Differential expression of circulating miRNAs after alemtuzumab induction therapy in lung transplantation

Abstract

Alemtuzumab is a monoclonal antibody targeting CD52, used as induction therapy after lung transplantation (LTx). Its engagement produces a long-lasting immunodepletion; however, the mechanisms driving cell reconstitution are poorly defined. We hypothesized that miRNAs are involved in this process. The expression of a set of miRNAs, cytokines and co-signaling molecules was measured with RT-qPCR and flow cytometry in prospectively collected serum samples of LTx recipients, after alemtuzumab or no induction therapy. Twenty-six LTx recipients who received alemtuzumab and twenty-seven matched LTx recipients without induction therapy were included in the analysis. One year after transplantation four miRNAs were differentially regulated: miR-23b (p = 0.05) miR-146 (p = 0.04), miR-155 (p < 0.001) and miR-486 (p < 0.001). Expression of 3 miRNAs changed within the alemtuzumab group: miR-146 (p < 0.001), miR-155 (p < 0.001) and miR-31 (p < 0.001). Levels of IL-13, IL-4, IFN-γ, BAFF, IL-5, IL-9, IL-17F, IL-17A and IL-22 were different one year after transplantation compared to baseline. In no-induction group, concentration of sCD27, sB7.2 and sPD-L1 increased overtime. Expression of miR-23b, miR-146, miR-486, miR-155 and miR-31 was different in LTx recipients who received alemtuzumab compared to recipients without induction therapy. The observed cytokine pattern suggested proliferation of specific B cell subsets in alemtuzumab group and co-stimulation of T-cells in no-induction group.

Conflict of interest statement

The authors declare no competing interests.

© 2022. The Author(s).

Figures

Figure 1

Expression difference of the analyzed dysregulated miRNAs at baseline and one year after induction therapy within the same groups. The Figure shows box-plots depicting miRNA expressions at the two timepoints. Both groups are represented on the graph next to each other. Mann–Whitney test was used to test the difference in expression between the two time points within the same group. In alemtuzumab group miR-155 and miR-146 are significantly upregulated compared to baseline levels. On the contrary, miR-31 is drastically downregulated. The boxes indicate interquartile range, the line across the boxes the median and the lower and upper margin of the vertical lines the minimum and maximum, respectively. *p = 0.05, **p < 0.01, *** p < 0.001.

Figure 2

Expression difference of the analyzed dysregulated miRNAs between the two study groups at one year after induction therapy. The Figure shows box-plots depicting miRNA expressions of the two groups at the same timepoint, 12 months after induction therapy. Mann–Whitney test was used to test the difference in expression between the two groups at the same timepoint. MiR-155, miR-146 and miR-23b are significantly upregulated compared to no-induction group. On the contrary, miR-486 is mildly downregulated. The boxes indicate interquartile range, the line across the boxes the median and the lower and upper margin of the vertical lines the minimum and maximum, respectively. *p = 0.05, **p < 0.01, ***p < 0.001.

Figure 3

Expression difference of the measured cytokines within control group at baseline and one year after induction therapy. The Figure shows box-plots depicting cytokine expressions at the two timepoints measured with flow cytometry. IL-13, IL-4, IL-5, IL-9, IL-17F, IL-22 are significantly downregulated compared to baseline levels. Interferon-γ, B cell activating factor (BAFF), sCD27, B7.2 and PD-L1 are significantly upregulated. The boxes indicate interquartile range, the line across the boxes the median and the lower and upper margin of the vertical lines the minimum and maximum, respectively. *p = 0.05, **p < 0.01, ***p < 0.001.

Figure 4

Expression difference of the measured cytokines within alemtuzumab group at baseline and one year after induction therapy. The Figure shows box-plots depicting cytokine expressions at the two timepoints measured with flow cytometry. IL-13, IL-4, IL-17A, IL-9, IL-22 are significantly downregulated compared to baseline levels. Interferon-γ and B cell activating factor (BAFF) are significantly upregulated. The boxes indicate interquartile range, the line across the boxes the median and the lower and upper margin of the vertical lines the minimum and maximum, respectively. *p = 0.05, **p < 0.01, ***p < 0.001.