Figure 7.

IL-32 expression promotes a more immature plasma cell phenotype

(A) Venn-diagram of overlapping significant genes (p <0.01) that were more highly expressed in WT cells compared with KO cells (comparing two INA-6 KO clones [KO1, KO2] with WT mock cells) and upregulated in IL-32 patients (comparing IL-32- expressing patients versus nonexpressing patients). See also Table S5.

(B) Gene expression of MME, IRF8, and SORL1 in patients expressing IL-32 (10th percentile) compared with nonexpressing (90th percentile) patients. Significance determined by limma in R.

(C) Gene expression of MME, IRF8, and SORL1 in INA-6 IL-32 KO1, KO2, and WT mock cells. Significance determined by limma in R with Benjamini-Hochberg-adjusted p-values. Data presented are mean cpm ± SD of two replicates.

(D) Evaluation of gene expression of markers associated with less differentiated stages of B cell maturation in CoMMpass IA13, comparing IL-32 expressing patients (upper 10th percentile) with nonexpressing patients (lower 90th percentile). Significance determined by limma in R. Boxplots show the median and 25th/75th quantiles and smallest/largest value within the 1.5 times interquartile rang.

(E) Scatterplot of genes associated with less differentiated stages of B cell maturation in single cells with (N = 142) and without (N = 346) IL32-expression (from single cell transcriptomics). p values were calculated using the FindMarkers function in Seurat by comparing the high and low IL32 groups.

(F) Surface expression of CD45 and CD38 in INA-6 KO and WT cells. Data are presented as median fluorescence intensity (MFI) from 3 independent experiments and significance determined by unpaired student's t test. Bare plots show mean ± SEM.

(G) Concentration of kappa light chain/cell detected in conditioned media from WT and KO cells as indicated. p values were calculated by the ratio paired t test. ns, not significant; ∗p ≤0.05, ∗∗p ≤0.01, ∗∗∗p ≤0.001, ∗∗∗∗p ≤0.0001.