- Email: [email protected]
Thymostimulin (TS) effects on immunological responses of cyclophosphamide treated newborn guinea pigs
300
64
T H Y M O S I N / T F X / S T D ~ L A T E S T - H E L P E R CELLS OF IMMUNOGLOBULIN/Ig/SYNTPLESIS inviTRO. A . G 6 r s k i , M . N o w a c z y k t G.Kowalskam E . S k o p i ~ s k a - R d ~ e w s k a t Z.GaclonK. The T r a n s p l a n t a t i o n Institute, W a r s a w M e d i c a l School, 02006 Warsaw, POLAND. We h a v e p r e v i q u s l ~ shown that an e x t r a c t from calf thymus similar to thy m o s i n f r a c t i o n V / T F X / e x e r t s an ~ m m u n o m o d u l a t l n g a c t i o n on the in vitro for m a t i o n of T - l y m p h o i d c o l o n i e s / T h y m u s 1 9 8 1 , 3 , 1 2 9 / . I n the present c o m m u n i c a tion we show that TFX is also active on Ig synthesis in vitro. H u m a n lymph o c y t e s were a c t i v a t e d in v i t r o w i t h T - d e p e n d e n t / ~ @ ~ / o r i n d e p e n d e n t / C o w a n / s t i m u l a n t s and the n u m b e r of B cells p o s i t i v e for i n t r a c e l l u l a r Ig scored with an aid of f l u o r e s c e n c e m i c r o s c o p e ~ f o l l o w l n g the s t a i n i n g with F I T C - l a b e l l e d a n t i h u m a n ~ - g l o b u l i n a n t i b o d i e s T TFX added to the cultures s t i m u l a ted the PWM, but not C o w a n - d r i v e n Ig s y n t h e s i s in vitro. Likewise, patients treated w i t h TFX had l y m p h o c y t e s that p r o d u c e d 2-3x more Ig in r e s p o n s e to PWM/but not C o w a n / t h a n did n o r m a l l y m p h o c y t e s . However, the a d d i t i o n of TFX to c u l t u r e s of l y m p h o c y t e s p r o d u c i n g m a r k e d l y e l e v a t e d n u m b e r s of Ig-positlve cells i n h i b i t e d Ig synthesis. P u r i f i e d and i r r a d i a t e d T cells of TFXtreated p a t i e n t s caused a m a r k e d l y e n h a n c e d h e l p e r effect w h e n mixed w i t h n o r m a l B cells and c u l t u r e d w i t h PWM. Thus, t h y m o s l n / T F X / e x e r t s an ~mmLu~om o d u l a t i n g a c t i o n on h u m a n a n t i b o d y s y n t h e s i s in vitro s t i m u l a t i n g T-hellmr cells but also p r e v e n t i n g e x c e s s i v e h u m o r a l responses. @ A g i f t of Kent Labs and Tago, Inc./USA/ THYMOSTIMULIN (TS) EFFECTS ON IM~IJNOLOGICAL RESPONSES OF CYCLOPHOSPHAMIDE TREATED NEWBORN GUINEA PIGS. R. Falchetti, C. Cafiero, U. Diorlo and L. Caprino Istltuto Farmacologlco Serono, Via Casillna 125 - Rome Italy Neonatal guinea pigs (i-3 days old) were treated with a single intraperitoneal dose (50 mg/kg) of Cy. Twenty-four hours later the Cy-treated animals received, daily for six days an intraperltoneal injection of TS, I and 5 mg/kg, or saline. Newborn guinea pigs receiving only Cy treatment showed a decrease in Terminal Deoxynucleotydyl Transferase (TdT) activity of thymocytes; a decrease in the percentage of E-rosette forming thymocytee and a significant elevation in both spleen and peripheral blood T-lymphocytes. Moreo vet a significant increase of the response to T-mitogens Phytoemagglutinin (PHA) and Concanavalin A (Con A) was obtained. TS administration to the Cy-treated animals induced a further marked decrease of TdT specific activity in the thymocytes and a statistically si gnificant increase in the percentage of E-rosette-forming cells present in the thymus. The percentage of T-lymphocytes in spleen and peripheral blood was significantly reduced when compared to that of Cy-treated animals, with values very close to those found in Cyuntreated controls. Similarly TS reduced the reactivity of spleen cells to PHA and Con A previously increased by Cy administration. These studies indicate that in vivo TS admini stration to newborn guinea pigs is effective in stimulating the recovery of the T-lympho~ cytes functions previously depressed by Cy administration. HEPATOSIN. G. Renoux~ M. Renouxand J.M. Guillaumin. Laboratoire d'Immunologie, Facult~ de M~decine, B.P. 3223, 37032 Tours Cedex, :~ance.
