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Kikkawa, S. et al.

A comparative analysis of the antigenic, structural, and functional properties of three different preparations of recombinant human interleukin-18.

We compared the structural and functional properties of three recombinant human interleukin-18 (rIL-18) preparations, commercially available ( Pep rIL-18) and prepared in our laboratory (active and inactive, according to their ability to potentiate IL-12-mediated interferon-gamma [ IFN-gamma] induction in lymphocytes). All three preparations showed multimer formation on SDS-PAGE/immunoblotting using monoclonal antibodies (mAb) against the inactive form of rIL-18. In contrast, only the 18-kDa bands were recognized in each sample by mAb against the active form of rIL-18. The amounts of multimers and the 18-kDa moiety of Pep rIL-18 resembled those of the inactive rather than the active form. Likewise, the reaction profile of Pep rIL-18 toward mAb was very similar to that of inactive but not active rIL-18 on sandwich ELISA. Pep rIL-18 potentiated IFN-gamma-inducing activity together with IL-12, but its potency was 100-fold less than that of the active rIL-18, and excess doses were required for its activity. The inactive rIL-18 showed virtually no IFN-gamma-inducing ability, but when reduced and reconstituted, it inhibited the IFN-gamma-inducing activity of active rIL-18. These results suggest that there are two categories of recombinant IL-18 that are structurally, functionally, and antigenically different, and the mAb 125-2H and 21 can discriminate these two IL-18 populations by recognizing the epitopes specifically expressed on active and inactive IL-18, respectively.[1]

References

A comparative analysis of the antigenic, structural, and functional properties of three different preparations of recombinant human interleukin-18. Kikkawa, S., Shida, K., Okamura, H., Begum, N.A., Matsumoto, M., Tsuji, S., Nomura, M., Suzuki, Y., Toyoshima, K., Seya, T. J. Interferon Cytokine Res. (2000) [Pubmed]

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