- Email: [email protected]
APO-1) int he recovery phase of experimental autoimmune encephalomyelitis
P o s t e r A b s t r a c t s / J o u r n a l o f N e u r o i m m u n o l o g y 9 0 (1998) 1 3 - 1 0 5
335 Differential Progression of CNS Pathology in Two Clinically Distinct Models o f M S W. Smith Begolka, C.L. Vanderlugt, S.M. Rahbe, S.D. Miller, Northwestern University, USA
61
338 A Role for H u m o r a l F a c t o r s i n D e m y e l i n a t i o n a n d Disease in E x p e r i m e n t a l A u t o i m m u n e N e u r i t i s in t h e Lewis R a t J. Taylgr, J. Pollard, Univ.of Sydney,Australia
Theiler's virus-induced demyelinating disease (TMEV-IDD) and relapsingre-,itting experimental autoimmune encephalomyelitis (R-EAE) are two clinically distinct models of multiple sclerosis. Our data suggests that the kinetics of cytoklne expression together with the nature of the persistent inflammatory infiltrate within the CNS contribute to the observed differences in clinical course. We show that in TMEV-infected SJL mice, mRNAs for IFN-y, TNF-a, IL-10 and IL-4 were detectable during the pre-clinical phase, increased throughout the time course, and correlated with a rich CD4 * and F4/80" infiltrate during disease. CNS samples from R-EAE-mice displayed a pattern for IFN-y and TNF-a that peaked during the acute phase and relapse(s), a pattern that also correlated with dramatic fluctuations in the CD4 + and F4/80 ÷ populations during disease progression. Further support for the role of IFN-7 and TNF-¢ in disease induction/progression was demonstrated by the lack these eytokines from the CNS of mice tolerized subsequent to disease priming. Lastly, IL-4 mRNA was undeteetable until remission(s), demonstrating a possible role of this cytokine in down-regulating ongoing disease, whereas IL-10 expression was identical to that ~een in TMEV-IDD, arguing against a regulatory role in either model.
Objective: to clarify the role of Immoral factors in demyelination and disease in experimental autoimmune neuritis (EAN) in the Lewis rat. Methodology: 3 different adoptive transfer models are being used: (i) serum from EAN rats is being injected directly into the sciatic nerve; (ii) EAN serum is being co-transfered with myelin sensitised LNC from EAN rats to empirically ioeasure relative contribution of cellular and humeral immunity to disease and demyelination in EAN: (iii) serum from EAN rats is being co-tmnsferred with a mild clinical dose of P2 T-cells to determine if humeral factors will augment demycliuation and disease. Antibodies to purified bovine P2, P0, PMP-22. SAG and two novel peripheral myelin proteins of 40kDa & 60kDa MW are being measured in actively induced EAN by ELISA to clarify the relationship of these antibodies to disease onset and course. Results: serum from EAN rats collected on day 45 post immunisation can augment and prolong disease in AT-EAN with P2 T-ceils. Conclusion: preliminary, results indicate that humeral factors do contribute to disease in Lewis rat EAN. Experiments described in the methods are being conducted to further assess these results.
336 Inhibition of Nitric Oxide Production Renders PVG Rats Highly Susceptible to EAE
339
W. Cowden, t:. Cullen, JCSMI~,Canberra,Australia, M. Staykovo, D. Willenborg, The CanberraHospital,Australia
PVG rats, which have a very low susceptibility to experimental autoimmune encephalomyelitis (EAE), were rendered highly susceptible to clinical disease when treated with N-methylarginine (NMA), an inhibitor of nitric oxide synthase (NOS). Following MBP-CFA, PVG rats developed higher serum levels of the surrogate markers of nitric oxide production, nitrite and nitrate (RNI), than did their Lewis counterparts. In vitro, the levels of RNI produced in IFN-y-stimulated spleen cell cultures for PVG rats were significantly higher than those for Lewis rats. A mechanism by which increased NO production might protect PVG rats against clinical EAE was suggested by the finding that lymph node cells, isolated from NMA-treated MBP-immunised PVG rats, proliferated in response to MBP at a rate approximately three times greater than those from MBPimmunised, saline treated rats. Thus, the greater number of MBP-specific T cells generated in the NOS inhibitor-treated versus untreated rats could account for their increased susceptibility to developing EAE. The findings in this study suggest that NO plays a role in protecting PVG rats against developing EAE.
