Monocyte Colony-stimulating Factor Research Articles

Adult Recurrent Respirator Papillomatosis

To determine a new therapeutic approach using granulocyte monocyte-colony-stimulating factor (GM-CSF) and pegylated interferon alpha 2a (Peg-IFNα-2a) as adjuvant therapy in patients with adult recurrent respiratory papillomatosis. Descriptive observational clinical trial. Departments of Otolaryngology and Immunology. Fourteen patients with adult recurrent respiratory papillomatosis were examined regarding medical history and number of operations before and after treatment. Voice disorder and glottal stop were evaluated using the Voice-Related Quality-of-Life instrument. Papilloma staging was determined using the Coltera/Derkay diagram. The patients received Peg-IFNα-2a at 180 mcg weekly for 6 months. In the third month, the patients began GM-CSF treatment at 400 mcg weekly for 2 months. The patients were observed for 12 months after treatment ended. Eleven patients met the study criteria; 3 patients had tracheotomies before treatment, and they were decannulated after treatment. Before treatment, the scale of voice quality ranged from 34 to 45 points (mean, 38.31). After treatment, the range was 12 to 35 points (mean, 21.09; P < .001). Prior to therapy, the glottal stop ranged from 50% to 90% (average, 62.27%). After therapy, the range decreased to 0% to 15% (mean, 4.63%; P < .001). The number of surgical interventions decreased. Two patients each had 1 surgical intervention after treatment began. A new adjuvant treatment based on immunogenetic mechanisms against human laryngeal papilloma virus, with expectations of reducing disease aggressiveness and the number of operations, avoids the risks of surgery. Peg-IFNα-2a and GM-CSF is an adjuvant therapy for treating adult recurrent respiratory papillomatosis.

Type 3 Muscarinic Receptors Modulate Macrophage Phenotype in the Presence of Th2 Cytokines

Purpose: Macrophages (Mφ) play a critical role in IBD. Resident Mφs within the intestinal lamina propria are exposed to many antigens, but are tolerant to stimulation and prevent inflammation (Heinsbroek & Gordon, 2009). We showed previously that stimulation of type 3 (M3R) muscarinic receptors (MR) encoded by Chrm3 promoted the development of pro-inflammatory classically activated Mφs (CAM) (McLean, et al, 2012). Whether M3R plays a role in the development of alternatively activated Mφs (AAM), which promote wound healing and tissue repair, is unclear. Our aim was to test the hypothesis that M3R plays a role in AAM development. Methods: Bone marrow cells were isolated from WT and Chrm3-/- mice and differentiated in the presence of monocyte colony stimulating factor (M-CSF) for 7 days. Flow cytometry was used to analyze cell populations. After 24 hour treatment with a muscarinic receptor-selective agonist, bethanechol (BTH), +/- the Th2 cytokine IL-4, cells were harvested and mRNA expression of the AAM marker Arg-1 assess by real-time PCR. Results: We confirmed that ˜95% of our cell population was positive for both CD11b and F4/80, indicating that these cells are Mφs. These Mφs express mRNA for M1R, M2R, M3R and M4R. In WT Mφs, BTH treatment significantly attenuated Arg-1 expression, indicating a constitutive bias against AAM development. Treatment of naïve WT Mφs with IL-4 significantly up-regulated Arg-1 expression; addition of BTH to IL-4 did not enhance this effect. In contrast, in Chrm3-/- Mφs up-regulation of Arg-1 in response to IL-4 was significantly greater than that observed in WT Mφs. Moreover, addition of BTH to IL-4 potentiated this up-regulation of Arg-1 in Chrm3-/- Mφs. Conclusion: Treatment with BTH alone attenuated the development of AAMs from naïve WT Mφs. As expected, treatment of WT Mφs with IL-4 induced the development of AAMs. In WT Mφs, addition of BTH to IL-4 did not modulate this effect. In M3R-deficient Mφs, IL-4 further polarized Mφs to the AAM phenotype. Notably, exposing M3R-deficient Mφs to both IL-4 & BTH further enhances AAM polarization. We previously reported that M3R expression exerts pro-inflammatory effects on Mφs. Our findings suggest that in the presence of IL-4, M3R activation attenuates the development of AAMs and, in the presence of IL-4, further polarizes M3R-deficient Mφs to an AAM phenotype, possibly by acting at M1R, M2R, and/or M4R. This effect is not observed in WT Mφs, likely due to M3R activity. These data further support a pro-inflammatory role of M3R. Pharmacologic attempts to manipulate Mφ phenotypes should focus on blocking M3R expression or activation to promote AAMs, a strategy that may reduce disease activity or maintain remission in IBD patients.

