THE THERAPEUTIC TREATMENT OF PEPITEM INCREASES THE PROTEIN EXPRESSION OF SIRT1 IN A MOUSE MODEL OF EXPERIMENTAL AUTOIMMUNE ENCEPHALOMYELITIS (EAE) AS A MODEL FOR HUMAN MS

Objective: To investigate the effect of the prophylactic and therapeutic treatment of the immunopeptide PEPITEM on the protein expression of SIRT1 in a mouse model of experimental autoimmune encephalomyelitis (EAE) as a model for human MS.   Methods: Using C57BL/6 female mice, we dosed the PEPITEM in the EAE model via intraperitoneal injections either prophylactically or therapeutically. The disease was induced using MOG35-55 and complete Freund's adjuvants augmented with pertussis toxin. The EAE score was recorded daily until the end of the experiment (21 days). A Western blot analysis was performed on the brain lysate to measure the protein concentration of SIRT1. Results: The therapeutic treatment with PEPITEM increased the protein expressions of SIRT1 on the EAE mice whereas the prophylactic injections did not affect the protein expression of SIRT1. Conclusion: Collectively, the therapeutic treatment with PEPITEM suggesting anti-inflammatory effect of PEPITEM on the brain damage in EAE mice which offers a novel and safe strategy for drug therapy in MS, opening new avenues for research and treatment

Characterising the PEPITEM pathway in patients with atherosclerosis

Atherosclerosis is an asymptomatic disease which is regarded as one of the most fatal diseases. However, the mechanism of the immune response is not well understood. There is accumulated evidence supporting the idea that inflammatory response initiates the disease. A new novel peptide has been discovered in our lab which down-regulates T cell recruitment during inflammation called PEPITEM (Peptide Inhibitor of Trans Endothelial Migration). We are interested in testing the action of PEPITEM on PBL isolated from atherosclerosis patients. We first demonstrated that PEPITEM did not affect the levels of adhesion of PBL from either diseased or healthy donors. Interestingly however, we did observe that PBL isolated from atherosclerosis patients adhere more readily than those isolated from healthy control subjects. Therefore, we studied the surface expression of certain adhesion molecules and chemokine receptors on the PBL of atherosclerosis patients. We found significantly higher surface expression of Beta-receptor family (Beta-1 and Beta-2) and PSGL-1 receptors in some PBL subsets in atherosclerosis patients. In addition, we looked at the effect of PEPITEM and adiponectin (AQ) treatment on the migration of PBL and we revealed for the first time based on our knowledge that there was no effect of treatment on PBL isolated from atherosclerosis patients. These observations will contribute to understanding the potential therapeutic applications of PEPITEM on atherosclerosis

Mycotic aortic aneurysm due to brucellosis

Brucellosis is a multisystem zoonotic disease. Mycotic aneurysm due to Brucella is rare and has no clear management approach. Here, we present two cases of mycotic aortic aneurysm due to Brucella. The first patient was treated with surgical resection of a symptomatic infrarenal abdominal aortic aneurysm combined with lifelong doxycycline and rifampicin. The second patient improved with conservative treatment including a 6-month course of antibiotics and regular clinical and radiologic monitoring. Through these cases, we hope to draw attention to this serious adverse effect of Brucella and the importance of management of its local arterial complications, especially in endemic areas

BM-MSCs alleviate diabetic nephropathy in male rats by regulating ER stress, oxidative stress, inflammation, and apoptotic pathways

Introduction: Diabetic nephropathy (DN), a chronic kidney disease, is a major cause of end-stage kidney disease worldwide. Mesenchymal stem cells (MSCs) have become a promising option to mitigate several diabetic complications.Methods: In this study, we evaluated the therapeutic potential of bone marrow-derived mesenchymal stem cells (BM-MSCs) in a rat model of STZ-induced DN. After the confirmation of diabetes, rats were treated with BM-MSCs and sacrificed at week 12 after treatment.Results: Our results showed that STZ-induced DN rats had extensive histopathological changes, significant upregulation in mRNA expression of renal apoptotic markers, ER stress markers, inflammatory markers, fibronectin, and intermediate filament proteins, and reduction of positive immunostaining of PCNA and elevated P53 in kidney tissue compared to the control group. BM-MSC therapy significantly improved renal histopathological changes, reduced renal apoptosis, ER stress, inflammation, and intermediate filament proteins, as well as increased positive immunostaining of PCNA and reduced P53 in renal tissue compared to the STZ-induced DN group.Conclusion: In conclusion, our study indicates that BM-MSCs may have therapeutic potential for the treatment of DN and provide important insights into their potential use as a novel therapeutic approach for DN

Mice model of iron overload (SB6.Cg-Tg(Thy1-YFPH)2Jrs/J) : study of immune function and autoimmunity

Access to thesis permanently restricted to Ball State community onlyBoth Immune cells and pathogenic microorganisms require iron for proliferation and multiplication. However, role of iron supplementation on immune function is still unclear. Studies show that iron-deficient mice are protected from developing Experimental Autoimmune Encephalomyelitis (EAE), an animal model of Multiple Sclerosis (MS) in humans. In this project, we developed a mice model of iron overload in (B6.Cg-Tg (Thy1-YFPH) 2Jrs/J mice). Seven mice were injected (ip), 100 μl iron dextran and seven with Phosphate buffered saline (PBS), five days/week for four weeks. Blood samples verified iron overload 170 versus 138μg/dl (P < 0.005). Flow Cytometry revealed high T-cells and low and CD8+ T-cell. Histological sections indicated perivascular immune cell infiltrations in the brain, but not in the spinal cord. Confocal microscopy of spinal cord sections showed myelinated axons with no breaks. The absence of demyelination and clinical signs, but high CD3+ with low CD4+ T-cells suggests an altered immune cell function in iron overload mice that needs further exploration.Thesis (M.S.)Department of Physiology and Health Scienc

Mycotic aortic aneurysm due to brucellosis

Brucellosis is a multisystem zoonotic disease. Mycotic aneurysm due to Brucella is rare and has no clear management approach. Here, we present two cases of mycotic aortic aneurysm due to Brucella. The first patient was treated with surgical resection of a symptomatic infrarenal abdominal aortic aneurysm combined with lifelong doxycycline and rifampicin. The second patient improved with conservative treatment including a 6-month course of antibiotics and regular clinical and radiologic monitoring. Through these cases, we hope to draw attention to this serious adverse effect of Brucella and the importance of management of its local arterial complications, especially in endemic areas

Subtractive panning for the isolation of monoclonal PEPITEM peptide antibody by phage display

Abstract Antibody phage display is a key tool for the development of monoclonal antibodies against various targets. However, the development of anti-peptide antibodies is a challenging process due to the small size of peptides for binding. This makes anchoring of peptides a preferred approach for panning experiments. A common approach is by using streptavidin as the anchor protein to present biotinylated peptides for panning. Here, we propose the use of recombinant expression of the target peptide and an immunogenic protein as a fusion for panning. The peptide inhibitor of trans-endothelial migration (PEPITEM) peptide sequence was fused to the Mycobacterium tuberculosis (Mtb) α-crystalline (AC) as an anchor protein. The panning process was carried out by subtractive selection of the antibody library against the AC protein first, followed by binding to the library to PEPITEM fused AC (PEPI-AC). A unique monoclonal scFv antibodies with good specificity were identified. In conclusion, the use of an alternative anchor protein to present the peptide sequence coupled with subtractive panning allows for the identification of unique monoclonal antibodies against a peptide target