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Interleukin-2 (IL-2) stimulates the in vivo reconstitution of natural killer (NK) activity by bone marrow cells
360
IN VITRO MIGRATION OF HUMAN LARGE GRANULAR LYMPHOCYTES: MODULATION BY LIMPHOKINES AND A
18
STREPTOCOCCAL PREPARATION. N. Polentarutti, B. Bottazzi, S. Rossini, F. Zanaboni and A. Mantovani Istituto di Ricerche Farmacologiche "Mario Negri", Milan, Italy Migration of large granular lymphocytes (LGL)-enriched peripheral blood mononuclear cells was studied using modified Boyden chambers and nitrocellulose filters (Bottazzi et al., J. Immunol., in press). Migratory capacity in response to casein or activated serum correlated with the frequency of LGL in the various fraction of Percoll gradients. Responsive cells were B73.1 + and OKT3-. Preincubation of LGL with [Interferon (IFN), recombinant Interleukin 2 (IL-2) and the streptococcal preparation OK432 enhanced the spontaneous motility of LGL. The responsiveness to chemoattractans was not increased. Small lymphocytes were not affected by preincubation with such stimuli. ~IFN, IL-2 and 0K432 added in the lower compartment of the chemotaxis chamber did not affect the migration into filters of LGL. The prompt migration of LGL in response to various stimuli would suggest that this population represents one of the first lines of resistance against noxious agents that can be modulated by biological response modifiers.
INTERLEUKIN-2 (IL-2) STI},~LATES THE IN VIV0 RECONSTITUTION OF NATURAL KILLER (NK) ACTIVITY BY BONE MARROW CELLS.
19
C. Riccardi, A. Oiampietri, L. Cannarile, E. Ayroldi and O. Migliorati. Institute of pharmacology, University of Perugia, 06100 Perugia, Italy. We have studied the effect of in vivo inoculation of IL-2 on the generation of splenic NK activity by bone marrow (BM~ progenitors. B6D2FI mice were lethally irradiated (950 rads) and grafted i.v. with 10xlO- anti-Thy.l plus complement-treated syngeneic BM cells 5-8 hrs after irradiation (IRR). The splenic NK activity was then evaluated at 4, 7, 9, II, 12 and 14 days after IRR and BM graft. Our data, exspressed as lytic units/spleen clearly show that after a dramatic decline on day 4th, NK activity is progressively reconstituted reaching the levels of untreated controls on day IIth. Treatment with IL-2 (10u/day ip) from day 0 trough day 3 after !RR and BM graft resulted in a premature reconstitution so that the NK activity of IL-2-treated mice was already measurable on day 7th and reached levels of reactivity on day 9th and llth significantly higher than that of medium-injected controls. These data demonstrate that IL-2 is able to induce in vivo the maturation of BM precursors of NK cells. This work was supported by "Progetto Finalizzato 0ncologia"~ Contract N. 84.00762.44, CNR Rome, Italy COLONY
STIMULATING
PACTOR'CSF)
P~ODUCED
BY NURINB
TUMOR
CELLS.
A.Pessina,M.G. Neri and A . M u s c h i a t o . I n s t i t u t e of M e d i c a l M i c r o b i o l o g y , U n i v e r s i t y of M i l a n , I T A L Y . S p o n t a n e o u s C S F p r o d u c t i o n b y m a n y tumors c u l t u r e d "in v i t r o " is \yell c o r r e f a t e d ,~.,ith the c h a n g e s o b s e r v e d "in v i v o " (e.g. : p e r i P h e r a l l e u k o c y t o s i s , i n c r e a s e of the GM-C.FU in the b o n e marro~I and in the spleen).Ho'~'ever,tumors c a u s i n g "in v i v o " s i m i l a r c h a n g e s , d o not p r o d u c e " i n v i t r o " sicn'.~icant q u a n t i t i e s o ~ CgF. W e c o m p a r e d the "in v i t r o " pi-o~ucti-~< o ~ ~ Y Ly a T - ~ e ' l l>nr,p h o m a ( E L 4 ) and a adre~1~-cortical t u m o r cell li!le[Y1) by t e s t i n 9 the C o l o ny S t ! m u l a t i n g A c t i v i t y ( C S A ) o~ t}~eir C o n d i t i o n e d Media(C}.~) on m u r i n e bone marro~f cells c l o n e d in soft a g a r We o b s e r v e d s s p o n t a n e o u s , h i g h level of C S A in the C M F r o m YI c e l l s ( Y I - C M ) ~sh~le no s i g n i f i c a n t C S A ~las F o u n d in the CI4 F r o m E L 4 c u l t ~ s . H o w e v e r , t h e s t i m u l a t i o n of E L 4 c e l l s '~rith PNA goa d e d the c e l l s to p r o d u c e h i g h a m o u n t s of CSP. Y I - C M and E L 4 - C N ( ~ r o m PHA s t i m u l a t e d c e l l s ) ,~Tere p r o c e s s e d by ~e] c h r o m a t o g r a p h y and the C S A of e l u ted Fractions s h o w e d a pea]< c o r r e s p o n d i n g to a M . W . o£ 2 0 - 2 5 , 0 0 0 for E L 4 C M and t,:,,o p e a k s ( n e a r to a M.W. o£ ! 0 0 , 0 0 0 and 2 9 , 0 0 0 ) for YI-CM.
20
IN VITRO MIGRATION OF HUMAN LARGE GRANULAR LYMPHOCYTES: MODULATION BY LIMPHOKINES AND A
18
STREPTOCOCCAL PREPARATION. N. Polentarutti, B. Bottazzi, S. Rossini, F. Zanaboni and A. Mantovani Istituto di Ricerche Farmacologiche "Mario Negri", Milan, Italy Migration of large granular lymphocytes (LGL)-enriched peripheral blood mononuclear cells was studied using modified Boyden chambers and nitrocellulose filters (Bottazzi et al., J. Immunol., in press). Migratory capacity in response to casein or activated serum correlated with the frequency of LGL in the various fraction of Percoll gradients. Responsive cells were B73.1 + and OKT3-. Preincubation of LGL with [Interferon (IFN), recombinant Interleukin 2 (IL-2) and the streptococcal preparation OK432 enhanced the spontaneous motility of LGL. The responsiveness to chemoattractans was not increased. Small lymphocytes were not affected by preincubation with such stimuli. ~IFN, IL-2 and 0K432 added in the lower compartment of the chemotaxis chamber did not affect the migration into filters of LGL. The prompt migration of LGL in response to various stimuli would suggest that this population represents one of the first lines of resistance against noxious agents that can be modulated by biological response modifiers.
INTERLEUKIN-2 (IL-2) STI},~LATES THE IN VIV0 RECONSTITUTION OF NATURAL KILLER (NK) ACTIVITY BY BONE MARROW CELLS.
19
C. Riccardi, A. Oiampietri, L. Cannarile, E. Ayroldi and O. Migliorati. Institute of pharmacology, University of Perugia, 06100 Perugia, Italy. We have studied the effect of in vivo inoculation of IL-2 on the generation of splenic NK activity by bone marrow (BM~ progenitors. B6D2FI mice were lethally irradiated (950 rads) and grafted i.v. with 10xlO- anti-Thy.l plus complement-treated syngeneic BM cells 5-8 hrs after irradiation (IRR). The splenic NK activity was then evaluated at 4, 7, 9, II, 12 and 14 days after IRR and BM graft. Our data, exspressed as lytic units/spleen clearly show that after a dramatic decline on day 4th, NK activity is progressively reconstituted reaching the levels of untreated controls on day IIth. Treatment with IL-2 (10u/day ip) from day 0 trough day 3 after !RR and BM graft resulted in a premature reconstitution so that the NK activity of IL-2-treated mice was already measurable on day 7th and reached levels of reactivity on day 9th and llth significantly higher than that of medium-injected controls. These data demonstrate that IL-2 is able to induce in vivo the maturation of BM precursors of NK cells. This work was supported by "Progetto Finalizzato 0ncologia"~ Contract N. 84.00762.44, CNR Rome, Italy COLONY
STIMULATING
PACTOR'CSF)
P~ODUCED
BY NURINB
TUMOR
CELLS.
A.Pessina,M.G. Neri and A . M u s c h i a t o . I n s t i t u t e of M e d i c a l M i c r o b i o l o g y , U n i v e r s i t y of M i l a n , I T A L Y . S p o n t a n e o u s C S F p r o d u c t i o n b y m a n y tumors c u l t u r e d "in v i t r o " is \yell c o r r e f a t e d ,~.,ith the c h a n g e s o b s e r v e d "in v i v o " (e.g. : p e r i P h e r a l l e u k o c y t o s i s , i n c r e a s e of the GM-C.FU in the b o n e marro~I and in the spleen).Ho'~'ever,tumors c a u s i n g "in v i v o " s i m i l a r c h a n g e s , d o not p r o d u c e " i n v i t r o " sicn'.~icant q u a n t i t i e s o ~ CgF. W e c o m p a r e d the "in v i t r o " pi-o~ucti-~< o ~ ~ Y Ly a T - ~ e ' l l>nr,p h o m a ( E L 4 ) and a adre~1~-cortical t u m o r cell li!le[Y1) by t e s t i n 9 the C o l o ny S t ! m u l a t i n g A c t i v i t y ( C S A ) o~ t}~eir C o n d i t i o n e d Media(C}.~) on m u r i n e bone marro~f cells c l o n e d in soft a g a r We o b s e r v e d s s p o n t a n e o u s , h i g h level of C S A in the C M F r o m YI c e l l s ( Y I - C M ) ~sh~le no s i g n i f i c a n t C S A ~las F o u n d in the CI4 F r o m E L 4 c u l t ~ s . H o w e v e r , t h e s t i m u l a t i o n of E L 4 c e l l s '~rith PNA goa d e d the c e l l s to p r o d u c e h i g h a m o u n t s of CSP. Y I - C M and E L 4 - C N ( ~ r o m PHA s t i m u l a t e d c e l l s ) ,~Tere p r o c e s s e d by ~e] c h r o m a t o g r a p h y and the C S A of e l u ted Fractions s h o w e d a pea]< c o r r e s p o n d i n g to a M . W . o£ 2 0 - 2 5 , 0 0 0 for E L 4 C M and t,:,,o p e a k s ( n e a r to a M.W. o£ ! 0 0 , 0 0 0 and 2 9 , 0 0 0 ) for YI-CM.
20