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A nervous system induced factor inducing cytokines in immune cells
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
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446
T Cell Response to MyelIn Basic Protein a n d CNS Inflammatory Activity D u r i n g the Course of Relapsing-remitting Multiple Sclerosis M. VergeUi, B. Mazzanti, E. Traggiai, A, Parigi, L. Massacesi, Univ.ofFlorence, Italy
Induction of Tolerance in Established AutoimmuneDisease with a Combinationof CD4-speeific Antibody and IntravenousAntigen
The aim of this study is to investigate the functional properties of myelin basic protein (MBP)-specific T-cells isolated at different times from the peripheral blood of MS patients who underwent a serial gadoliniumenhanced magnetic resonance imaging (GD-MRI) study. GD-MRI has been proved to be a good tool to monitor disease activity and to study the natural history of MS. Peripheral blood mononuclear cells (PBMC) were collected monthly from MS patients at the same time they underwent GD-MRI examination. At each time point, MBP-specific T cell lines (TCLs) were generated by using a modified split-well technique. TCLs have been characterized in terms of epitope specificity, cytotoxic activity and cylokine production. Episodic changes in the frequency, peptide specificity and functional repertoire of MBPspecific ]'-cells were observed in several patients. The relationships between these changes and the development of new lesions in the CNS have been analyzed. These data may help in understanding the rote of MBP-specific T-cells in the patbogenesis of multiple sclerosis as well as the relevance of phenomena such as epitope spreading in the disease.
444 Differential Expressionof B7 CostimulatoryMoleculesin Human PolymorphonuelearLeucocytes
J. O'Neill, G. Pryce, D. Baker, UCL, ILK.
Both CD4 depleting and non-depleting mAb inhibit the progression of established, actively-induced chronic relapsing experimental allergic encephalomyelitis in ABH mice. Disease eventually returned following cessation ofmAb administration, the onset of which could be accelerated by a further antigen-rechallenge in adjuvant. This served to reactivate primed cells. Intravenous (i.v.) antigen (antigen-coupled cells) administration rapidly induced unresponsiveness and was effective when injected even after the onset of clinical signs. In contrast to soluble antigen, the use of cell vehicles maintain circulating levels of antigen bound to the cell surface. These induced inhibition in an MHC non-restricted manner, as xenogeneic and allogeneic cells were effective. A combination of CD4-specific mAb followed by intravenous antigen could resist disease re-induction by antigen rechallenge. This was depletion and timing dependent, where tolerance induction was best achieved when i.v. antigen was delivered 1 week following depletion. Such a combination induced marked unresponsiveness when administered to post-acute remission animals. This significantly inhibited the development of relapse whereas control animals demonstrated multiple relapses. The data suggest that it may be possible to re-induce antigen-specific tolerance in established autoimmune disease following transient T cell de-bulking of primed T cells, and may have relevance to the treatment of human autoimmune disease, such as multiple sclerosis
447 The Effect of a Novel NO-NSAID on ChronicNeuroinflammation L.M. Baker. B. Hauss-Wcgrzyniak, G.L. Wenk, UniversityofArizona, USA
A. Windhagen. S. Mania, A. Gebert, I. Ferger, U. Wurster, F. Heidenreich, Medical SchoolHannover,Germany
Polymorphonuclear leukocytes (PMN) are important phagocytic cells during acute inflammatory responses. Upon activation PMNs secrete cytokines and oxygen metabolitcs that might be involved in the regulation o f the acquired i m m u n e response. Since PMN infiltrates are found in lesions of T-cell mediated a u t o i m m u n e diseases, we wished to further investigate PMN/Tcell interactions. W e show here that peripheral blood P M N s constitutively express the T cell costimulatory molecule B7-1, but not B7-2, in the cytoplasm as detected by FACS analysis and immunocytochemistry. Expression o f B7-1 m R N A in P M N s increased upon stimulation with LPS and G M - C S F as detected by RT-PCR. Importantly 137-1 and B7-2 molecules were both expressed on the cell surface of PMNs isolated from patients with i n f l a m m a t o r y disease. Our findings suggest that P M N s are involved in the regulation of the adaptive i m m u n e response by differential expression o f B7 molecules.
