T cell dependent antibody response (TDAR) assay

Hooke has extensive experience measuring the T cell dependent antibody response (TDAR). This assay is commonly run in C57BL/6 and B6C3F1 mice to evaluate the effects of test compounds on antibody production. The assay is also used for immunotoxicity assessment.

To induce TDAR, mice are immunized with keyhole limpet hemocyanin (KLH), a highly immunogenic antigen, which induces both T cell-mediated (cellular) and B cell-mediated (humoral) immune responses. This assay can be used to evaluate antibody production, germinal center formation, antibody class switching, and immunotoxicity. Haptenated KLH (for example, NP-KLH) is also used to evaluate the TDAR response.

The advantage of KLH as a model antigen is that it induces a T cell-dependent antibody response without the use of an adjuvant.

Treatment with test compounds starts at immunization, and antibody production is typically measured between 7 and 28 days post-immunization. Typically, both IgM and IgG antibodies are measured in serum.

Induction of TDAR in C57BL/6 mice

Induction of anti-KLH response in C57BL/6 mice

Representative anti-KLH results from T cell-dependent antibody response (TDAR) assay

anti-KLH IgM typical results anti-KLH IgG typical results

C57BL/6 mice were immunized with KLH or PBS and received vehicle (PBS) daily for 14 days. Serum was collected at 7 (blue) and 14 (pink) days post-immunization. Results represent anti-KLH IgM and IgG in units/mL +/- SEM. Student's T-test was calculated between each vehicle and KLH pair. *** p<0.001

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References

Swaminathan A et al, Br J Clin Pharmacol 78:1135 (2014)
White K et al, J Immunotoxicology 4:2, 153 (2007), DOI: 10.1080/15476910701337688