The nuclear membrane leukotriene synthetic complex is a signal integrator and transducer

Leukotrienes (LTs) are lipid-signaling molecules derived from arachidonic acid (AA) that initiate and amplify inflammation. To initiate LT formation, the 5-lipoxygenase (5-LO) enzyme translocates to nuclear membranes, where it associates with its scaffold protein, 5-lipoxygenase-activating protein (FLAP), to form the core of the multiprotein LT synthetic complex. FLAP is considered to function by binding free AA and facilitating its use as a substrate by 5-LO to form the initial LT, LTA(4). We used a combination of fluorescence lifetime imaging microscopy, cell biology, and biochemistry to identify discrete AA-dependent and AA-independent steps that occur on nuclear membranes to control the assembly of the LT synthetic complex in polymorphonuclear leukocytes. The association of AA with FLAP changes the configuration of the scaffold protein, enhances recruitment of membrane-associated 5-LO to form complexes with FLAP, and controls the closeness of this association. Granulocyte monocyte colony-stimulating factor provides a second AA-independent signal that controls the closeness of 5-LO and FLAP within complexes but not the number of complexes that are assembled. Our results demonstrate that the LT synthetic complex is a signal integrator that transduces extracellular signals to modulate the interaction of 5-LO and FLAP.

Polymorphonuclear Neutrophils in Osteomyelitis: Link to Osteoclast Generation and Bone Resorption

Chronic and persistent inflammatory processes in bones may lead to severe erosions with consequent functional impairment sometimes requiring amputation of the limb. To explore the relationship between inflammation and bone erosion, biopsies of patients with osteomyelitis due to arterial occlusive disease or to diabetes mellitus were examined (n=31). Histologically, inflammation and bone erosion were confirmed. In the eroded bones the number of osteoclasts correlated with the abundance of infiltrated polymorphonuclear neutrophils (PMN), which were highly activated as shown by expression of MHC class II. For functional characterisation of the infiltrating PMN, patients with implant-associated osteomyelitis, a condition associated with persistent bacterial infection and bone destruction, were recruited. The cells were recovered from infected sites and examined ex vivo. These PMN expressed MHC class II and produced interleukin (IL)-8, a further indication of PMN activation. To assess a possible link between infiltrating PMN and bone erosion, we tested the effect of IL-8 on osteoclast generation in vitro. CD14+ monocytes derived from the peripheral blood of healthy individuals were cultivated with monocyte colony stimulating factor (M-CSF) and IL-8. Within 3 days, a translocation of the transcription factor NFATcl into the nucleus was seen, and by 10 to 20 days multinucleated cells with typical osteoclast morphology appeared that expressed tartrate-resistant acid phosphatase (TRAP) and cathepsin K. Moreover, the cells were able to resorb bone, proving that IL-8 was able to induce the differentiation of monocytes to osteoclasts. Because IL-8 is a major cytokine produced by activated PMN, we propose that in the course of persistent infection infiltrating PMN contribute to induction of osteoclast formation, thus providing a link between inflammation and bone erosion.