Chronic brain inflammation is produced by continuous infusion of lipopolysaccharide (LPS) into the ventricular space of the rat's brain. Activated gila are the main source oflL-I after an infusion of LPS. IL-I can produce a variety of responses by inducing the release of prostaglandins which cause inflammation. Nonsteroidal anti-inflammatory drugs (NSAIDs) are widely used to prevent the deleterious effects associated with inflammation but their administration is limited by their negative side effect on the gastrointestinal tract. Nitcoflurbiprofen (NFP) is a novel NSAID that demonstrated anti-inflammatory efficacy and good gastrointestinal tolerability. Chronic LPS infusion produced an extensive inflammation within the brain, particularly within temporal lobe regions. Daily peripheral administration of NFP significantly attenuated the neuroinflammation produced by LPS, as indicated by the decreased reactive state of both microglia and astrocytes. The results of the present study suggest that NFP represents a novel class of compounds that have anti-inflammatory properties but show a more favorable toxicity profile than the classical NSAIDs.
445
448
The Role of NK Receptors in G a m m a - d d t a T Cell Cytotoxieity in Multiple Sclerosis
A N e r v o u s System I n d u c e d F a c t o r I n d u c i n g Cytokines in I m m u n e Cells M. Bakhiet. Y. Liu, KatolinskaInstitute, Stockholm,Sweden
R. Zeine, Universityof Otlawa, Canada, F. Mandy, NatwnalLaboratoryfor Analyt~cal Cytology (HealthCanada), M.S. Freedm an, Unwersityof Ottawa, Canada
We have previously described the cytotoxicity mechanisms utilized by" peripheral blood- and CSF-derived yST cells from patients with MS, and demonstrated that reagents which interfere with the perforin-based and Fasbased cytotoxicity pathways can be used to inhibit yST cell-mediated lysis of target cells, including in vitro cultured human oligodendrocytes. Here we investigate the roles of NK cell markers and receptors (NKR) in the regulation ofyST cell cytotoxicity in MS. We focus on CD56 and the inhibitory NKR, CD94, whose levels of expression have been shown to correlate with the cytotoxic potential of human T cell clones, yST cells were isolated by culture on anti-TCRy8 precoated plates and expanded with IL-2 (blood) and IL-2/IL-4 (CSF). Target lines included Daudi and Jurkat cells. Cytotoxicity was measured using a 5 hr 5tCr-release assay. Flow cytometric analysis of our yST cell populations revealed heterogeneity with 30% to 45% CD56* and over 60% CD94 + subsets. Further subfractionation by sorting will allow assessment of cytotoxicity potential within subsets. Treatment of CD94 + 78 T cells with anti-CD94 Abs is expected to downregulate their cytotoxieity.
We have recently described a novel trypanokine (TL'IT), a factor secreted by trypanosomes, that modulates the cytokine network of the host for the parasite benefit. Here we investigated the early events after subcutaneous infection of rats with Trypanosoma brucei or challenge with TLTF, Furthermore, cross-talk between the central nervous system (CNS) and the immune system during these early events was studied. Two hours after infection, IFN-g is produced in the spleen due to a signal transmitted from inoculation site via the splenic nerve. This signal induced IFN-g production via a soluble factor. To characterize this factor, hybrids between splenocytes taken I rain after infection were made and tested for IFN-g-inducing activity. Consequently, an immune system-released activating factor (ISRAF), a glycoprotein with approximately 23 kD, was obtained. Surgical denervation of the spleen abrogated ISRAF and the ensuing IFN-g production, reduced parasitemia and prolonged survival of the rats. In vivo application of ISRAF showed enhanced parasite growth. ISRAF also reversed the immunosuppression that occurs late during the infection. Hereby, a novel nervous system-induced factor inducing cytokines in immune ceils is presented.
