Protective effect of Rhododendron weyrichii flower extract against UVB-induced proinflammatory cytokine production in human keratinocytes

Ultraviolet B (UVB) radiation is harmful to the skin and induces cytokine release from keratinocytes leading toinflammatory skin disorders. Previous studies have shown that chronic exposure to UVB radiation increases tumornecrosis factor (TNF)-α and interleukin (IL)-6 secretion through various signaling pathways, resulting in skininflammation and increased risk of skin cancer. The present study was undertaken to investigate the protective effectsof Rhododendron weyrichii flower (RWF) extracts against UVB damage of immortalized human keratinocytes(HaCaT). To determine the anti-inflammatory effects of RWF, we examined UVB-induced proinflammatory cytokineproduction in HaCaT cells in the presence or absence of RWF extract, using enzyme-linked immunosorbent assay(ELISA). The results indicated that the RWF extract inhibited the production of proinflammatory molecules suchas IL-6 and TNF-α, but not IL-8, in UVB-irradiated HaCaT cells. These results demonstrate that RWF potentiallyprotects against UVB-induced skin inflammation. In addition, using high-performance liquid chromatography (HPLC)fingerprinting, kaempferol (0.335 ppm) and astragalin (2.569 ppm) were identified and quantified as RWF extractconstituents. Moreover, we tested the potential application of RWF extracts as a cosmetic treatment by performinghuman skin primary irritation tests. In these tests, the RWF extracts did not induce adverse reactions. Based on theseresults, we suggest that RWF extracts be considered anti-inflammatory candidates for pharmaceutical and/or cosmeticapplications.

Rhamnogalacturonan II is a Toll-like receptor 4 agonist that inhibits tumor growth by activating dendritic cell-mediated CD8+ T cells

We evaluated the effectiveness of rhamnogalacturonan II (RG-II)-stimulated bone marrow-derived dendritic cells (BMDCs) vaccination on the induction of antitumor immunity in a mouse lymphoma model using EG7-lymphoma cells expressing ovalbumin (OVA). BMDCs treated with RG-II had an activated phenotype. RG-II induced interleukin (IL)-12, IL-1beta, tumor necrosis factor-alpha (TNF-alpha) and interferon-gamma (IFN-gamma) production during dendritic cell (DC) maturation. BMDCs stimulated with RG-II facilitate the proliferation of CD8+ T cells. Using BMDCs from the mice deficient in Toll-like receptors (TLRs), we revealed that RG-II activity is dependent on TLR4. RG-II showed a preventive effect of immunization with OVA-pulsed BMDCs against EG7 lymphoma. These results suggested that RG-II expedites the DC-based immune response through the TLR4 signaling pathway.

The Mycobacterium avium subsp. paratuberculosis fibronectin attachment protein, a toll-like receptor 4 agonist, enhances dendritic cell-based cancer vaccine potency

In this study, we showed the direct interaction between Mycobacterium avium subsp. paratuberculosis fibronectin attachment protein (FAP) and toll-like receptor4 (TLR4) via co-localization and binding by using confocal microscopy and co-immunoprecipitation assays. FAP triggered the expression of pro- and anti-inflammatory cytokines in a TLR4-dependent manner. In addition, FAP-induced cytokine expression in bone marrow-derived dendritic cells (BMDCs) was modulated in part by glycogen synthase kinase-3 (GSK-3). FAP-induced expression of CD80, CD86, major histocompatibility complex (MHC) class I, and MHC class II in TLR4+/+ BMDCs was not observed in TLR4-/- BMDCs. Furthermore, FAP induced DC-mediated CD8+ T cell proliferation and cytotoxic T lymphocyte (CTL) activity, and suppressed tumor growth with DC-based tumor vaccination in EG7 thymoma murine model. Taken together, these results indicate that the TLR4 agonist, FAP, a potential immunoadjuvant for DC-based cancer vaccination, improves the DC-based immune response via the TLR4 signaling pathway.

Optimization and Limitation of Calcium Ionophore to Generate DCs from Acute Myeloid Leukemic Cells / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: Calcium ionophore (CI) is used to generate dendritic cells (DCs) from progenitor cells, monocytes, or leukemic cells. The aim of this study was to determine the optimal dose of CI and the appropriate length of cell culture required for acute myeloid leukemia (AML) cells and to evaluate the limitations associated with CI. MATERIALS AND METHODS: To generate leukemic DCs, leukemic cells (4 x 10(6) cells) from six AML patients were cultured with various concentrations of CI and/or IL-4 for 1, 2 or 3 days. RESULTS: Potent leukemic DCs were successfully generated from all AML patients, with an average number of 1.2 x 10(6) cells produced in the presence of CI (270 ng/ml) for 2 days. Several surface molecules were clearly upregulated in AML cells supplemented with CI and IL-4, but not CD11c. Leukemic DCs cultured with CI had a higher allogeneic T cell stimulatory capacity than untreated AML cells, but the addition of IL-4 did not augment the MLR activity of these cells. AML cells cultured with CI in the presence or absence of IL-4 showed increased levels of apoptosis in comparison to primary cultures of AML cells. CONCLUSION: Although CI appears to be advantageous in terms of time and cost effectiveness, the results of the present study suggest that the marked induction of apoptosis by CI limits its application to the generation of DCs from AML cells.