Korean Red Ginseng Saponin Fraction Downregulates Proinflammatory Mediators in LPS Stimulated RAW264.7 Cells and Protects Mice against Endotoxic Shock

Korean red ginseng has shown therapeutic effects for a number of disease conditions. However, little is known about the antiinflammatory effect of Korean red ginseng saponin fraction (RGSF) in vitro and in vivo. Therefore, in this study, we showed that RGSF containing 20(S)-protopanaxadiol type saponins inhibited nitric oxide production and attenuated the release of tumor necrotic factor (TNF)-α, interleukin (IL)-6, granulocyte monocyte colony stimulating factor (GMCSF), and macrophage chemo-attractant protein-1 in lipopolysaccharide (LPS) stimulated murine macrophage RAW264.7 cells. Moreover, RGSF down-regulated the mRNA expressions of inducible nitric oxide synthase, cyclooxyginase-2, IL-1β, TNF-α, GMCSF, and IL-6. Furthermore, RGSF reduced the level of TNF-α in the serum and protected mice against LPS mediated endotoxic shock. In conclusion, these results indicated that ginsenosides from RGSF and their metabolites could be potential sources of therapeutic agents against inflammation.

FMS Kinase Inhibitors: Current Status and Future Prospects

FMS, first discovered as the oncogene responsible for Feline McDonough Sarcoma, is a type III receptor tyrosine kinase that binds to the macrophage or monocyte colony-stimulating factor (M-CSF or CSF-1). Signal transduction through that binding results in survival, proliferation, and differentiation of monocyte/macrophage lineage. Overexpression of CSF-1 and/or FMS has been implicated in a number of disease states such as the growth of metastasis of certain types of cancer, in promoting osteoclast proliferation in bone osteolysis, and many inflammatory disorders. Inhibition of CSF-1 and/or FMS may help treat these pathological conditions. This article reviews FMS gene, FMS kinase, CSF-1, IL-34, and their roles in bone osteolysis, cancer biology, and inflammation. Monoclonal antibodies, FMS crystal structure, and small molecule FMS kinase inhibitors of different chemical scaffolds are also reviewed.

Distribution of granulocyte–monocyte colony-stimulating factor and its receptor α-subunit in the adult human brain with specific reference to Alzheimer’s disease

Granulocyte-monocyte colony-stimulating factor (GM-CSF) is a member of the hematopoietic growth factor family, promoting proliferation and differentiation of hematopoietic progenitor cells of the myeloid lineage. In recent years, GM-CSF has also proved to be an important neurotrophic factor in the central nervous system (CNS) via binding to the GM-CSF receptor (GM-CSF R). Furthermore, studies on rodent CNS revealed a wide distribution of both the major binding α-subunit of the GM-CSF R (GM-CSF Rα) and its ligand. Since respective data on the expression pattern of these two molecules are still lacking, the present study has been designed to systematically analyze the protein expression of GM-CSF and GM-CSF Rα in the human brain, with particular emphasis on their regulation in Alzheimer's disease (AD). One major finding is that both GM-CSF and GM-CSF Rα were ubiquitously but not uniformly expressed in neurons throughout the CNS. Protein expression of GM-CSF and GM-CSF Rα was not restricted to neurons but also detectable in astrocytes, ependymal cells and choroid plexus cells. Interestingly, distribution and intensity of immunohistochemical staining for GM-CSF did not differ among AD brains and age-matched controls. Concerning GM-CSF Rα, a marked reduction of protein expression was predominantly detected in the hippocampus although a slight reduction was also found in various cortical regions, thalamic nuclei and some brainstem nuclei. Since the hippocampus is one of the target regions of neurodegenerative changes in AD, reduction of GM-CSF Rα, with consecutive downregulation of GM-CSF signaling, may contribute to in the progressive course of neurodegeneration in AD.

