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Cytotoxic T lymphocyte-mediated cell death in paraneoplastic sensory neuronopathy with anti-Hu antibody
102
Poster Abstracts / Journal of Neuroimmunology 90 (1998) 13-105
581 Reactive Changes of Thymocytes and Retieuloepithelium during Autotransplantationof the Thymus
584 Systemic lntedeukin-12 Displays Antitumor Activity in the Mouse Central Nervous System
A.A. Stadnikov, O.Y. Nickolaeva. StateMedJcalAcademy, Drenbur~ Russia
K. Shim i ~ , H. Kishim a, Osaka UniversityMedicalSchool, Japan, Y. Miyao, Suita Municipal Hospital, Japan, K. Tamurn, Osaka University Medical School, Japan, M. Tamura, Minoh MunicipaI Hospital, Japan, M. Sasaki, Osaka University Medical School, Japan
Lazarenko's method of the tissues cultivation in rive was used.
It was performed in dO natsre nongrel female rats using histological and experimental methods. On the first stage (I0 rats) surgical approach to aidiastinum with subsequent extraction of the thunus were maked. After the success in thyluS isolation the inplantation pices of the organ with celloidias fragments were performed under skin of the experimental animals (30 rats). Extirpation and investigation of the inplantates were made in 2.4.6.8.10,12 and 20 days, ?ha main morphological changes (nell adaptation and death,processesof proliferation, growth and cytodiffcremtiation,iutercellular relationships, capacity of histo- and organmtypic potency) mere demonstrated. Prolonged survival of the reticulmepithcli0eytes, their active proliferation, new
formation lynphocytes clones were estimated. This method is effective for investigations of the processes of the thinic tissue sad ceils adaptation, reparative regeneration of thymocytes and epitheliocytes, their reorganization and remodelling in the inflamati0n regions,
In various systemic cancers, interleukin-12 (IL-12) induces antatumor immunity mediated by T-lymphocytes and natural killer ceils. To determine whether 1L-12 Ires untitumur activity, against malignant gliomas in the central nervous system (CNS). which is considered as an immunologicully privileged site. we treated mice with meningeal gliomatosis by intraperitoneal (ip.) or intrathecal (i.t.) administration of recombinant murine [L-12. Although untreated mice revealed symptoms such as body weight loss or paraplegia due to the meningeal gliomatosis within 8 days folhiwing tumor inoculation, 80% of the mace treated with IL-12 at 05 u g i.p. were cured. Many lymphocyles. mostly CD4" and CD8" cells, iuliltmted to the tumors of IL-12-treated mice. The nmnbers of these cells increased in the cervical lymph nodes, into which the cerebrospinal fluid drains, and there they secreted a considerable amount of interferon-g. Mice cured by IL-12 rejected subcutaneous or i.t. rechallenge with their original glioma ceils, but the same mice were not able to reject other sy_ngeneic mmor cells. These results indicate that the immune system recognizes malignant glioma cells in the subamchnoid space of the CNS and that s3"stemic IL-12 may produce effective antitumor activity and lung-lasting tumor-specific immunity.
582 Protective Effect of Naked DNA Immunizationin Experimental AutoimmuneEncephalitis
585 Cytotoxic T Lymphocyte-mediatedCell Death in Paraneoplastic Sensory Neuronopathywith Anti-HuAntibody
J, Nowizka, MedicaIAcademy of Lodz, Poland, (2. Kowal, Albert Einstein College of Medicine, New York, USA, A. Walczak, A. Glabinski, A. Jurewicz, K. Selmaj, MedicaIAcademy ofLodz, Poland
K. Tanaka. T. Inuzuka, R. Nakano, S. Tsuji, BrainResearchlnstitute, Ntigata University, Japan, M. Tanaka, DepartmentofNeurology & ClinicaI Research Center, National West,N iigata Central Hospital, Japan
Naked DNA vaccination offers several therapeutical possibilities from inducing to down-regulation of immune responses. We have evaluated the effect of DNA vaccination with pRc-CMV plasmid containing PLP gene in SJL-J mice. Injection of the plasmid construct resulted in PLP gene expression in murine muscles as measured by Western blotting. We next assessed how DNA immunization effect development of EAE induced with PLP. The results have shown that in mice immunized three weeks prior with pRc/CMV-PLP tendency to more safer EAE was observed. However, in mice immunized four months prior and sensitized with PLP, the EAE course was significantly ameliorated as assessed by lower clinical scores and shorter disease duration. The results suggest that naked DNA immunization might have protective effect against EAE.
