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Measurement of interleukin-6 production by monocytes for in vitro safety testing of hemoglobin solutions
W K Bleeker1, E M de Groot, P J den Boer
1Central Laboratory of The Netherlands Red Cross Blood Transfusion Service, Amsterdam.
Abstract:
Induction of interleukin-6 (IL-6) production by isolated human mononuclear blood cells was taken as in vitro model for the induction of inflammatory reactions. The model was very sensitive to bacterial endotoxin (detection limit < 10pg/ml). Hemoglobin (Hb) solutions, prepared under non-sterile conditions also induced IL-6 production, which correlated with a positive reaction in the Limulus assay. Purification of the Hb solutions with a detergent prevented IL-6 production, showing that pure Hb itself does not activate the monocytes. We conclude that this assay is a useful and sensitive test of contamination with components that can induce inflammatory reactions, especially microbial products.
Insights
This study shows that a new assay using human blood cells can detect inflammatory triggers like bacterial endotoxins and contaminants in hemoglobin solutions. The test is sensitive and useful for identifying microbial products that cause inflammation.
Area of Science:
- Immunology
- Biochemistry
Background:
- Interleukin-6 (IL-6) is a key cytokine in inflammatory responses.
- Assessing inflammatory reactions in vitro requires sensitive and reliable models.
- Bacterial endotoxins are potent inducers of inflammation.
Purpose of the Study:
- To establish an in vitro model for detecting inflammatory reaction inducers.
- To evaluate the sensitivity of the model to bacterial endotoxin and hemoglobin contaminants.
- To determine if purified hemoglobin can induce inflammation.
Main Methods:
- Isolated human mononuclear blood cells were used to model inflammation.
- The model's response to bacterial endotoxin and hemoglobin solutions was measured by IL-6 production.
- Hemoglobin solutions were tested before and after purification with detergent.
- The Limulus assay was used to detect endotoxins.
Main Results:
- The in vitro model demonstrated high sensitivity to bacterial endotoxin (detection limit < 10 pg/ml).
- Hemoglobin solutions prepared under non-sterile conditions induced IL-6 production, correlating with a positive Limulus assay.
- Purification of hemoglobin solutions eliminated IL-6 induction, indicating pure hemoglobin does not activate monocytes.
Conclusions:
- The developed assay is a sensitive and useful tool for detecting contaminants that induce inflammatory reactions.
- The assay is particularly effective for identifying microbial products.
- This model can help ensure the safety of biological preparations by detecting inflammatory triggers.