Conventional Test Method:

Rabbit Pyrogen Test (RPT) (USP 151)

Alternative Test Method:

Name & Description:

PBMC IL - 6 In Vitro Pyrogen Test

This test is based on the principle that white blood cell monocytes differentiate into macrophages and release inflammatory cytokines when exposed to bacterial endotoxin. The test exposes human peripheral blood mononuclear cells (PBMC) to the test substance and measures cytokine release (IL-6) using an immunoassay.

Validation Status:

EU: Validated by ECVAM for the detection of pyrogenicity mediated by Gram-negative endotoxin (2006)

US: Found suitable by ICCVAM for use to detect Gram-negative endotoxin in human parenteral drugs on a case-by-case basis, subject to validation for each specific product to demonstrate equivalence to the RPT (2008)

Regulatory Status:

No information

Effect or Potential Effect on Animal Use:

Replacement
(when used to test for Gram-negative endotoxins)

References:

• ECVAM (2006) ESAC Statement on the validity of in-vitro pyrogen tests.
• Hoffmann S., Peterbauer A., Schindler S., Fennrich S., Poole S., Mistry Y., Montag-Lessing T., Spreitzer I., Löschner B., van Aalderen M., Bos R., Gommer M., Nibbeling R., Werner-Felmayer G., Loitzl P., Jungi T., Brcic M., Brügger P., Frey E., Bowe G., Casado J., Coecke S., de Lange J., Mogster B., Naess L.M., Aaberge I.S., Wendel A. & Hartung T. (2005) International validation of novel pyrogen tests based on human monocytoid cells. Journal of Immunological Methods 298: 161-173.
• NICEATM-ICCVAM (2007). Independent Peer Review Panel Report: Five In Vitro Test Methods Proposed for Assessing Potential Pyrogenicity of Pharmaceuticals and Other Products.
• NICEATM-ICCVAM (2008) ICCVAM Background Review Document: Validation Status of Five In Vitro Test Methods Proposed for Assessing Potential Pyrogenicity of Pharmaceuticals and Other Products. 778pp. Research Triangle Park NC: National Institutes of Health (NIH).
• NICEATM-ICCVAM (2009) In Vitro Pyrogen Test Methods: Overview and Recommendations
• Poole S., Mistry Y., Ball C., Gaines Das R.E., Opie L.P., Tucker G. & Patel M. (2003) A rapid ‘one-plate’ in vitro test for pyrogens. Journal of Immunological Methods 274: 209-220.

Information Last Reviewed:

May 2012