T cell clonality and characteristics of CDR3 sequences in TCR clonotypes bearing AV18-1 and BV8-2.

<p>T cell clonality was analyzed by CDR3 size spectratyping and nucleotide acid sequences were determined in TCR clonotypes bearing AV18-1 and BV8-2. RNA was extracted from footpads of Pd-induced allergy mice at 7 days (n=5) and nucleotide acid sequences of CDR3 were determined in 49-52 cDNA clones per mouse. Representative profiles of CDR3 size spectratyping and deduced amino acid sequences of the CDR3 bearing AV18-1 (A) and BV8-2 (B) are shown. (A) For AV18-1, a polyclonal peak pattern (multiple peaks with Gaussian distribution) was obtained from lymph nodes of control mice. In contrast, Pd-induce allergy mice exhibited oligoclonal peak patterns with a dominant peak at the same position among five mice. Identical CDR3 sequences (†, ¶, *, Ω, Φ, ‡, Ψ) were obtained from the footpads of different mice. (B) For BV8-2, Pd-induced allergy mice showed oligoclonal peak patterns but they were different among the mice. CDR3 sequence analysis also showed multiple predominant clones presented in each mouse but they did not share CDR3 sequences among different mice. All experiments were performed in triplicate.