093-0140 1/5
C l i n . e x p . I m m u n o l . ( 1 9 8 8 ) 7 4 4 5 4 - 4 5 8
MHC C l a s s I and C l a s s I - l i k e gene p r o d u c t e x p r e s s i o n
b y m a l i g n a n t T c e l l s : r e l a t i o n s h i p s b e t w e e n
CD1a, HLA-ABC and f 2 - m i c r o g l o b u l i n
R . A . JONES, C . S . SCOTT, F . E . KATZ J . A . CHILDt D e p a r t m e n t o f H a e m a t o l o g y , C o o k r i d g e H o s p i t a l , L e e d s .
* D e p a r t m e n t o f H a e m a t o l o g y / O n c o l o g y , G r e a t Ormond S t . H o s p i t a l , L o n d o n a n d t D e p a r t m e n t o f H a e m a t o l o g y , L e e d s G e n e r a l
I n f i r m a r y , L e e d s .
( A c c e p t e d f o r p u b l i c a t i o n 2 9 J u l y 1 9 8 8 )
SUMMARY
B e t a 2 - m i c r o g l o b u l i n ( f 1 2 m ) f o r m s t h e i n v a r i a n t l i g h t c h a i n o f t h e M H C - e n c o d e d HLA-ABC a n d t h e
n o n - M H C - e n c o d e d CDI m o l e c u l e s . W h i l e HLA-ABC (MHC C l a s s I m o l e c u l e s a re v i r t u a l l yu b i q u i t i o u s i n t i s s u e d i s t r i b u t i o n , CD1 d e t e r m i n a n t s b y c o n t r a s t are more r e s t r i c t e d . We h a v e
a s s e s s e d , b y i n d i r e c t i m m u n o e n z y m e a s s a y , t h e r e l a t i v e m e m b r a n e d e n s i t i e s o f t h e s e m o l e c u l e s on
m a l i g n a n t t h y m i c a n d p o s t - t h y m i c T c e l l s . I t was f o u n d t h a t t h e T c e l l s o f mature p o s t - t h y m i c
p r o l i f e r a t i o n s e x p r e s s e d s i g n i f i c a n t l y more f 2 m - a s s o c i a t e d p r o t e i n , p r e d o m i n a n t l y HLA-ABC i n
n a t u r e , t h a n t h y m i c - A L L b l a s t s . T h i s p a r a l l e l s t h e s i t u a t i o n f o u n d i n n o r m a l p e r i p h e r a l T c e l l s a n d
t h y m o c y t e s . I n c o n t r a s t t o p o s t - t h y m i c T c e l l s , t h y m i c - A L L b l a s t s s h o w e d c o n s i d e r a b l e case t o case
v a r i a t i o n w i t h r e s p e c t t o n o n - H L A - a s s o c i a t e d f2m a n d , o f p a r t i c u l a r i n t e r e s t , n o t a l l o f t h i s excess
132m c o u l d b e a c c o u n t e d f o r b y CD 1 a . We t h e r e f o r e c o n c l u d e t h a t o t h e r f 3 2 m - c o n t a i n i n g m o l e c u l e s
may b e e x p r e s s e d on t h y m i c - A L L b l a s t s a n d p o s s i b l y a l s o on p o s t - t h y m i c l e u k a e m i c T c e l l s . I n
a d d i t i o n , i t was f o u n d t h a t T c e l l s f r o m CD4+ cases o f p o s t - t h y m i c p r o l i f e r a t i o n s e x p r e s s e d more
2 m - a s s o c i a t e d d e t e r m i n a n t s t h a n o t h e r T c e l l s , w h e t h e r o f e i t h e r n o r m a l or m a l i g n a n t o r i g i n , a n d
t h a t c e r t a i n p o s t - t h y m i c m a l i g n a n c i e s express s i g n i f i c a n t l y i n c r e a s e d l e v e l s o f f 2 m - a s s o c i a t e d p r o t e i n
r e l a t i v e t o n o r m a l p e r i p h e r a l T - c e l l s . T h i s i s i n d i r e c t c o n t r a s t t o t h e s i t u a t i o n seen i n many s o l i dm a l i g n a n c i e s .
K e y w o r d s T c e l l l e u k a e m i a CDlI32m HLA-ABC
INTRODUCTION
T h e 1 2 kD p r o t e i n b e t a 2 - m i c r o g l o b u l i n ( f 3 2 m ) c o n s t i t u t e s t h e
l i g h t c h a i n o f s e v e r a l s t r u c t u r a l l y r e l a t e d g l y c o p r o t e i n s . T h e
m o s t w i d e s p r e a d o f t h e s e a r e t h e MHC-encoded 4 5 kD HLA-
ABC (MHC C l a s s I m o l e c u l e s ( C r e s s w e l l e t a l . , 1 9 7 4 ; P e t e r s o n ,R a s k L i n d b l o m , 1 9 7 4 ) . T h r e e o t h e r J 3 2 m - c o n t a i n i n g H L A - l i k e
g l y c o p r o t e i n s o f 4 3 - 4 9 kD f o r m t h e f i r s t c l u s t e r o f d i f f e r e n t i a -
t i o n ( C D l a , CDlb a n d C D l c ) a n d a r e n o t M H C - e n c o d e d
( M c M i c h a e l G o t c h , 1 9 8 7 ; M i l s t e n e t a l . , 1 9 8 7 ) .F u n c t i o n a l l y , HLA-ABC m o l e c u l e s a r e i n v o l v e d i n MHC-
r e s t r i c t e d T c e l l r e c o g n i t i o n e v e n t s ( Z i n k e r n a g e l D o h e r t y ,1 9 7 9 ; M c M i c h a e l , 1 9 8 0 ) a n d a r e e x p r e s s e d o n t h e s u r f a c e o f
m o s t n u c l e a t e d c e l l s ( D a a r e t a l , 1 9 8 4 ; N a t a l i e t a l . , 1 9 8 4 ) . CD1
m o l e c u l e s , b y c o n t r a s t , h a v e r e s t r i c t e d t i s s u e d i s t r i b u t i o n a n d i t
h a s b e e n s u g g e s t e d t h a t t h e y may b e h o m o l o g u e s o f m u r i n e T I
a n d o r Qa a n t i g e n s , d e f i n i n g f u n c t i o n a l l y d i s c r e t e l y m p h o c y t e
C o r r e s p o n d e n c e : R . A . J o n e s , D e p a r t m e n t o f H a e m a t o l o g y , C o o k -
r i d g eH o s p i t a l , L e e d s L S 1 6 6 Q B , UK .
