- Email: [email protected]
In vivo administration of cytokines in murine bone marrow chimeras
WS12-6
*s
IMMUNOTHERAPY OF COLON CANCER METASTASES:ROLE OF TNF AND ILl. J.F. JEANNIN, P.LAGADEC, H.PELLETIER, INSERM, Faculty of medicine, Dijon, France. The emergence of metastases is the most devastating aspect of colon cancer. We have developped models of colon cancer metastases in rat. In the next experiments colon cancer cells were obtained from a chemically induced colon adenocarcinoma in BD IX rats. They were cultivated in vitro and cloned. A clone of cells giving numerous solid nodules and not ascitic tumors, when injected i.p. was selected.The trearnent started from day 14 after the tumor cell injection, when the rats heated numerous nodules from 1 to 5 mm. The rats were killed and autopsied on day 42. Endotoxins and Lentinan significantly reduced the growth of carcinomatoses. They induced the disappearance of tumors in 27 out of the 110 rats treated with endotoxins and in11 out of the 30 rats treated with lentinan. OK 432 was ineffective. Macrophages being expected to play a role in tumor rejection, their activity was tested after their in vitro avtivation. Endotoxins and OK 432 induced direct macrophages-mediated tumor cell lysis (34% and 31% respectively, p<0.001) whereas lentinan was without effect. Enclotoxins and OK 432 induced the production of TNF (cytotoxicity of 75% and 90% respectively on wehi 164 clone 13 cells, p<0.001) whereas lentinan was without effect. Both endotoxins and OK induced the production of ILl (ILl activity of supernatants 124% and 138% respectively, p<0.001), lentinan was weakly effective only in the presence of indomethacin
(48%, p<0.01). Therefore there was no correlation between the therapeutic effect of endotoxins, lentinan or OK 432 and their ability to activate macrophages in vitro especially to induce the production of TNF and ILl.
WS12-7
*7
CHANGES OF IMMUNOLOGIC PARAMETERS FOLLOWING ADMINISTRATION OF RECOMBINANT INTERLEUKIN 2 IN PATIENTS WITH CHRONIC TYPE B HEPATITIS M. OnJi, S. Yamaguchi, H. Miyaoka, F. Bandou, N. Horiike, K. Michitaka and Y. Ohta Third Department of Internal Medicine, Ehime University School of Medicine, Ehime Japan Ten patients with chronic type B hepatitis were treated with recombinant interleukin 2 (rlL 2). Acute effects of rlL 2 administration on lymphoid cells included a rapid decrease in lymphocytes, especially in cytotoxic T cells and natural killer cells. These acute effects lasted within 24 hours. The same phenomenons were observed at 28 day of rlL 2 administration. The number of lymphocytes and CD4 positive cells was increased after rlL 2 administration for 28 days (p 0.01). Natural killer cells espcially CDI6 positive, Leu7 negative cells were increased (p 0.05). Beta-2 microglobulin of mononuclear cells was increased through the production of gamma-interferon during administration of rlL 2. Serum soluble IL 2 receptors were distinctly increased during administration of rlL 2. These effects may favor the elimination of virusinfected hepatocytes', and these phenomenons support the augumentation of cytotoxic T cell function in chronic type B hepatitis.
