Identification and studies of MHC class III genes in animal models of rheumatoid arthritis

Rheumatoid arthritis is an autoimmune disorder in which our own immune system attacks our own body tissues by mistake. It is a chronic inflammatory disease which primarily affects the joints in the body. The damage is caused in the joints in the form of inflammations [1]. The main parts affected by Rheumatoid arthritis are the peripheral joints which include finger joints, wrists, toes and knees. Though this disease is prevalent for a long time the cause of the disease is not known. It is believed that the genes present in the Major Histocompatibility Complex region might contribute to the onset of rheumatoid arthritis in one way or another [2] [3] [4]. Various animal models are used to study the between rheumatoid arthritis and the genes in Major Histocompatibility Complex. DA strains of rat are the widely used animal models for rheumatology studies. In our laboratory it is observed that rats carrying MHC class III congenic fragment, DA.1HR56 develop less severe diseases in our animal models of rheumatoid arthritis. The animal models used in our lab are pristine‐induced arthritis (PIA) and oil‐induced arthritis (OIA), both of which are T‐cell mediate disease models. Based on testing different subcongenic fragments, we conclude that this protective phenotypes comes from a 220‐kb region in chromosome 20 starting from 3649561 to 3864755; comprising approximately more than 20 genes like TNF, Lta, Ltb, Aif1, Bat2 and Lst1, a lot of which are potential candidate for immune regulation. Some of these are much better investigated while some are not so well‐known. Analyzing the 20 genes in the congenic region for polymorphism and its comparative gene expression studies between DA strain and DA.1HR56 will help us to corner out the gene or group of gene which leads to this protective phenotype. Based on these data, functional studies of selected genes will be undertaken

Effekt av Matrix‐M på uttryck av generna för STING, IFN‐β, och osteopontin i vävnad från gris

Matrix‐M is the adjuvant component in traditional ISCOM vaccines. The adjuvant when used along with antigen increases the efficiency of the vaccines by inducing a balanced Th1/Th2 response and a long‐lasting antibody response. Transcriptomal studies in the pig show that the majority of upregulated genes during the early inflammatory response to Matrix‐M alone are Interferon regulated genes (Ahlberg, Lovgren Bengtsson, Wallgren, & Fossum, 2012). The present study aimed to establish qPCR assays for detection of gene expression of stimulator of interferon genes (STING), osteopontin (OPN) and interferon‐beta (IFN‐β) in porcine tissue. STING is a protein localized in the endoplasmatic reticulum that on activation is believed to enhance the production of Type I interferons (Sun et al., 2009). Overexpression of OPN is believed to play a key role in inducing the production of Interferon‐α and other Type 1 interferons by plasmacytoid dendritic cells (Shinohara et al., 2006). A comparative gene expression study of STING and OPN in pigs treated with Matrix‐M revealed that one or both of the genes were up‐regulated in pigs with increased expression of the IFN‐β gene 24 hours after administration of Matrix‐M

Investigation of calcium-dependent activity and conformational dynamics of zebra fish 12-lipoxygenase

Background A 12-lipoxygenase in zebra fish (zf12-LOX) was found to be required for normal embryonic development and LOXs are of great interest for targeted drug designing. In this study, we investigate the structural-functional aspects of zf12-LOX in response to calcium. Methods A soluble version of zf12-LOX was created by mutagenesis. Based on multiple sequence alignment, we mutated the putative calcium-responsive amino acids in N-PLAT domain of soluble zf12-LOX. Using a series of biophysical methods, we ascertained the oligomeric state, stability, structural integrity and conformational changes of zf12-LOX in response to calcium. We also compared the biophysical properties of soluble zf12-LOX with the mutant in the absence and presence of calcium. Results Here we provide a detailed characterization of soluble zf12-LOX and the mutant. Both proteins exist as compact monomers in solution, however the enzyme activity of soluble zf12-LOX is significantly increased in presence of calcium. We find that the stimulatory effect of calcium on zf12-LOX is related to a change in protein structure as observed by SAXS, adopting an open-state. In contrast, enzyme with a mutated calcium regulatory site has reduced activity-response to calcium and restricted large re-modeling, suggesting that it retains a closed-state in response to calcium. Taken together, our study suggests that Ca2 +-dependent regulation is associated with different domain conformation(s) that might change the accessibility to substrate-binding site in response to calcium. General significance The study can be broadly implicated in better understanding the mode(s) of action of LOXs, and the enzymes regulated by calcium in general

Additional file 1 of Discovery of a small molecule that inhibits Bcl-3-mediated cyclin D1 expression in melanoma cells

Additional file 1: Supplementary Figure 1