p o p u l a t i o n s ( F l a h e r t y , 1 9 8 1 ; K n o w l e s , 1 9 8 4 ) . T h e CDla a n t i -
g e n i s e x p r e s s e d o n t h y m o c y t e s i n r e c i p r o c a l q u a n t i t y t o HLA-
ABC ( J o n e s , S c o t t C h i l d , 1 9 8 8 ) , a n d i s a l s o e x p r e s s e d o n
d e r m a l d e n d r o c y t e s a n d t h e b l a s t s f r o m a p r o p o r t i o n o f c a s e s o f
t h y m i c - a c u t e l y m p h o b l a s t i c l e u k a e m i a ( A L L ) ( B o u m s e l l e t a l . ,
1 9 8 6 ) , a n d may f u n c t i o n a s a r e c e p t o r f o r e x t r a c e l l u l a r / 2 m( K e f f o r d e t a l . , 1 9 8 4 ) .
I t i s w e l l d o c u m e n t e d t h a t m a l i g n a n t c e l l s i n s o m e s o l i d
t u m o u r s e x h i b i t d e f e c t i v e e x p r e s s i o n o f f 2 m - a s s o c i a t e d d e t e r m i -
n a n t s c o m p a r e d w i t h t h e i r n o r m a l c o u n t e r p a r t s ( F l e m i n g e t a l . ,
1 9 8 1 ; T u r b i t t M a c k i e , 1 9 8 1 ; N a t a l i e t a l . , 1 9 8 4 ) , b u t l i t t l e i s
known a b o u t t h e s i t u a t i o n i n l y m p h o i d m a l i g n a n c i e s . I n a
p r e v i o u s r e p o r t , we d e t a i l e d t h e e x p r e s s i o n o f 1 2 m , HLA-ABC
a n d C D 1 a a n t i g e n s b y n o r m a l t h y m i c a n d p e r i p h e r a l b l o o d T
c e l l s u b p o p u l a t i o n s ( J o n e s , S c o t t C h i l d , 1 9 8 8 ) a s p a r t o f a
w i d e r i n v e s t i g a t i o n i n t o t h e e x p r e s s i o n a n d s e c r e t i o n o f f 2 m by
l y m p h o i d t i s s u e . We h a v e n ow e x t e n d e d t h i s i n v e s t i g a t i o n t o
i n c l u d e t h e T c e l l m a l i g n a n c i e s , i n w h i c h 3 2 m , HLA-ABC a n d
CD a a n t i g e n e x p r e s s i o n w a s e x a m i n e d , a s a s s e s s e d b y i n d i r e c t
i m m u n o e n z y m e a s s a y , b y t h e l y m p h o i d c e l l s f r o m 3 9 c a s e s o f T
c e l l m a l i g n a n c y .
4 5 4
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E x p r e s s i o n o f f 2 m - a s s o c i a t e d m o l e c u l e s o n T c e l l s
T a b l e 1 . A n t i b o d i e s u s e d
A n t i b o d y S p e c i f i c i t y S u b c l a s s S o u r c e R e f e r e n c e
B 2 . 6 2 . 2 f l 2 m I g G 1 ARS* L i e b e u f e t a l 1 9 8 1W 6 / 3 2 HLA-ABC c o r e I g G 2 a S e r o t e c B a r n s t a b l e e t a l 1 9 7 8N A 1 / 3 4 CDIa a n t i g e n I g G 2 a S e r o t e c M c M i c h a e l e t a l 1 9 7 9H R P x - r a m l g t M o u s e I g P o l y c l o n a l D a k o
* k i n d l y d o n a t e d b y Dr A . R . S a n d e r s o n .
t H o r s e r a d i s h p e r o x i d a s e c o n j u g a t e d r a b b i t a n t i - m o u s e I g .
T a b l e 2 . C l a s s i f i c a t i o n o f TdT- p o s t - t h y m i c d i s o r d e r s
D i s o r d e r CD3 CD4 CD5 CD7 CD 8 CD16 CD25
T-PLL V + + + V - VT-NHL + +(-) + + -(+) - VS S + + + - - - -ATLL + + + - - - +
LDGL +-
-(+)-(+) + V V
C a s e s w e r e c l a s s i f i e d by c o m b i n e d m o r p h o l o g i c a l a n d i m m u -n o l o g i c a l c h a r a c t e r i s t i c s .
MATERIALS AND METHODS
T c e l l m a l i g n a n c i e sA t o t a l o f 3 9 T c e l l m a l i g n a n c i e s w e r e e x a m i n e d , c l a s s i f i e d a s
e i t h e r TdT+ l e u k a e m i a s ( t h y m i c - A L L ; n = 2 0 ) o r TdT- p o s t -t h y m i c d i s o r d e r s ( n = 1 9 ) . D i a g n o s e s w e r e e s t a b l i s h e d a c c o r d i n gt o s t a n d a r d i m m u n o l o g i c a l c r i t e r i a .