Circulating cytokine concentrations are not associated with major depressive disorder in a community-based cohort

ObjectiveThe objective was to test the hypotheses that cytokine levels are elevated in community-residing persons at high risk for obstructive sleep apnea with major depressive disorder (MDD) compared to nondepressive persons and that cytokine levels show stronger correlations with somatic than psychological symptoms of depression. MethodA case–control study within the cross-sectional Akershus Sleep Apnea Project was performed. Two controls matched for age, gender, metabolic syndrome and obstructive sleep apnea were drawn for each case of MDD. ResultsGroup comparisons revealed no significant difference in the levels of 17 cytokines [interleukin-1β, -2,-4, -5, -6, -7, -8, -10, -12(p70), -13 and -17; tumor necrosis factor-α; interferon-γ; granulocyte colony-stimulating factor; granulocyte–monocyte colony-stimulating factor; macrophage chemoattractant protein-1 and monocyte inhibitory protein-1β] between persons with (n=34) and without MDD (n=68). There was no association between cytokines levels and MDD in multivariate regression analyses. The concentration of interleukin-4 was significantly more positively correlated with psychological than somatic symptoms (r=0.046 vs. −0.143, respectively, P=0.024), while no different correlations were observed for other cytokines. ConclusionThe cytokine levels were not elevated in MDD, and cytokine levels were not more strongly associated with somatic than psychological symptoms of depression. The depression-specific effect on inflammation may be weak in community-based samples with prevalent somatic comorbidity.

Resveratrol, wine, and atherosclerosis.

This review emphasizes the effects of resveratrol on factors involved in the mechanism of atherosclerosis and risk factors for atherosclerosis. The effects of wine and resveratrol on atherosclerosis are also discussed. Resveratrol is a potent antioxidant and an anti-inflammatory agent. It reduces the expression of cell adhesion molecules, monocyte colony stimulating factors, matrix metalloproteinases, and growth factors; and inhibits platelet aggregation and vascular smooth muscle cell proliferation. It reduces the serum levels of total cholesterol, triglycerides (TG), and raises high-density lipoprotein cholesterol, inhibits expression of C-reactive protein and lowers the levels of advanced glycation end products and its receptor in the vascular tissue. It lowers the risk factors for plaque rupture. Epidemiological data show that moderate consumption of alcohol has an inverse association with carotid atherosclerosis while high consumption has a positive association with carotid atherosclerosis. Wine reduces the extent of atherosclerosis in animal model. The antiatherosclerotic effect of wine is mainly due to it resveratrol content. Resveratrol reduces the extent of atherosclerosis in animal model of atherosclerosis (apolipoprotein [Apo] E-deficient and Apo E(-/-)/low-density lipoprotein receptor-deficient mice and macrophage). In rabbit model of atherosclerosis, both reduction and acceleration of atherosclerosis have been reported with resveratrol. There are no data for regression and slowing of progression of atherosclerosis. Robust clinical trials for suppression of atherosclerosis are lacking. In conclusion, resveratrol has potential but experimental studies in depth and robust clinical trials are lacking for this agent to be of any value in the primary and secondary prevention of coronary and peripheral artery disease.

Childhood Herpes Zoster-Triggered Guttate Psoriasis

Psoriasis is commonly triggered or exacerbated by various stress factors, certain drugs, or streptococcal throat infections. Viral infections such as HIV, CMV, chikungunya, or herpes simplex virus are very uncommon triggers for psoriasis. Cases of varicella-triggered psoriasis are exceptional. A 7-year-old boy with a previous history of guttate psoriasis presented with generalized acute guttate psoriasis shortly after an extensive herpes zoster infection affecting the first and second left lumbar dermatomes. INF-alpha and granulocyte monocyte colony stimulating factor influence peripheral monocytes to transform into INF-dendritic cells (DC's), similar to those involved in psoriasis. These INF-DC's express toll-like receptors 7 and 8, which are responsive to viral single stranded RNA. Hence, viral infections and interferon (INF)-alpha may play a role in triggering psoriasis.

