- Email: [email protected]
TSP
157
P13.05 SERUM PROM PATIENTS WITH I D I O P A T H I C SENSORY NEURONOPATHY INHIBITS NEURITE OUTGROWTH FROM EMBRYONIC DORSAL ROOT GANGLIA G,W, van Diik. J.H.J. Wokke, N.C. N ~ and P.R. B&" Research Lab., Dept. of Neurology, Rudolf Magnus Institute of Neurosciences Introduction: S¢~ory neurouupathy is characterized by sensory loss, sensory ataxia, ereflexia and normal muscle power. If no cusmes of sensory neuronol~hy (pmaneoplastic, toxic, Sj6~'ea's syndrome) are found, we are dealing with idiopathic sensory nmmmopathy (ISN). ISN is probably imm,,emediated. Lymphocytic infiltmton in dorael root ganglia (DRG) of patients i m p l i c ~ these ganglia as the primary site of the disease process. We investigated the inltuence of serum from patients with ISN on embryonic DRO neurite outgrowth in vitro. Methods: DRG celia from 15-day-old embryonic rats were cultured and incuhated for 48 h. Neurite outgrowth was assayed with a neurofilament ELISA. All serum dilutious (1-, 5-, 10%) were tested in five fold (nffiS). Results: Serum from ISN patients 0ignificanlly inhibited neurite outgrowth with increasing se~um co,castration. Sentm of healthy, age-matched conUol0 stimulat~ neurite outgrowth with increasing conuentratiou. Senna from patients with other neurological diseases or with a well-known autoimmune dion, s (Sjfgren's 0yndrome) could not be diffenmtiated from serom of controls. Conclmien: These results suggest that substanc,~ in serum from patients with ISN do play a role in the pathogenesis of the disease. Indirect immonohistochemistry and immonobints will be done to see if immonoglobulins of patiants bind to sections of rat and human DRG.
P13.06 CDS+ B cells (B-1 cells) secreting anti-peripheral nerve myelin antibodies in patients with mono¢inaal gammopathy and demyellaating polyneuropathy Ekerfeh C, Emerudh J, Vrethem M, l~pazqzneat of Neurology and Clinical Immunology, University Hospital, S-581 85 Link0ping, Sweden. B cells expressing the CD5 mm'ker, so called B-I cells, belong to a subpopalaLion of B cells with autoreactive ~ . Increased frequeuees of B-1 ceils have been reported in aatoimmuno diseases, In patients with monocinnal gammopathy and demyelinating polyncuropathy, the Mcomponent often consists of auto-antibodies reacting against myelin components. We therefore investigated if B-1 cells may be involved in the production of anti-myelin antibodies.ln 4 of 13 patients with monoclonal gammopathy and polynanropathy we found a very high proportion of presumed autoreactive B-1 cells, expressed as CDI9+CD5+ in CD19+, significantly correlating with high levels of anti myelin antibodies. The proportion of B-lcells correlated significandy to the amount of anti-myelin antibedi~ (P
P17.04
W14.06
CNS VASCULAR PERIGY'FES HAVE IMMUNE POTENTIAL
Restricted TCR Va and VS usage by HTlV-specific CDg* CTL from peripheral blood of patients with HAM/TSP
R. Bainbanov, R. Washington and P. Dore-Duffv Dept. of Neurology, Wayne State Sch. of Meal., Detroit, Michigan. Introduction: Pericytas (PC) and endothelial cells (EC) are involved in complex regulatow events which maintain vascular hemontasis. These events are likely to be important in neuroimmune networks centered at the blood brain barrier (BBB) in inflammation, infection and trauma. We have, questioned whether PC may have immune potential. Matorlala end Method=: PC were isolated from purified rat cerebral microvasseis. Expression of macrophage cell surface markers, adhesion molecules and the transferrin receptor (fiR) were examined using immunoffuorescent techniques. Antigen expression was analyzed by laser and flow cytometry. Phagocytosis experiments were performed using newly devised methods involving flow cytometry. Results: PC express Fc receptors and the rat marker OX-42. Little staining is seen for ED-I. PC express muscle specific a~in and fiR. PC respond to ¢ytokines and express VCAM-t, and MHC C~ass II antigen. PC phagocytize in an Fcdependent manner. PC are important to sustained expression of EC Eselectin, Conclusion: Pericytas have immune potential. They have macmphage like characteristics, and are likely to function in antigen presentation.