T h e p o s t - t h y m i c d i s o r d e r s w e r e s u b d i v i d e d , a s d e t a i l e d i n
T a b l e 2 , i n t o T p r o l y m p h o c y t i c l e u k a e m i a ( T - P L L ; n = 3 , T c e l ln o n - H o d g k i n s l y m p h o m a ( T - N H L ; n = 4 ) , a d u l t T c e l l l e u k a e -m i a / l y m p h o m a ( A T L L ; n = 1 , S e z a r y S y n d r o m e ( S S ; n = 4 ) a n dl y m p h o p r o l i f e r a t i v e d i s e a s e o f g r a n u l a r l y m p h o c y t e s (LDGL;n = 7 . Of t h i s l a t t e r g r o u p , a l l c a s e s h a d l a r g e g r a n u l a r
l y m p h o c y t e ( L G L ) m o rp h o l o gy , a n d t h e T c e l l s f r o m f o u r o u t o f
s e v e n c a s e s w e r e C D 1 6 ( L e u I I b ) + .P e r i p h e r a l b l o o d o r b o n e m a r r o w m o n o n u c l e a r c e l l s w e r e
i s o l a t e d u s i n g d e n s i t y g r a d i e n t c e n t r i f u g a t i o n ( L y m p h o p r e p ;N y e g a a r d ) , a n d r e s u s p e n d e d i n H B S S / B S A . T c e l l c o m p o n e n t se x c e e d e d 85 i n e a c h c a s e , a s d e t e r m i n e d b y m u l t i p l e c r i t e r i a ,w i t h a t l e a s t o n e o f t h e f o l l o w i n g a p p l i c a b l e : i n d i r e c t r o s e t t i n gw i t h m o n o c l o n a l a n t i b o d i e s a g a i n s t T c e l l d e t e r m i n a n t s
( M a c K a r i l l e t a l . , 1 9 8 7 ) , n u c l e a r Td T e x p r e s s i o n , f o c a l a c i dp h o s p h a t a s e r e a c t i o n I c a s e ) , o r p e r i p h e r a l l e u c o c y t e c o u n t i ne x c e s s o f 1 0 0 x 1 0 6 / m l c o u p l e d w i t h CD14 a n d CD19 d e t e r m i -n a n t s < 5 ( 2 c a s e s ) . HLA-Dr ( l a ) w a s a l s o d e t e r m i n e d i n e a c hc a s e , a n d c o n s i d e r e d p o s i t i v e w h e n > 30 HLA-Dr+ c e l l s w e r ep r e s e n t .
Q u a n t i t a t i o n o f c e l l s u r f a c e f 3 2 m - a s s o c i a t e d m e m b r a n e d e t e r m i -n a n t s b y i n d i r e c t s a t u r a t i o n b i n d i n g a s s a y
T h i s m e t h o d i s d e s c r i b e d i n d e t a i l e l s e w h e r e ( J o n e s , S c o t t C h i l d , 1 9 8 8 ) b u t , b r i e f l y , c o n s i s t e d o f i n c u b a t i n g t e s t c e l l s w i t hm o n o c l o n a l a n t i b o d i e s a t s a t u r a t i o n , r e m o v a l o f t h e u n b o u n dc o m p o n e n t b y w a s h i n g , a n d r e s u s p e n d i n g t h e c e l l s i n s a t u r a t i n g
c o n c e n t r a t i o n so f
H R P x - l a b e l l e d r a b b i t a n t i - m o u s e I g G . A f t e r
w a s h i n g a w a y u n b o u n d s e c o n d a n t i b o d y , t h e c e l l s w e r e a s s a y e df o r b o u n d p e r o x i d a s e u s i n g 2 1 mm o - p h e n y l e n e d i a m i n e i nc i t r a t e / p h o s p h a t e b u f f e r pH 5 * 0 c o n t a i n i n g h y d r o g e n p e r o x i d ea t 2 9 mm. T h e r a t e o f c h a n g e i n a b s o r b a n c e a t 4 9 2 n m w a s
d et e rm i n e d a n d p e r o x i d a s e a c t i v i t i e s e x p r e s s e d a s U / 1 0 6 c e l l s ,w h e r e 1 U was d e f i n e d a s a r a t e o f c h a n g e i n A 4 9 2 o f 0 0 1 / m i nu n d e r s t a n d a r d a s s a y c o n d i t i o n s .
S t a t i s t i c a l a n a l y s i sTo t e s t t h e s i g n i f i c a n c e o f d i f f e r e n c e b e t w e e n t w o p o p u l a t i o n st h e M a n n - W h i t n e y u - t e s t was u s e d , o r f o r p a i r e d d a t a , t h eW i l c o x o n m a t c h e d p a i r s s i g n e d - r a n k s t e s t . C o r r e l a t i o n a n a l y s i swa s p e r f o r m e d u s i n g S p e a r m a n s r a n k c o r r e l a t i o n c o e f f i c i e n t
r j . A l l t h e a b o v e t e s t s a r e s u i t a b l e f o r n o n - p a r a m e t r i c d a t a .
RESULTS
C e l l - s u r f a c e f l 2 m a n d HLA-ABC a n t i g e n e x p r e s s i o n w a s d e t e r -m i n e d b y i m m u n o e n z y m e a s s a y on t h e l y m p h o i d c e l l s f r o m 3 9
c a s e s o f T c e l l m a l i g n a n c y , a n d CDla a n t i g e n e x p r e s s i o n wa s
a l s o d e t e r m i n e d i n 2 5 o f t h e s e . I n a r e p l i c a t e s t u d y ( n = 2 0 ) ,
e s t i m a t i o n o f / 3 2 m : H L A - A B C r a t i o g a v e a v a l u e f o r 1 s . d . o f0 0 8 , a n d t h e r e f o r e c a s e s w i t h r a t i o s g r e a t e r t h a n s . d . a b o v e u n i t i . e . > 1 1 6 : 1 w e r e c o n s i d e r e d s i g n i f i c a n t l y i n c r e a s e d . T h e t w o -
s t a g e d e t e r m i n a t i o n o f c e l l - s u r f a c e f2m d e n s i t y g a v e a c o -
e f f i c i e n t o f v a r i a t i o n ( C V ) o f 1 2 8 .
Mean t 2 m a n d HLA-ABC v a l u e s f o r t h e TdT+ t h y m i c - A L Lb l a s t s ( n = 2 0 ) w e r e 1 7 6 s . e . 2 6 ) a n d 1 4 9 s . e . 2 - 2 ) ) U / 1 0 6 c e l l sr e s p e c t i v e l y ( T a b l e 3 . I n c o n t r a s t , TdT- l e u k a e m i c T c e l l s f r o m
p o s t - t h y m i c d i s o r d e r s ( n = 1 9 ) ( T a b l e 4 ) e x p r e s s e d s i g n i f i c a n t l ymore ( P < 0 0 0 1 ) f 3 2 m a n d HLA-ABC d e t e r m i n a n t s , w i t h meanv a l u e s o f 5 2 9 s . e . 6 2 ) a n d 5 0 6 s . e . 6 - 4 ) U / 1 0 6 c e l l s r e s p e c t i v e l y .