Effective P r e v e n t i o n a n d T h e r a p y o f E x p e r i m e n t a l Allergic Encephalomyelitis b y M y e o p h e n o l a t e G. Trail. N. Carter, S. Hodgkinsoll, Universityof New South Wales,Australia
Myeophecolate Mofetil (MM) is a derivative of Mycophenolie acid which inhibits inosine monophosphate dehydrogenase, an essential enzyme for purina synthesis in lymphocytes, but not other cells. MM is an effective and safe drug in transplantation. We examined the capacity of MM to prevent EAE, the animal model that resembles multiple sclerosis (MS). Oral treatment of MM {30mg/kg/day) was given from day 0-14 in Lewis tats immunized with MBP in CFA. Control animals were given the MM vehicle dally in the same volume. Animals were scored daily. Representativeanimals were sampled for FACS analysis, immunohiatology and auto-antibody analysis. The control group had a mean incidence of disease of 100% with a mean clinical score of 2.2 compared to the treated group (0-14) with a disease incidenee of15% and a mean score of O.l p< O.O01). This wea accompanied by a elgnifieant reduction in auto-antibody titers. No infiltration of T-cells or macrophages was observed on immunostaielng of brain stem sections in treated animals but an abundant infiltrate was observed in sham controls. FACS analysis of splenocytas and lymphocytes showed no changes in T and B-cell poputations. Animals given MM (30mg/kg/day) at onset of disease onset at 12 days post immunization, had accelerated
recovery and leas severe peak of disease compared to stiam treated controls, with a mean clinical score of 1.5 (p
337
340
A Potential Role for FAS ( C P 9 5 / A P O - 1 ) in the R e c o v e r y P h a s e of E x p e r i m e n t a l A u t o i m m u n e Encephalomyelitis G.C. Suvannaveih. S.D. Miller, Northwestern UniversityMedical School, USA
A n t i b o d i e s M e d i a t e C e r e b r a l G r a f t Versus H o s t Disease C. Solaro, E. Capello, M. CiUi, U. Curdellino, D. Giuati, L. Roccatagliata, A. Bacigalupo, G. Abbruzzese, G.L. Manuardi, A. Uccelli, Universityof Geneva, Italy
EAE is a CD4 ÷ Thl inflammatory disease model for multiple sclerosis (MS). SJL mice that are induced with EAE often exhibit a relapsing-remitting disease course, one that is similar to that seen in MS patients. While many of the salient features of this model have been elucidated, the exact meehanisms of disease remission in these animals remain poorly understood. Fas is a member of the TNF receptor family and has been shown to be a critical mediator of cellular apoptosis and activation-induced cell death (AICD). One possibility to account for the spontaneous remission is that A1CD of disease-initiating T cells occurs. To investigate this possibility, and in particular, the potential role of Fas, Fasdeficient SJL mice were immunized with PLPI39-151, the immanodominant epitope on the PLP protein. Whereas control Fas +/+ wild-type SJL and Fas +/mice exhibited the characteristic relapsing-remitting disease course with each animal entering remission following the acute phase of disease, a significant number of sick Fas 4- never entered remission. In fact, these Fas -/- animals never recovered, as they eventually suecambed to the disease. In comparison, no Fas +/- or Fas +/+ mice died. These results suggest that Fas/FasL interactions may be critical in regulating events following the acute phase of disease and may represent an intrinsic regulatory mechanism for the remission from disease that is observed in normal SJL mice. (Supported by NIH grant NS-30871-04)
Graft versus host disease (GVHD) frequently follows heterologous bone marrow lransplantation (BMT) and is mediated by T-cells and antibodies targeting self-antigens. Because of the presence of the blood-brain-barrier (BBB), tim cerebral nervous system is rarely involved by GVHD. A 24 years old male (PA), who underwent BMT for aplastic anemia, developed generalized intractable myoclonus after the occurrence of head trauma two years later. Based on file presence of a single clonal band of lgG in the CSF, pulses of cyclophosphamide together with plasma-exchange were performed with tempontry recovery. To demonstrate that ~mtibodies isolated from this patient might be encephalitogenic, 15 Spregue Dowley rats were immunized using either PA's purified IgG, PA's CSF or CSF from patients with other neurological diseases, administered iotra-carotid following a pulse with 25% mannitol to open the BBB. Four out of 5 animals anmtmized with PA's CSF developed involan 'tury movements within 24 hours. Pathology of these rats showed early meningeal inflammation and mononuclear infiltrates within the basal ganglia. Animals from the other two groups did not develop clinical signs and pathology was negative. These findings support the hypothesis that clonal antilxxlies within the CNS mediates a myoclonic form of GVHD.
