ABSTRACT
BACKGROUND: Even though allergies are an important health issue, wide manufacturer-dependent differences in the detected amounts of allergen-specific IgE (sIgE) have repeatedly been found. These discrepancies hinder diagnostics and research into clinically significant cutoff points for life-threatening symptoms. METHODS: To evaluate whether the reported differences have led to changes in diagnostic testing, we analyzed data from six years of round robin testing (RRT, also known as proficiency testing) at the Gesellschaft zur Förderung der Qualitätssicherung in medizinischen Laboratorien e.V. (Society for Promoting Quality Assurance in medical Laboratories) for the important allergen sources bee venom, wasp venom, and birch pollen. The results of the four main suppliers of in vitro diagnostic sIgE testing were compared in a pseudo-anonymized form using overlay images of box plot graphs for the semiquantitative data and allergen class results. Coefficients of variation (CV) were obtained to study the development of interlaboratory comparability. RESULTS: We found that the large differences between the manufacturer collectives remained constant between January 2010 and April 2015 without any real improvement. The CVs were good for two of the four analyzed suppliers, one was marginal and one above the quality level of 20%. CONCLUSION: The numerous publications that have found discrepancies in the sIgE results of the different suppliers did not change the status quo within the last six years. Unfortunately, this is unlikely to change until there is a characterized standard material with known values of sIgE.
ABSTRACT
BACKGROUND: Cervical carcinoma cells including those infected with the oncogenic human papilloma virus (HPV) and several cervical carcinoma cell lines show a strong expression of the CD40 receptor, unlike benign cervical epithelial cells infected with HPV. The functional relevance of this up-regulated expression in the tumor is not fully understood. Nevertheless, it might offer a unique possibility to target those malignant cells due to the antiviral and antitumoral effects of the CD40/CD40 ligand (CD40L) interactions. AIM: In vitro assessment of the effect of CD40L on HPV 18-P105 promoter activity and the subsequent release of IL-6 by the promoter transfected HeLa(CD40) cells, which express CD40 constitutively. MATERIAL AND METHODS: Transfection of HeLa(CD40) cells was achieved by electroporation after optimizing the parameters by the pCMV-ß-Gal vector and ß-Gal stain. Transfected HeLa(CD40) cells were challenged with BHK(CD40L) and TNFα, in addition to BHK(wt) and medium alone as controls. HPV18-P105 promoter activity was demonstrated by luciferase reporter gene assay while IL-6 was assessed by ELISA. RESULTS: CD40/CD40L interactions and TNFα treatment significantly reduced HPV18-P105 promoter activity (56.0 ± 10.2% and 64.1 ± 9.1% vs. control, respectively; p < 0.001). Likewise, IL-6, which is a sensitive cytokine of CD40 activation, was significantly increased in HeLa(CD40) cells in the same experiments (2.7 fold after stimulation with BHK(CD40L) and 5.2 fold after stimulation with TNFα vs. control; p < 0.01 and p < 0.001, respectively). CONCLUSION: It is likely that the CD40/CD40L interactions and TNFα are effective against cervical carcinomas by repressing transcriptional activity of HPV promoter. This can result in new adjuvant treatments.
