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Differences in verotoxin neutralizing activity of therapeutic immunoglobulins and sera from healthy controls

Unterschiede der verotoxinneutralisierenden Aktivität von therapeutisch anwendbaren Immunoglobulinen und Seren von Gesunden

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Summary

Intestinal infection byEscherichia coli O157 and other verotoxin (VT) producingE. coli has been increasingly recognized as an important factor for the causation of classic (enteropathic) hemolytic uremic syndrome (HUS) and hemorrhagic colitis (HC). Toxins most frequently involved are VT1 and VT2. As with other toxin-mediated diseases, administration of immunoglobulin (Ig) may be beneficial. However, little is known about the immune response elicited by the toxin(s), and the prevalence of VT neutralizing antibodies in the healthy population. We studied the capacity of seven Igs and a commercial plasma preparation to neutralize four different VTs (VT1, VT2, VT2c and VT2e). The results were compared with the neutralization titers (NT50%) of normal human serum samples from various age groups. Plasma products and normal sera were separated by protein G affinity chromatography to investigate the factor(s) responsible for VT neutralization. All Igs neutralized VT1 (8 to 96 NT50%). None of them inhibited VT2, VT2c or VT2e effectively. In contrast, none of 40 pediatric, and only one of 20 adult control sera (starting dilution 1 : 4) neutralized VT1 (25 NT50%). All 60 samples as well as the plasma preparation blocked VT2 (22 to 446 NT50%, median 137), but not VT2c and VT2e. The VT1 neutralizing activity was eluted with the IgG fraction. The VT2 neutralizing activity was not bound by protein G, but was recovered in the IgG-free effluent. In conclusion, therapeutic Igs significantly neutralize VT1, but are largely ineffective against other VTs. In contrast, all control sera inhibited VT2, but rarely VT1. Different principles, notably IgG and non-IgG (probably non-immunoglobulin) factors, respectively, appear to be responsible for the reduction of VT1 and VT2 cytotoxicityin vitro. Patients with VTEC disease are rarely expected to benefit from the use of presently available Igs.

Zusammenfassung

Enterale Infektionen durch Verotoxin-(VT-)produzierendeEscherichia coli werden zunehmend als Ursache des klassischen hämolytisch-urämischen Syndroms (HUS) erkannt. Das HUS ist häufig mit VT2 allein, seltener mit VT1 und VT2 assoziiert. Wie bei anderen bakteriell-toxischen Erkrankungen könnten Immunglobuline (Ig) von therapeutischem Nutzen sein. Die VT-induzierte Immunantwort und die Prävalenz neutralisierender Antikörper in der gesunden Bevölkerung sind jedoch nur unzureichend bekannt. Wir untersuchten die VT-Neutralisationstiter (NT50%) von sieben Ig- und einem kommerziellen Plasmapräparat gegenüber VT1, VT2, VT2c und VT2e und verglichen sie mit normalen Serumproben (NHS) aus verschiedenen Altersstufen. Zur näheren Charakterisierung der neutralisierenden Aktivität wurden das Plasmapräparat und NHS über eine Protein-G-Säule chromatographisch aufgetrennt. Alle Ig-neutralisierten VT1 (8–96 NT50%), jedoch nicht VT2, VT2c oder VT2e. Andererseits wurde VT1 von keinem der 40 pädiatrischen und nur von einem von 20 adulten Kontrollseren neutralisiert (25 NT50%; Anfangsverdünnung 1 : 4). Alle 60 Serumproben und die Plasmakonserve blockierten VT2 (22–446 NT50%, Median 137), aber nicht VT2c oder VT2e. Die VT1-neutralisierende Aktivität wurde mit der IgG-Fraktion eluiert, während die VT2-neutralisierende Aktivität im IgG-freien Durchlauf lokalisiert wurde. Therapeutische Ig neutralisieren VT1in vitro, sind aber weitgehend unwirksam gegen andere VT. Im Gegensatz dazu inhibieren Seren aller Altersgruppen VT2, aber selten VT1. Wahrscheinlich sind unterschiedliche Wirkprinzipien für die Neutralisation von Verotoxinen durch normale Seren verantwortlich, nämlich IgG für VT1 und eine Nicht-Immunglobulinfraktion für VT2. Bei der Mehrzahl der HUS-Patienten ist von der Gabe derzeit verfügbarer Ig-Präparate ein VT-spezifischer therapeutischer Effekt nicht zu erwarten.

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  1. M. Bitzan

    Present address: Dept. of Microbiology, The Hospital for Sick Children, 555 University Avenue, M5G 1X8, Toronto, Canada

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  1. Universitäts-Kinderklinik, Martinistraße 52, W-2000, Hamburg 20, Germany

    M. Bitzan,  M. Klemt,  R. Steffens &  D. E. Müller-Wiefel

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Bitzan, M., Klemt, M., Steffens, R. et al. Differences in verotoxin neutralizing activity of therapeutic immunoglobulins and sera from healthy controls. Infection 21, 140–145 (1993). https://doi.org/10.1007/BF01710530

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