TGF-β1-dependent L1CAM expression has an essential role in macrophage-induced apoptosis resistance and cell migration of human intestinal epithelial cells

Patients with chronic inflammatory bowel disease (IBD) have an increased risk to develop colorectal cancer (CRC) particularly after long duration of the disease. Chronic inflammation of the intestinal mucosa is characterized by a marked enrichment of immune cells such as macrophages as well as by high expression of cytokines and growth factors including transforming growth factor-beta 1 (TGF-β1). The adhesion molecule L1CAM mediates chemoresistance and migration of tumor cells and is elevated in CRC tissues being associated with metastatic spread and poor prognosis for the patients. In this study, we examine the role of TGF-β1-induced L1CAM expression and macrophages in malignant transformation of intestinal epithelial cells. We demonstrate that TGF-β1 stimulation leads to a Slug-dependent upregulation of L1CAM expression already in the colonic intestinal epithelial cell line NCM460 thereby enhancing cell motility and apoptosis resistance. Accordingly, NCM460 cells acquired a migratory and apoptosis-resistant phenotype if transfected with L1CAM. Immunohistochemistry of colonic biopsies revealed considerable L1CAM expression in intestinal epithelial cells in tissues from IBD patients but not in normal colonic tissues. Moreover, L1CAM expression increased with duration of disease being associated with the presence of CD33+ macrophages. Coculture with macrophages generated from monocyte colony-stimulating factor (MCSF)-treated monocytes led to the upregulation of Slug and L1CAM in NCM460 cells thereby elevating cell motility and apoptosis resistance. Pharmacological inhibition of TGF-β1 signalling abolished expression of Slug and L1CAM in cocultured NCM460 cells resulting in decreased cell migration and apoptosis resistance. In conclusion, these data provide new insights into the mechanisms by which IBD promotes malignant transformation of intestinal epithelial cells and underscore the role of L1CAM and macrophages in this scenario.

Autocrine CSF-1 and CSF-1 Receptor Coexpression Promotes Renal Cell Carcinoma Growth

Renal cell carcinoma is increasing in incidence but the molecular mechanisms regulating its growth remain elusive. Coexpression of the monocytic growth factor colony-stimulating factor (CSF)-1 and its receptor CSF-1R on renal tubular epithelial cells (TEC) will promote proliferation and antiapoptosis during regeneration of renal tubules. Here, we show that a CSF-1-dependent autocrine pathway is also responsible for the growth of renal cell carcinoma (RCC). CSF-1 and CSF-1R were coexpressed in RCCs and TECs proximally adjacent to RCCs. CSF-1 engagement of CSF-1R promoted RCC survival and proliferation and reduced apoptosis, in support of the likelihood that CSF-1R effector signals mediate RCC growth. In vivo CSF-1R blockade using a CSF-1R tyrosine kinase inhibitor decreased RCC proliferation and macrophage infiltration in a manner associated with a dramatic reduction in tumor mass. Further mechanistic investigations linked CSF-1 and epidermal growth factor signaling in RCCs. Taken together, our results suggest that budding RCC stimulates the proximal adjacent microenvironment in the kidney to release mediators of CSF-1, CSF-1R, and epidermal growth factor expression in RCCs. Furthermore, our findings imply that targeting CSF-1/CSF-1R signaling may be therapeutically effective in RCCs.

Successful treatment of Trichosporon fungemia in a patient with refractory acute myeloid leukemia using voriconazole combined with liposomal amphotericin B

Trichosporon fungemia is a rare and fatal fungal infection that occurs in patients with prolonged neutropenia associated with hematologic malignancies. A 21-year-old male developed Trichosporon fungemia during remission induction therapy for acute myeloid leukemia (AML). Although two courses of induction therapy failed to induce a remission of AML, combination therapy with voriconazole and liposomal amphotericin B (L-AmB) followed by monocyte colony-stimulating factor ameliorated the Trichosporon fungemia and enabled the patient to receive reduced-intensity bone marrow transplantation (BMT) from his human leukocyte antigen-A one-locus mismatched mother. The patient achieved a durable remission after BMT without exacerbation of Trichosporon fungemia. The combination therapy with voriconazole and L-AmB may therefore be useful in controlling Trichosporon fungemia associated with prolonged neutropenia after remission induction therapy for AML.