Hepatosln is a heat-resistant, endotoxin-free peptide (M.W. circa 5 Kd) synthetized by liver cells, and particularly when cocultured wi~h sodium diethyldithiocarbamate, DTC. In vitro, pgs of Hepatosin induced 18 to 25 % Thy-1 + cells from nu/nu spleen cells in a short-term induction assays, whereas even 10 ~g left unchange~d B cell numbers in a 19-hr incubation assay. Hepatoein was equally able to recruit T cells from bone-marrow cells, and even in nude mice. Similar results were obtained on human cells, evidencing Eepatosin was active across the species barrier. Propanolol did not modify these effects, evidencing Hepatosin is not an ubiquitin-like product. In vivo, 0.1 to 10 ~g Eepatosin induced in n_~/nu mice the recruitment o£ functicnally active T cells, and stimulated to augmented responses C57BI/6 mice, muresponsive to most immunomodulators. Administration of C.I ~g to NZB mice reduced spleen weight and the I~M primary antibody response to E~RBC and increased the specific IgG-response. Present data suggest that Hepatosin might be the already hypothetized pre-th~ic event which allows thymic hormones to further induce preco~mitted cells to further differentiation.
64
T H Y M O S I N / T F X / S T D ~ L A T E S T - H E L P E R CELLS OF IMMUNOGLOBULIN/Ig/SYNTPLESIS inviTRO. A . G 6 r s k i , M . N o w a c z y k t G.Kowalskam E . S k o p i ~ s k a - R d ~ e w s k a t Z.GaclonK. The T r a n s p l a n t a t i o n Institute, W a r s a w M e d i c a l School, 02006 Warsaw, POLAND. We h a v e p r e v i q u s l ~ shown that an e x t r a c t from calf thymus similar to thy m o s i n f r a c t i o n V / T F X / e x e r t s an ~ m m u n o m o d u l a t l n g a c t i o n on the in vitro for m a t i o n of T - l y m p h o i d c o l o n i e s / T h y m u s 1 9 8 1 , 3 , 1 2 9 / . I n the present c o m m u n i c a tion we show that TFX is also active on Ig synthesis in vitro. H u m a n lymph o c y t e s were a c t i v a t e d in v i t r o w i t h T - d e p e n d e n t / ~ @ ~ / o r i n d e p e n d e n t / C o w a n / s t i m u l a n t s and the n u m b e r of B cells p o s i t i v e for i n t r a c e l l u l a r Ig scored with an aid of f l u o r e s c e n c e m i c r o s c o p e ~ f o l l o w l n g the s t a i n i n g with F I T C - l a b e l l e d a n t i h u m a n ~ - g l o b u l i n a n t i b o d i e s T TFX added to the cultures s t i m u l a ted the PWM, but not C o w a n - d r i v e n Ig s y n t h e s i s in vitro. Likewise, patients treated w i t h TFX had l y m p h o c y t e s that p r o d u c e d 2-3x more Ig in r e s p o n s e to PWM/but not C o w a n / t h a n did n o r m a l l y m p h o c y t e s . However, the a d d i t i o n of TFX to c u l t u r e s of l y m p h o c y t e s p r o d u c i n g m a r k e d l y e l e v a t e d n u m b e r s of Ig-positlve cells i n h i b i t e d Ig synthesis. P u r i f i e d and i r r a d i a t e d T cells of TFXtreated p a t i e n t s caused a m a r k e d l y e n h a n c e d h e l p e r effect w h e n mixed w i t h n o r m a l B cells and c u l t u r e d w i t h PWM. Thus, t h y m o s l n / T F X / e x e r t s an ~mmLu~om o d u l a t i n g a c t