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Inhibition of experimental autoimmune encephalomyelitis (EAE) in Lewis rats by SK&F 86002—A new anti-inflammatory compound
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LOW-DOSE CYCLOSPORIN A INDUCES RELAPSING EAE IN THE LEWIS RAT C.D. Dijkstra, C.J.A. de Groot, J.C. Koetsier, I. Matthaei, C.H. Polman and T. S~inia Departments of Histology and Neurology, Free University, Amsterdam, The Netherlands Workshop topic: Immunotherapy and autoimmune diseases It is generally recognized that Cyclosporin A (CsA) in a sufficiently high dose (for example 25 mg/kg daily) can inhibit the development of experimental allergic encephalomyelitis (EAE) in Lewis rats (for review see Nussenblat et al. Proc. Allergy 1986, 38:167-172). Since preliminary data indicate that the clinical usefullness of the drug in these doses is limited because of its nephrotoxicity, we decided to study the effect of CsA in much lower doses on actively induced EAE in a group of 30 Lewis rats. To our surprise in all animals prophylactically treated with CsA (by subcutaneous injection of 2 mg/kg, 3 times a week for 3 weeks) a severe clinical relapse occurred about one week after recovery from the first attack. In most anlmal~ even a secon~ (and third) re]apsP was seen b~fore sner~f~r~rion at 50 days post-immunization, relapses in general being clinically severe and long lasting. This exceedingly high relapse rate significantly differs from that in the control group (no relapses at all) and from that normally expected in Lewis rats (McFarlin et al. J. Immunol. 1974, |]3:712-715). Since the mechanism responsible for this observation is not clear yet, further analysis of the effects of immunomodulating treatment in EAE is urgently needed. Send correspondence to: Dr. C.D. Dijkstra Department of Histology Medical Faculty Free University P.O. Box 7161 1007 MC Amsterdam The Netherlands.
Inhibition of Experimental Autoimmune Encephalomyelitis (EAE) in Lewis Rats bv SK&F 86002 - A New Anti-lnflammatory Compound. M.J. DiMartino, C.E. Wolff, G. Campbell and N. Hanna, Smith Kline and French Laboratories, Swedeland, PA.
Experimental Autoimmune Encephalomyelitis, is an immune-mediated inflammatory disease which can be prevented by immunosuppressive agents (e.g., methotrexate) or corticosteroids but not by currently used non-steroidal anti-inflammatory drugs (e.g., indomethacin).
SK&F86002
[6-(4-fluorophenyl)2,3-Dihydro-5(4-Pyridinvl)imidazo(2,1-b)thiazole] is a dual cvclooxvgenase and lipoxygenase i n h i b i t o r which is effective in inhibiting immune and non immune-mediatedinflammatory responses (Agents and Actions, 20: 113, 1987),
In the present study we evaluated SK&F 86002 for i t s a b i l i t y to
45 i n h i b i t EAE induced in female Lewis rats.
Daily oral administration of SK&F
86D02, i n i t i a t e d during the induction period of EAE, produced a dose related i n h i b i t i o n in the incidence of hindleg paralysis (ED50 ~ 20 mg/Kg/dav). complete i n h i b i t i o n was achieved at 60 mg/Kg/day.
A
In contrast, i ndomethacin
administered at high anti-inflammatory dose levels (2 mg/Kg/day, p.o.) did not affect the incidence or duration of hindleg paralysis.
Moreover, SK&F 86002
administered to rats with established EAE at a dose of 60 mg/Kg/day, p.o. completely suppressed relapsing hindleg paralysis.
Similar effects were
produced by methotrexate (0.3 mg/kg/day, p.o.) and prednisolone (20 mg/kg/day, p.o.); whereas, indomethacin ( 2 mg/kg/day, p.o.} either had no effect or enhanced the incidence of relapsing hindleg paralvsis.
These results suggest
that dual i n h i b i t o r s of cyclooxygenase and lipoxygenase enzymes, such as SK&F B6002, exhibit a steroid-liKe pharmacological p r o f i l e , and by i n h i b i t i n g immune-mediated inflammatory responses may provide effective therapy for chronic inflammatory diseases with autoimmune etiology.
EXPRESSION AND MODULATION OF TUMOR-ASSOCIATED HUMAN GLIOMA CELL CULTURES B. Dipasquale, M. Colombatti, M. Gerosa *, G. Tridente.
P. Lorenzi,
ANTIGENS
G. Stevanoni,
Institute of Immunological Sciences and * Department University of Verona, 37134 Verona, Italy. TOPIC:
NEURAL CELL MARKERS
IN
P. Germano,
of Neurosurgery,
IN HEALTH AND DISEASE.