Thujone inhibits lung metastasis induced by B16F-10 melanoma cells in C57BL/6 mice

The antimetastatic potential of thujone, a naturally occurring monoterpene, was evaluated. Metastasis was induced in C57BL/6 mice by injecting highly metastatic B16F-10 melanoma cells through the lateral tail vein. Administration of thujone (1 mg·(kg body weight)(-1)), prophylactically and simultaneously with tumor induction, inhibited tumor nodule formation in the lungs by 59.45% and 57.54%, respectively, with an increase in the survival rate (33.67% and 32.16%) of the metastatic tumor bearing animals. These results correlated with biochemical parameters such as lung collagen hydroxyproline, hexosamine and uronic acid contents, serum sialic acid and γ-glutamyl transpeptidase levels, and histopathological analysis. Treatment with thujone downregulated the production of proinflammatory cytokines such as tumor necrosis factor-α, interleukin (IL)-1β, IL-6, and granulocyte-monocyte colony-stimulating factor. Thujone administration downregulated the expression of matrix metalloproteinase (MMP)-2, MMP-9, extracellular signal-regulated kinase (ERK)-1, ERK-2, and vascular endothelial growth factor (VEGF) and also upregulated the expression of nm-23, tissue inhibitor of metalloproteinase (TIMP)-1, and TIMP-2 in the lung tissue of metastasis-induced animals. Treatment with thujone inhibited the activity of MMP-2 and MMP-9 in gelatin zymographic analysis. Thujone treatment significantly inhibited the invasion of B16F-10 melanoma cells across the collagen matrix in a Boyden chamber. Thujone also inhibited the adhesion of tumor cells to collagen-coated microtire plate wells and the migration of B16F-10 melanoma cells across a polycarbonate filter in vitro. These results indicate that Thujone can inhibit the lung metastasis of B16F-10 cells through inhibition of tumor cell proliferation, adhesion, and invasion, as well as by regulating expression of MMPs, VEGF, ERK-1, ERK-2, TIMPs, nm23, and levels of proinflammatory cytokines and IL-2 in metastatic animals.

Diverse patterns of cyclooxygenase‐independent metalloproteinase gene regulation in human monocytes

BACKGROUND AND PURPOSEMatrix metalloproteinase (MMP) production from monocyte/macrophages is implicated in matrix remodelling and modulation of inflammation. However, knowledge of the patterns and mechanisms of gene regulation of MMPs and their endogenous tissue inhibitors (TIMPs) is fragmentary. MMP up-regulation may be a target for cyclooxygenase (COX) and prostaglandin (PG) receptor inhibition, but the extent and mechanisms of COX-independent MMP up-regulation are unclear.EXPERIMENTAL APPROACHWe studied MMP mRNA expression and selected protein levels in human peripheral blood monocytes before and after adhesion, upon stimulation with bacterial lipopolysaccharide (LPS), PGE2 or forskolin and after culturing with monocyte colony-stimulating factor on plastic or human fibronectin for up to 7 days.KEY RESULTSMonocyte adherence for 2 h transiently up-regulated COX-2, MMP-1, MMP-7 and MMP-10 mRNAs, and persistently up-regulated MMP-2, MMP-9, MMP-14 and MMP-19 mRNAs. LPS, PGE2 or forskolin selectively increased MMP-1, MMP-9, MMP-10, MMP-12 and MMP-14 mRNAs. LPS increased PGE2 production through COX but up-regulated MMP levels independently of COX. Differential dependence on inhibition of p42/44 and p38 mitogen-activated protein kinases, c-jun N-terminal kinase and inhibitor of κB kinase2 paralleled the diverse patterns of MMP stimulation by LPS. Differentiation on plastic increased mRNA levels of MMP-7, MMP-9, MMP-12 and MMP-14 and TIMP-2 and TIMP-3 independently of COX; fibronectin accelerated MMP but not TIMP up-regulation.CONCLUSIONS AND IMPLICATIONSAdhesion, LPS stimulation and maturation of human monocytes lead to selective, COX-independent MMP and TIMP gene regulation, which is a potential target for selective inhibition by signalling kinase inhibitors.