WS12-8
*8
IN V I V O A D M I N I S T R A T I O N OF C Y T O K I N E S IN M U R I N E B O N E M A R R O W C H I M E R A S M. I m a m u r a , H. F u j i m o t o , T. F u k u h a r a , S. H a s h i n o , M. K o b a y a s h i , M. K a s a i , K. S a k u r a d a , a n d T. M i y a z a k i 3rd Deot. Int. M e d . , H o k k a i d o Univ. Sch. M e d . , S a p p o r o , J a p a n immunodeficiency associated with graft-versus-host d i s e a s e is a m a j o r o b s t a c l e that m u s t be o v e r c o m e . We h a v e e x a m i n e d w h e t h e r in v i v o a d m i n i s t r a t i o n of r I L 2 o r r G - C S F can a c t i v a t e i m m u n o r e a c t i v i t y and o b v i a t e i m m u nodeficiency. 5 , 0 0 0 u n i t s of r I L 2 or 5 - 1 0 ~ g of r G - C S F w a s i n j e c t e d i.p. i n t o c h i m e r a s for 7 - i 0 d a y s f r o m day 7-14. Mitogen-induced responses were e x a m i n e d by c u l t u r i n g 5 x i 0 5 s p l e e n c e l l s f r o m n o r m a l or c h i m e r i c m i c e w i t h Con A, PHA, LPS, P W M f o r 2 days. A ! l o r e a c t i v e M L R s w e r e e x a m i n e d by cult u r i n g 5 x 1 0 5 s p l e e n c e l l s f r o m n o r m a l or c h i m e r i c m i c e w i t h the same n u m b e r of X - i r r a d i a t e d s p l e e n c e l l s f r o m n o r m a l or c h i m e r i c m i c e for 5 days. Thereafter, °H-TdR incorporation was counted. C y t o t o x i c a c t i v i t y w a s det e r m i n e d by a 5 1 C r r e l e a s e assay. Mitogen-induced r e s p o n s e s of s p l e e n c e l l s w e r e i n c r e a s e d in the c h i m e r a s i n j e c t e d w i t h r I L 2 or r G - C S F . Allor e a c t i v e M L R s w e r e a l s o i n c r e a s e d in ( B A L B / c ~ C 3 H / H e ) c h i m e r a s but not in (CBA/J ~ C3H/He)chimeras. A l t h o u g h c y t o t o x i c c e l l s w e r e not g e n e r a t e d in the s p l e e n of ( B A L B / c ~ C 3 H / H e ) c h i m e r a s , t h e s e s r e s u l t s i n d i c a t e that r i L 2 or r G - C S F s h o u l d be c a u t i o u s l y u s e d in b o n e m a r r o w t r a n s p l a n t a t i o n .
104
*s
IMMUNOTHERAPY OF COLON CANCER METASTASES:ROLE OF TNF AND ILl. J.F. JEANNIN, P.LAGADEC, H.PELLETIER, INSERM, Faculty of medicine, Dijon, France. The emergence of metastases is the most devastating aspect of colon cancer. We have developped models of colon cancer metastases in rat. In the next experiments colon cancer cells were obtained from a chemically induced colon adenocarcinoma in BD IX rats. They were cultivated in vitro and cloned. A clone of cells giving numerous solid nodules and not ascitic tumors, when injected i.p. was selected.The trearnent started from day 14 after the tumor cell injection, when the rats heated numerous nodules from 1 to 5 mm. The rats were killed and autopsied on day 42. Endotoxins and Lentinan significantly reduced the growth of carcinomatoses. They induced the disappearance of tumors in 27 out of the 110 rats treated with endotoxins and in11 out of the 30 rats treated with lentinan. OK 432 was ineffective. Macrophages being expected to play a role in tumor rejection, their activity was tested after their in vitro avtivation. Endotoxins and OK 432 induced direct macrophages-mediated tumor cell lysis (34% and 31% respectively, p<0.001) whereas lentinan was without effect. Enclotoxins and OK 432 induced the production of TNF (cytotoxicity of 75% and 90% respectively on wehi 164 clone 13 cells, p<0.001) whereas lentinan was without effect. Both endotoxins and OK induced the production of ILl (ILl activity of supernatants 124% and 138% respectively, p<0.001), lentinan was weakly effective only in the presence of indomethacin
(48%, p<0.01). Therefore there was no correlation between the therapeutic effect of endotoxins, lentinan or OK 432 and their ability to activate macrophages in vitro especially to induce the production of TNF and ILl.
