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Humanized Mediator Release Assay as a Read-Out for Allergen Potency
JoVE Journal
Immunology and Infection
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JoVE Journal Immunology and Infection
Humanized Mediator Release Assay as a Read-Out for Allergen Potency

Humanized Mediator Release Assay as a Read-Out for Allergen Potency

DOI:
10.3791/62702-v

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10:22 min

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June 29, 2021

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, , ,
1Department of Biosciences,University of Salzburg

Chapters

  • 00:04Introduction
  • 01:04Complement Inactivation of Human Sera
  • 02:05Humanized-RBL Cell Culture
  • 04:07Passive Sensitization of Humanized-RBL Cells
  • 04:57Antigen-stimulated Degranulation and Mediator Release
  • 06:19β-hexosaminidase Activity Measurement
  • 07:21Results: Effect of Allergen on β-hexosaminidase Release
  • 09:38Conclusion

Summary

Automatic Translation

Automatic Translation

Here we present the mediator release assay, using a rat basophilic leukemia cell line transfected with the human IgE receptor, to simulate the degranulation of effector cells typically observed in type 1 allergic reactions. This method investigates the biological activity of allergens in a highly sensitive, reproducible, and tailorable manner.

Tags

Keywords: Humanized Mediator Release AssayAllergen PotencyType 1 Allergic ReactionsRecombinant AllergenPurified AllergenAllergen ExtractIgE ReactivityCross-reactivityAllergen Immunotherapy (AIT) Treatment EfficacyAg8 CellsHumanized RBL CellsCell CultureCell SuspensionCell Counting96-well Plate

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