335 Differential Progression of CNS Pathology in Two Clinically Distinct Models o f M S W. Smith Begolka, C.L. Vanderlugt, S.M. Rahbe, S.D. Miller, Northwestern University, USA
61
338 A Role for H u m o r a l F a c t o r s i n D e m y e l i n a t i o n a n d Disease in E x p e r i m e n t a l A u t o i m m u n e N e u r i t i s in t h e Lewis R a t J. Taylgr, J. Pollard, Univ.of Sydney,Australia
Theiler's virus-induced demyelinating disease (TMEV-IDD) and relapsingre-,itting experimental autoimmune encephalomyelitis (R-EAE) are two clinically distinct models of multiple sclerosis. Our data suggests that the kinetics of cytoklne expression together with the nature of the persistent inflammatory infiltrate within the CNS contribute to the observed differences in clinical course. We show that in TMEV-infected SJL mice, mRNAs for IFN-y, TNF-a, IL-10 and IL-4 were detectable during the pre-clinical phase, increased throughout the time course, and correlated with a rich CD4 * and F4/80" infiltrate during disease. CNS samples from R-EAE-mice displayed a pattern for IFN-y and TNF-a that peaked during the acute phase and relapse(s), a pattern that also correlated with dramatic fluctuations in the CD4 + and F4/80 ÷ populations during disease progression. Further support for the role of IFN-7 and TNF-¢ in disease induction/progression was demonstrated by the lack these eytokines from the CNS of mice tolerized subsequent to disease priming. Lastly, IL-4 mRNA was undeteetable until remission(s), demonstrating a possible role of this cytokine in down-regulating ongoing disease, whereas IL-10 expression was identical to that ~een in TMEV-IDD, arguing against a regulatory role in either model.
Objective: to clarify the role of Immoral factors in demyelination and disease in experimental autoimmune neuritis (EAN) in the Lewis rat. Methodology: 3 different adoptive transfer models are being used: (i) serum from EAN rats is being injected directly into the sciatic nerve; (ii) EAN serum is being co-transfered with myelin sensitised LNC from EAN rats to empirically ioeasure relative contribution of cellular and humeral immunity to disease and demyelination in EAN: (iii) serum from EAN rats is being co-tmnsferred with a mild clinical dose of P2 T-cells to determine if humeral factors will augment demycliuation and disease. Antibodies to purified bovine P2, P0, PMP-22. SAG and two novel peripheral myelin proteins of 40kDa & 60kDa MW are being measured in actively induced EAN by ELISA to clarify the relationship of these antibodies to disease onset and course. Results: serum from EAN rats collected on day 45 post immunisation can augment and prolong disease in AT-EAN with P2 T-ceils. Conclusion: preliminary, results indicate that humeral factors do contribute to disease in Lewis rat EAN. Experiments described in the methods are being conducted to further assess these results.
336 Inhibition of Nitric Oxide Production Renders PVG Rats Highly Susceptible to EAE
339
W. Cowden, t:. Cullen, JCSMI~,Canberra,Australia, M. Staykovo, D. Willenborg, The CanberraHospital,Australia
PVG rats, which have a very low susceptibility to experimental autoimmune encephalomyelitis (EAE), were rendered highly susceptible to clinical disease when treated with N-methylarginine (NMA), an inhibitor of nitric oxide synthase (NOS). Following MBP-CFA, PVG rats developed higher serum levels of the surrogate markers of nitric oxide production, nitrite and nitrate (RNI), than did their Lewis counterparts. In vitro, the levels of RNI produced in IFN-y-stimulated spleen cell cultures for PVG rats were significantly higher than those for Lewis rats. A mechanism by which increased NO production might protect PVG rats against clinical EAE was suggested by the finding that lymph node cells, isolated from NMA-treated MBP-immunised PVG rats, proliferated in response to MBP at a rate approximately three times greater than those from MBPimmunised, saline treated rats. Thus, the greater number of MBP-specific T cells generated in the NOS inhibitor-treated versus untreated rats could account for their increased susceptibility to developing EAE. The findings in this study suggest that NO plays a role in protecting PVG rats against developing EAE.
