A Specific Blood Signature Reveals Higher Levels of S100A12: A Potential Bladder Cancer Diagnostic Biomarker Along With Urinary Engrailed-2 Protein Detection.

Urothelial carcinoma of the urinary bladder (UCB) or bladder cancer remains a major health problem with high morbidity and mortality rates, especially in the western world. UCB is also associated with the highest cost per patient. In recent years numerous markers have been evaluated for suitability in UCB detection and surveillance. However, to date none of these markers can replace or even reduce the use of routine tools (cytology and cystoscopy). Our current study described UCB's extensive expression profile and highlighted the variations with normal bladder tissue. Our data revealed that JUP, PTGDR, KLRF1, MT-TC, and RNU6-135P are associated with prognosis in patients with UCB. The microarray expression data identified also S100A12, S100A8, and NAMPT as potential UCB biomarkers. Pathway analysis revealed that natural killer cell mediated cytotoxicity is the most involved pathway. Our analysis showed that S100A12 protein may be useful as a biomarker for early UCB detection. Plasma S100A12 has been observed in patients with UCB with an overall sensitivity of 90.5% and a specificity of 75%. S100A12 is highly expressed preferably in high-grade and high-stage UCB. Furthermore, using a panel of more than hundred urine samples, a prototype lateral flow test for the transcription factor Engrailed-2 (EN2) also showed reasonable sensitivity (85%) and specificity (71%). Such findings provide confidence to further improve and refine the EN2 rapid test for use in clinical practice. In conclusion, S100A12 and EN2 have shown potential value as biomarker candidates for UCB patients. These results can speed up the discovery of biomarkers, improving diagnostic accuracy and may help the management of UCB.

Novel human recombinant antibodies against Mycobacterium tuberculosis antigen 85B.

BACKGROUND: Tuberculosis is the leading cause of death due to bacterial infections worldwide, mainly caused by Mycobacterium tuberculosis. The antigen 85 complex comprises a set of major secreted proteins of M. tuberculosis, which are potential biomarkers for diagnostic. RESULTS: In this work, the first human single chain fragment variable (scFv) antibodies specific for the tuberculosis biomarker 85 B were selected by phage display from naïve antibody gene libraries (HAL7/8). Produced as scFv-Fc in mammalian cells, these antibodies were further characterized and analysed for specificity and applicability in different tuberculosis antigen detection assays. Sandwich detection of recombinant 85 B was successful in enzyme linked immunosorbent assay (ELISA), lateral flow immunoassay and immunoblot. Whereas detection of M. tuberculosis cell extracts and culture filtrates was only possible in direct ELISA and immunoblot assays. It was found that the conformation of 85 B, depending on sample treatment, influenced antigen detection. CONCLUSIONS: Recombinant antibodies, selected by phage display, may be applicable for 85 B detection in various assays. These antibodies are candidates for the development of future point of care tuberculosis diagnostic kits. Using 85 B as a biomarker, the antigen conformation influenced by sample treatment is important.

Serum anti-mycobacterial heat shock proteins antibodies in sarcoidosis and tuberculosis.

Sarcoidosis (SA) is a multisystem granulomatous disorder of unknown etiology. Infectious organisms, e.g. Mycobacterium tuberculosis, genetic factors and autoimmunity are considered as etiologic agents. Pathologic similarities between SA and tuberculosis (TB) suggest M. tuberculosis heat shock proteins (Mtb-hsp) as causative factors. To test the "mycobacterial" origin of SA, we evaluated the presence of serum anti-Mtb-hsp70, -Mtb-hsp65 and -Mtb-hsp16 antibodies in SA and TB. Analysis of anti-Mtb-hsp antibodies was carried out in 37 patients with SA, 29 patients with TB and 18 healthy individuals by ELISA. Our results show a significantly higher occurrence of anti-Mtb-hsp70 antibodies in SA and TB patients than in controls. The anti-Mtb-hsp65 and -Mtb-hsp16 antibodies occured significantly more often in TB than in controls and SA. We found significantly higher percentages of anti-Mtb-hsp70 and -Mtb-hsp65 antibodies in Stage II compared to Stage I of SA. Analysis of anti-Mtb-hsp antibodies levels revealed significantly higher anti-Mtb-hsp70 antibodies level in SA and TB than in controls. Significantly higher frequency and levels of anti-Mtb-hsp70 than anti-Mtb-hsp65 and -Mtb-hsp16 antibodies was found in SA only. In summary, the frequency and level of anti-Mtb-hsp70 antibodies were comparable between SA and TB but were significantly higher compared to controls and other tested Mtb-hsp. These data may suggest a role for Mtb-hsp70 protein in the pathogenesis of sarcoidosis.