P1-062 GAS-THERAPY IN RHEUMATOID ARTHRITIS (RA) TREATMENT: WHEN WEST MEETS EAST

It is possible to regulate the activity of connective tissue (CT) by acupuncture by acting on to ãacupointsÓ (sites of greatest CT accumulation). High concentrations of acetylcholine - neutrotransmitter of cholinergic system were found within the bondary of most acupuncture points. It is suggested that autonomic nervous system, vagus nerve can be influenced by acupuncture. This treatment increases the efficiency of transport of gasotransmitters: such as carbon monoxide, nitric oxide and hydrogen sulfide which are produced endogenously, play a roles neurotransmitters and inhibits energy production, conversely to exogenously oxygen enhanced this reaction. The famous ãQi energyÓ would be nothing else than a state of molecular equilibrium ( redox potential) that may shift towards either activation (oxygen * Yang) or inhibition (gasotransmitters * Yin).

Redox state can regulate inflammatory cells differentiation. In synovial fluid neutrophils differentiation toward to highly-specialized ãmicrophagesÓ was observed. These cells are responsible for major injury in the joint and can disrupt the capacity of endothelial progenitor cells to differentiate into mature endothelial cells (bloking of neovascularization of ãpannusÓ). Moreover, neutrophiles might be responsible for controlling migration and activation of lymphocytes and monocytes. Targeting neutrophils seems to be very promising therapeutic metod.

Taken together, understanding of the complex interactions between CT and vagus nerve modulation (ãacupunctureÓ), gases neurotransmitters and redox status of inflammatory cells (ãQi energyÓ) provide valuable opportunities for the development of novel therapeutic strategies for RA. Once again western medicine reveals knowledge that has been known in an eastern medicine ages before.

Hepatocellular carcinoma is a leading cause of cancer-related death worldwide. Hep88 mAb, an antibody against tumor-associated antigen in hepatocellular carcinoma cell line established from a Thai patient, has been previously shown to induce cancer cell death. These dying cells exhibited morphological characteristics of apoptosis-like programmed cell death (PCD), including cytoplasmic vacuolization and dilatation of mitochondria and endoplasmic reticulum. In the present study, effects of Hep88 mAb on cell cycle progression and apoptosis induction in human hepatocellular carcinoma cell line, HepG2, were examined. Growth-inhibitory activity of Hep88 mAb on HepG2 cell line was associated with G2/M cell cycle arrest. In addition, Hep88 mAb induced a significant increase in the sub-G1 peak, indicating an apoptotic cell population. The apoptosis-inducing effects of Hep88 mAb were further investigated using Annexin V-FITC/propidium iodide staining and flow cytometry. Early apoptotic cells (Annexin V single positive cells) were increased by Hep88 mAb treatment in a dose-and time-dependent manner. Although it has been suggested that apoptosis-like PCD is involved in anti-tumor activity of Hep88 mAb, the results from this study demonstrated that Hep88 mAb induced Hep G2 cell line to undergo cell cycle arrest and subsequent apoptosis. Therefore, further investigation of the cellular and molecular mechanisms underlying anti-cancer activities of Hep88 mAb as well as the potential uses as an anti-neoplastic agent against hepatocellular carcinoma is required.

Aberrant T cell responses mediate a variety of autoimmune diseases, including rheumatoid arthritis, systemic lupus erythematosus, multiple sclerosis and ulcerative colitis. Immunosuppressants are widely used for the treatment of T cell-mediated autoimmune diseases. However, because of the toxicity and tolerance of these drugs, it is important to keep looking for novel immunosuppressants. Here, a series of novel benzothiazole derivatives were synthesized and screened for the immunosuppressive activity. A water-soluble benzothiazole derivative, BD926 had the most effective activity among these compounds. Treatment with BD926 significantly inhibited mouse and human T cell proliferation stimulated either by anti-CD3/anti-CD28 mAbs or by alloantigen in a dose-dependent manner measured by flow cytometry in vitro. We did not observe obvious cytotoxicity of BD926 against human resting na•ve T cells, IL-4 treated-activated T cells and fibroblast-like synoviocyte determined by CCK-8 assay. Furthermore, BD926 did not inhibit IL-2, IL-4 and IL-10 secretion, but inhibited IFN-g, IL-6 and IL-17 production measured by ELISA and induced cell cycle arrest at G₀/G₁ phase in activated T cells determined by flow cytometry. Interestingly, AKT and p70S6K phosphorylation was not inhibited by BD926 in IL-2-induced T cells assessed by western blot. Finally, administration of BD926 mitigated T cell-mediated delayed-type hypersensitivity in mice in a dose-dependent manner in vivo. Collectively, these data suggest that BD926 inhibits T cell proliferation and may be used as a lead compound for the design and development of new immunosuppressant for the intervention of autoimmune diseases.