C o m p a r i s o n b y W i l c o x o n m a t c h e d p a i r s t e s t s h o w e d t h a tt h y m i c - A L L b l a s t s e x p r e s s e d s i g n i f i c a n t l y m o r e ( P < 0 - 0 ) f J 2 m
t h a n HLA-ABC. T h e r e w a s n o s i g n i f i c a n t d i f f e r e n c e b e t w e e nf 2 m a n d HLA-ABC e x p r e s s i o n f o r t h e T c e l l s f r o m p o s t - t h y m i cd i s o r d e r s , h o w e v e r . T h i s i s f u r t h e r i l l u s t r a t e d b y t h e f i n d i n g o f
s i g n i f i c a n t l y d i f f e r e n t ( P < 0 0 5 ) mean t h 2 M : HLA-ABC r a t i o sf o r t h y m i c - A L L b l a s t s ( m e a n 1 - 2 6 : 1 ; r a n g e 0 - 8 4 : 1 t o 1 - 8 3 : 1 )c o m p a r e d t o p o s t - t h y m i c l e u k a e m i c T c e l l s ( m e a n 1 0 6 : 1 ; r a n g e
0 - 8 7 : 1 t o 1 2 6 : 1 . T h e s e t w o s e t s o f d a t a i n d i c a t e t h a t o t h e r f 2 m -c o n t a i n i n g m o l e c u l e s i n a d d i t i o n t o HLA-ABC a r e p r e s e n t ont h e b l a s t s f r o m s o m e c a s e s o f t h y m i c - A L L . I n t h i s c o n t e x t , i t w a s
a p p a r e n t t h a t CD a e x p r e s s i o n , a s a f u n c t i o n o f t h e t o t a l t 2 Mc o m p o n e n t , w a s s i g n i f i c a n t l y h i g h e r ( P < 0 0 0 1 ) o n t h y m i c - A L Lb l a s t s ( m e a n 0 - 1 4 , s . e . 0 - 0 5 ) c o m p a r e d t o p o s t - t h y m i c l e u k a e m i cT c e l l s ( m e a n 0 - 0 1 , s . e . 0 - 0 ) . S p e c i f i c a l l y , o n t h e b l a s t s f r o m s i x
o u t o f 1 4 c a s e s o f t h y m i c - A L L , p r o p o r t i o n a t e CDI a e x p r e s s i o n
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R . A . J o n e s e t a l .
w a s e q u a l t o o r i n e x c e s s o f 0 1 , a n d o f t h e s e , f o u r e x h i b i t e d12m:HLA-ABC r a t i o s i n e x c e s s o f 1 1 6 : 1 . I n t h e p o s t - t h y m i c
l e u k a e m i c T c e l l s , b y c o n t r a s t , p r o p o r t i o n a t e CD l a e x p r e s s i o ni n e x c e s s o f 0 - 1 w a s s e e n i n n o n e o f I I c a s e s , a n d f l 2 m : HLA-
ABC r a t i o a b o v e [ 1 6 : 1 w a s s e e n i n o n e o u t o f 1 1 . T h i s a p p a r e n t
c o r r e l a t i o n b e t w e e n p r o p o r t i o n a t e CDla e x p r e s s i o n a n d n o n -
HLA f 2 m w a s t e s t e d b y S p e a r m a n s r a n k c o r r e l a t i o n a n a l y s i sa n d f o u n d t o b e s i g n i f i c a n t a t P < 0 0 5 r , = 0 4 4 ) . I t c a n b e s e e n
h o w e v e r t h a t i n many c a s e s o f t h y m i c - A L L a n d p o s t - t h y m i cp r o l i f e r a t i o n s , t h e a p p a r e n t e x c e s s 3 2 m e x p r e s s i o n i s n o t w h o l l y
a c c o u n t e d f o r b y e x p r e s s i o n o f t h e CD a a n t i g e n , p o s s i b l e
r e a s o n s f o r w h i c h a r e d i s c u s s e d b e l o w .
No s i g n i f i c a n t d i f f e r e n c e e x i s t e d b e t w e e n f 2 m e x p r e s s i o n b y
t h e l e u k a e m i c T c e l l s f r o m HLA-Dr+ ( m e a n 5 4 2 , s . e . 1 0 1 U / 1 0 6
c e l l s ; n = 1 0 ) c o m p a r e d t o HLA-DR- ( m e a n 5 1 5 , s . e . 7 - 4 U / 1 0 6
c e l l s ; n = 9 ) p o s t - t h y m i c d i s o r d e r s . H o w e v e r , w h e n c o m p a r i n g
t h e s e d i s o r d e r s a s a f u n c t i o n o f C D 4 o r C D8 e x p r e s s i o n , T c e l l s
f r o m t h e CD4+ g r o u p w e r e f o u n d t o e x p r e s s s i g n i f i c a n t l y h i g h e r
( P < 0 0 2 5) l e v e l s o f f 2 m ( m e a n 6 4 3 , s . e . 9 7 U / 1 0 6 c e l l s ; n = 1 0 )
c o m p a r e d t o t h o s e f r o m t h e CD 8 + g r o u p ( m e a n 4 1 1 , s . e . 5 - 8 U /1 0 6 c e l l s ; n = 8 .
