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A comparative study of Mono Mac 6 cells, isolated mononuclear cells and Limulus amoebocyte lysate assay in pyrogen
L Moesby1, S Jensen, E W Hansen
1The Royal Danish School of Pharmacy, Department of Pharmacology, 2 Universitetsparken, 2100, Copenhagen, Denmark. lm@mail.dfh.dk
Abstract:
Pyrogen induced secretion of interleukin 6 (IL-6) in Mono Mac 6 (MM6) cells was measured. The ability of the MM6 cell culture to detect pyrogens was compared to the Limulus amoebocyte lysate (LAL) test and isolated mononuclear cells (MNC). The detection limit of MM6 for lipopolysaccharide (LPS) and Staphylococcus aureus was comparable to that of MNC. Aspergillus niger and Candida albicans induced IL-6 in isolated MNC, but not in MM6. The detection limit for Salmonella typhimurium in the MM6 assay was comparable to that of the LAL assay. As expected, S. aureus and C. albicans did not show any LAL activity. A. niger and Influenza virus showed some activity in the LAL test, but could not be detected by MM6 cells. In conclusion, the MM6 assay is a good supplement to the current pyrogen assays for detection of LPS, S. aureus and S. typhimurium, but the MM6 assay could not detect A. niger, C. albicans and Influenza virus.
Insights
The Mono Mac 6 (MM6) cell assay effectively detects lipopolysaccharide (LPS) and certain bacteria, supplementing existing pyrogen tests. However, it cannot detect fungal or viral pyrogens like Aspergillus niger or Influenza virus.
Area of Science:
- Biotechnology
- Immunology
- Microbiology
Background:
- Pyrogens are fever-inducing substances that necessitate sensitive detection methods.
- Current pyrogen detection assays include the Limulus amoebocyte lysate (LAL) test and assays using isolated mononuclear cells (MNC).
- The Mono Mac 6 (MM6) cell line offers a potential alternative for pyrogen detection.
Purpose of the Study:
- To evaluate the efficacy of the MM6 cell assay in detecting various pyrogens.
- To compare the pyrogen detection capabilities of MM6 cells with established LAL and MNC assays.
- To determine the detection limits of MM6 cells for specific pyrogenic substances.
Main Methods:
- Cultured MM6 cells were exposed to pyrogens, and interleukin-6 (IL-6) secretion was measured.
- The detection limits of MM6 cells for lipopolysaccharide (LPS), Staphylococcus aureus, Salmonella typhimurium, Aspergillus niger, Candida albicans, and Influenza virus were assessed.
- Results were compared against those obtained from the LAL assay and isolated MNC.
Main Results:
- MM6 cells demonstrated comparable detection limits to MNC for LPS and S. aureus.
- MM6 cells did not detect IL-6 induction by A. niger and C. albicans, unlike MNC.
- The detection limit of MM6 for S. typhimurium was comparable to the LAL assay.
- A. niger and Influenza virus showed LAL activity but were not detected by MM6 cells.
Conclusions:
- The MM6 assay serves as a valuable supplement for detecting LPS, S. aureus, and S. typhimurium.
- The MM6 assay's limitations include its inability to detect pyrogens such as A. niger, C. albicans, and Influenza virus.
- MM6 cells offer a specific but not universal pyrogen detection capability compared to LAL and MNC assays.