583 Suppression of M u r i n e
Experimental Autoimmune Encephalomyelitis b y IL-2 T a r g e t e d Fusion Toxin D A B 3 8 9 I L - 2 S.M. Phillins. S.A. Zekavat, M.A. Ramadan All, P.J. Perrin, Univ. ofPenn., USA, P. Bachs, Seragen, Inc., USA
DABjsglL-2 is a diptheria toxin-lL-2 chimeric protein. DAB~sg[L-2 targets activated T cells which express the high affinity IL-2 receptor (CD25) and kills them. Because activated cells are targeted with this strategy, the antigenic T cell epitope does not need to be defined. This is particularly important when considering treatments for diseases such as MS, where an immune component is accepted but the identity of the autoantigen is not known. We therelbre investigated the in vitro and in vivo of DAB3sglL-2 to suppress immunopathotogy in marine EAE. DAB3sglL-2 suppressed IL-2, lectin mitogen, and MBP induced lymphocyte proliferation, CD25 expression, IL-2, and IFN-? production. In vivo, DAB3891L-2 suppressed the development of active and adoptively transferred EAE. DAB3sglL-2 modestly reduced total CD4 +, CD25 + cells at the systemic level and little effect on CD8 + cells. In contrast, DAB3sglL-2 pro|bundly suppressed the accumulation of CD4 +, CD8 ÷ and CD25 + lymphocytes and macrophages within the CNS lesions. These results suggest that DAB389IL-2. with its minimal side effects, may have potential for the genetically independent targeted reduction of immunopathology in human MS.
Paraneoplastic sensory neuronopathy (PSN) has been shown to harbor characteristic anti-neuronal autoantibody "anti-Hu" in their sara and cerebrnspinal fluid. Creation of animal models exhibiting clinical or pathological features seen in PSN by means of passive transfer of anti-Hu positive lgG has not been achieved. Although anti-Hu antibody was shown to induce neuronal cell lysis in vitro, this result has not been reproduced so far. Since prominent T cell infihration is seen in the central nervous system and posterior spinal ganglion oftbe patients with anti-Hu syndrome, we studied cytotoxic T cell (CTL) activity in peripheral mononuclear cells from a patient with PSN harboring anti-Hu antibody. The activated CD8+ T cells from the patient's venous blood were shown to lyse her own fibroblasts which were incubated with interferon-'/to induce HLA class 1 molecules on their surface and the recombinant HuD protein was injected into the cells by mieroinjector. This is the first report showing the existence of CTL activity in a patient with PSN~
586 CytotoxicT CellsAgainsta Peptidefrom Yo Protein in Patients w i t h P a r a n e o p l a s l l c Cerebellar D e g e n e r a t i o n w i t h Anti-Yo A n t i b o d y M. Tanaka, National WestNiigata Centre Hospital, Japan, K. Tanaka, BrainResearch Institute, Niigata University, Japan, S. Tokiguchi, Ojays General Hospital, Japan, K. Shinozawa, Saiseikai-Kaw aguchi GeneralHospital, Japan, S. Tsuji, Brain Reaearch Institute, Nflgata University, Japan
High anti-Yo antibody titer is present in some patients with paraneoplastic cerebellar degeneration, but there is no evidence that the antibody causes Purkinje cell loss. We examined cytotoxie T cell (CTL) activity against a peptide of Yo protein that was defined by the HLA-based approach called reverse immunogenetics. The peripheral venous blood of three patients showed CTL activity against a peptide of Yo protein combined with autologous fibroblasts as the target after activation with Yo peptide. In paraneoplastic cerebellar degeneration, CTL therefore may be involved in the loss of Purkinje cells that present Yo protein.
Poster Abstracts / Journal of Neuroimmunology 90 (1998) 13-105
581 Reactive Changes of Thymocytes and Retieuloepithelium during Autotransplantationof the Thymus
584 Systemic lntedeukin-12 Displays Antitumor Activity in the Mouse Central Nervous System
A.A. Stadnikov, O.Y. Nickolaeva. StateMedJcalAcademy, Drenbur~ Russia
K. Shim i ~ , H. Kishim a, Osaka UniversityMedicalSchool, Japan, Y. Miyao, Suita Municipal Hospital, Japan, K. Tamurn, Osaka University Medical School, Japan, M. Tamura, Minoh MunicipaI Hospital, Japan, M. Sasaki, Osaka University Medical School, Japan
Lazarenko's method of the tissues cultivation in rive was used.
It was performed in dO natsre nongrel female rats using histological and experimental methods. On the first stage (I0 rats) surgical approach to aidiastinum with subsequent extraction of the thunus were maked. After the success in thyluS isolation the inplantation pices of the organ with celloidias fragments were performed under skin of the experimental animals (30 rats). Extirpation and investigation of the inplantates were made in 2.4.6.8.10,12 and 20 days, ?ha main morphological changes (nell adaptation and death,processesof proliferation, growth and cytodiffcremtiation,iutercellular relationships, capacity of histo- and organmtypic potency) mere demonstrated. Prolonged survival of the reticulmepithcli0eytes, their active proliferation, new
formation lynphocytes clones were estimated. This method is effective for investigations of the processes of the thinic tissue sad ceils adaptation, reparative regeneration of thymocytes and epitheliocytes, their reorganization and remodelling in the inflamati0n regions,
In various systemic cancers, interleukin-12 (IL-12) induces antatumor immunity mediated by T-lymphocytes and natural killer ceils. To determine whether 1L-12 Ires untitumur activity, against malignant gliomas in the central nervous system (CNS). which is considered as an immunologicully privileged site. we treated mice with meningeal gliomatosis by intraperitoneal (ip.) or intrathecal (i.t.) administration of recombinant murine [L-12. Although untreated mice revealed symptoms such as body weight loss or paraplegia due to the meningeal gliomatosis within 8 days folhiwing tumor inoculation, 80% of the mace treated with IL-12 at 05 u g i.p. were cured. Many lymphocyles. mostly CD4" and CD8" cells, iuliltmted to the tumors of IL-12-treated mice. The nmnbers of these cells increased in the cervical lymph nodes, into which the cerebrospinal fluid drains, and there they secreted a considerable amount of interferon-g. Mice cured by IL-12 rejected subcutaneous or i.t. rechallenge with their original glioma ceils, but the same mice were not able to reject other sy_ngeneic mmor cells. These results indicate that the immune system recognizes malignant glioma cells in the subamchnoid space of the CNS and that s3"stemic IL-12 may produce effective antitumor activity and lung-lasting tumor-specific immunity.
582 Protective Effect of Naked DNA Immunizationin Experimental AutoimmuneEncephalitis
585 Cytotoxic T Lymphocyte-mediatedCell Death in Paraneoplastic Sensory Neuronopathywith Anti-HuAntibody
J, Nowizka, MedicaIAcademy of Lodz, Poland, (2. Kowal, Albert Einstein College of Medicine, New York, USA, A. Walczak, A. Glabinski, A. Jurewicz, K. Selmaj, MedicaIAcademy ofLodz, Poland
K. Tanaka. T. Inuzuka, R. Nakano, S. Tsuji, BrainResearchlnstitute, Ntigata University, Japan, M. Tanaka, DepartmentofNeurology & ClinicaI Research Center, National West,N iigata Central Hospital, Japan
Naked DNA vaccination offers several therapeutical possibilities from inducing to down-regulation of immune responses. We have evaluated the effect of DNA vaccination with pRc-CMV plasmid containing PLP gene in SJL-J mice. Injection of the plasmid construct resulted in PLP gene expression in murine muscles as measured by Western blotting. We next assessed how DNA immunization effect development of EAE induced with PLP. The results have shown that in mice immunized three weeks prior with pRc/CMV-PLP tendency to more safer EAE was observed. However, in mice immunized four months prior and sensitized with PLP, the EAE course was significantly ameliorated as assessed by lower clinical scores and shorter disease duration. The results suggest that naked DNA immunization might have protective effect against EAE.
