Using DR52c/Ni2+ mimotope tetramers to detect Ni2+ reactive CD4+ T cells in patients with joint replacement failure

Abstract

T cell mediated hypersensitivity to nickel (Ni2+) is one of the most common causes of allergic contact dermatitis. Ni2+ sensitization may also contribute to the failure of Ni2+ containing joint implants, and revision to non-Ni2+ containing hardware can be costly and debilitating. Previously, we identified Ni2+ mimotope peptides, which are reactive to a CD4+ T cell clone, ANi2.3 (Vα1, Vβ17), isolated from a Ni2+ hypersensitive patient with contact dermatitis. This T cell is restricted to major histocompatibility complex class II (MHCII) molecule, Human Leukocyte Antigen (HLA)-DR52c (DRA, DRB3*0301). However, it is not known if Ni2+ induced T cell responses in sensitized joint replacement failure patients are similar to subjects with Ni2+ induced contact dermatitis. Here, we generated DR52c/Ni2+ mimotope tetramers, and used them to test if the same Ni2+ T cell activation mechanism could be generalized to Ni2+ sensitized patients with associated joint implant failure. We confirmed the specificity of these tetramers by staining of ANi2.3 T cell transfectomas. The DR52c/Ni2+ mimotope tetramer detected Ni2+ reactive CD4+ T cells in the peripheral blood mononuclear cells (PBMC) of patients identified as Ni2+ sensitized by patch testing and/or a positive Ni2+ LPT. When HLA-typed by a DR52 specific antibody, three out of four patients were DR52 positive. In one patient, Ni2+ stimulation induced the expansion of Vβ17 positive CD4+ T cells from 0.8% to 13.3%. We found that the percentage of DR52 positivity and Vβ17 usage in Ni2+ sensitized joint failure patients are similar to Ni sensitized skin allergy patients. Ni2+ independent mimotope tetramers may be a useful tool to identify the Ni2+ reactive CD4+ T cells.

The T Cell Repertoires from Nickel Sensitized Joint Implant Failure Patients

“…This technique is being increasingly used to identify pathogenic T cells in many different immune-mediated diseases [ 28 , 29 ]. In this study, we showed several highly expanded T cell clones by sequencing data, and demonstrated the preferential usage of TRBV19, TRBV5-1, and TRBV20-1, consistent with previous studies on Ni 2+ contact dermatitis [ 11 , 16 , 18 , 20 ]. Our study suggests, then, that the certain population of Ni 2+ -reactive T cells in joint implant failure may be shared with the T cells responsible for Ni 2+ induced contact dermatitis.…”

“…PBMC preparation and storage were performed according to previously established methods [ 18 ]. In addition, 10 7 PBMC cells were thawed from P7 and P9 patients, cultured in enriched MEM as previously described [ 32 ], and supplied with 10% heat-inactivated premium FBS (VWR, PA, USA, product 97068-085, lot 214B17).…”

“…We previously found that around 1% of peripheral blood T cells from several Ni-sensitized joint failure patients interact with the DR52c-mimotope. In addition, the percentage of TRBV19 CD4 T cells increases from 0.76% to 13.3% when stimulated with Ni 2+ in vitro [ 18 ]. This suggests that the TRBV19 TCR may also play an important role in inducing Ni 2+ sensitization in periprosthetic tissue after arthroplasty.…”

Identification and Characterization of Circulating Naïve CD4+ and CD8+ T Cells Recognizing Nickel

“…For instance, TRBV27, TRBV28, and TRBV6.5 were also detected in nickel-specific T cells isolated from allergic patients (18, 25). Moreover, it was demonstrated that TRBV19 gene's over-expression was correlated with the individuals reactivity to nickel (17, 18, 24, 51). However, we could not conclude that nickel priming induces enrichment and/or preferentially expansion of TCRβ clones carrying TRBV19 since this gene was highly represented prior to stimulation.…”