T cell epitope recognition involved in the low-responsiveness to a region of hen egg lysozyme (46-61) in C57BL/6 mice

Abstract

The predominant T cell epitope of hen egg lysozyme (HEL) in high-responder C3H mice has been previously identified as the HEL 46-61 region. In contrast, this region is poorly recognized by T cells from low-responder C57BL/6 mice upon immunization with HEL. In previous studies, we have demonstrated that several C57BL/6 derived T cell hybridomas reactive to this epitope and other HEL epitopes preferentially recognize phosphorylcholine (PC)-conjugated HEL over unconjugated HEL. To understand the mechanisms involved in this difference of T cell recognition, we have further analysed the reactivity of T cells and T cell hybridomas from low-responder C57BL/6 mice. T cells from HEL-immunized mice were preferentially reactive to HEL 47-60. These results suggest a potential deficiency in generating an appropriate T cell epitope from the 46-61 region of native HEL in low-responder C57BL/6 mice. The minimal T cell epitope of this region was defined as HEL 51-60 using the PCH4.1 T hybridoma clone. This minimal epitope represents a single amino acid shift from the minimal epitope of HEL high-responder C3H mice (HEL 52-61). Various peptides representing this region were synthesized with single alanine substitutions at each position. The residues at positions 51, 52, 53 and 57 of HEL appear to be involved in Ia binding and the residues at 55 and 56 in contracting the TCR. T cell reactivity to HEL 51-61 peptides with various substitutions at position 61 strongly suggest that primarily the size of the C-terminal residue interferes with binding to the Ia molecules of low-responder mice. In addition, substitutions of the TCR contacting residues at positions 55 and 56 with similar residues (isoleucine-->leucine or leucine-->isoleucine) significantly increased the T cell reactivity, suggesting a low reactivity with the native residues. Therefore, the requirement of many residues in the T cell epitope for interaction with Ia, the necessity for additional Ag processing to facilitate Ia binding, and the low affinity of the TCR contacting residues may together render C57BL/6 mice unresponsive to the HE 46-61 region.