Seol J.-Y.,Roswell Park Cancer Institute |
Mihich E.,Roswell Park Cancer Institute |
Berleth E.S.,Roswell Park Cancer Institute |
Berleth E.S.,Immco Diagnostics
Cytokine | Year: 2015
Tumor Necrosis Factor α (TNFα) induces both the apoptotic pathway and anti-apoptotic factors. Incubation of human dermal fibroblasts with TAPF (TNF Apoptosis Protection Fraction) protects them from apoptosis induced by the subsequent addition of TNF and cycloheximide (CHX). TAPF does not protect against apoptosis induced by CHX in combination with either TRAIL (TNF related apoptosis inducing ligand) or an agonistic Fas antibody, or against apoptosis induced by the chemotherapeutic agent doxorubicin. Incubation with TAPF does not affect the quantity of TNF that binds to the cell. TAPF prevents TNF-induced cleavage of caspases 8, 9, 3 and 7 and the apoptotic substrate PARP (poly-ADP ribose polymerase), but has no effect when these molecules are induced by an agonistic Fas antibody. TAPF induces rapid phosphorylation of the NF-κB/p65 (nuclear factor-κB) transcription factor at serine 536 which is indicative of its activation. TAPF increases the expression of cFLIP (cellular FLICE-inhibitory protein) which is a potent inhibitor of apoptosis that acts by preventing the cleavage of caspase 8. This increase in cFLIP is coincident with protection from TNF-induced apoptosis. Decreasing cFLIP levels using shRNA (short hairpin RNA) decreases protection by TAPF. TAPF also induced the anti-apoptotic A20 protein. These data indicate that TAPF protects human dermal fibroblasts from TNF-induced apoptosis by induction of cFLIP and subsequent inhibition of caspase 8 cleavage. © 2015 Elsevier Ltd.
Immco Diagnostics | Date: 2012-03-27
Diagnostic reagents and kits for clinical or medical laboratory use; laboratory testing for autoimmune diseases.
Agency: Department of Health and Human Services | Branch: | Program: SBIR | Phase: Phase I | Award Amount: 99.98K | Year: 2007
DESCRIPTION (Provided by applicant): Autoimmune hearing loss (AHL) is a subset of sensorineural hearing loss (SNHL) in which there is a sudden onset, or rapidly progressing and/or fluctuating hearing loss that often progresses to bilateral involvement and devastating profound hearing impairment. This is a treatable disorder if diagnosed early; however, no accurate diagnostic test for AHL exists. The development of an accurate test depends on the identification of target antigens. Multiple lines of evidence implicate antibodies to a 68 kDa antigen in AHL. Measurement of these autoantibodies will help physicians properly identify AHL patients and thereby contribute to prompt and proper administration of treatment. Moreover, modifying treatment based on monitoring antibody levels should prevent unnecessary toxicity from the immunosuppressive drugs commonly used to treat AHL. An accurate diagnostic test for AHL will considerably reduce the morbidity and socioeconomic burden associated with diagnosis, treatment and consequences of disease progression in the affected population. Current tests available for diagnosis of AHL are compromised by low sensitivity and specificity. For lack of a better diagnostic tool, Western blot for hsp 70 is currently the most widely used test. There is an absolute need for a precise test to aid physicians treating patients with idiopathic hearing loss and the AHL subset. Recently, our team has identified the 68kDa antigen as choline transporter-like protein 2 (CTL2). This prominent and abundant protein is expressed on the surface of inner ear supporting cells (IESC). Monoclonal antibodies against CTL2 have been shown to induce hearing loss in an experimental guinea pig model. Preliminary studies indicate CTL2 reactive antibodies may be a proximal cause of autoimmune deafness and can be used as a marker for diagnosis of AHL in humans. Monoclonal antibodies directed against CTL2 bind to IESC with a distinctive "wineglass" pattern as evidenced by in vitro immunofluorescence. Patients with AHL frequently have antibodies in the serum that bind to IESC with the same distinctive pattern as anti-CTL2 monoclonal antibodies. We have demonstrated that patients exhibiting such patterns of antibody binding are more likely to respond to treatment with corticosteroids. We propose to develop a simple, reliable, high throughput test to detect anti-CTL2 antibodies in patients with rapidly progressive hearing loss. We postulate that detection of these antibodies will improve sensitivity and specificity significantly. The proposed CTL2 ELISA represents a significant advance in technology to support diagnosis of AHL. This assay will outperform any other test commercially available for this intended use. Relevance of Research to the Public Health: The CTL2 ELISA will fulfill the needs of patients, physicians and the marketplace for an assay to help determine the course of treatment and monitor response to therapy. This will result in decreased morbidity, improved outcomes and minimize economic burden of the affected population. Furthermore, acceptance of the CTL2 ELISA as a valid and reliable aid for diagnosis of AHL will greatly expand the market for serological testing. Idiopathic Hearing Loss is a subset of Sensorineural Hearing Loss (SNHL), a disorder affecting millions of Americans and representing one of the most common and severe disabilities. Patients with Idiopathic Hearing Loss are often suspected of suffering from Autoimmune Hearing Loss, a treatable condition if diagnosed early, and proscribed immunosuppressive treatment. Current diagnostic methods are inadequate to support proper diagnosis AHL; our team proposes to develop a simple, yet precise test to meet this need.
Immco Diagnostics | Date: 2014-07-09
A testing device has a base and multiple strips connected to the base. Multiple antigens are placed on each strip. The testing device or strips may be affixed to a sheet under a shield. Fluid can be applied to the testing device or the strips. The testing device can be used for biochemical testing, such as line immunoassay (LIA) testing.
Xuan J.,University of Sichuan |
Xuan J.,Medical Director Kidney and Pancreas Transplantation Erie County Medical Center |
Shen L.,State University of New York at Buffalo |
Malyavantham K.,Immco Diagnostics |
And 3 more authors.
BMC Oral Health | Year: 2013
Background: Evidence in imaging studies suggests that there may be differences in glandular involvement in Sjogren's syndrome (SS) depending on the stage of the disease. No detailed histological studies are available to show if there are any such difference in glandular involvement at various time periods and stages of SS. This cross sectional study examines the inflammatory changes in mouse models of SS at various ages.Methods: The histological changes in major salivary and lacrimal glands were studied at ages of 3, 6, 9, 12, 15 and 18 months in both sexes in well characterized mouse models of SS, non-obese diabetes mouse and Interleukin-14 alpha-transgenic mice.Results: Our results indicate that early inflammation concurrently occur in submandibular and lacrimal glands around the age of 6 weeks. Parotid glands are involved much later in the course of SS with less severe inflammation. Sublingual glands are rarely involved.Conclusions: Our conclusions are that SS may be an organ specific disease with early inflammation occurring in submandibular and lacrimal glands, followed by the parotid. Non organ specific events occur in later courses of the disease. The understanding of the disease progression is important in tailoring early local therapeutic interventions before complete destruction of salivary and lacrimal glands. © 2013 Xuan et al.; licensee BioMed Central Ltd.