Induction of Myeloma Cell Line-specific Cytotoxic T Lymphocytes using Monocyte-derived Dendritic Cells Pulsed with Myeloma Cell Line Lysates

BACKGROUND: In multiple myeloma (MM), the idiotype (ID) determinant of the paraprotein has been used for immunotherapy using dendritic cells (DCs). However, ID-specific immune responses showed limited clinical responses after the Id vaccination. Therefore, an alternative approach using DCs pulsed with other tumor antigens is required. METHODS: We investigated the possibility of immunotherapy for MM using myeloma cell line-specific cytotoxic T lymphocytes (CTLs), that were stimulated in vitro by monocyte-derived DCs pulsed with the myeloma cell line ysates. CD14+ cells isolated from the peripheral blood of HLA-A0201+ healthy donors were cultured in the presence of GM-CSF and IL-4. On day 6, the immature DCs were pulsed with the myeloma cell line lysates (IM-9: HLA0201+ and ARH-77: HLA0201+), and then maturation of DCs was induced by the addition of TNF- alpha for 2 days. CTL lines were generated by a 2 time stimulation with DCs to the autologous CD3+ T cells. RESULTS: DCs pulsed with myeloma cell lysates showed the production of IL-12p70, but less than that of unpulsed DCs. CTLs lines stimulated with the DCs pulsing, for the myeloma cell line lysates, showed potent cytotoxic activities against autologous target cells, but not against HLA-A2-cell lines (RPMI-8226). Mature DCs pulsed with the myeloma cell line lysates showed a higher stimulatory capacity for autologous CTL when compared with mature non-pulsed DCs. CONCLUSION: These results suggest that DCs pulsed with the myeloma cell line lysates can generate potent myeloma cell line-specific CTLs for the myeloma cell-based immunotherapeutic approach in MM.

Selective Expansion of Natural Killer Cells from Peripheral Blood Mononuclear Cells by K562 Cell Line and IL-2

BACKGROUND: Several attempts have been made to expand human NK cells from peripheral blood mononuclear cells (PBMCs). This study examined the selective expansion of NK cells using interleukin 2 (IL-2) plus the K562 cell line, the expression of the NK cell receptors, and the cytotoxic activity. METHODS: The PBMCs from seven healthy volunteers were cultured in a medium containing the IL-2 plus the K562 cell line for 14 days. The expression of the activating and inhibitory receptors on the resting NK cells and the 72 hr-expanded NK cells were analyzed. A flow cytometric cytotoxic assay was used to determined the killing activity of the non-expanded NK cells and the 7 day-expanded NK cells against the K562 target cells. RESULTS: The NK cells from PBMCs expanded 4.5-fold after 7 days, and contained 56.5% CD3-CD56+ cells. The IL-2 or IL-2 plus K562 increased the expression levels of CD158b (MFI, mean florescence intensity), CD158e1/e2 (MFI), and NKp44 (MFI), while it decreased the expression levels of NKp30 (%), CD16 (MFI), and 2B4 (MFI). The non-expanded NK cells lysed 9.0% and 27.6% of the K562 target cells in the 1 : 1 and 5 : 1 effector and target ratio, respectively, and the 7-day expanded NK cells lysed 36.9% and 57.2% of the K562 target cells, respectively. CONCLUSION: The selective expansion of CD3-CD56+ NK cells occurred only during 7 days of culture. IL-2 or IL-2 plus the K562 cells altered the expression of various activating and inhibitory receptors of NK cells, and the cytotoxicity of the expanded NK cells was higher than in the non-expanded cells.

Chronic Exertional Compartment Syndrome in Behcet's Disease : A case report

We experienced a patient who presented recurrent episode of pain, swelling, and weakness of right lower leg. She was diagnosed as Behcet's disease 1 year ago. The symptom was developed after exercise and was relieved by rest. After several different examinations and differential diagnoses, we confirmed the diagnosis as chronic exertional compartment syndrome on the basis of measuring intramuscular compartmental pressure. The patient was recommended to take a rest and she got a shoe-modification, prescription of ankle- foot orthosis, and physical therapy. She experienced symptom relief after that. Chronic exertional compartment syndrome is uncommon disease caused by reversible ischemia secondary to increased pressure within a closed osseofascial compartment after exercise producing expansion of muscle volume.

The Effect of Peripheral Vascular Disease on Diabetic Neuropathy

OBJECTIVE: To evaluate the effect of peripheral vascular disease (PVD) on diabetic neuropathy with the use of Doppler ultrasound and electrodiagnostic study. METHOD: One hundred fifty one patients with diabetes mellitus underwent nerve conduction studies. PVD was diagnosed when ankle-brachial index (ABI) was 0.9 and less and also toe-brachial index (TBI) was 0.7 and less. Electrophysiologically normal group was subdivided into non- PVD group (A1) and PVD group (A2). Diabetic neuropathy group was subdivided into non-PVD group (B1) and PVD group (B2). The frequency of diabetic neuropathy and the difference of amplitude, conduction velocity, and F wave latency within A groups and B groups were investigated. RESULTS: Diabetic neuropathy was significantly correlated with PVD (p<0.05). There was no definite difference of electrophysiologic parameters between A1 and A2 groups. B1 group showed significantly reduced amplitude of sensory nerve action potential (SNAP) in sural nerve compared with B2 group (p<0.05). In all patients, the amplitude of SNAP in sural nerve was related with duration of diabetes and TBI by multiple linear regression analysis. CONCLUSION: This study supports the influence of PVD on diabetic neuropathy and suggests vascular abnormality in patients with diabetic neuropathy may result in predominantly axonal injury rather than demyelinating injury.

Anatomical Considerations of the Lateral Femoral Cutaneous Nerve Related to Nerve Conduction Study

OBJECTIVE: Lateral femoral cutaneous nerve (LFCN) conduction study is an objective measure for the diagnosis of meralgia paresthetica. Sensory nerve action potential of LFCN is not frequently evoked because of anatomical variations around inguinal area. The purpose of this study is to support the diagnosis of meralgia paresthetica by considering anatomical variations of LFCN in Korean adult cadavers. METHOD: Eighteen lower limbs of total nine adult cadavers were studied. Men were five and women were four. The points that LFCN or the main branch of LFCN met the imaginary line from anterior superior iliac spine (ASIS) to pubic tubercle and to lateral border of patella were recorded, respectively and distances from ASIS to those points were measured. RESULTS: The distance from ASIS to the point that LFCN or the main branch of LFCN met the imaginary line from ASIS to pubic tubercle and to lateral border of patella was respectivlely 1.36+/-0.68 cm (minimal 0.2, maximal 3.0) and 10.74+/-5.68 (minimal 3.3, maximal 20.1) cm. There was no significant distance difference between men and women. CONCLUSION: This study showed anatomic variations of LFCN around ASIS and femoral part. This knowledge may help LFCN conduction study for the diagnosis of meralgia paresthetica.

Trp-Lys-Tyr-Met-Val-Met stimulates phagocytosis via phospho-lipase D-dependent signaling in mouse dendritic cells

Dendritic cells (DCs) play a key role in activating the immune response against invading pathogens as well as dying cells or tumors. Although the immune response can be initiated by the phagocytic activity by DCs, the molecular mechanism involved in this process has not been fully investigated. Trp-Lys-Tyr-Met-Val-Met-NH2 (WKYMVM) stimulates the activation of phospholipase D (PLD) via Ca2+ increase and protein kinase C activation in mouse DC cell line, DC2.4. WKYMVM stimulates the phagocytic activity, which is inhibited in the presence of N-butanol but not t-butanol in DC2.4 cells. Furthermore, the addition of phosphatidic acid, an enzymatic product of PLD activity, enhanced the phagocytic activity in DC2.4 cells. Since at least two of formyl peptide receptor (FPR) family (FPR1 and FPR2) are expressed in DC2.4 as well as in mouse bone marrow-derived dendritic cells, this study suggests that the activation of FPR family by WKYMVM stimulates the PLD activity resulting in phagocytic activity in DC2.4 cells.

Dorsally Migrated Sequestrated Lumbar Disc Herniation with Cauda Equina Syndrome: Two Cases Report / 대한척추외과학회지

We have experienced 2 patients with dorsally migrated sequestrated lumbar disc herniation with cauda equina syndrome and obtained a complete motor and sensory recovery after early decompressive laminectomy and discectomy with or without fusion in all cases. The purpose of this paper is to report two unusual cases of lumbar disc herniation that we experienced, and to emphasize the importance of early decompressive surgery for the treatment of cauda equina syndrome accompanied with dorsally migrated sequestrated lumbar disc herniation.