Immunochemical analysis of Lewis rat antisera to the synthetic encephalitogenic peptide S49

Eugene D. Day, George A. Hashim, Nicholas Potter, Kenneth J. Lazarus

Research output: Contribution to journalArticle

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Abstract

Discrete populations of anti-S49 antibodies were found in the antisera of Lewis rats recovered from S49-induced experimental allergic encephalomyelitis (EAE). A potent inducer of EAE in Lewis rats, S49 is a synthetic peptide representing residues 69-84 of bovine myelin basic protein but with deletions at Gly-77 and His-78 to form an analogue of guinea pig or rat 69-84, GSLPQKAQRPQDENG. Each population within a given antiserum, as identified by Scatchard and Sipsian window analysis, was found to exhibit reactivity for a different S49 determinant, and the affinities of each population were relatively restricted and discontinuous. The high affinity populations (107-108 M-1) were cross-reactive with YS8 (YGSLPQKAQGHRPQDENG) in equilibrium competitive inhibition reactions whereas the low affinity populations (105-106 M-1) were reactive only with S49 and YS49 among a panel of peptide analogues. Of the YS8 cross-reactive antibodies the highest affinity (108 M-1) were also cross reactive with S81 (YGSLPQKAQGHRPQDEG) but not S49 (69-84-Gly), thus emphasizing the need for Tyr-68 for format stability of the determinant involved. The other YS8 cross-reactive population (107 M-1) was completely reactive with S49 but totally unreactive with S81 in equilibrium reactions, thus emphasizing the requirement for Asn-84 but not Tyr-68 for the determinant's topographic stability. Peptides shorter than S49 from the N-terminal end, but retaining the sequences AQRPQDEN or SQRSQDEN (suspected residence of minimal encephalitogenic determinants), reacted only under conditions of two-step non-equilibrium competitive inhibition assays. Such reactions would occur only at very low affinity (<105 M-1) with the anti-S49 antibodies. It was hypothesized that the encephalitogenic T-cell determinant for Lewis rats, although permitting B-cell responses at very low affinity, may exclude high affinity responses in susceptible animals.

Original languageEnglish (US)
Pages (from-to)1587-1603
Number of pages17
JournalNeurochemical Research
Volume10
Issue number12
DOIs
StatePublished - Dec 1 1985
Externally publishedYes

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Rats
Immune Sera
Peptides
Population
Antibodies
Autoimmune Experimental Encephalomyelitis
Myelin Basic Protein
T-cells
Anti-Idiotypic Antibodies
Assays
Animals
Antibody Affinity
Cells
Guinea Pigs
B-Lymphocytes
T-Lymphocytes

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Cellular and Molecular Neuroscience

Cite this

Immunochemical analysis of Lewis rat antisera to the synthetic encephalitogenic peptide S49. / Day, Eugene D.; Hashim, George A.; Potter, Nicholas; Lazarus, Kenneth J.

In: Neurochemical Research, Vol. 10, No. 12, 01.12.1985, p. 1587-1603.

Research output: Contribution to journalArticle

Day, Eugene D. ; Hashim, George A. ; Potter, Nicholas ; Lazarus, Kenneth J. / Immunochemical analysis of Lewis rat antisera to the synthetic encephalitogenic peptide S49. In: Neurochemical Research. 1985 ; Vol. 10, No. 12. pp. 1587-1603.
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abstract = "Discrete populations of anti-S49 antibodies were found in the antisera of Lewis rats recovered from S49-induced experimental allergic encephalomyelitis (EAE). A potent inducer of EAE in Lewis rats, S49 is a synthetic peptide representing residues 69-84 of bovine myelin basic protein but with deletions at Gly-77 and His-78 to form an analogue of guinea pig or rat 69-84, GSLPQKAQRPQDENG. Each population within a given antiserum, as identified by Scatchard and Sipsian window analysis, was found to exhibit reactivity for a different S49 determinant, and the affinities of each population were relatively restricted and discontinuous. The high affinity populations (107-108 M-1) were cross-reactive with YS8 (YGSLPQKAQGHRPQDENG) in equilibrium competitive inhibition reactions whereas the low affinity populations (105-106 M-1) were reactive only with S49 and YS49 among a panel of peptide analogues. Of the YS8 cross-reactive antibodies the highest affinity (108 M-1) were also cross reactive with S81 (YGSLPQKAQGHRPQDEG) but not S49 (69-84-Gly), thus emphasizing the need for Tyr-68 for format stability of the determinant involved. The other YS8 cross-reactive population (107 M-1) was completely reactive with S49 but totally unreactive with S81 in equilibrium reactions, thus emphasizing the requirement for Asn-84 but not Tyr-68 for the determinant's topographic stability. Peptides shorter than S49 from the N-terminal end, but retaining the sequences AQRPQDEN or SQRSQDEN (suspected residence of minimal encephalitogenic determinants), reacted only under conditions of two-step non-equilibrium competitive inhibition assays. Such reactions would occur only at very low affinity (<105 M-1) with the anti-S49 antibodies. It was hypothesized that the encephalitogenic T-cell determinant for Lewis rats, although permitting B-cell responses at very low affinity, may exclude high affinity responses in susceptible animals.",
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