Effective P r e v e n t i o n a n d T h e r a p y o f E x p e r i m e n t a l Allergic Encephalomyelitis b y M y e o p h e n o l a t e G. Trail. N. Carter, S. Hodgkinsoll, Universityof New South Wales,Australia
Myeophecolate Mofetil (MM) is a derivative of Mycophenolie acid which inhibits inosine monophosphate dehydrogenase, an essential enzyme for purina synthesis in lymphocytes, but not other cells. MM is an effective and safe drug in transplantation. We examined the capacity of MM to prevent EAE, the animal model that resembles multiple sclerosis (MS). Oral treatment of MM {30mg/kg/day) was given from day 0-14 in Lewis tats immunized with MBP in CFA. Control animals were given the MM vehicle dally in the same volume. Animals were scored daily. Representativeanimals were sampled for FACS analysis, immunohiatology and auto-antibody analysis. The control group had a mean incidence of disease of 100% with a mean clinical score of 2.2 compared to the treated group (0-14) with a disease incidenee of15% and a mean score of O.l p< O.O01). This wea accompanied by a elgnifieant reduction in auto-antibody titers. No infiltration of T-cells or macrophages was observed on immunostaielng of brain stem sections in treated animals but an abundant infiltrate was observed in sham controls. FACS analysis of splenocytas and lymphocytes showed no changes in T and B-cell poputations. Animals given MM (30mg/kg/day) at onset of disease onset at 12 days post immunization, had accelerated
recovery and leas severe peak of disease compared to stiam treated controls, with a mean clinical score of 1.5 (p
337
340
A Potential Role for FAS ( C P 9 5 / A P O - 1 ) in the R e c o v e r y P h a s e of E x p e r i m e n t a l A u t o i m m u n e Encephalomyelitis G.C. Suvannaveih. S.D. Miller, Northwestern UniversityMedical School, USA
A n t i b o d i e s M e d i a t e C e r e b r a l G r a f t Versus H o s t Disease C. Solaro, E. Capello, M. CiUi, U. Curdellino, D. Giuati, L. Roccatagliata, A. Bacigalupo, G. Abbruzzese, G.L. Manuardi, A. Uccelli, Universityof Geneva, Italy
EAE is a CD4 ÷ Thl inflammatory disease model for multiple sclerosis (MS). SJL mice that are induced with EAE often exhibit a relapsing-remitting disease course, one that is similar to that seen in MS patients. While many of the salient features of this model have been elucidated, the exact meehanisms of disease remission in these animals remain poorly understood. Fas is a member of the TNF receptor family and has been shown to be a critical mediator of cellular apoptosis and activation-induced cell death (AICD). One possibility to account for the spontaneous remission is that A1CD of disease-initiating T cells occurs. To investigate this possibility, and in particular, the potential role of Fas, Fasdeficient SJL mice were immunized with PLPI39-151, the immanodominant epitope on the PLP protein. Whereas control Fas +/+ wild-type SJL and Fas +/mice exhibited the characteristic relapsing-remitting disease course with each animal entering remission following the acute phase of disease, a significant number of sick Fas 4- never entered remission. In fact, these Fas -/- animals never recovered, as they eventually suecambed to the disease. In comparison, no Fas +/- or Fas +/+ mice died. These results suggest that Fas/FasL interactions may be critical in regulating events following the acute phase of disease and may represent an intrinsic regulatory mechanism for the remission from disease that is observed in normal SJL mice. (Supported by NIH grant NS-30871-04)
Graft versus host disease (GVHD) frequently follows heterologous bone marrow lransplantation (BMT) and is mediated by T-cells and antibodies targeting self-antigens. Because of the presence of the blood-brain-barrier (BBB), tim cerebral nervous system is rarely involved by GVHD. A 24 years old male (PA), who underwent BMT for aplastic anemia, developed generalized intractable myoclonus after the occurrence of head trauma two years later. Based on file presence of a single clonal band of lgG in the CSF, pulses of cyclophosphamide together with plasma-exchange were performed with tempontry recovery. To demonstrate that ~mtibodies isolated from this patient might be encephalitogenic, 15 Spregue Dowley rats were immunized using either PA's purified IgG, PA's CSF or CSF from patients with other neurological diseases, administered iotra-carotid following a pulse with 25% mannitol to open the BBB. Four out of 5 animals anmtmized with PA's CSF developed involan 'tury movements within 24 hours. Pathology of these rats showed early meningeal inflammation and mononuclear infiltrates within the basal ganglia. Animals from the other two groups did not develop clinical signs and pathology was negative. These findings support the hypothesis that clonal antilxxlies within the CNS mediates a myoclonic form of GVHD.