I. Elovaere,U. Utz, S. Smith, T. Lchky, S, Jucobson Neum/mmunolouyBranch,NIH,Bethnsda,MD20892, USA Objective: To determineT cell receptor (TCR)Vo end VI~ chain usagein HTLVoepeeificcytotexic ]ymphocytes (CTL) obtained from peripheral blood of patients with HTLV-l-ossociated myelepsthy/tropiszl spastic pzrepzreeLs(HAM/TSP). Methods: To choroctwizeTCR repertoire,CDB" lypmphocytestrom peripheralMood were cloned in limiting dilution, and the resulting we/Is were screened for HTLV-l-epecific precursor CTL activity. The RNA wee iseietet] from HLA-A2end HLA-lt14-restricted HTLVtax 11-19 end tax 9055-specific C08" CTL I'mee,gad eDNAwas analyzedby PCR amplificationusing Vo end VB chain family-specific oligonudeetideprimers. Results: In e petiont with early HAM/TSP,HLA-A2.restricted tax 11-19-epecific CTL lines used preferentially Vo2 (9/9) and Vo3 (4~). This was combinedwith preferentialusage of V~ (3/5). Among tax 11-19-epecific call lines trom two HLA-A2 patients with longer duration end severe pureporesis,the CTLs used preferentiallyVo2 (6/9) and/orVoI2 (5/9} or Vo7 (2/4), respectively. Their TCR V8 usagewas preferentiallyrestricted to V61 or V812. The TCR Vo end V8 usage of HLA-814 HAM/TSPpatient woe limited but heterogeneous. Diecuonion o ~ conclusions: HTLV-t tax 11-19-epecificCD8° CTL obtainedtrom different HLAA2 HAMffSP patients expresse restricted heterogeneityin TCR Vo end V6 chain usagewith little or no overlapbetween individuals.HTLV-l-speeificCTLusing restricted TCR Vo and V9 repertoires may play s critical role in the developmentof HAM/TSP.The immuonthcrapeuticstrategies that focus on dim/notingthese cells could be clinically significant. Tho informationobtainedfrom such e therapy could be extended to other diseases of retroviral origin.
P08.05 Dendritic cells and fulliculo-stelinte cells in emterior pituitaries W. Altaerts 1, D.M. Fluitsma 3, E.C.M. Hoefsmit 3, P.H.M. Jeucken 1, H. Morreau 2, F.T. Bosman 2 and H.A. Drexhage 1
t lmmunology Department, 2pathology Department, Erasmu§ University, P.O, Box 1738, 3000 DR Rotterdam, the Netherlands, endJCell Biology and Immunology Department, Free University Amsterdam, Van der Boechorststraat 7, 1081 BT Amsterdam, the Netherlands Immunolsbeling of cryo-sections of anterior pituitaries, using moabs against markers of the monocyte/dandritic cell/macrophage lineage, revealed readily detectable cell numbers. Using cryo-immuno-electron microscopy (anti-MHC-class II) of anterior pituitaries, we were able to distinguish MHC-class II-expreaning cells that resembled "classical" lymphoid dendritic cells (DC) and MHC-class II-expressing cells that corresponded to "classical" follieulo-stellate (FS) cells. Using double immunolabeling of cryo-sections of these rat AP with gold labels, we demonstrated a small, but definite overlap between the t w o populations. At the light microscopical level, $100 -+ FS cells and DC expressing CD45 were found to occupy predominantly different tissue compartments, namely the epithelial parenchyme cords end connective tissue compartments respectively, though there was some overlap. The present data are in favour of a close relationship and a partial overlap between the populations of pituitary lymphoid DC and the earlier described FS cells. Since FS cells are known to act as regulatorcells for endocrine pituitary responses, a similar function of DC can be visualized as well. Evidence will be given that lymphoid DC are indeed able to regulate pituitary endocrine responses.
P04.03 Genetic susceptibility to Multiple Sclerosis: an HLA class II polumorphlsm study In 29 italian famllles. M. Eoli. A. Salmaggi. L. La Mantia. M. Zaffaroni, C. Milanese. Neurological Institute C. Besta, Milan and S. Antonio Abbate, Hospital, Gallarate, Italy Several epidemiological observations suggest that genetic factor may influence susceptibility to Multiple Sclerosis (MS) according to a polygenic mode of inheritance. Population studies showed that HLA complex conUbutes to the genetic susceptibility to MS in Caucasian populations, but conflicting results emerged from family studies. To evaluate a possible role of HLA complex as a major determinant in MS susceptibility we selected 29 Italian families with two or more first degree relatives affected by MS. All samples were genotyped for class II alleles (DRy, DQa and DQ~) by polymerase chain reaction (PCR) amplificationand sequence specific oligoprobe hybridization. The contribution of HLA complex to MS susceptibility was evaluated by a) intrafamiiy association study, b) linkage analysis assuming a dominant or a recessive mode of inheritance with varying penetrances, c) identty by descent analysis. The results demonstred the contribution of HLA genes to MS susceptibility.