C o m p a r i n g t h e d a t a f r o m t h i s s t u d y w i t h t h a t f r o m t h e s t u d yo f n o r m a l T c e l l s ( J o n e s , S c o t t C h i l d , 1 9 8 8 ; d a t a s u m m a r i s e d
T a b l e 3 . C e l l - s u r f a c e f 3 2 m , HLA-ABC a n d CD la e x p r e s s i o n b y TdT+
t h y m i c ALL b l a s t s
f 2 m HLA-ABC CDla f 2 m : H L A *
T a b l e 4 . C e l l - s u r f a c e I ? 2 m , HLA-ABC a n d CD l a e x p r e s s i o n TdT- p o s t -t h y m i c T - c e l l s
I a /2m HLA-ABC CD1a f i 2 m : H L A *
N o r m a l PB T c e l l s t ( m e a n + s . e . )
CD4+8- 2 2 - 2 + 1 - 2 2 1 - 8 + 1 - 3
CD-8+ 3 0 - 4 + 1 - 5 3 0 - 2 + 2 - 0
CD4+ p o s t - t h y m i c T c e l l s2 12 2
2 3
2 4
2 5
2 6
2 7
2 8
2 9
3 0
T-PLL
T PLL
S S
S s
S s
S s
T-NHLT-NHL
T-NHL
ATLL
CD8+ p o s t - t h y m i c T c e l l s l3 1 T-NHL
3 2 LDGL
3 3 LDGL 3 4 LDGL
3 5 LDGL +
3 6 LDGL +
3 7 LDGL +
3 8 LDGL +
3 9 T - P L L §
7 9 0 0 0
3 2 - 0
3 0 0
4 6 - 0
4 7 - 54 1 5
7 8 - 0
465
46 5
9 6 - 0
2 8 - 07 4 - 0
290
425
330
40 0
2 7 - 0
5 5 - 0
3 4 - 0
730
3 4 - 0
1 2 3 - 5
3 9 - 0
40 0
3 6 - 0900
4 1 0
4 4 . 5
1 0 4 - 5
2 7 - 55 9 - 0
2 9 - 0
4 4 - 0
2 8 5
3 8 - 02 5 5
5 1 0
3 4 - 0
ND 103+004
ND 1 - 0 3 +0 0 5
1 - 0 ( 0 - 0 1 )
ND
0 - 7 ( 0 - 0 )
ND
2 5 ( 0 ° 0 5 )
0 - 2 ( 0 0 )
ND
ND
1 0 ( 0 0 2 )
0 0 ( 0 0 )
0 - 6 ( 0 0 2 )ND
ND
0 - 7 ( 0 - 0 1 )ND
0 0 ( 0 0 )0 - 2 ( 0 1 )
1 - 7 ( 0 - 0 3 )ND
1 - 0 8
0 - 9 4
1 - 0 51 - 1 8
1 - 1 81 - 1 5
0 - 8 7
1 1 3
1 - 0 4
0 - 9 2
1 - 0 2
1 - 2 5
1 - 0 0
0 - 9 7
1 - 1 6
1 - 0 51 - 0 61 0 8
1 - 0 0
Normal t h y m o c y t e s f ( m e a n + ± s . e . )
CDla+ 1 6 - 4 + 2 - 0 1 7+04
CDl a- 1 7 - 6 + 1 - 7 1 6 - 0 + 1 - 9
M a l i g n a n t t h y m i c - A L L b l a s t s
C a s e 1 1 0 - 5 1252 65 7 - 0
3 1 9 - 5 2 1 - 0
4 3 6 - 5 3 8 - 0
5 2 6 - 0 2 6 - 0
6 1 1 - 5 1 1 0
7 2 6 - 0 2 5 - 0
8 1 5 - 5 1 4 - 0
9 1 6 - 0 1 4 - 0
1 0 1 6 - 5 1 4 5
1 1 7 . 5 65
1 2 2 2 - 0 1 8 5
1 3 7 . 5 6 0
1 4 4 7 - 5 3 4 0
1 5 6 - 5 4 . 5
1 6 3 4 5 2 2 - 0
1 7 5 - 0 30
1 8 1 9 - 0 1 1 - 01 9 7 - 0 40
2 0 110 6 - 0
1 5 7 1 - 9
05+0-2
0 - 7 ( 0 - 0 7 )0 1 ( 0 0 2 )
0 7 ( 0 - 0 4 )10 ( 0 0 3 )
ND
0 8 ( 0 - 0 7 )ND
ND
1 - 7 ( 0 1 1 )
1 1 ( 0 - 0 7 )
1 - 0 ( 0 - 1 3 )0 0 ( 0 0 )
0 - 2 ( 0 - 0 3 )4 - 7 ( 0 - 1 )
1 ( 0 2 3 )2 0 - 0 ( 0 5 8 )ND
9 - 6 ( 0 - 5 1 )ND
ND
0 - 8 40 - 9 3
0 - 9 3
0 - 9 6
1 - 0 0
1 - 0 4
1 - 0 4
1 I 1 1
1 - 1 4
1 1 4
1 - 1 5
1 - 1 9
1 - 2 5
1 - 4 0
1 - 4 4
1 - 5 71 - 6 7
1 - 7 31 - 7 5
1 - 8 3
R e s u l t s a r e r e l a t i v e f i 2 m , HLA-ANC a n d CD la a n t i g e n d e n s i t i e s ,
s h o w n a s e n z y m e ( p e r o x i d a s e ) u n i t s / 1 0 6 c e l l s ( m e a n o f d u p l i c a t e a s s a y s ) .
F i g u r e s i n p a r e n t h e s e s i n d i c a t e CD I a e x p r e s s i o n a s a p r o p o r t i o n o f t o t a l
f l 2 m e x p r e s s i o n . ND n o t d e t e r m i n e d .
* 2 m : HLA-ABC r a t i o
t C e l l - s u r f a c e ( J 2 m a n d HLA-ABC e x p r e s s i o n b y n o r m a l CD4+8-( n = 1 0 ) a n d CD4- 8 + ( n = 1 0 ) p e r i p h e r a l b l o o d T - c e l l s a s d e t e r m i n e d
p r e v i o u s l y ( J o n e s , S c o t t C h i l d , 1 9 8 8 ) .
