Journal: Communications Medicine
Article Title: The clinical significance of the Mediterranean fever gene MEFV variants in Castleman disease
doi: 10.1038/s43856-026-01392-1
Figure Lengend Snippet: A Sanger sequencing confirmed the presence of MEFV variants in accordance with WES. Sequence alignment was performed using the BioEdit 7.0.9 software. B Pedigree chart of affected iMCD-TAFRO adolescence, diagnosed with iMCD-TAFRO at 15 years old, rectangle in red. C Quantitative PCR showed that the expression of MEFV mRNA in peripheral blood mononuclear cells (PBMCs) decreased as the number of mutations increased. GAPDH was used as an internal control. N = 3, PBMCs of individuals were collected and tested in triplicate. Means and standard deviations were shown. Turkey was used for multiple comparisons following one-way ANOVA analysis. * p = 0.013, *** p < 0.001. D LPS plus ATP stimulation induced cell aggregation of PBMCs. More remarkable aggregations were observed as the number of MEFV mutations increased, as the MEFV E148Q/E148Q-P369S/WT-R408Q/R408Q PBMCs of the TAFRO patient showed the most severe aggregations. LPS, lipopolysaccharide. ATP, Adenosine Triphosphate. Scale bar size 100×. E Quantitative PCR revealed a sharp increase in IL-6 and IL-1βmRNA levels following LPS plus ATP stimulus. MEFV variants further amplified the up-regulation of IL-6 and IL-1β, as cells from a TAFRO patient bearing MEFV E148Q/E148Q-P369S/WT-R408Q/R408Q achieved the highest mRNA expression level. N = 3, stimulation experiments were performed in triplicate. Means and standard deviations were shown. Turkey was used for multiple comparisons following one-way ANOVA analysis. ** P = 0.008, *** P < 0.001, NS not significant P = 1.000. F Cytokines assay using Luminex demonstrated inflammatory activation in PBMCs with MEFV variants, particularly in the TAFRO patient. N = 2, means were shown. VEGF vascular endothelial growth factor. G Colchicine inhibited LPS-induced cytokine release in PBMCs of TAFRO. N = 2, means were shown. TNF-α tumor necrosis factor-α. H Pie chart showing the high prevalence of MEFV variations in 37 CD. I Details of the MEFV variants in 37 CD. UCD Unicentric Castleman Disease, aMCD asymptomatic Multicentric Castleman Disease, iMCD idiopathic Multicentric Castleman Disease, NOS Not Otherwise Specified, IPL Idiopathic Plasmacytic Lymphadenopathy, TAFRO thrombocytopenia, anasarca, fever, reticulin fibrosis, organomegaly. J E148Q, P369S, and R408Q ranked the top 3 variants of MEFV among 6 variants identified in 37 CD. The allele frequency of MEFV variants from East Asian populations was shown in parallel. The binomial test was used for comparison. E148Q *** P < 0.001, P369S ** P = 0.002, R408Q *** P < 0.001, L110P ** P = 0.006, R202Q P = 0.601, G304R P = 0.564. NS not significant. a, data from gnomAD v4.1.0. K MEFV E148Q-P369S-R408Q-positive CD exhibited a more severe disease course compared with the negative ones. Means were shown. For those that conform to the normal distribution, the independent samples two-sided t -test was used; otherwise, the Mann–Whitney U test was adopted. WBC ** P = 0.001, Hb ** P = 0.002, PLT * P = 0.018, CRP * P = 0.049, ALB * P = 0.014, eGFR * P = 0.014. WBC white blood cell, Hb hemoglobin, PLT platelet, CRP C-reactive protein, ALB albumin, eGFR estimated Glomerular Filtration Rate.
Article Snippet: Sequence alignment was performed using the BioEdit 7.0.9 software.
Techniques: Sequencing, Software, Real-time Polymerase Chain Reaction, Expressing, Control, Amplification, Luminex, Activation Assay, Comparison, MANN-WHITNEY, Filtration