Requirement of the Acquired Immune System in Successful Cancer Chemotherapy With cis-Diamminedichloroplatinum (II) in a Syngeneic Mouse Tumor Transplantation Model

We examined the involvement of acquired immune response in the therapeutic effect of systemic tumor chemotherapy with cis-diamminedichloroplatinum (II) (cis-DDP) using a mouse syngeneic cancer transplantation model. The therapeutic effect observed in BALB/c mice was negligible in athymic BALB/c mice. The therapeutic effect became obvious when athymic mice were immunologically reconstituted with splenocytes containing CD4 T cells of BALB/c mice. The BALB/c mice in which tumor chemotherapy was successful exhibited cell line-specific tumor immunity to the second tumor transplantation. Splenocytes of these mice exhibited cell line-specific cytotoxicity. After systemic cis-DDP treatment, an increase in apoptotic cells and macrophage infiltration was observed in the tumor mass. In the same lesion, an increase in the levels of high-mobility group box protein and its intracellular translocation from the nucleus to the cytoplasm were observed. In a chase study on the infiltrated macrophage with labeled tumor cells, the translocation of tumor cell components to regional lymph nodes was observed after the chemotherapy. The peripheral monocytosis induced by administrations of monocyte colony-stimulating factor augmented the therapeutic efficacy of cis-DDP treatment. These indicate that the phagocytosis of apoptotic tumor cells exhibiting the adjuvant protein by infiltrated macrophages and the subsequent antigen presentation by the macrophages to T cells take place, and that the acquired immune response to the tumor cells as the consequence plays an essential role in the therapeutic effect of cis-DDP.

MHC Class I–Presented Tumor Antigens Identified in Ovarian Cancer by Immunoproteomic Analysis Are Targets for T-Cell Responses against Breast and Ovarian Cancer

The purpose of this study is to test whether peptide epitopes chosen from among those naturally processed and overpresented within MHC molecules by malignant, but not normal cells, when formulated into cancer vaccines, could activate antitumor T-cell responses in humans. Mixtures of human leukocyte antigen A2 (HLA-A2)-binding ovarian cancer-associated peptides were used to activate naive T cells to generate antigen-specific T cells that could recognize ovarian and breast cancers in vitro. Combinations of these peptides (0.3 mg of each peptide or 1 mg of each peptide) were formulated into vaccines in conjunction with Montanide ISA-51 and granulocyte monocyte colony stimulating factor which were used to vaccinate patients with ovarian and breast cancer without evidence of clinical disease in parallel pilot clinical trials. T cells specific for individual peptides could be generated in vitro by using mixtures of peptides, and these T cells recognized ovarian and breast cancers but not nonmalignant cells. Patient vaccinations were well tolerated with the exception of local erythema and induration at the injection site. Nine of the 14 vaccinated patients responded immunologically to their vaccine by inducing peptide-specific T-cell responses that were capable of recognizing HLA-matched breast and ovarian cancer cells. Mixtures of specific peptides identified as naturally presented on cancer cells and capable of activating tumor-specific T cells in vitro also initiate or augment immune responses toward solid tumors in cancer patients.

Acidity increases the uptake of native LDL by human monocyte-derived macrophages

The extracellular pH is locally decreased in advanced atherosclerotic lesions, particularly in lipid-rich areas of the lesions. Since accumulation of LDL-derived cholesterol and formation of foam cells are key processes in atherogenesis, we tested here the effects of acidic pH on the uptake of native LDL. First, human monocytes were differentiated into macrophages in the presence of granulocyte–monocyte-colony stimulating factor (GM-CSF) after which native LDL was applied to the monocyte-derived macrophages at pH 5.5, 6.5, or 7.5 and the binding and uptake of LDL by macrophages were determined. The lower the pH was, the higher was the binding and uptake of LDL by macrophages. Also, acidic pH was found to increase the production of cell surface proteoglycans by macrophages and binding of LDL to the glycosaminoglycan chains of the proteoglycans. The acidity-induced increase in the uptake of LDL by macrophages could be inhibited by pretreating the cells with heparinase and chondroitinase as well as by inhibiting the production of proteoglycans with NaClO 3. Thus, the observed increase in the uptake of native LDL to macrophages appears to depend on the increased ability of LDL to bind to cell surface proteoglycans at acidic pH. Taken together, our present results indicate that acidity increases the effective concentration of LDL on macrophage surfaces by increasing the amount of cell surface proteoglycans and by enhancing the binding of LDL to them and so promotes LDL uptake with ensuing foam cell formation.

IL-22 promotes osteoclastogenesis in rheumatoid arthritis through induction of RANKL from synovial fibroblasts (117.25)

Abstract This study aims to determine the regulatory role of interleukin(IL)-22 on the expression of receptor activator of NF-kB ligand(RANKL) and the induction of osteoclastogenesis in rheumatoid arthritis(RA). The concentrations of IL-22 and RANKL in sera and synovial fluids(SF)of RA patients were measured by ELISA. After RA synovial fibroblasts were treated with rhIL-22, the expression of RANKL mRNA and protein was determined using real-time PCR,western blot,and immunostaining.Human monocytes were cocultured with IL-22-prestimulated RA synovial fibroblasts and monocyte-colony stimulating factor(M-CSF), and then osteoclastogenesis was determined by counting of TRAP-positive multinucleated cells. Serum and SF IL-22 concentration in RA patients was higher than in osteoarthritis patients and healthy subjects. When RA synovial fibroblasts were treated with rhIL-22, the expression of RANKL mRNA and protein was upregulated in a dose-dependent manner. The IL-22-induced RANKL expression was significantly down-regulated by inhibition of p38 MAPK/NF-kB and JAK-2/STAT3. When human monocytes were cultured with IL-22-prestimulated RA synovial fibroblasts in the absence of RANKL, the monocytes were differentiated into osteoclasts, which was blocked by the inhibition of p38 MAPK/NF-kB and JAK-2/STAT3.Our results suggest that IL-22 up-reugulates RANKL expression in RA synovial fibroblasts and it also promotes osteoclastogenesis, which are mediated by p38 MAPK/NF-kB and JAK-2/STAT3 pathways.

Enhanced expression of cytokines/chemokines in cerebrospinal fluids in mumps meningitis in children

The mumps virus is frequently the causative agent in aseptic meningitis and mumps has still prevailed in Japan. We compared data obtained from patients with mumps meningitis and patients with aseptic meningitis caused by other viruses in order to identify mumps meningitis-specific cytokine/chemokine alterations in cerebrospinal fluid (CSF). We elucidated the cytokine/chemokine network based on the cytokine/chemokine profiles in CSF from children with mumps meningitis and meningitis due to other viral infections using multiplex cytokine measurement. Seventeen cytokines/chemokines, namely interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-7, IL-8, IL-10, IL-12 (p70), IL-13, IL-17, interferon (IFN)-γ, tumor necrosis factor (TNF)-α, granulocyte colony-stimulating factor (G-CSF), granulocyte monocyte colony-stimulating factor (GM-CSF), monocyte chemoattractant protein-1 (MCP-1) and macrophage inflammatory protein-1β (MIP-1β), were measured simultaneously in CSF supernatants from eight children with mumps meningitis, 11 children with other types of viral meningitis and eight children with fever without neurological complications such as convulsion. We found that IL-8, IL-10, IL-12, IL-13 and IFN-γ showed a statistically significant increase in CSF from mumps meningitis when compared to other types of viral meningitis and fever without neurological complications. Mumps meningitis may induce a distinct immunological response when compared with other types of viral meningitis.