Chemokine mRNA expression induced by IPP in IPP-expanded Vδ2⁺ T cell lines as determined by RPA.

Vδ2⁺ T cell lines were established by culturing PBMCs from a healthy donor for 4 weeks in vitro, following a single stimulation with IPP (IPP 1 × ). A population of the same PBMCs was also maintained in IL-2 alone (IL-2). The cells that had been stimulated 4 weeks previously were then stimulated again with IPP (IPP 2 × ), and RNA was extracted at 4, 12, 24, and 48 hours post-stimulation. (A) The result of the RPA analysis for chemokine mRNA expression using a multiprobe RPA system (hCK5). The undigested probe set (U) and the digested probe set (D) are shown in the first 2 lanes respectively, and the control RNA (ctr) provided with the kit is shown in the extreme right-hand lane. (B) Quantitative analysis of the bands by phosphoimaging are shown and are expressed as a ratio of the gene of interest to the sum of the housekeeping genes L32 and GAPDH. Data shown are representative of 3 different healthy donors.