From owner-cellbiol@net.bio.net Sun Jan 19 22:00:00 1997 Path: biosci!rutgers!gatech!csulb.edu!hammer.uoregon.edu!arclight.uoregon.edu!feed1.news.erols.com!howland.erols.net!surfnet.nl!Hermes.nki.nl!pcb72.nki.nl!user From: kirberg@nki.nl (Jorg Kirberg) Newsgroups: bionet.molbio.methds-reagnts,bionet.immunology,bionet.cellbiol Subject: Re: Antibodies to Hen Egg Lysozyme Date: Mon, 20 Jan 1997 18:22:13 +0100 Organization: nki Lines: 104 Message-ID: References: <1997Jan14.153933.5812@galileo.cc.rochester.edu> <5bvu2f$re0@cantine.wu-wien.ac.at> NNTP-Posting-Host: pcb72.nki.nl Xref: biosci bionet.molbio.methds-reagnts:53691 bionet.immunology:10714 bionet.cellbiol:6449 In article <5bvu2f$re0@cantine.wu-wien.ac.at>, bunka@i112pc09.vu-wien.ac.at (Sebastian Bunka) wrote: > In article <1997Jan14.153933.5812@galileo.cc.rochester.edu>, > ttha@uhura.cc.rochester.edu (Tom Thatcher) writes: > > I'm looking for polyclonal and/or monoclonal antibodies against hen > > egg lysozyme. So far I've found one commercial source of a polyclonal > > (Biodesign) and no sources of monoclonals. Any help would be appreciated. > > > To my best knowledge there is no (commercial available) monoclonal. > > But another good source of polyclonal rabbit-anti-HEL, IgG fraction > is: Rockland, Gilbertsville, PA; Order-Nr. 200-4172. (Don't have > the phone number anymore ...). > > I've found the IgG fraction quite good (don not buy the whole serum). > > Much better was immunizing a sheep w/ HEL !! (Well, I don't > have any more of that serum - in case you ask ;-)) > -- > Dear Sebastian and Tom, as there are transgenic mice with a HEL specific B cell receptor, there must be somewhere the hybridoma from which the genes were taken. That will be a good source for your monoclonal: %A Eris, J. M. %A Basten, A. %A Brink, R. %A Doherty, K. %A Kehry, M. R. %A Hodgkin, P. D. %D 1994 %T Anergic self-reactive B cells present self antigen and respond normally to CD40-dependent T-cell signals but are defective in antigen-receptor-mediated functions %B Proc Natl Acad Sci U S A %V 91 %N 10 %P 4392-6 %X B-cell tolerance to soluble protein self antigens such as hen egg lysozyme (HEL) is mediated by clonal anergy. Anergic B cells fail to mount antibody responses even in the presence of carrier-primed T cells, suggesting an inability to activate or respond to T helper cells. To investigate the nature of this defect, B cells from tolerant HEL/anti-HEL double-transgenic mice were incubated with a membrane preparation from activated T-cell clones expressing the CD40 ligand. These membranes, together with interleukin 4 and 5 deliver the downstream antigen-independent CD40-dependent B-cell-activating signals required for productive T-B collaboration. Anergic B cells responded to this stimulus by proliferating and secreting antibody at levels comparable to or better than control B cells. Furthermore, anergic B cells presented HEL acquired in vivo and could present the unrelated antigen, conalbumin, targeted for processing via surface IgD. In contrast, the low immunoglobulin receptor levels on anergic B cells were associated with reduced de novo presentation of HEL and a failure to upregulate costimulatory ligands for CD28. These defects in immunoglobulin-receptor-mediated functions could be overcome in vivo, suggesting a number of mechanisms for induction of autoantibody responses. %O May 10 %0 Journal Article %A Hartley, S. B. %A Crosbie, J. %A Brink, R. %A Kantor, A. B. %A Basten, A. %A Goodnow, C. C. %D 1991 %T Elimination from peripheral lymphoid tissues of self-reactive B lymphocytes recognizing membrane-bound antigens %B Nature %V 353 %N 6346 %P 765-9 %X The long-standing hypothesis that tolerance to self antigens is mediated by either elimination or functional inactivation (anergy) or self-reactive lymphocytes is now accepted, but little is known about the factors responsible for initiating one process rather than the other. In the B-cell lineage, tolerant self-reactive cells persist in the peripheral lymphoid organs of transgenic mice expressing lysozyme and anti-lysozyme immunoglobulin genes, but are eliminated in similar transgenic mice expressing anti-major histocompatibility complex immunoglobulin genes. By modifying the structure of the lysozyme transgene and the isotype of the anti-lysozyme immunoglobulin genes, we demonstrate here that induction of anergy or deletion is not due to differences in antibody affinity or isotype, but to recognition of monomeric or oligomeric soluble antigen versus highly multivalent membrane-bound antigen. Our findings indicate that the degree of receptor crosslinking can have qualitatively distinct signalling consequences for lymphocyte development. Yours, jorg --------------------------------------------------------------------- Joerg Kirberg Tel. 0031 - 20 - 512 19 98 The Netherlands Cancer Institute FAX 0031 - 20 - 512 20 11 Div. Molecular Genetics (H4) E_mail kirberg@nki.nl Plesmanlaan 121 NL - 1066 CX Amsterdam The Netherlands .