Skip to main content

The T-Dependent Antibody Response to Keyhole Limpet Hemocyanin in Rodents

  • Protocol
  • First Online:
Immunotoxicity Testing

Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 598))


Central to the evaluation of potential immunotoxicants is the concept that measurement of multiple parameters is required for the determination of toxicity toward the immune system. A carefully considered integration of endpoints involved in the immune response should be used to determine an immunotoxic effect. A functional evaluation, specifically the rodent T-cell-dependent antibody response (TDAR) model developed for regulated immunotoxicity evaluations, has been established to detect potential immunotoxicity, especially immunosuppression, caused by chemicals and novel pharmaceuticals in development. This chapter provides an overview and detailed procedures involved in the TDAR assay that measures the immune response (i.e., antibody production) to an introduced antigen (i.e., keyhole limpet hemocyanin (KLH)) in rats or mice treated with a chemical (e.g., a known immunotoxicant and/or a new drug candidate). The TDAR model of competent immune function requires the participation of multiple effector cells such as antigen presenting cells, T lymphocytes, and B lymphocytes to produce the final product, the antigen-specific antibody response. Thus, alterations in the level of antibody production to the specific antigen may reflect effects on any or all of the cell populations involved in TDAR.

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
USD 84.99
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 169.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 109.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions


  1. Dean JH, House RV, Luster MI (2007) Immunotoxicology: effects of and response to drugs and chemicals. In: Wallace Hayes A (ed) Principles and methods of toxicology, 5th edn. Taylor and Francis, Philadelphia, pp 1761–1793

    Google Scholar 

  2. Gore ER, Gower J, Kurali E, Sui J-L, Bynum J, Ennulat D, Herzyk DJ (2004) Primary antibody response to keyhole limpet hemocyanin in rat as a model for immunotoxicity evaluation. Toxicology 197:23–35

    Article  CAS  PubMed  Google Scholar 

  3. Temple L, Kawabata TT, Munson AE, White KL (1993) Comparison of ELISA and plaque-forming cell assays for measuring the humoral immune response to SRBC in rats and mice treated with benzo(a)pyrene or cyclophosphamide. Fund Appl Toxicol 21:412–419

    Article  CAS  Google Scholar 

  4. Kim CJ, Berlin JA, Bugelski PJ, Haley P, Herzyk DJ (2007) Comparison of immune functional tests using T-dependent antigens in immunotoxicology studies: a meta-analysis. Per Exp Clin Immunotoxicol 1:60–73

    Google Scholar 

  5. International Conference on harmonisation of technical requirements for registration of pharmaceuticals for human use (2006) ICH harmonised tripartite guideline: immunotoxicity studies for human pharmaceuticals S8. (

Download references


The authors thank Mark Thompson and Lyudmila Denisova for their technical assistance and Cindy Pauley for establishing the described ELISA methods for the detection of anti-KLH antibodies at Merck and Co., Inc.

Author information

Authors and Affiliations


Corresponding author

Correspondence to Danuta J. Herzyk .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2010 Humana Press, a part of Springer Science+Business Media, LLC

About this protocol

Cite this protocol

Plitnick, L.M., Herzyk, D.J. (2010). The T-Dependent Antibody Response to Keyhole Limpet Hemocyanin in Rodents. In: Dietert, R. (eds) Immunotoxicity Testing. Methods in Molecular Biology™, vol 598. Humana Press.

Download citation

  • DOI:

  • Published:

  • Publisher Name: Humana Press

  • Print ISBN: 978-1-60761-400-5

  • Online ISBN: 978-1-60761-401-2

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics