Aca 2013 Abstract Export Plenary Session: Plenary Session 1: Novel aspect of therapeutics of autoimmune diseases

Allergy to insulin is rare due to the introduction of human insulin and its analogues, and currently reported in less than 1-2% of diabetic patients. Although, immunologic responses to insulin in the form of IgG and IgE antibodies are common clinical problems related to these immunologic reactions are uncommon. Usually, the allergic reactions are limited to the site of insulin injection and systemic, potentially life-threatening reactions such as anaphylaxis have been reported infrequently. However, treatment of diabetics with insulin allergy during pregnancy could be a real challenge for the treating physician in countries with the lake of experience with the use of new insulin alogs. Treatment options for insulin allergy include symptomatic therapy with antihistamines, use of an alternative insulin preparation, use of oral hypoglycaemic agents, insulin desensitization using small doses of insulin subcutaneously, and pancreatic transplantation for severe resistant cases. However, management in pregnant women is more difficult as treatment options are limited. A 29-year-old pregnant woman with type 1 diabetes developed generalized erythematous skin rash with marked pruritus following insulin administration. She has a strong family history of diabetes; her mother, all her sisters and her brother are diabetics, her mother has allergy to insulin (protamine). Due to insulin allergy, the patientÕs diabetes was controlled by switching to insulin Lispro with no adverse effects. Her antenatal follow up was regular but needed frequent visits; she had a successful pregnancy outcome with no congenital abnormalities in the baby.

Background and aims:The dysfunction of central tolerance can cause the development of autoimmune disease, and microRNA (miR)-181a plays a critical role in central tolerance. Higher miR-181a expression could induce the T cell receptor (TCR) signaling, which might induce higher expression of activated T cell. In addition, higher miR-181a expression also could affect the interlukin (IL)-2 expression which influenced negative selection of T cell. Therefore, we designed a hospital-based case-control study to evaluate the effects of miR-181a and IL-2 in the development of ankylosing spondylitis (AS).

The current study explored the relationship between lineage^-CD4^-CD127⁺ cells and lymphocyte infiltration in rheumatoid arthritis. A lineage^-CD4^-CD127⁺ cell is positive for CD117 and ROR-gamma and known as a lymphoid tissue inducer cell, which is critical for secondary and tertiary lymphoid tissue development and maintenance of CD4 T cell-dependent immune responses. Rheumatoid arthritis is chronic systemic inflammation disease that destroys joint cartilages, bones and tissues. Some rheumatoid arthritis patients show that lymphocytes infiltrate to the damaged joint and form lymphoid tissue-like tertiary lymphoid tissue structures in synovial tissue. The factors involved in the formation of normal lymphoid tissues are expressed in abnormal lymphoid tissues of rheumatoid arthritis synovial membrane such as CXCL13, CCL19, CCL21, CCR7, CXCR5, lymphotoxin-α/β, and IL-7.

Introduction:

Adipose-derived stem cells (ASCs) have immunomodulatory properties and are intensively studied as a potential therapeutical method to treat rheumatoid arthritis (RA). ASCs immunosuppresive function may be affected by adipocytokines released by intra-articular adipose tissue. The aim of this study was to evaluate the influence of high/middle molecular weight adiponectin isoform (HMW/MMW) and TNFα on immunomodulatory function of ASCs from RA and osteoarthritis (OA) patients.

Materials and methods:

Infrapatellar fat pad was obtained from RA and OA patients. ASCs were isolated and cultured with/without HMW/MMW (10μg/ml), TNFα (10ng/ml), IFNγ (10ng/ml). Expression of indoleamine 2,3-dioxygenase (IDO), heme oxygenase 1 (HO-1) genes (Real Time PCR) and IL-6, IL-8, VEGF, TGFβ, IL-1Ra concentration in culture supernatants (ELISA) were measured.

