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.
Typical results from T cell-dependent antibody response (TDAR) assay
Below are typical results from a TDAR assay using cells from C57BL/6 mice.
Baseline represents results from 16 mice with serum collected 1 day prior to immunization (Day -1). The KLH group includes 10 mice immunized with KLH, and the PBS group includes 10 mice immunized with PBS.
Both groups received i.v. vehicle daily beginning on Day 0. Serum was collected on the indicated days post-immunization.
A two-tailed Student's t test was calculated between the PBS and KLH groups for each day that serum was collected. *p<0.05, **p<0.01, ***p<0.001.
200 µg KLH immunization, Day 0 + 200 µg KLH booster, Day 14
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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
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