We have studied the expression of tumor-associated antigens by im~unocytochemistry (ICC) and flow cytometry (FACS analysis) in six cell lines derived from human glioblastoma multiforme that have been established in our laboratory. A wide morphologic eterogeneity between these lines was observed, with large plurinucleated cells and small round cells coexisting in the same culture. The proliferation-associated antigen Ki-67 was detected in the plurinucleated cell population of 2 cell lines, whereas the same cells of 2 other lines were clearly negative; nearly 80% of small round cells were Ki-67 positive. Two glioma-associated antigens (GAA) defined by MoAbs GE2 and CG12 were expressed in a highly eterogeneous mannier as far as % of positive cells and cell surface Ag density (FACS analysis). GAA were instead expressed intracellularly in all cell lines tested (ICC and FACS analysis). GAA were drastically reduced under conditions that reduced ceilular proliferation (treatment with phorbol esthers, high cell
LOW-DOSE CYCLOSPORIN A INDUCES RELAPSING EAE IN THE LEWIS RAT C.D. Dijkstra, C.J.A. de Groot, J.C. Koetsier, I. Matthaei, C.H. Polman and T. S~inia Departments of Histology and Neurology, Free University, Amsterdam, The Netherlands Workshop topic: Immunotherapy and autoimmune diseases It is generally recognized that Cyclosporin A (CsA) in a sufficiently high dose (for example 25 mg/kg daily) can inhibit the development of experimental allergic encephalomyelitis (EAE) in Lewis rats (for review see Nussenblat et al. Proc. Allergy 1986, 38:167-172). Since preliminary data indicate that the clinical usefullness of the drug in these doses is limited because of its nephrotoxicity, we decided to study the effect of CsA in much lower doses on actively induced EAE in a group of 30 Lewis rats. To our surprise in all animals prophylactically treated with CsA (by subcutaneous injection of 2 mg/kg, 3 times a week for 3 weeks) a severe clinical relapse occurred about one week after recovery from the first attack. In most anlmal~ even a secon~ (and third) re]apsP was seen b~fore sner~f~r~rion at 50 days post-immunization, relapses in general being clinically severe and long lasting. This exceedingly high relapse rate significantly differs from that in the control group (no relapses at all) and from that normally expected in Lewis rats (McFarlin et al. J. Immunol. 1974, |]3:712-715). Since the mechanism responsible for this observation is not clear yet, further analysis of the effects of immunomodulating treatment in EAE is urgently needed. Send correspondence to: Dr. C.D. Dijkstra Department of Histology Medical Faculty Free University P.O. Box 7161 1007 MC Amsterdam The Netherlands.
Inhibition of Experimental Autoimmune Encephalomyelitis (EAE) in Lewis Rats bv SK&F 86002 - A New Anti-lnflammatory Compound. M.J. DiMartino, C.E. Wolff, G. Campbell and N. Hanna, Smith Kline and French Laboratories, Swedeland, PA.
Experimental Autoimmune Encephalomyelitis, is an immune-mediated inflammatory disease which can be prevented by immunosuppressive agents (e.g., methotrexate) or corticosteroids but not by currently used non-steroidal anti-inflammatory drugs (e.g., indomethacin).
SK&F86002
[6-(4-fluorophenyl)2,3-Dihydro-5(4-Pyridinvl)imidazo(2,1-b)thiazole] is a dual cvclooxvgenase and lipoxygenase i n h i b i t o r which is effective in inhibiting immune and non immune-mediatedinflammatory responses (Agents and Actions, 20: 113, 1987),
In the present study we evaluated SK&F 86002 for i t s a b i l i t y to
45 i n h i b i t EAE induced in female Lewis rats.
Daily oral administration of SK&F
86D02, i n i t i a t e d during the induction period of EAE, produced a dose related i n h i b i t i o n in the incidence of hindleg paralysis (ED50 ~ 20 mg/Kg/dav). complete i n h i b i t i o n was achieved at 60 mg/Kg/day.
A
In contrast, i ndomethacin
administered at high anti-inflammatory dose levels (2 mg/Kg/day, p.o.) did not affect the incidence or duration of hindleg paralysis.
Moreover, SK&F 86002
administered to rats with established EAE at a dose of 60 mg/Kg/day, p.o. completely suppressed relapsing hindleg paralysis.
Similar effects were
produced by methotrexate (0.3 mg/kg/day, p.o.) and prednisolone (20 mg/kg/day, p.o.); whereas, indomethacin ( 2 mg/kg/day, p.o.} either had no effect or enhanced the incidence of relapsing hindleg paralvsis.
These results suggest
that dual i n h i b i t o r s of cyclooxygenase and lipoxygenase enzymes, such as SK&F B6002, exhibit a steroid-liKe pharmacological p r o f i l e , and by i n h i b i t i n g immune-mediated inflammatory responses may provide effective therapy for chronic inflammatory diseases with autoimmune etiology.
EXPRESSION AND MODULATION OF TUMOR-ASSOCIATED HUMAN GLIOMA CELL CULTURES B. Dipasquale, M. Colombatti, M. Gerosa *, G. Tridente.
P. Lorenzi,
ANTIGENS
G. Stevanoni,
Institute of Immunological Sciences and * Department University of Verona, 37134 Verona, Italy. TOPIC:
NEURAL CELL MARKERS
IN
P. Germano,
of Neurosurgery,
IN HEALTH AND DISEASE.
We have studied the expression of tumor-associated antigens by im~unocytochemistry (ICC) and flow cytometry (FACS analysis) in six cell lines derived from human glioblastoma multiforme that have been established in our laboratory. A wide morphologic eterogeneity between these lines was observed, with large plurinucleated cells and small round cells coexisting in the same culture. The proliferation-associated antigen Ki-67 was detected in the plurinucleated cell population of 2 cell lines, whereas the same cells of 2 other lines were clearly negative; nearly 80% of small round cells were Ki-67 positive. Two glioma-associated antigens (GAA) defined by MoAbs GE2 and CG12 were expressed in a highly eterogeneous mannier as far as % of positive cells and cell surface Ag density (FACS analysis). GAA were instead expressed intracellularly in all cell lines tested (ICC and FACS analysis). GAA were drastically reduced under conditions that reduced ceilular proliferation (treatment with phorbol esthers, high cell