Prolactin Increases the Frequency of Follicular T Helper Cells with Enhanced IL21 Secretion and OX40 Expression in Lupus-Prone MRL/lpr Mice

Figure 2

PRL receptor expression on T_naïve and T_FH cells. Splenocytes from 9- and 18-week-old C57BL/6 and MRL/lpr mice were stained with anti-CD4, anti-CXCR5, and anti-PD1 for T_FH cells (as shown in Figure 1(c)) and with anti-CD4, anti-CD44, and anti-CD62L for T_naïve cells, then cells were stained with an anti-PRL receptor antibody. (a) Demonstration of the gating strategy for the flow cytometry analysis of T_naïve cells. Doublets were excluded by gating on FSC-H×FSC-A, lymphocytes were identified on the basis of their scatter properties (FSC-A×SSC-A plot), and live cells were gated in the Ghost Red⁻. The gate of CD4⁺ T_naïve was selected (CD62L⁺ CD44⁻). (b) Expression of the PRL receptor is reported as the fold change in receptor expression with respect to PRL receptor expression in T_naïve cells. (c) Representative histogram of PRL receptor expression in MRL/lpr mouse cells. The measurement was carried out in duplicate in six mice per group. Pooled data are presented as the ; using ANOVA. (d) T_naïve and T_FH cells from 18-week-old MRL/lpr mice were purified by Sort, and the isoform of the PRL receptor was determined by real-time (RT-) PCR. The murine breast cancer cell line EpH4 1424 was used as a positive control for the expression of the long and short PRL receptor isoforms (not shown). Two different experiments were performed; in each experiment, a pool of cells isolated from three mice was used.

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