583 Suppression of M u r i n e
Experimental Autoimmune Encephalomyelitis b y IL-2 T a r g e t e d Fusion Toxin D A B 3 8 9 I L - 2 S.M. Phillins. S.A. Zekavat, M.A. Ramadan All, P.J. Perrin, Univ. ofPenn., USA, P. Bachs, Seragen, Inc., USA
DABjsglL-2 is a diptheria toxin-lL-2 chimeric protein. DAB~sg[L-2 targets activated T cells which express the high affinity IL-2 receptor (CD25) and kills them. Because activated cells are targeted with this strategy, the antigenic T cell epitope does not need to be defined. This is particularly important when considering treatments for diseases such as MS, where an immune component is accepted but the identity of the autoantigen is not known. We therelbre investigated the in vitro and in vivo of DAB3sglL-2 to suppress immunopathotogy in marine EAE. DAB3sglL-2 suppressed IL-2, lectin mitogen, and MBP induced lymphocyte proliferation, CD25 expression, IL-2, and IFN-? production. In vivo, DAB3891L-2 suppressed the development of active and adoptively transferred EAE. DAB3sglL-2 modestly reduced total CD4 +, CD25 + cells at the systemic level and little effect on CD8 + cells. In contrast, DAB3sglL-2 pro|bundly suppressed the accumulation of CD4 +, CD8 ÷ and CD25 + lymphocytes and macrophages within the CNS lesions. These results suggest that DAB389IL-2. with its minimal side effects, may have potential for the genetically independent targeted reduction of immunopathology in human MS.
Paraneoplastic sensory neuronopathy (PSN) has been shown to harbor characteristic anti-neuronal autoantibody "anti-Hu" in their sara and cerebrnspinal fluid. Creation of animal models exhibiting clinical or pathological features seen in PSN by means of passive transfer of anti-Hu positive lgG has not been achieved. Although anti-Hu antibody was shown to induce neuronal cell lysis in vitro, this result has not been reproduced so far. Since prominent T cell infihration is seen in the central nervous system and posterior spinal ganglion oftbe patients with anti-Hu syndrome, we studied cytotoxic T cell (CTL) activity in peripheral mononuclear cells from a patient with PSN harboring anti-Hu antibody. The activated CD8+ T cells from the patient's venous blood were shown to lyse her own fibroblasts which were incubated with interferon-'/to induce HLA class 1 molecules on their surface and the recombinant HuD protein was injected into the cells by mieroinjector. This is the first report showing the existence of CTL activity in a patient with PSN~
586 CytotoxicT CellsAgainsta Peptidefrom Yo Protein in Patients w i t h P a r a n e o p l a s l l c Cerebellar D e g e n e r a t i o n w i t h Anti-Yo A n t i b o d y M. Tanaka, National WestNiigata Centre Hospital, Japan, K. Tanaka, BrainResearch Institute, Niigata University, Japan, S. Tokiguchi, Ojays General Hospital, Japan, K. Shinozawa, Saiseikai-Kaw aguchi GeneralHospital, Japan, S. Tsuji, Brain Reaearch Institute, Nflgata University, Japan
High anti-Yo antibody titer is present in some patients with paraneoplastic cerebellar degeneration, but there is no evidence that the antibody causes Purkinje cell loss. We examined cytotoxie T cell (CTL) activity against a peptide of Yo protein that was defined by the HLA-based approach called reverse immunogenetics. The peripheral venous blood of three patients showed CTL activity against a peptide of Yo protein combined with autologous fibroblasts as the target after activation with Yo peptide. In paraneoplastic cerebellar degeneration, CTL therefore may be involved in the loss of Purkinje cells that present Yo protein.