Figure 5.

Patient clustering by top-weighted immunological parameters in patients with juvenile-onset SLE compared with healthy controls

(A) Top-weighted immunological parameters of patients with juvenile-onset SLE (appendix pp 11–12) were stratified using k-means clustering. Immunophenotype is standardised within each column by Z score and plotted as a heat map, representing the relationship to the mean of the group (red represents relatively high frequency and blue represents relatively low frequency). Each row represents a patient with juvenile-onset SLE. Four groups of patients were recognised with distinct immune cell profiles. (B) Scatter dot plots displaying top-weighted immunological parameters between the k-means clustered groups. Mean (error bars indicate SE) was calculated with one-way ANOVA, and p values were calculated with Tukey's multiple comparisons test. The dashed lines represent the mean for the healthy control population for each cell type. (C) sPLS-DA plot showing the clustering of the validated top-weighted immunological parameters in patients with juvenile-onset SLE between k-means clustered juvenile-onset SLE groups. Individual distribution points and confidence ellipses (ovals) are plotted for each group. (D) Using this analysis, the weighting of each cell type in component 1 and 2 is displayed, where the inner circle is the 0·5 cutoff. (E) Box and whisker plots displaying baseline measures over 3–7 years of follow-up of clinical measures of disease activity between the k-means clustered groups of patients with juvenile-onset SLE. (F) Average measure over 3–7 years of follow-up of clinical measures of disease activity between the k-means clustered groups of patients with juvenile-onset SLE. Mean (error bars indicate SE) was calculated with one-way ANOVA, and p values were calculated with Tukey's multiple comparisons test. Dashed lines represent the clinical cutoff for active disease in the C3 plot and the assigned cutoff associated with active lupus in the SLEDAI-2K plot. (G) Box and whisker plot displaying longitudinal disease activity, using the same dataset from panel F, assessed as LLDAS. Mean (error bars indicate SE) was calculated with one-way ANOVA, and p values were calculated with Tukey's multiple comparisons test. (H) Individual patient trajectory of SLEDAI-2K and C3 over 15 clinical encounters displayed as spaghetti plots. Each line represents one patient with juvenile-onset SLE. Smoothing lines were added to indicate the trend of juvenile-onset SLE groups from previous k-means clustering. C3=complement component C3. iNKT=invariant natural killer T cells. LLDAS=lupus low disease activity state. SLE=systemic lupus erythematosus. SLEDAI-2K=systemic lupus erythematosus disease activity index 2000. sPLS-DA=sparse partial least squares-discriminant analysis.