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Case Study

July 08, 2016

Cytokine release assay

Cytokine measurement within a 14-day intravenous toxicity study with a humanized immunoglobulin in rats

From the animals, 8 hours after the first administration blood samples were collected to performe cytokine measurement.

The cytokine measurement was performed with a rat cytokine 10-plex panel assay kit using Luminex®xMAP® system.

Cytokine determination was statistically analyzed by the SAS package.

Results:

The level of several cytokines (GM-CSF, IL-10, IL-6, TNF-alpha) was below the quantification limit in both vehicle- and test item-treated animals, hence the change in their level could not be evaluated. The concentration values of three cytokines IFN‑gamma, IL-1 beta and IL-4) were estimated by extrapolation of the evaluation software. Concentration values in the respective quantification range were observed only in case of IL-2 and IL-12.

Although IL-12 level showed differences between the genders in both vehicle- and test item-treated animals, the effect of the treatment upon its level in both gender was negligible, the difference remained similar between the genders after the treatment with test item. The serum level of IL-2 showed changes in the test item-treated animals (in both gender), but the changes were inconclusive (decrease in males but increase in females after the treatment with test item) and not significant due to the high standard deviation within the treatment groups.

The change in the level of IFN-gamma was significant (p<0.01 for male and p<0.05 for female) after the treatment with the test item, but the concentration values are extrapolated and cannot be considered as a reliable basis for the judgment of a significant effect caused by the treatment.

The serum level of IL-1 beta decreased only in females after the treatment with test item, but this change was not statistically significant. The IL-4 level did not show any change upon the treatment with test item.

Conclusion:

None of the cytokines in the panel showed significant change upon the treatment with the test item comparing to the respective cytokine level in serum from animals treated with the vehicle.

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