Article ID | Journal | Published Year | Pages | File Type |
---|---|---|---|---|
8417664 | Journal of Immunological Methods | 2014 | 10 Pages |
Abstract
There is a growing interest in the monitoring of complement activation, not only in clinical settings but also in experimental models. However, for rodents only a limited number of tools are available to assess complement activity and activation. Here we describe three ELISAs for the measurement of rat classical (CP), MB-lectin (LP) and alternative (AP) pathway activities in serum and plasma. Moreover, we optimised a soluble C5b-9 (sC5b-9) ELISA for the detection of low level complement activation in rat. We determined the conditions for correct sample handling and showed that the assays had low inter- and intra-assay variation. We applied these assays to monitor complement activation in an experimental rat model of renal ischemia/reperfusion injury. We did not observe major complement consumption following reperfusion in CP or LP, and only minor AP consumption at 24Â h post reperfusion. However, MBL depletion prior to ischemia/reperfusion using a monoclonal antibody, transiently and specifically inhibited 75% of LP activity and ameliorated the AP consumption at 24Â h. To further assess complement activation during renal IRI, we monitored serum sC5b-9 and found that it was only significantly increased 72Â h post-reperfusion, but not when rats were pre-treated with anti-MBL or after sham surgery. In conclusion the described assays enable sensitive, reproducible and comprehensive assessment of complement activation in experimental rat models.
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Authors
J. Kotimaa, P. van der Pol, S. Leijtens, N. Klar-Mohammad, G. Schilders, M.R. Daha, H. Rutjes, C. van Kooten,