P13.05 SERUM PROM PATIENTS WITH I D I O P A T H I C SENSORY NEURONOPATHY INHIBITS NEURITE OUTGROWTH FROM EMBRYONIC DORSAL ROOT GANGLIA G,W, van Diik. J.H.J. Wokke, N.C. N ~ and P.R. B&" Research Lab., Dept. of Neurology, Rudolf Magnus Institute of Neurosciences Introduction: S¢~ory neurouupathy is characterized by sensory loss, sensory ataxia, ereflexia and normal muscle power. If no cusmes of sensory neuronol~hy (pmaneoplastic, toxic, Sj6~'ea's syndrome) are found, we are dealing with idiopathic sensory nmmmopathy (ISN). ISN is probably imm,,emediated. Lymphocytic infiltmton in dorael root ganglia (DRG) of patients i m p l i c ~ these ganglia as the primary site of the disease process. We investigated the inltuence of serum from patients with ISN on embryonic DRO neurite outgrowth in vitro. Methods: DRG celia from 15-day-old embryonic rats were cultured and incuhated for 48 h. Neurite outgrowth was assayed with a neurofilament ELISA. All serum dilutious (1-, 5-, 10%) were tested in five fold (nffiS). Results: Serum from ISN patients 0ignificanlly inhibited neurite outgrowth with increasing se~um co,castration. Sentm of healthy, age-matched conUol0 stimulat~ neurite outgrowth with increasing conuentratiou. Senna from patients with other neurological diseases or with a well-known autoimmune dion, s (Sjfgren's 0yndrome) could not be diffenmtiated from serom of controls. Conclmien: These results suggest that substanc,~ in serum from patients with ISN do play a role in the pathogenesis of the disease. Indirect immonohistochemistry and immonobints will be done to see if immonoglobulins of patiants bind to sections of rat and human DRG.
P13.06 CDS+ B cells (B-1 cells) secreting anti-peripheral nerve myelin antibodies in patients with mono¢inaal gammopathy and demyellaating polyneuropathy Ekerfeh C, Emerudh J, Vrethem M, l~pazqzneat of Neurology and Clinical Immunology, University Hospital, S-581 85 Link0ping, Sweden. B cells expressing the CD5 mm'ker, so called B-I cells, belong to a subpopalaLion of B cells with autoreactive ~ . Increased frequeuees of B-1 ceils have been reported in aatoimmuno diseases, In patients with monocinnal gammopathy and demyelinating polyncuropathy, the Mcomponent often consists of auto-antibodies reacting against myelin components. We therefore investigated if B-1 cells may be involved in the production of anti-myelin antibodies.ln 4 of 13 patients with monoclonal gammopathy and polynanropathy we found a very high proportion of presumed autoreactive B-1 cells, expressed as CDI9+CD5+ in CD19+, significantly correlating with high levels of anti myelin antibodies. The proportion of B-lcells correlated significandy to the amount of anti-myelin antibedi~ (P
P17.04
W14.06
CNS VASCULAR PERIGY'FES HAVE IMMUNE POTENTIAL
Restricted TCR Va and VS usage by HTlV-specific CDg* CTL from peripheral blood of patients with HAM/TSP
R. Bainbanov, R. Washington and P. Dore-Duffv Dept. of Neurology, Wayne State Sch. of Meal., Detroit, Michigan. Introduction: Pericytas (PC) and endothelial cells (EC) are involved in complex regulatow events which maintain vascular hemontasis. These events are likely to be important in neuroimmune networks centered at the blood brain barrier (BBB) in inflammation, infection and trauma. We have, questioned whether PC may have immune potential. Matorlala end Method=: PC were isolated from purified rat cerebral microvasseis. Expression of macrophage cell surface markers, adhesion molecules and the transferrin receptor (fiR) were examined using immunoffuorescent techniques. Antigen expression was analyzed by laser and flow cytometry. Phagocytosis experiments were performed using newly devised methods involving flow cytometry. Results: PC express Fc receptors and the rat marker OX-42. Little staining is seen for ED-I. PC express muscle specific a~in and fiR. PC respond to ¢ytokines and express VCAM-t, and MHC C~ass II antigen. PC phagocytize in an Fcdependent manner. PC are important to sustained expression of EC Eselectin, Conclusion: Pericytas have immune potential. They have macmphage like characteristics, and are likely to function in antigen presentation.
