Objective. To increase the therapeutic activity of glucocorticoids in experimental arthritis by encapsulation in long-circulating polyethylene glycol liposomes, which have shown the ability to preferentially accumulate in inflamed joints after intravenous administration.Methods. Rats with adjuvant-induced arthritis (AIA) were treated intravenously with liposomal and free prednisolone phosphate (PLP) a few days after the first signs of disease. The effect on paw inflammation scores during the weeks after treatment was evaluated. Liposome biodistribution and joint localization were investigated by labeling the preparation with radioactive 111 In-oxine. By studying PLP encapsulated in other types of liposomes, which show a distinctive tissue distribution pattern and reduced accumulation in inflamed joints, the importance of targeted delivery to inflamed joints for achieving an increased therapeutic effect was illustrated.Results. Liposomal PLP proved to be highly effective in the rat AIA model. A single injection of 10 mg/kg resulted in complete remission of the inflammatory response for almost a week. In contrast, the same dose of unencapsulated PLP did not reduce inflammation, and only a slight effect was observed after repeated daily injections. Evidence was found that preferential glucocorticoid delivery to the inflamed joint was the key factor explaining the observed strong therapeutic benefit obtained with the liposomal preparation, while other possible mechanisms, such as splenic accumulation or prolonged release of prednisolone in the circulation, were excluded.Conclusion. Targeted delivery using longcirculating liposomes is a promising, novel means to successfully intervene in arthritis with glucocorticoid therapy.

Reestablishing self-tolerance in autoimmunity is thought to depend on self-reactive regulatory T cells (Tregs). Exploiting these antigen-specific regulators is hampered by the obscure nature of disease-relevant autoantigens. We have uncovered potent disease-suppressive Tregs recognizing Heat Shock Protein (Hsp) 70 self-antigens, enabling selective activity in inflamed tissues. Hsp70 is a major contributor to the MHC class II ligandome. Here we show that a conserved Hsp70 epitope (B29) is present in murine MHC class II and that upon transfer, B29-induced CD4 + CD25 + Foxp3 + T cells suppress established proteoglycan-induced arthritis in mice. These self-antigen–specific Tregs were activated in vivo, and when using Lymphocyte Activation Gene-3 as a selection marker, as few as 4,000 cells sufficed. Furthermore, depletion of transferred Tregs abrogated disease suppression. Transferred cells exhibited a stable phenotype and were found in joints and draining lymph nodes up to 2 mo after transfer. Given that ( i ) B29 administration by itself suppressed disease, ( ii ) our findings were made with wild-type (T-cell receptor nontransgenic) Tregs, and ( iii ) the B29 human homolog is presented by HLA class II, we are nearing translation of antigen-specific Treg activation as a promising intervention for chronic inflammatory diseases.

Objective. Stress proteins, such as members of the heat-shock protein (HSP) family, are up-regulated by cells in inflamed tissue and can be viewed functionally as "biomarkers" for the immune system to monitor inflammation. Exogenous administration of stress proteins has induced immunoregulation in various models of inflammation and has also been shown to be effective in clinical trials in humans. This study was undertaken to test the hypothesis that boosting of endogenous HSP expression can restore effective immunoregulation through T cells specific for stress proteins.Methods. Stress protein expression was manipulated in vivo and in vitro with a food component (carvacrol), and immune recognition of stress proteins was studied.Results. Carvacrol, a major compound in the oil of many Origanum species, had a notable capacity to coinduce cellular Hsp70 expression in vitro and, upon intragastric administration, in Peyer's patches of mice in vivo. As a consequence, carvacrol specifically promoted T cell recognition of endogenous Hsp70, as demonstrated in vitro by the activation of an Hsp70-specific T cell hybridoma and in vivo by amplified T cell responses to Hsp70. Carvacrol administration also increased the number of CD4؉CD25؉FoxP3؉ T cells, systemically in the spleen and locally in the joint, and almost completely suppressed proteoglycan-induced experimental arthritis. Furthermore, protection against arthritis could be transferred with T cells isolated from carvacrol-fed mice.Conclusion. These findings illustrate that a food component can boost protective T cell responses to a self stress protein and down-regulate inflammatory disease, i.e., that the immune system can respond to diet.

Although intercellular transfer of cell surface molecules has been observed between several cells of the immune system, the physiological relevance of this phenomenon remained obscure. Until now the transfer of molecules between antigen-presenting cells (APC) and T cells has been described as a unidirectional process from APC to T cells. However, here we show that T cells in turn donate molecules to APC, and that T cell-derived vesicles can mediate this transfer. The transferred proteins are incorporated into the APC as active molecules. Our data provide evidence that T cells use intercellular molecule transfer to mediate cell contact-dependent regulation of T cell responses via modulation of the APC.

