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Characteristics of toxin-neutralization by anti-tetanus human monoclonal antibodies directed against the three functional domains [A], [B], and [C] of the tetanus toxin molecule and a reliable method for evaluating the protective effects of monoclonal antibodies

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Abstract

Five anti-tetanus human monoclonal antibodies (MAbs) produced by hybrid cell lines we established previously were characterized. Their abilities to neutralize tetanus toxinin vitro and to protect mice against challenge with toxin were studied by observing the changes in the progress of symptoms in mice. Immunostaining showed that MAbs MAb-G4 and G2 recognized the N-terminal domain, [A] and the C-terminal domain, [C] of the tetanus toxin molecule, respectively, while MAbs MAb-G1, G3 and G6 recognized its middle domain, [B]. Enzyme-linked immunosorbent assay showed that the binding affinity of MAb-G3 was 2.9 X 1010 M−1 and those of the other MAbs were as high as approximately 1011 M−1. Inin vitro neutralization experiments, at sufficient doses all the MAbs as single reagents protected mice completely against the effect of tetanus toxin. However, at lower doses than those sufficient to rescue mice, the kinetic patterns of progress of symptoms with the individual MAbs differed with each other and, except for MAb-G4, were different from that of anti-tetanus human polyclonal antibody. They suppressed the development and/or slowed the rate of progress of symptoms for over 96 h and delayed death of the mice. We propose that the comparison of the minimum survival dose with that of human polyclonal antibody of known international units is a reliable method for estimating the actual protective activity of a MAb. Intravenous (IV) injection of doses of individual MAbs or their mixtures at over 0.03 IU per mouse protected mice from subsequent challenge with 20 MLD of tetanus toxin. Moreover, mice could be rescued by IV injection of individual MAbs or their mixture at doses equivalent to 0.03 IU per mouse, even 10 h after intramuscular (IM) injection of 4 MLD of tetanus toxin. The importance of these findings for evaluating the protective effects of anti-tetanus MAbs for clinical use is discussed.

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Authors and Affiliations

  1. Department of Tuberculosis Research-Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka, 565, Suita-Osaka, Japan

    M. Matsuda, N. Sugimoto & Y. Ma

  2. Morinaga Institute of Biological Science, 2-1-1 Shimosueyoshi-Tsurumi-ku, 230, Yokohama Kanagawa, Japan

    M. Kamei & S. Hashizume

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  1. M. Matsuda
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  2. M. Kamei
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Matsuda, M., Kamei, M., Sugimoto, N. et al. Characteristics of toxin-neutralization by anti-tetanus human monoclonal antibodies directed against the three functional domains [A], [B], and [C] of the tetanus toxin molecule and a reliable method for evaluating the protective effects of monoclonal antibodies. Eur J Epidemiol 8, 1–8 (1992). https://doi.org/10.1007/BF02427384

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