I. Elovaere,U. Utz, S. Smith, T. Lchky, S, Jucobson Neum/mmunolouyBranch,NIH,Bethnsda,MD20892, USA Objective: To determineT cell receptor (TCR)Vo end VI~ chain usagein HTLVoepeeificcytotexic ]ymphocytes (CTL) obtained from peripheral blood of patients with HTLV-l-ossociated myelepsthy/tropiszl spastic pzrepzreeLs(HAM/TSP). Methods: To choroctwizeTCR repertoire,CDB" lypmphocytestrom peripheralMood were cloned in limiting dilution, and the resulting we/Is were screened for HTLV-l-epecific precursor CTL activity. The RNA wee iseietet] from HLA-A2end HLA-lt14-restricted HTLVtax 11-19 end tax 9055-specific C08" CTL I'mee,gad eDNAwas analyzedby PCR amplificationusing Vo end VB chain family-specific oligonudeetideprimers. Results: In e petiont with early HAM/TSP,HLA-A2.restricted tax 11-19-epecific CTL lines used preferentially Vo2 (9/9) and Vo3 (4~). This was combinedwith preferentialusage of V~ (3/5). Among tax 11-19-epecific call lines trom two HLA-A2 patients with longer duration end severe pureporesis,the CTLs used preferentiallyVo2 (6/9) and/orVoI2 (5/9} or Vo7 (2/4), respectively. Their TCR V8 usagewas preferentiallyrestricted to V61 or V812. The TCR Vo end V8 usage of HLA-814 HAM/TSPpatient woe limited but heterogeneous. Diecuonion o ~ conclusions: HTLV-t tax 11-19-epecificCD8° CTL obtainedtrom different HLAA2 HAMffSP patients expresse restricted heterogeneityin TCR Vo end V6 chain usagewith little or no overlapbetween individuals.HTLV-l-speeificCTLusing restricted TCR Vo and V9 repertoires may play s critical role in the developmentof HAM/TSP.The immuonthcrapeuticstrategies that focus on dim/notingthese cells could be clinically significant. Tho informationobtainedfrom such e therapy could be extended to other diseases of retroviral origin.
P08.05 Dendritic cells and fulliculo-stelinte cells in emterior pituitaries W. Altaerts 1, D.M. Fluitsma 3, E.C.M. Hoefsmit 3, P.H.M. Jeucken 1, H. Morreau 2, F.T. Bosman 2 and H.A. Drexhage 1
t lmmunology Department, 2pathology Department, Erasmu§ University, P.O, Box 1738, 3000 DR Rotterdam, the Netherlands, endJCell Biology and Immunology Department, Free University Amsterdam, Van der Boechorststraat 7, 1081 BT Amsterdam, the Netherlands Immunolsbeling of cryo-sections of anterior pituitaries, using moabs against markers of the monocyte/dandritic cell/macrophage lineage, revealed readily detectable cell numbers. Using cryo-immuno-electron microscopy (anti-MHC-class II) of anterior pituitaries, we were able to distinguish MHC-class II-expreaning cells that resembled "classical" lymphoid dendritic cells (DC) and MHC-class II-expressing cells that corresponded to "classical" follieulo-stellate (FS) cells. Using double immunolabeling of cryo-sections of these rat AP with gold labels, we demonstrated a small, but definite overlap between the t w o populations. At the light microscopical level, $100 -+ FS cells and DC expressing CD45 were found to occupy predominantly different tissue compartments, namely the epithelial parenchyme cords end connective tissue compartments respectively, though there was some overlap. The present data are in favour of a close relationship and a partial overlap between the populations of pituitary lymphoid DC and the earlier described FS cells. Since FS cells are known to act as regulatorcells for endocrine pituitary responses, a similar function of DC can be visualized as well. Evidence will be given that lymphoid DC are indeed able to regulate pituitary endocrine responses.
P04.03 Genetic susceptibility to Multiple Sclerosis: an HLA class II polumorphlsm study In 29 italian famllles. M. Eoli. A. Salmaggi. L. La Mantia. M. Zaffaroni, C. Milanese. Neurological Institute C. Besta, Milan and S. Antonio Abbate, Hospital, Gallarate, Italy Several epidemiological observations suggest that genetic factor may influence susceptibility to Multiple Sclerosis (MS) according to a polygenic mode of inheritance. Population studies showed that HLA complex conUbutes to the genetic susceptibility to MS in Caucasian populations, but conflicting results emerged from family studies. To evaluate a possible role of HLA complex as a major determinant in MS susceptibility we selected 29 Italian families with two or more first degree relatives affected by MS. All samples were genotyped for class II alleles (DRy, DQa and DQ~) by polymerase chain reaction (PCR) amplificationand sequence specific oligoprobe hybridization. The contribution of HLA complex to MS susceptibility was evaluated by a) intrafamiiy association study, b) linkage analysis assuming a dominant or a recessive mode of inheritance with varying penetrances, c) identty by descent analysis. The results demonstred the contribution of HLA genes to MS susceptibility.