WS12-7
*7
CHANGES OF IMMUNOLOGIC PARAMETERS FOLLOWING ADMINISTRATION OF RECOMBINANT INTERLEUKIN 2 IN PATIENTS WITH CHRONIC TYPE B HEPATITIS M. OnJi, S. Yamaguchi, H. Miyaoka, F. Bandou, N. Horiike, K. Michitaka and Y. Ohta Third Department of Internal Medicine, Ehime University School of Medicine, Ehime Japan Ten patients with chronic type B hepatitis were treated with recombinant interleukin 2 (rlL 2). Acute effects of rlL 2 administration on lymphoid cells included a rapid decrease in lymphocytes, especially in cytotoxic T cells and natural killer cells. These acute effects lasted within 24 hours. The same phenomenons were observed at 28 day of rlL 2 administration. The number of lymphocytes and CD4 positive cells was increased after rlL 2 administration for 28 days (p 0.01). Natural killer cells espcially CDI6 positive, Leu7 negative cells were increased (p 0.05). Beta-2 microglobulin of mononuclear cells was increased through the production of gamma-interferon during administration of rlL 2. Serum soluble IL 2 receptors were distinctly increased during administration of rlL 2. These effects may favor the elimination of virusinfected hepatocytes', and these phenomenons support the augumentation of cytotoxic T cell function in chronic type B hepatitis.
WS12-8
*8
IN V I V O A D M I N I S T R A T I O N OF C Y T O K I N E S IN M U R I N E B O N E M A R R O W C H I M E R A S M. I m a m u r a , H. F u j i m o t o , T. F u k u h a r a , S. H a s h i n o , M. K o b a y a s h i , M. K a s a i , K. S a k u r a d a , a n d T. M i y a z a k i 3rd Deot. Int. M e d . , H o k k a i d o Univ. Sch. M e d . , S a p p o r o , J a p a n immunodeficiency associated with graft-versus-host d i s e a s e is a m a j o r o b s t a c l e that m u s t be o v e r c o m e . We h a v e e x a m i n e d w h e t h e r in v i v o a d m i n i s t r a t i o n of r I L 2 o r r G - C S F can a c t i v a t e i m m u n o r e a c t i v i t y and o b v i a t e i m m u nodeficiency. 5 , 0 0 0 u n i t s of r I L 2 or 5 - 1 0 ~ g of r G - C S F w a s i n j e c t e d i.p. i n t o c h i m e r a s for 7 - i 0 d a y s f r o m day 7-14. Mitogen-induced responses were e x a m i n e d by c u l t u r i n g 5 x i 0 5 s p l e e n c e l l s f r o m n o r m a l or c h i m e r i c m i c e w i t h Con A, PHA, LPS, P W M f o r 2 days. A ! l o r e a c t i v e M L R s w e r e e x a m i n e d by cult u r i n g 5 x 1 0 5 s p l e e n c e l l s f r o m n o r m a l or c h i m e r i c m i c e w i t h the same n u m b e r of X - i r r a d i a t e d s p l e e n c e l l s f r o m n o r m a l or c h i m e r i c m i c e for 5 days. Thereafter, °H-TdR incorporation was counted. C y t o t o x i c a c t i v i t y w a s det e r m i n e d by a 5 1 C r r e l e a s e assay. Mitogen-induced r e s p o n s e s of s p l e e n c e l l s w e r e i n c r e a s e d in the c h i m e r a s i n j e c t e d w i t h r I L 2 or r G - C S F . Allor e a c t i v e M L R s w e r e a l s o i n c r e a s e d in ( B A L B / c ~ C 3 H / H e ) c h i m e r a s but not in (CBA/J ~ C3H/He)chimeras. A l t h o u g h c y t o t o x i c c e l l s w e r e not g e n e r a t e d in the s p l e e n of ( B A L B / c ~ C 3 H / H e ) c h i m e r a s , t h e s e s r e s u l t s i n d i c a t e that r i L 2 or r G - C S F s h o u l d be c a u t i o u s l y u s e d in b o n e m a r r o w t r a n s p l a n t a t i o n .
104