Basic encephalitogenic protein: A simplified purification on sulphoethyl-sephadex.
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Biomedical subjects
Publications and source records attributed to M Sela.
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Primary antibody response against the dinitrophenyl group has been elicited in vitro after the stimulation of normal mouse spleen explants with 2,4-dinitrophenyl (DNP)-hemocyanin or alpha-DNP-poly-L-lysine (PLL). Antibodies were detected in the culture medium by the inactivation of DNP-T4 phage. The specificity of the reaction was manifested by the lack of the capacity of the medium to inactivate the unmodified bacteriophage and by the inhibition of the inactivation of DNP-T4 with DNP-lysine.
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The results reported establish a cellular basis for the inverse relationship between the net electrical charge of immunogens and of the antibodies provoked by them. Glass bead columns were used to reduce the number of immunocompetent spleen cells preferentially reactive with more acidic immunogens. After a single immunization, titers of antibodies to an acidic dinitrophenylated copolymer of tyrosine, glutamic acid, and lysine (DNP-901) elicited in recipient mice by filtered spleen cells were significantly lower than those generated by unfiltered cells. After secondary stimulation, the major portion of the antibodies provoked by the acidic antigen was found in the more acidic fraction eluted from DEAE-Sephadex, in contrast to the more basic antibodies, of the same specificity, generated by unfiltered spleen cells. Results obtained by transplanting a limiting number of spleen cells indicate a depletion in the number of precursor cells reactive with dinitrophenyl on DNP-901 after glass bead chromatography, whereas there was no change in the response to dinitrophenyl on a basic copolymer, DNP-912, containing the same amino acids in different molar ratios.
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