i o n on h u m a n a n t i b o d y s y n t h e s i s in vitro s t i m u l a t i n g T-hellmr cells but also p r e v e n t i n g e x c e s s i v e h u m o r a l responses. @ A g i f t of Kent Labs and Tago, Inc./USA/ THYMOSTIMULIN (TS) EFFECTS ON IM~IJNOLOGICAL RESPONSES OF CYCLOPHOSPHAMIDE TREATED NEWBORN GUINEA PIGS. R. Falchetti, C. Cafiero, U. Diorlo and L. Caprino Istltuto Farmacologlco Serono, Via Casillna 125 - Rome Italy Neonatal guinea pigs (i-3 days old) were treated with a single intraperitoneal dose (50 mg/kg) of Cy. Twenty-four hours later the Cy-treated animals received, daily for six days an intraperltoneal injection of TS, I and 5 mg/kg, or saline. Newborn guinea pigs receiving only Cy treatment showed a decrease in Terminal Deoxynucleotydyl Transferase (TdT) activity of thymocytes; a decrease in the percentage of E-rosette forming thymocytee and a significant elevation in both spleen and peripheral blood T-lymphocytes. Moreo vet a significant increase of the response to T-mitogens Phytoemagglutinin (PHA) and Concanavalin A (Con A) was obtained. TS administration to the Cy-treated animals induced a further marked decrease of TdT specific activity in the thymocytes and a statistically si gnificant increase in the percentage of E-rosette-forming cells present in the thymus. The percentage of T-lymphocytes in spleen and peripheral blood was significantly reduced when compared to that of Cy-treated animals, with values very close to those found in Cyuntreated controls. Similarly TS reduced the reactivity of spleen cells to PHA and Con A previously increased by Cy administration. These studies indicate that in vivo TS admini stration to newborn guinea pigs is effective in stimulating the recovery of the T-lympho~ cytes functions previously depressed by Cy administration. HEPATOSIN. G. Renoux~ M. Renouxand J.M. Guillaumin. Laboratoire d'Immunologie, Facult~ de M~decine, B.P. 3223, 37032 Tours Cedex, :~ance.
Hepatosln is a heat-resistant, endotoxin-free peptide (M.W. circa 5 Kd) synthetized by liver cells, and particularly when cocultured wi~h sodium diethyldithiocarbamate, DTC. In vitro, pgs of Hepatosin induced 18 to 25 % Thy-1 + cells from nu/nu spleen cells in a short-term induction assays, whereas even 10 ~g left unchange~d B cell numbers in a 19-hr incubation assay. Hepatoein was equally able to recruit T cells from bone-marrow cells, and even in nude mice. Similar results were obtained on human cells, evidencing Eepatosin was active across the species barrier. Propanolol did not modify these effects, evidencing Hepatosin is not an ubiquitin-like product. In vivo, 0.1 to 10 ~g Eepatosin induced in n_~/nu mice the recruitment o£ functicnally active T cells, and stimulated to augmented responses C57BI/6 mice, muresponsive to most immunomodulators. Administration of C.I ~g to NZB mice reduced spleen weight and the I~M primary antibody response to E~RBC and increased the specific IgG-response. Present data suggest that Hepatosin might be the already hypothetized pre-th~ic event which allows thymic hormones to further induce preco~mitted cells to further differentiation.