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The thioester containing proteins of the vertebrates, complement components C3 and C4, and alpha-2-macroglobulin
$54
]-he Scientific & Social Program: Vth ISDCl Co,,gfess
C6 HAPTEN PROVIDES A NEGATIVE SIGNAL TO ANTIGEN-SENSZTVE B LYMPHOCgTES IN TROUT. *S.L.Kaattari & J.L.Stegeman. Department of Microbiology, Oregon State University, Corvallis, OR 97331-3804, lISA. Previous studies have been interpreted to suggest thac mouovalent hapten can only passively inhibit the induction of an antigen--specific B cell response. This inhibition putatively occurs by competitive inhibition with antigen for the immunoglobulin receptor. Evidence from our laboratory suggests that such passive inhibition cannot solely be responsible for the hapten-specific elimination of the antibody response. Co-incubation of TNP-Zysine has been found to prevent, not only the TNP-specific antibody response to TNP-lipopolysaccharide, but also the anti-TNP response induced by the polyclonal activator, unconjugated lipopolysaccharide. The use of polyclonally activating concentrations of the unconjugated LPS theoretically unlinks the anti-TNP response from the anti-TNP receptor. Thus, since induction of anti-TNP antibodies occurs strictly through activation via the LPS receptor, inhibition by conincubation with the TNP-lysine hapten indicates that interaction of the anti-TNP receptor with hapten must induce a negative signal. This inhibition was found to occur only early during induction (within the first two days of culture). These kinetic studies reveal that the induction of this negative (tolergenic?) signal can only occur prior or during the early inductive phase of the antibody response.
C7 THE THIOESTER C O N T A I N I N G PROTEINS OF THE VERTEBRATE5 COMPLEMENT COMPONENTS C3 AND C4, AND ALPHA-2-MACROGI~OBUI IN *Alister W Dodds ~, Anthony C Willisa, R Paul Levine b and S-K Alex Law ~ d MRC lmmunochemistry Unit, University of Oxford, Oxford, OXI 3QU, LJK. bDept of Genetics Washington University Medical School, St Louis, MO, USA. Using anion exchange c h r o m a t o g r a p h y on Q-Sepharose of plasma or serum mliowed by radioactive methylamiue incorporation and autoradiography of SDS-PAGE gels it is possible to detect and identify the major mammalian thioester containing proteins (Auerbach, H.S., Burger, R., Dodds, A.W. and Colten, H.R. 1990 J. Clin. Invest. 86, 96-106). We have extended this study to include examples of all of the major vertebrate classes. In all of the mammals studied to date three major proteins are apparent, with thioester containing chains of approximately 18(I, 110 and 95 kDa corresponding to c*2M, C3 and C4 respectively, though in some species minor bands or doublets, which could represent differences in post-translational modification or the presence ot more than one gene, are evident. In the reptiles, birds and amphibians studied the situation is similar to that seen in mammals. In the bony fish two patterns are apparent. The arrangement in cod is similar to that seen in the mammals while salmon and catfish appear to be similar to the plaice (Starkey, P.M. 1983 An. N. Y. Acad. Sci. 421,112-118), in lacking a single chain o.2M of 180 kDa, having instead molecules more closely resembling C3, together with molecules which in the case of the catfish have been identified as C4 and probably' (73 by N terrninal sequence determination. In the shark there are molecules with thioester containing chains simdar in M. Wt. to mammalian c~2M and (73, though to date no C4 like molecules have been detected, in the lowest vertebrate studied, the lamprey, three thioester containing bands are apparenL similar to c~2M and C4 and another molecule with a thioester containing chain of approximately 85 kDa. Supported by a Collaborative Research Grant from NATO.
]-he Scientific & Social Program: Vth ISDCl Co,,gfess
C6 HAPTEN PROVIDES A NEGATIVE SIGNAL TO ANTIGEN-SENSZTVE B LYMPHOCgTES IN TROUT. *S.L.Kaattari & J.L.Stegeman. Department of Microbiology, Oregon State University, Corvallis, OR 97331-3804, lISA. Previous studies have been interpreted to suggest thac mouovalent hapten can only passively inhibit the induction of an antigen--specific B cell response. This inhibition putatively occurs by competitive inhibition with antigen for the immunoglobulin receptor. Evidence from our laboratory suggests that such passive inhibition cannot solely be responsible for the hapten-specific elimination of the antibody response. Co-incubation of TNP-Zysine has been found to prevent, not only the TNP-specific antibody response to TNP-lipopolysaccharide, but also the anti-TNP response induced by the polyclonal activator, unconjugated lipopolysaccharide. The use of polyclonally activating concentrations of the unconjugated LPS theoretically unlinks the anti-TNP response from the anti-TNP receptor. Thus, since induction of anti-TNP antibodies occurs strictly through activation via the LPS receptor, inhibition by conincubation with the TNP-lysine hapten indicates that interaction of the anti-TNP receptor with hapten must induce a negative signal. This inhibition was found to occur only early during induction (within the first two days of culture). These kinetic studies reveal that the induction of this negative (tolergenic?) signal can only occur prior or during the early inductive phase of the antibody response.
C7 THE THIOESTER C O N T A I N I N G PROTEINS OF THE VERTEBRATE5 COMPLEMENT COMPONENTS C3 AND C4, AND ALPHA-2-MACROGI~OBUI IN *Alister W Dodds ~, Anthony C Willisa, R Paul Levine b and S-K Alex Law ~ d MRC lmmunochemistry Unit, University of Oxford, Oxford, OXI 3QU, LJK. bDept of Genetics Washington University Medical School, St Louis, MO, USA. Using anion exchange c h r o m a t o g r a p h y on Q-Sepharose of plasma or serum mliowed by radioactive methylamiue incorporation and autoradiography of SDS-PAGE gels it is possible to detect and identify the major mammalian thioester containing proteins (Auerbach, H.S., Burger, R., Dodds, A.W. and Colten, H.R. 1990 J. Clin. Invest. 86, 96-106). We have extended this study to include examples of all of the major vertebrate classes. In all of the mammals studied to date three major proteins are apparent, with thioester containing chains of approximately 18(I, 110 and 95 kDa corresponding to c*2M, C3 and C4 respectively, though in some species minor bands or doublets, which could represent differences in post-translational modification or the presence ot more than one gene, are evident. In the reptiles, birds and amphibians studied the situation is similar to that seen in mammals. In the bony fish two patterns are apparent. The arrangement in cod is similar to that seen in the mammals while salmon and catfish appear to be similar to the plaice (Starkey, P.M. 1983 An. N. Y. Acad. Sci. 421,112-118), in lacking a single chain o.2M of 180 kDa, having instead molecules more closely resembling C3, together with molecules which in the case of the catfish have been identified as C4 and probably' (73 by N terrninal sequence determination. In the shark there are molecules with thioester containing chains simdar in M. Wt. to mammalian c~2M and (73, though to date no C4 like molecules have been detected, in the lowest vertebrate studied, the lamprey, three thioester containing bands are apparenL similar to c~2M and C4 and another molecule with a thioester containing chain of approximately 85 kDa. Supported by a Collaborative Research Grant from NATO.