1 LDGL c a s e s n u m b e r e d 3 3 , 3 4 , 3 6 a n d 3 8 w e r e CD16 ( l e u I 1 b ) +
§ C o - e x p r e s s e d b o t h CD 4 a n d C D8 d e t e r m i n a n t s .
i n T a b l e s 3 a n d 4 ) , we f o u n d t h a t t i 2 m e x p r e s s i o n b y TdT+
CDla+ t h y m i c - A L L b l a s t s ( m e a n 2 2 8 , s . e . 4 7 ; n = 9 ) was n o t
s i g n i f i c a n t l y d i f f e r e n t t o t h a t o f TdT+ CDl a+ n o r m a l t h y m o -
c y t e s ( m e a n 1 6 4 , s . e . 2 0 ; n = 1 4 . S i m i l a r l y , t h e r e w a s o
s i g n i f i c a n t d i f f e r e n c e b e t w e e n t h e l e v e l o f e x p r e s s i o n on T c e l l s
f r o m CD8+ p o s t - t h y m i c d i s o r d e r s ( m e a n 4 1 1 , s . e . 5 8 ; n = 8 )
a n d t h e n o r m a l p e r i p h e r a l CD4-8+ s u b p o p u l a t i o n ( m e a n 3 0 4 ,
s . e . 1 5 ; n = 1 0 . H o w e v e r , w h e n t h e l e v e l o f,2m e x p r e s s i o n b y Tc e l l s f r o m CD4+ p o s t - t h y m i c d i s o r d e r s ( m e a n 6 4 3 , s . e . 9 7 ;
n = 1 0 ) w a s c o m p a r e d t o t h a t o f t h e n o r m a l p e r i p h e r a l CD4+8-s u b p o p u l a t i o n ( m e a n 2 2 2 , s . e . 1 - 2 ; n= 1 0 , s i g n i f i c a n t l y h i g h e r( P < 0 - 0 0 1 ) e x p r e s s i o n was f o u n d .
R e s u l t s a r e r e l a t i v e f 3 2 m , HLA-ABC a n d CDla a n t i g e n d e n s i t i e s ,s h o w n a s e n z y m e ( p e r o x i d a s e ) u n i t s / 1 0 6 c e l l s ( m e a n o f d u p l i c a t e a s s a y s ) .F i g u r e s i n p a r e n t h e s e s i n d i c a t e CD I a e x p r e s s i o n a s a p r o p o r t i o n o f t o t a l
/ 3 2 m e x p r e s s i o n . ND n o t d e t e r m i n e d .
* f l 2 m : H L A - A B C r a t i o .
t C e l l - s u r f a c e f l 2 m , HLA-ABC a n d CDla e x p r e s s i o n b y n o r m a l
CDla + ( n = 1 4 ) a n d CDIa- ( n = l 1 ) t h y m o c y t e s a s d e t e r m i n e d p r e -
v i o u s l y ( J o n e s , S c o t t
C h i l d , 1 9 8 8 ) .
DISCUSSION
T h i s s t u d y i s a n e x t e n s i o n o f a p r e v i o u s i n v e s t i g a t i o n ( J o n e s ,
S c o t t C h i l d , 1 9 8 8 ) w h i c h e x a m i n e d t h e e x p r e s s i o n o f /2m,
HLA-ABC a n d C D l a a n t i g e n s o n m o n o c l o n a l a n t i b o d y -
d e f i n e d n o r m a l t h y m i c a n d p e r i p h e r a l b l o o d T c e l l s u b p o p u l a -
t i o n s . I n t h e p r e s e n t s t u d y o f 3 9 cases o f T c e l l m a l i g n a n c y , we
f o u n d t h a t t h e
e x p r e s s i o no f
f l 2 m - a s s o c i a t e d p r o t e i nwas
4 5 6
L4
4
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E x p r e s s i o n o f f 3 2 m - a s s o c i a t e d m o l e c u l e s o n T c e l l s
s i g n i f i c a n t l y h i g h e r o n t h e T c e l l s f r o m p o s t - t h y m i c d i s o r d e r sc o m p a r e d w i t h t h y m i c - A L L b l a s t s . T h i s p a r a l l e l s t h e s i t u a t i o ni n n o r m a l l y m p h o i d t i s s u e , w h e r e / 2 m e x p r e s s i o n b y p e r i p h e r a lT c e l l s i s s i g n i f i c a n t l y h i g h e r t h a n t h a t o f t h y m o c y t e s ( J o n e s ,S c o t t C h i l d , 1 9 8 8 ) .
I t h a s b e e n p r o p o s e d t h a t T-CLL ( d e s i g n a t e d LDGL i n t h i s
s t u d y ; P a n d o l f i , 1 9 8 6 ) a n d S e z a r y c e l l s c o r r e s p o n d , i n t e r m s o f
i m m u n o l o g i c a l d i f f e r e n t i a t i o n , t o s u b s e t s o f n o r m a l p e r i p h e r a l
T c e l l s ( B o u m s e l l e t a l . , 1 9 8 1 ) . Mean f i 2 m e x p r e s s i o n b y t h e Tc e l l s f r o m t h e s e s u b g r o u p s ( n = 1 1 ) w a s 5 1 - 4 s . e . 8 8 ) U / 1 0 6 c e l l s ,
w h i c h w a s s i g n i f i c a n t l y h i g h e r (P<0 0 0 1 ) t h a n mean f 2 m
e x p r e s s i o n b y t h e c e l l s f r o m n o r m a l CD4+ a n d CD8+ s u b p o p u -
l a t i o n s ( m e a n 2 6 - 3 , s . e . 1 3 ) .We h a v e a l s o s h o w n t h a t l e u k a e m i c TdT- CD4+ T c e l l s
e x p r e s s s i g n i f i c a n t l y more f 2 m - a s s o c i a t e d p r o t e i n t h a n l e u k a e -m i c TdT-CD8+ T c e l l s . T h i s i s a n i n t e r e s t i n g o b s e r v a t i o n a n d i s
t h e r e v e r s e o f t h e s i t u a t i o n i n n o r m a l l y m p h o i d t i s s u e , w h e r e