79
443
446
T Cell Response to MyelIn Basic Protein a n d CNS Inflammatory Activity D u r i n g the Course of Relapsing-remitting Multiple Sclerosis M. VergeUi, B. Mazzanti, E. Traggiai, A, Parigi, L. Massacesi, Univ.ofFlorence, Italy
Induction of Tolerance in Established AutoimmuneDisease with a Combinationof CD4-speeific Antibody and IntravenousAntigen
The aim of this study is to investigate the functional properties of myelin basic protein (MBP)-specific T-cells isolated at different times from the peripheral blood of MS patients who underwent a serial gadoliniumenhanced magnetic resonance imaging (GD-MRI) study. GD-MRI has been proved to be a good tool to monitor disease activity and to study the natural history of MS. Peripheral blood mononuclear cells (PBMC) were collected monthly from MS patients at the same time they underwent GD-MRI examination. At each time point, MBP-specific T cell lines (TCLs) were generated by using a modified split-well technique. TCLs have been characterized in terms of epitope specificity, cytotoxic activity and cylokine production. Episodic changes in the frequency, peptide specificity and functional repertoire of MBPspecific ]'-cells were observed in several patients. The relationships between these changes and the development of new lesions in the CNS have been analyzed. These data may help in understanding the rote of MBP-specific T-cells in the patbogenesis of multiple sclerosis as well as the relevance of phenomena such as epitope spreading in the disease.
444 Differential Expressionof B7 CostimulatoryMoleculesin Human PolymorphonuelearLeucocytes
J. O'Neill, G. Pryce, D. Baker, UCL, ILK.
Both CD4 depleting and non-depleting mAb inhibit the progression of established, actively-induced chronic relapsing experimental allergic encephalomyelitis in ABH mice. Disease eventually returned following cessation ofmAb administration, the onset of which could be accelerated by a further antigen-rechallenge in adjuvant. This served to reactivate primed cells. Intravenous (i.v.) antigen (antigen-coupled cells) administration rapidly induced unresponsiveness and was effective when injected even after the onset of clinical signs. In contrast to soluble antigen, the use of cell vehicles maintain circulating levels of antigen bound to the cell surface. These induced inhibition in an MHC non-restricted manner, as xenogeneic and allogeneic cells were effective. A combination of CD4-specific mAb followed by intravenous antigen could resist disease re-induction by antigen rechallenge. This was depletion and timing dependent, where tolerance induction was best achieved when i.v. antigen was delivered 1 week following depletion. Such a combination induced marked unresponsiveness when administered to post-acute remission animals. This significantly inhibited the development of relapse whereas control animals demonstrated multiple relapses. The data suggest that it may be possible to re-induce antigen-specific tolerance in established autoimmune disease following transient T cell de-bulking of primed T cells, and may have relevance to the treatment of human autoimmune disease, such as multiple sclerosis
447 The Effect of a Novel NO-NSAID on ChronicNeuroinflammation L.M. Baker. B. Hauss-Wcgrzyniak, G.L. Wenk, UniversityofArizona, USA
A. Windhagen. S. Mania, A. Gebert, I. Ferger, U. Wurster, F. Heidenreich, Medical SchoolHannover,Germany
Polymorphonuclear leukocytes (PMN) are important phagocytic cells during acute inflammatory responses. Upon activation PMNs secrete cytokines and oxygen metabolitcs that might be involved in the regulation o f the acquired i m m u n e response. Since PMN infiltrates are found in lesions of T-cell mediated a u t o i m m u n e diseases, we wished to further investigate PMN/Tcell interactions. W e show here that peripheral blood P M N s constitutively express the T cell costimulatory molecule B7-1, but not B7-2, in the cytoplasm as detected by FACS analysis and immunocytochemistry. Expression o f B7-1 m R N A in P M N s increased upon stimulation with LPS and G M - C S F as detected by RT-PCR. Importantly 137-1 and B7-2 molecules were both expressed on the cell surface of PMNs isolated from patients with i n f l a m m a t o r y disease. Our findings suggest that P M N s are involved in the regulation of the adaptive i m m u n e response by differential expression o f B7 molecules.