Lasting B cell persistence depends on survival signals that are transduced by cell surface receptors. In this study, we describe a novel biological mechanism essential for survival and homeostasis of normal peripheral mature B cells and chronic lymphocytic leukemia cells, regulated by the heparin-binding cytokine, midkine (MK), and its proteoglycan receptor, the receptor-type tyrosine phosphatase ζ (RPTPζ). We demonstrate that MK initiates a signaling cascade leading to B cell survival by binding to RPTPζ. In mice lacking PTPRZ, the proportion and number of the mature B cell population are reduced. Our results emphasize a unique and critical function for MK signaling in the previously described MIF/CD74-induced survival pathway. Stimulation of CD74 with MIF leads to c-Met activation, resulting in elevation of MK expression in both normal mouse splenic B and chronic lymphocytic leukemia cells. Our results indicate that MK and RPTPζ are important regulators of the B cell repertoire. These findings could pave the way toward understanding the mechanisms shaping B cell survival and suggest novel therapeutic strategies based on the blockade of the MK/RPTPζ-dependent survival pathway.

Background. Hyper-IgE syndrome (HIES) is multisystem disorder which is related to increase in the serum IgE, dermatitis, eosinophilia, and recurrent pulmonary or skin infections. Patients with HIES have increased risk of autoimmune diseases.

Background and aims: autoimmune thyroid disease (AITD) represent a group of disorders of organ function that occur as a result of immune responses directed against its own thyroid follicular cells in the presence of circulating antibodies specific for thyroid antigens, following with activation of cellular immune response, releasing of cytokines, lymphocyte infiltration and cell destruction. AITD includes two main clinical entities: Graves' disease and Hashimoto thyroiditis. They affect often women of reproductive age. During healthy pregnancy predominant is Th2 over Th1 immune response, which explains the improvement of autoimmune disease during pregnancy, while after delivery because of the changes in Th1/Th2 ratios often leads to deterioration of AITD. Regulatory network of NKT and Tregs seems to play an important role in mediating maternal tolerance to fetus.

Methods: We investigated the role of NKT and Tregs in presence of ATD in pregnancy and in postpartum period in women with hormonal status determination and with positive titer of thyroid antibodies and autoantibodies and compared them with healthy pregnant and not pregnant women, using flow cytometry analysis.

Results: cells of innate immunity: NKT and Tregs are increased in healthy pregnancies, in pregnancies with hypo-and hyperthyroidism, and postpartum in examine groups. NKT cells are decreased in pregnant women with positive antibodies indicating a lost of NKT cellsÕ protective effect in pregnancy with positive antibodies.

Conclusion: Pregnancy and early postpartum period have a great effect on interaction of innate immunity and the function of the thyroid gland. During AITD changes are more pronounced, especially postpartum.
P2-005 DIFFICULT LUPUS NEPHRITIS IN PREGNANCY
M. Lazo¹, J.J. Lichauco<sup>1


1</sup>Section of Rheumatology, St. Luke's Medical Center, Quezon City, Philippines

Introduction: Systemic lupus erythematosus (SLE) is an autoimmune disease that occurs frequently in women of childbearing age. Although fertility is not compromised, pregnancies among SLE patients are commonly associated with complications. We describe a case of a 30 year old Filipino female who had an unplanned pregnancy during active lupus nephritis (Type IV). She was treated with monthly Methylprednisolone pulse therapy but eventually delivered via caesarian section (C- section) due to intrauterine growth retardation (IUGR) at 35 weeks AOG.

Case report: A 30 year-old female (G3P3) was diagnosed with Type IV Lupus Nephritis in 2005. She received induction treatment with IV Cyclophosphamide for 6 months followed by Mycophenolate mofetil at 2 gms/day for maintenance treatment. She continued to have active disease and subsequently received 2 doses of Rituximab 1 gm IV with 500 mg/m² IV Cyclophosphamide. She achieved clinical remission for 3 months but had an unplanned pregnancy during recurrent active kidney disease. Proteinuria with active urinary sediments were observed despite IV pulse methylprednisolone therapy. At 35 weeks AOG, abdominal ultrasound showed asymmetric IUGR requiring C-section.