cd30 human instant elisa kit  (Thermo Fisher)

Journal: PLoS Pathogens

Article Title: Immunosuppressive FK506 treatment leads to more frequent EBV-associated lymphoproliferative disease in humanized mice

doi: 10.1371/journal.ppat.1008477

Figure Lengend Snippet: A) RNA expression profiling of the human B cell transcriptome of mice from three independent experiments with EBV − FK506 – (n = 5, clear), EBV − FK506 + (n = 6, orange), EBV + FK506 – (n = 7, purple) and EBV + FK506 + (n = 8, magenta). Principal component plot: Sample similarity in a 2D projection by multi-dimensional scaling. Spurious batch effects of unknown origin and HFL donor-specific effects were corrected by surrogate variable analysis. The two principal components represent 60% of the total variance in the data set. B) Volcano plots: The horizontal line corresponds to FDR = 0.01 and genes below this line are labeled ‘not significant’. The two vertical lines correspond to a 2-fold change in expression and genes outside this range are labelled ‘strong’. The legend refers to ‘not’—not significant and weak effect; ‘signif & strong’—significant and strong effect; ‘signif’—significant but weak effect; ‘strong’—not significant but strong effect. C) Normalized AmpliSeq expression levels of CXCL10 , CD28 , IL6 and TNFRSF8 (CD30) are depicted for individual mice with mean ± SEM and FDR-corrected p-value summaries (*: FDR-p<0.05, **: FDR-p<0.01, ***: FDR-p<0.001.) D) Upset plot identifies private, shared and promiscuous DEGs among groups. The vertical bar chart indicates the intersection size of transcripts shared between certain groups defined by solid points below the chart. The total number of DEGs for each condition is depicted in the horizontal bar chart. See also – Figs and and Tables.

Article Snippet: Commercially available ELISA kits (CD30 Human Instant ELISA Kit, ThermoFisher Scientific; lens epithelial protein (LENEP) ELISA Kit, MyBiosource; PRR4 ELISA Kit (Proline Rich 4, Lacrimal), Cloud-Clone Corp.) were used to determine the concentration of sCD30, PRR4 and LENEP in the serum of individual mice or patients and in the supernatants of cultured LCLs according to the manufacturer’s instructions.

Techniques: RNA Expression, Labeling, Expressing

Journal: PLoS Pathogens

Article Title: Immunosuppressive FK506 treatment leads to more frequent EBV-associated lymphoproliferative disease in humanized mice

doi: 10.1371/journal.ppat.1008477

Figure Lengend Snippet: A-C) Protein concentration of sCD30 was measured in serum samples with ELISA. A) Serum was obtained at the day of sacrifice from animals from two independent experiments. B) Post hoc stratification of FK506-treated mice with or without macroscopically visible tumors with EBV-infection status indicated by black (EBV + ) or clear (EBV – ) symbols. A-B) Mean ± SEM, MWT. Dashed lines indicate quantification thresholds. C) sCD30 was measured in the sera of 31 healthy children undergoing elective tonsillectomy, 21 children that presented with non-EBV associated fever (Febrile non-EBV, EBV VCA-IgM negative) and 13 pediatric PTLD patients around diagnosis (PTLD Acute) and again at least 36 months later (PTLD Recovered) (See and Tables). Dashed line represents the median of the healthy controls. MWT and Wilcoxon matched-pairs test. D) Representative immunohistochemistry staining of CD30 (red) and PAX5 (brown), or CD30 (red) and EBNA2 (brown), in splenic sections or tumor tissue of huNSG-A2 mice. Scale bars: 50μm. Quantification of CD30 + (n = 7) and CD30 + EBNA2 + cells (n = 6) per mm 2 in tumor sections and non-tumorous spleen tissue of EBV-infected mice is depicted for two independent experiments. E) Cytokine concentration was measured in the serum of EBV-PTLD patients (n = 8) at diagnosis (acute) and again at recovery time point at least 36m post-diagnosis and in FK506-treated EBV − (n = 8) and EBV + (n = 10) experimental mice, composite data from two independent experiments. Upper panel: Heatmaps and hierarchical clustering of pediatric patients or huNSG-A2 mice based on serum cytokine levels (IFNγ, TNFα, IL-6, IL-8 and IL-10) at diagnosis (red) and recovery (blue) for PTLD (left) and on the day of sacrifice for huNSG-A2 mice (right) treated with FK506 (blue) and additionally EBV-infected (red). Tumor presence versus absence in huNSG-A2 mice is indicated in black and grey respectively. Lower panel: Cytokine concentration (median (IQR) with min and max range as whiskers) in the serum of EBV-PTLD patients and in FK506-treated EBV − and EBV + experimental mice. Wilcoxon matched-pairs test and MWT respectively. *P < 0.05, **P < 0.01, ***P < 0.001. See also and and Tables.

Article Snippet: Commercially available ELISA kits (CD30 Human Instant ELISA Kit, ThermoFisher Scientific; lens epithelial protein (LENEP) ELISA Kit, MyBiosource; PRR4 ELISA Kit (Proline Rich 4, Lacrimal), Cloud-Clone Corp.) were used to determine the concentration of sCD30, PRR4 and LENEP in the serum of individual mice or patients and in the supernatants of cultured LCLs according to the manufacturer’s instructions.

Techniques: Protein Concentration, Enzyme-linked Immunosorbent Assay, Infection, Biomarker Discovery, Immunohistochemistry, Staining, Concentration Assay