Serial peri-operative plasma and serum samples from 55 horses suffering from acute abdominal disease and presented for surgical intervention were assayed for the presence of endotoxins (lipopolysaccharides; LPS), tumor necrosis factor (TNF) and interleukin-6 (IL-6). At study entry, venous blood was collected for blood cultures. Intra-operatively collected ascitic fluid samples were assayed for the presence of LPS. The clinical course of the disease was documented. Four horses were excluded from the study. At study entry, 21 of the 51 horses (41%) had increased platelet-rich plasma LPS concentrations, i.e. ≥ 5 ng/l (mean 35.5; range 7-197 ng/l), in 34 horses (67%) IL-6 concentrations were increased, i.e. exceeding 35 ng/l (mean 364; range 36—1762 ng/l). Detectable TNF was present in 5 horses (10%); 3 of them died spontaneously during surgery. In all TNF positive samples, markedly increased LPS and IL-6 concentrations were detected. Study entry IL-6 concentrations were significantly higher in non-surviving (mean 394; range < 20—1762 ng/l) than in surviving horses (mean 116; range < 20—894 ng/l; P < 0.0002). This was most evident when non-surviving horses with inflamed bowel disease were considered (mean 1096; range 650—1762 ng/l; P < 0.0001). Positive ascitic fluid LPS concentrations (≥ 3 ng/l) were encountered in 18 horses. There was no significant correlation between the LPS concentrations in ascites and plasma. Study entry concentrations of IL-6 were significantly correlated with LPS concentrations (r = 0.62; P < 0.001) and were inversely correlated with platelet counts (r = -0.46; P < 0.002). The magnitude of a calculated LPS-cytokine score strongly correlated with mortality (P < 0.001). Receiver operating characteristic (ROC) curve analysis showed that the IL-6 assay had reasonable accuracy for the prediction of unfavorable outcome (i.e. the area under the curve equalled 0.72), in contrast to the LPS assay. The results indicate that LPS, TNF and IL-6 are predominantly released in the systemic circulation of horses suffering from inflamed and ischemic bowel disease. IL-6 concentrations have predictive value for unfavorable outcome and the simultaneous presence of increased LPS, TNF and IL-6 concentrations is especially associated with a poor clinical condition and outcome.The clinical signs culminating in equine acute abdominal disease range widely, from mild symptoms to fulminant shock with profound metabolic and respiratory acidosis, hypotension, and hypovolemia with death ocat RYERSON UNIV on June 18, 2015 ini.sagepub.com Downloaded from

Previous studies in mouse models of autoimmune diabetes and encephalomyelitis have indicated that the selective delivery of self-antigen to the endocytic receptor DEC205 on steady-state dendritic cells (DCs) may represent a suitable approach to induce Ag-specific immune tolerance. In this study, we aimed to examine whether DEC205+ DC targeting of a single immunodominant peptide derived from human cartilage proteoglycan (PG) can promote immune tolerance in PG-induced arthritis (PGIA). Besides disease induction by immunization with whole PG protein with a high degree of antigenic complexity, PGIA substantially differs from previously studied autoimmune models not only in the target tissue of autoimmune destruction but also in the nature of pathogenic immune effector cells. Our results show that DEC205+ DC targeting of the PG peptide 70–84 is sufficient to efficiently protect against PGIA development. Complementary mechanistic studies support a model in which DEC205+ DC targeting leads to insufficient germinal center B cell support by PG-specific follicular helper T cells. Consequently, impaired germinal center formation results in lower Ab titers, severely compromising the development of PGIA. Overall, this study further corroborates the potential of prospective tolerogenic DEC205+ DC vaccination to interfere with autoimmune diseases, such as rheumatoid arthritis.

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Adjuvant arthritis (AA) is an induced form of (sub)chronic arthritis. Strains of rats have a varying genetic susceptibility to AA, whereas mice generally are not susceptible. AA is most easily induced with mycobacteria suspended in oil, although in some strains of rats it can be induced with oily adjuvants in the absence of mycobacteria. The disease is a T cell-mediated autoimmune arthritis that is frequently used to study immunological aspects of rheumatoid arthritis (RA) and other arthritic or inflammatory diseases in humans. Furthermore, it is used as a model for developing and testing antiinflammatory drugs. There are no particularly well-defined autoantigens in AA; in this respect, the model resembles spontaneous arthritic diseases in humans. In all susceptible rat strains, the inflammatory process of AA is self remitting, although usually the disease is severe and leads to permanent joint malformations, including ankylosis; a time line for AA development is presented. This unit describes the induction and evaluation of AA and the preparation of adjuvant used to induce AA.