Chronic brain inflammation is produced by continuous infusion of lipopolysaccharide (LPS) into the ventricular space of the rat's brain. Activated gila are the main source oflL-I after an infusion of LPS. IL-I can produce a variety of responses by inducing the release of prostaglandins which cause inflammation. Nonsteroidal anti-inflammatory drugs (NSAIDs) are widely used to prevent the deleterious effects associated with inflammation but their administration is limited by their negative side effect on the gastrointestinal tract. Nitcoflurbiprofen (NFP) is a novel NSAID that demonstrated anti-inflammatory efficacy and good gastrointestinal tolerability. Chronic LPS infusion produced an extensive inflammation within the brain, particularly within temporal lobe regions. Daily peripheral administration of NFP significantly attenuated the neuroinflammation produced by LPS, as indicated by the decreased reactive state of both microglia and astrocytes. The results of the present study suggest that NFP represents a novel class of compounds that have anti-inflammatory properties but show a more favorable toxicity profile than the classical NSAIDs.
445
448
The Role of NK Receptors in G a m m a - d d t a T Cell Cytotoxieity in Multiple Sclerosis
A N e r v o u s System I n d u c e d F a c t o r I n d u c i n g Cytokines in I m m u n e Cells M. Bakhiet. Y. Liu, KatolinskaInstitute, Stockholm,Sweden
R. Zeine, Universityof Otlawa, Canada, F. Mandy, NatwnalLaboratoryfor Analyt~cal Cytology (HealthCanada), M.S. Freedm an, Unwersityof Ottawa, Canada
We have previously described the cytotoxicity mechanisms utilized by" peripheral blood- and CSF-derived yST cells from patients with MS, and demonstrated that reagents which interfere with the perforin-based and Fasbased cytotoxicity pathways can be used to inhibit yST cell-mediated lysis of target cells, including in vitro cultured human oligodendrocytes. Here we investigate the roles of NK cell markers and receptors (NKR) in the regulation ofyST cell cytotoxicity in MS. We focus on CD56 and the inhibitory NKR, CD94, whose levels of expression have been shown to correlate with the cytotoxic potential of human T cell clones, yST cells were isolated by culture on anti-TCRy8 precoated plates and expanded with IL-2 (blood) and IL-2/IL-4 (CSF). Target lines included Daudi and Jurkat cells. Cytotoxicity was measured using a 5 hr 5tCr-release assay. Flow cytometric analysis of our yST cell populations revealed heterogeneity with 30% to 45% CD56* and over 60% CD94 + subsets. Further subfractionation by sorting will allow assessment of cytotoxicity potential within subsets. Treatment of CD94 + 78 T cells with anti-CD94 Abs is expected to downregulate their cytotoxieity.
We have recently described a novel trypanokine (TL'IT), a factor secreted by trypanosomes, that modulates the cytokine network of the host for the parasite benefit. Here we investigated the early events after subcutaneous infection of rats with Trypanosoma brucei or challenge with TLTF, Furthermore, cross-talk between the central nervous system (CNS) and the immune system during these early events was studied. Two hours after infection, IFN-g is produced in the spleen due to a signal transmitted from inoculation site via the splenic nerve. This signal induced IFN-g production via a soluble factor. To characterize this factor, hybrids between splenocytes taken I rain after infection were made and tested for IFN-g-inducing activity. Consequently, an immune system-released activating factor (ISRAF), a glycoprotein with approximately 23 kD, was obtained. Surgical denervation of the spleen abrogated ISRAF and the ensuing IFN-g production, reduced parasitemia and prolonged survival of the rats. In vivo application of ISRAF showed enhanced parasite growth. ISRAF also reversed the immunosuppression that occurs late during the infection. Hereby, a novel nervous system-induced factor inducing cytokines in immune ceils is presented.