i 2 m e x p r e s s i o n b y t h e p e r i p h e r a l b l o o d CD4- 8 + s u b p o p u l a t i o ni s s i g n i f i c a n t l y h i g h e r ( P < 0 - O 0 1 ) t h a n t h e CD4+8- s u b p o p u l a -
t i o n ( J o n e s , S c o t t C h i l d , 1 9 8 8 ) . I t i s a p p a r e n t t h a t p o s t - t h y m i c
CD4+ l e u k a e m i c T c e l l s e x p r e s s s i g n i f i c a n t l y m o r e f 2 m - a s s o -
c i a t e d m o l e c u l e s t h a n a l l o t h e r t y p e s o f n o r m a l o r m a l i g n a n t Tc e l l . We a r e n o t a w a r e o f t h e p r e c i s e s i g n i f i c a n c e o f t h i s i n c r e a s e dl e v e l o f e x p r e s s i o n o f f l 2 m - a s s o c i a t e d m o l e c u l e s , r e l a t i v e t on o r m a l p e r i p h e r a l T c e l l s , i n c e r t a i n T l i n e a g e l y m p h o i d
p r o l i f e r a t i o n s , bu t i t c l e a r l y c o n t r a s t s w i t h t h e s i t u a t i o n s e e n i n
many s o l i d t u m o u r s w h e r e p r o g r e s s i o n t o m a l i g n a n c y may b e
a s s o c i a t e d w i t h d e c r e a s e d c e l l - s u r f a c e MHC C l a s s I a n t i g e n s( F l e m i n g e t a l . , 1 9 8 1 ; T u r b i t t M a c k i e , 1 9 8 1 ; N a t a l i e t a l . ,
1 9 8 4 ) , a p r o c e s s w h i c h may f a c i l i t a t e e v a s i o n o f T c e l l c y t o t o x i -c i t y ( F e s t e n s t e i n S c h m i t , 1 9 8 1 ; G o o d i n g , 1 9 8 2 ) . I t h a s r e c e n t l yb e e n r e p o r t e d , h o w e v e r , t h a t s u s c e p t i b i l i t y t o NK a c t i v i t y v a r i e si n v e r s e l y w i t h HLA-ABC e x p r e s s i o n o n t h e t a r g e t c e l l ( H a r a l -B e l l a n e t a l . , 1 9 8 6 ) , a n d t h e r e f o r e c e l l s w h i c h e v a d e T c e l l
c y t o t o x i c i t y b y v i r t u e o f r e d u c e d MHC C l a s s I a n t i g e n e x -p r e s s i o n may b e c o m e t a r g e t s f o r t h e n a t u r a l k i l l e r c e l l m e d i a t e d
p a t h w a y . T h e c l i n i c a l s i g n i f i c a n c e o f t h e s e f i n d i n g s r e q u i r e sf u r t h e r i n v e s t i g a t i o n .
A p r e v i o u s s t u d y ( H o k l a n d e t a l . , 1 9 8 2 ) r e p o r t e d h i g h
d e n s i t y m e m b r a n e f 2 m ( b u t w i t h n o c o r r e s p o n d i n g i n c r e a s e i nHLA-ABC) i n 25 o f p e r i p h e r a l l y m p h o c y t e s , a n d t h i s
a p p e a r e d t o c o r r e l a t e w i t h n a t u r a l k i l l e r ( N K ) a c t i v i t y . T h e
m a j o r i t y o f NK c e l l s a r e a s s o c i a t e d w i t h t h e c h a r a c t e r i s t i c LGL
m o r p h o l o g y a n d l a b e l w i t h CD 1 6 a n t i b o d i e s ( L a n i e r , P h i l l i p s
W a r n e r , 1 9 8 6 ) , a n d i t c a n b e s e e n f r o m T a b l e 4 t h a t t h e T c e l l s
f r o m t h e f o u r c a s e s o f CD 1 6 + LDGL d o n o t h a v e a n a p p a r e n t
i n c r e a s e i n e i t h e r f l 2 m e x p r e s s i o n o r f l 2 m : H L A - A B C r a t i o ,
r e l a t i v e t o n o r m a l p e r i p h e r a l T c e l l s . We a r e c u r r e n t l y e x a m i n -i n g t h e n o r m a l p e r i p h e r a l C D 4 - 8 - 1 4 - 1 9 - s u b p o p u l a t i o n ,
w h i c h c o n t a i n s a l a r g e CD16+ c o m p o n e n t , t o d e t e r m i n e t h el e v e l o f e x p r e s s i o n o f f 2 m - a s s o c i a t e d m o l e c u l e s .
No s i g n i f i c a n t d i f f e r e n c e w a s d e t e c t e d i n t h e l e v e l o f
e x p r e s s i o n o f t i M - a s s o c i a t e d d e t e r m i n a n t s b e t w e e n T c e l l s f r o m
' a c t i v a t e d ' ( H L A - D r + ) a n d ' n o n - a c t i v a t e d ' ( H L A - D r - ) s t a g e so f p o s t - t h y m i c d i s e a s e , s u g g e s t i n g t h a t t h e l e v e l o f e x p r e s s i o n o f
t h e s e m o l e c u l e s i s n o t a c t i v a t i o n - r e l a t e d . T h i s i s i n c o n t r a s t t ot h e s i t u a t i o n i n v i t r o , w h e r e b y m i t o g e n i c s t i m u l a t i o n o f p e r i p h -e r a l T c e l l s i n d u c e s a m a r k e d i n c r e a s e i n t h e e x p r e s s i o n o f 3 2 m -a s s o c i a t e d p r o t e i n ( D o r v a l e t a l . , 1 9 7 7 i n p a r a l l e l w i t h HLA-Dr
i n d u c t i o n ( A c c o l l a , M o r e t t a C a r r e l , 1 9 8 4 ) .
