Induction of experimental allergic encephalomyelitis in Lewis rats with purified synthetic peptides: delineation of antigenic determinants for encephalitogenicity, in vitro activation of cellular transfer, and proliferation of lymphocytes.

Abstract

Four highly purified synthetic peptides encompassing segments of the 68-86 region [for the numbering system used, see Eylar, E. H., Brostoff, S., Hashim, G., Caccam, J. & Burnett, P. (1971) J. Biol. Chem. 246, 5770-5784] of myelin basic protein (MBP), a region known to induce experimental allergic encephalomyelitis (EAE) in Lewis rats, were used to define and compare structure-function relationships between the primary structure of the 68-86 sequence and the three following biological activities: induction of EAE in Lewis rats, stimulation of T lymphocytes in vitro as measured by augmented cellular transfer of EAE to syngeneic recipients, and lymphocyte proliferation, as measured by [3Jthymidine incorporation. Guinea pig (GP) MBP was approximately 60 or 1500 times more active than the GP68-84 (Y G S L P Q K S Q R S Q D E N; single-letter amino acid abbreviations) or the modified bovine (MB) 68-84 (Y G S L P Q K A Q R P Q D E N) peptides for induction of EAE, respectively. Furthermore, lymphocytes primed with either GPMBP, GP68,84, or MB68-84 crossreacted in vitro with either GPMBP, GP68-84, or MB68-84 for activation of lymphocyte transfer activity. In contrast, lymphocytes primed with either GP68-84 or MB68-84 exhibited antigen-specific proliferation in vitro exclusively in response to either GP or MB sequences, respectively. Neither GP75-84 (S Q R S Q D E N) nor GP75-86 (S Q R S Q D E N P V) induced EAE, activated lymphocytes for EAE transfer, or stimulated lymphocyte proliferation under conditions and doses tested. We conclude that (i) structurally distinct determinants, reflecting existence of functionally independent classes of antigen receptors, specify encephalitogenic and proliferative responses of primed lymphocytes and (it) determinants for EAE induction, cellular transfer of EAE, and lymphocyte proliferation include amino acid residues in the 68-74 (Y G S L P Q K) sequence ofGPMBP.Experimental allergic encephalomyelitis (EAE) is a cellmediated autoimmune response directed against autologous central nervous system myelin. Disease manifestations include clinical neurologic signs and perivascular infiltration of inflammatory mononuclear cells within the central nervous system (1, 2). Sensitization with myelin basic protein (MBP), a major protein constituent of central nervous system compact myelin, emulsified in complete Freund's adjuvant, will produce the full clinical and histological picture of EAE in a wide variety of animal species. In Lewis rats, sensitization with MBP in complete Freund's adjuvant induces the generation of lymphocytes in peripheral lymphoid organs with the capacity to adoptively transfer EAE into syngeneic Lewis recipients (3, 4). When lymphocytes sensitized to MBP/ complete Freund's adjuvant are cultured in medium containing MBP for 3-4 days, they exhibit an '100-fold increase in EAE transfer activity, as evidenced by transfer of the paralytic signs of EAE with as few as 107 activated lymphocytes, and a vigorous antigen-specific proliferative response, as evidenced by [3...

Inhibition of Experimental Allergic Encephalomyelitis with an Antibody that Recognizes a Novel Antigen Expressed on Lymphocytes, Endothelial Cells, and Microglia

“…For these studies, cell lines were established that were specific for an encephalitogenic peptide (EP) of myelin-basic protein (Mannie et al, 1985). The EP was dissolved in PBS and emulsified with an equal volume of complete Freund'a adjuvant (CFA) supplemented with 5.0 mg/ml Mycobacterium tuberculosis H37RA (DIFCO, Detroit, Michigan) for a final concentration of 1 mg/ml of EP.…”

AChlamydia pneumoniae-Specific Peptide Induces Experimental Autoimmune Encephalomyelitis in Rats

“…The former is an acute inflammatory disease with scant demyelination (2), whereas demyelination is a prominent feature of MS (3,4). MBP-reactive T cells in MS patients are predominantly directed toward a sequence contained within residues 84 -102 (17,30,31), whereas the dominant encephalitogenic epitope for LEW rats is composed of MBP 68 -86 (22,23). The 84 -102 peptide contains the sequence KNIVTPRTPPP, and our Blast search also turned up a chlamydial gene, Cpn0442, specifying a protein containing the sequence KNLFPPYEPPP, which conceivably could activate human MBP-reactive T cells.…”

“…Stimulation indices were considered significant only if they exceeded background by at least 3-fold. 68 -86 In the LEW rat, the dominant encephalitogenic MBP epitope is comprised of aa 68 -86 of guinea pig MBP (22,23). However, EAE is autoimmune because the disease can be induced with self (rat) whole MBP or rat MBP 68 -86 peptide (Tables I and II).…”

“…5b). Immunological specificity was demonstrated by the lack of proliferation in response to an irrelevant nonencephalitogenic peptide (MBP [11][12][13][14][15][16][17][18][19][20][21][22][23][24][25][26][27][28][29][30] or MBP [31][32][33][34][35][36][37][38][39][40][41][42][43][44][45][46][47][48][49][50] ). T cells derived from unimmunized rats showed no proliferative response to any of the peptides tested (not shown).…”

Experimental Autoimmune Encephalomyelitis

“…There are relapsing progressive EAE models that can be induced with Theiler`s Murine Encephalitis Virus (TMEV) ( (Mannie et al, 1985;McFarlin et al, 1975) Rat DA Spinal cord in IFA or MOG 1-125 in CFA or IFA Relapsing/progressive (Lorentzen et al, 1995;Weissert et al, 1998b) Mouse PL/J MBP, MBP Ac1-11 (or Ac1-9) in CFA + PT Acute monophasic (Zamvil et al, 1986) There are rodent EAE models that can be induced by passive transfer of T cells. These models are not subject of this chapter and are therefore not outlined in detail.…”