Overview and Recommendations
ICCVAM is currently evaluating the
usefulness and limitations of the following five in vitro pyrogen test
methods, which were submitted by the European Centre for the Validation of
Alternative Methods (ECVAM):
- The Human Whole Blood/IL-1 In Vitro Pyrogen Test (WB/IL-1)
- The Human Whole Blood/ IL-1 In Vitro Pyrogen Test Using
Cryopreserved Human Whole Blood (cryo WB/IL-1)
- The Human Whole Blood/IL-6 In Vitro Pyrogen Test (WB/IL-6)
- In Vitro Pyrogen Test Using Human PBMCs (PBMC/IL-6)
- An Alternative In Vitro Pyrogen Test Using the Human Monocytoid
Cell Line MONO MAC6 (MM6/IL-6)
The proposed in vitro pyrogen test methods involve the measurement of
cytokine levels (either IL-1β or IL-6) from human blood cells or a human monocytoid cell
line. Increased cytokine release is used as a biomarker of a pyrogenic response. The reliability
and relevance of each test method was evaluated with pyrogen-free parenteral
pharmaceuticals spiked with different concentrations of an endotoxin standard. Using a prediction
model developed based on in vivo rabbit data, results from each test method were compared
to the ''true status'' of the samples.
A public meeting of an independent scientific peer review Panel was convened on
February 6, 2007 to evaluate the validation status of these five in vitro pyrogen test methods.
These test methods are proposed as a substitute for the Rabbit Pyrogen Test (RPT) for the identification
of Gram-negative endotoxin on a case-by-case basis, subject to product-specific validation.
The peer review report from this meeting, entitled
Peer Review Panel Report: Current Validation Status of
Five In Vitro Pyrogen Test Methods, is now available.
Draft ICCVAM Test Method Recommendations
- Draft Test Method Uses and Draft Performance Standards -
PDF
- Draft Recommended Protocols
- The Human Whole Blood (WB)/IL-1 In Vitro Pyrogen Test:
Application of Cyropreserved Human WB -
PDF
- An Alternative In Vitro Pyrogen Test Using the Monocytoid
Cell Line Mono Mac 6 (MM6)/IL-6 -
PDF
- The Human Peripheral Blood Mononuclear Cell (PBMC)/IL-6 In Vitro
Pyrogen Test -
PDF
- The Human WB/IL-1 In Vitro Pyrogen Test -
PDF
- The Human WB/IL-6 In Vitro Pyrogen Test -
PDF
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