We c o u l d d e t e c t n o s i g n i f i c a n t d i f f e r e n c e i n t h e l e v e l o f
e x p r e s s i o n o f t 2 M - a s s o c i a t e d m o l e c u l e s b e t w e e n TdT+CDla+t h y m i c - A L L b l a s t s a n d TdT+CDla+ t h y m o c y t e s . H o w e v e r ,
t 2 m e x p r e s s i o n i n t h e s e l a t t e r c e l l s i s a l m o s t e x c l u s i v e l y CD a -
a s s o c i a t e d , w i t h o n l y a v e r y m i n o r HLA-ABC c o m p o n e n t ,
w h e r e a s TdT+CDla+ t h y m i c - A L L b l a s t s c o - e x p r e s s b o t h
CDla a n d HLA-ABC, a n d ma y t h e r e f o r e r e p r e s e n t a d i f f e r e n ts t a g e o f d i f f e r e n t i a t i o n t o t h e TdT+CDla+HLA-ABC- t h y m o -
c y t e ( B r a d s t o c k e t a l . , 1 9 8 0 ) . We c o u l d n o t c o m p a r e t h ee x p r e s s i o n o f t h 2 M - a s s o c i a t e d p r o t e i n b y t h e TdT+ CDla-
t h y m i c - A L L b l a s t s w i t h t h e i r c o r r e s p o n d i n g n o r m a l t h y m o c y t e
p h e n o t y p e s , s i n c e CDla- t h y m o c y t e s c o m p r i s e d l e s s t h a n 1 0
o f t h e w h o l e t h y m o c y t e f r a c t i o n , a n d o f t h e s e , a b o u t 90 w e r e
a l s o T d T - , r e p r e s e n t i n g a p h e n o t y p e m o r e c l o s e l y c o r r e s p o n d -i n g t o t h a t o f t h e m a t u r e p e r i p h e r a l T c e l l . B y e x a m i n i n g t h er a t i o o f f l 2 m t o HLA-ABC, i t i s p o s s i b l e t o a s s e s s t h e p o t e n t i a le x p r e s s i o n o f n o n HLA-ABC m o l e c u l e s . We h a v e s h o w n t h a te x c e s s ( n o n - H L A - a s s o c i a t e d ) f l 2 m i s p r e s e n t i n s i g n i f i c a n tq u a n t i t y o n t h e s u r f a c e o f b l a s t s f r o m s o m e c a s e s o f t h y m i c -ALL, a n d t h a t t h e r e i s a c o r r e l a t i o n b e t w e e n t h i s e x c e s s , a sd e t e r m i n e d b y f i 2 m : H L A - A B C r a t i o , a n d CDla e x p r e s s i o n ( a s a
p r o p o r t i o n o f t o t a l f l 2 m . I n s o m e o f t h e s e c a s e s , h o w e v e r , t h ea p p a r e n t non-HLA-ABC a s s o c i a t e d f 3 2 m i s n o t w h o l l y
a c c o u n t e d f o r b y CD a e x p r e s s i o n , a n d i t i s t h e r e f o r e p o s s i b l et h a t o t h e r t h 2 M - c o n t a i n i n g m o l e c u l e s a r e p r e s e n t . I n t h i s r e s p e c t ,f o u r d i s t i n c t f 2 m - c o n t a i n i n g g p 4 3 - 4 9 , 1 2 s u r f a c e a n t i g e n s o t h e r
t h a n HLA-ABC h a v e b e e n i d e n t i f i e d ( K n o w l e s B o d m e r ,1 9 8 2 ; O l i v e , D u b r e u i l M a w a s , 1 9 8 4 ; A m i o t e t a l . , 1 9 8 6 ) t h r e eo f w h i c h h a v e s i n c e b e e n a s c r i b e d t o t h e CD1 c l u s t e r ( M c M i -
c h a e l G o t c h , 1 9 8 7 ) . I t i s l i k e l y t h a t s o m e o f t h e s e m o l e c u l e s i na d d i t i o n t o CD I a may b e p r e s e n t o n t h e T c e l l s f r o m t h y m i c a n d
p o s s i b l y a l s o p o s t - t h y m i c d i s o r d e r s a n d may t h e r e f o r e a c c o u n t
f o r t h e a p p a r e n t non-HLA a n d n o n - C D l a - a s s o c i a t e d 3 2 m . Wea r e c ur re n tl y e x a m i n i n g t h e e x p r e s s i o n o f o t h e r t 2 M - a s s o c i a t e d
m o l e c u l e s i n m a l i g n a n t B a n d T c e l l s .I n c o n c l u s i o n t h e r e f o r e , we h a v e s h o w n t h a t TdT- l e u k a e -
m i c T c e l l s e x p r e s s s i g n i f i c a n t l y m o r e t 2 M - a s s o c i a t e d d e t e r m i -
n a n t s t h a n TdT+ t h y m i c - A L L b l a s t s , w h i c h p a r a l l e l s t h e
s i t u a t i o n i n n o r m a l l y m p h o i d t i s s u e . No s i g n i f i c a n t l y d i f f e r e n tl e v e l o f e x p r e s s i o n o f t h e s e m o l e c u l e s w a s d e t e c t e d b e t w e e n
t h y m i c - A L L b l a s t s a n d t h y m o c y t e s , o r b e t w e e n TdT-CD8+l e u k a e m i c T c e l l s a n d CD4-8+ n o r m a l p e r i p h e r a l T c e l l s . Wed i d h o w e v e r d e t e c t s i g n i f i c a n t l y e l e v a t e d e x p r e s s i o n i n
TdT-CD4+ l e u k a e m i c T c e l l s c o m p a r e d t o p e r i p h e r a l
CD4+8-T c e l l s a n d TdT-CD8+ l e u k a e m i c T c e l l s , a n d a l s o i nt h e T c e l l s f r o m t h e S e z a r y s y n d r o m e a n d LDGL g r o u p
c o m p a r e d t o t h e n o r m a l ( p h e n o t y p i c a l l y s i m i l a r ) p e r i p h e r a l T
c e l l . F i n a l l y , we h a v e d e m o n s tr a t e d a c o r r e l a t i o n b e t w e e na p p a r e n t non-HLA-ABC a s s o c i a t e d f l 2 m a n d CDla e x p r e s s i o n ,
a n d a l s o f o u n d e v i d e n c e p o i n t i n g t o a n a d d i t i o n a l f 2 m -
c o n t a i n i n g c o m p o n e n t .
ACKNOWLEDGMENTS
We w i s h t o e x p r e s s o u r g r a t i t u d e t o Dr D a r i o C a m p a n a , D e p t . o f
I m m u n o l o g y , R o y a l F r e e H o s p i t a l , L o n d o n , a n d t h e C l i n i c a l H a e m a t o -
l o g i s t s o f t h e Y o r k s h i r e R e g i o n f o r s u p p l y i n g much o f t h e m a t e r i a l , a n d
Dr A . R . S a n d e r s o n f o r s u p p l y i n g t h e a n t i - f l 2 m a n t i b o d y . T h i s w o r k w a s
s u p p o r t e d b y t h e F r i e n d s o f t h e L e u k a e m i a U n i t , a n d t h e S p e c i a lT r u s t e e s o f t h e G e n e r a l I n f i r m a r y , L e e d s .
4 5 7
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4 5 8 R . A . J o n e s e t a l .
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human t h y m o c y t e s w i t h d i f f e r e n t e x p r e s s i o n o n l e u k a e m i a l i n e s .
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