Results:

Conclusion:

HMW/MMW and TNFα influence function of ASCs from RA and OA. ASCs from both diseases seem to be dissimilar regarding their secretory activity and genes expression suggesting differences in their immunomodulatory properties.

Funding: grant no. 2011/01/N/NZ5/00932, National Science Center, Poland.

Background and aims: Rheumatoid arthritis (RA) is the most common inflammatory arthritis and a major cause of various disabilities. Involvement of the NF-kB pathway in RA pathogenesis is well established. In the inflamed synovium of rheumatoid arthritis patients, this is an overexpression of NF-kB, which is causes the transactivation of various genes for pro-inflammatory cytokines, chemokines, adhesion molecules, MMPs, and inducible nitric oxide (iNOS). Therefore, NF-kB plays a central role in the pathogenesis of RA by amplifying inflammation and causing joint damage. Currently, the clinical drugs used to treat RA, consisting of NSAID and steroids, can help relieve pain and reduce inflammation; however, the general effectiveness of these drugs have been unsatisfactory. Other therapeutic modalities like TNFα inhibitors and IL-1 receptor antagonists are expensive and associated with serious side effects. Therefore, the development of novel therapeutic strategies is urgently needed for the treatment of rheumatoid arthritis. Results: Recently, we found that MKP-1, a MAP kinase phosphatase, negatively regulates the NF-kB-dependent inflammatory response. In addition, we have found that Rolipram, which is already in late phase III clinical trials for COPD, is a potent anti-arthritis agent by enhancing expression of MKP-1, a critical endogenous negative regulator. Negative feedback regulation is an effective therapeutic strategy developed organically through evolution that can combat an overactive inflammatory response without causing serious adverse effects. Conclusions: Our studies will provide new insights into the molecular regulatory mechanisms underlying inflammation and provide a novel therapeutic approach in a broad sense for treatment of autoimmune diseases.

Results: Patients with positive rheumatoid factor are 39 (62%), anti cyclic citrullinated protein antibody positive are 32 (51%) and active disease are 17 (27%). Interstitial lung disease is found in 22 (35%) patients, pleural effusion are 3 (5%) patients and 7 (11%) patients bronchiectasi. 17 (27%) patients have restrictive ventilator insufficiency, 4 (6%) patients have obstructive ventilator insufficiency. We found correlation between pulmonary injuries with the severity of disease and the abnormality of immunological laboratory tests.

Rheumatoid arthritis (RA) fibroblasts participate in tissue destruction by producing interleukin-6, and metalloproteinases which are involved in extracellular matrix degradation, and osteoclast-mediated bone resorption. The present study was design to investigate the effects of NHAG (analog of glucosamine) on RA-DFBs. Primary cultured DFBs of RA patient were cultured overnight in complete DMEM. Next day media was aspirated and DMEM without FBS was added. The cells were pre-treated with NHAG (correspondent doses of 4, 20, 40, 60, 80, 100µM) for 4 hrs and then stimulated with TNF-α (5 ng/ml) for 24 hours at 37¡C with 5% CO₂. After incubation, supernatants were collected, centrifuged and used for determination of IL-6, MMP-2 and MMP-9. NHAG at the dose of 40 - 80 mM, exhibited dose dependent inhibition of IL-6 production by RA-DFBs. The gel-zymography revealed that NHAG markedly reduced the proactivated MMP-2 and MMP-9 in RA-DFBs at the dose of 80 and 100 mM. Further increase in the NHAG dose was avoided because at higher concentration, the cell viability was compromised. In our previous in vivo studies, NHAG has demonstrated a potent anti-arthritic activity. The present study further supports our earlier findings indicating that some of the beneficial effect of NHAG seen in our rodent arthritic model might be due to the inhibition of IL-6, and MMP-2 and MMP-9 as it was observed in in vitro study with RA-DFBs cells. This effect, coupled with its multiple inhibitory effects on T lymphocyte functions, might account for the significant reduction in the rate of disease progression in arthritic model treated with NHAG.