IL-10 Production In Patients Research Articles

Cytokine profiles in peripheral blood mononuclear cells from patients with cystic echinococcosis

IntroductionCystic echinococcosis (CE) is a chronic zoonotic disease caused by the larval stage of Echinococcus granulosus (E. granulosus), which affects domestic and wild carnivores as the definitive host and ungulates as intermediate hosts. In intermediate hosts, both Th1 and Th2 cells are involved in the immune responses to an echinoccocal infection. This study aimed to investigate production of IL-4, IL-10, and IFN-γ cytokines in peripheral blood mononuclear cells (PBMCs) of CE patients before and after surgical treatment. MethodsTo evaluate cytokine production in response to E. granulosus antigens, we investigated IL-4, IL-10, and IFN-γ production in PBMCs of 20 CE patients in response to hydatid cyst fluid antigen (HCF-Ag) before and after surgical treatment using ELISA. ResultsThe mean IL-4 production from HCF-Ag stimulated PBMCs was significantly decreased (p < 0.05), while IFN-γ was significantly increased in HCF-Ag stimulated PBMCs in patients after surgery (p = 0.005).Furthermore, our results showed that there is no significant difference between IL-10 production in patients before and after treatment (p = 0.562). ConclusionsOur data Indicated production of IL-4 in cultured PBMCs of CE patients stimulated with HCF-Ag was decreased significantly. While, production of IFN-γ was increased significantly in responses to HCF Ag after surgery. We concluded that the evaluation of IL-4 and IFN-γ in HCF-Ag stimulated PBMCs of CE patients should be considered as a useful marker in the follow up of patients with cystic echinococcosis.

Low frequency of IL-10+ B cells in patients with atherosclerosis is related with inflammatory condition.

Background and aimsB cells involvement in animal models of atherosclerosis has been unequivocally established. However, the role of these cells in patients with atherosclerosis is almost unknown. Besides the production of antibodies, B cells can also exhibit regulatory functions mainly through IL-10. Here, we characterized human B cell subsets, their production of IL-10 in patients with atherosclerosis and their potential association with inflammation.MethodsPatients with confirmed atherosclerotic events and controls with low cardiovascular risk were included. B cells subsets were determined in mononuclear cells (PBMC) using flow cytometry. PBMC were cultured ex vivo (5 h) and in vitro (48 h) to determine IL-10+ B cells and in some cases TNF-α+ and IFN-γ+ CD4+ T cells. The inflammatory state of the participants was determined through high sensitivity C reactive protein levels.ResultsIncrease in percentage and number of plasmablasts was observed in patients with atherosclerosis compared with controls. A decreased frequency of IL-10+ B cells was observed in patients, both in ex vivo and in vitro cultures. This decrease was detected in transitional, memory, and plasmablast subsets. Interestingly, the reduction of IL-10+ B cells negatively and significantly correlated with the inflammatory condition of the studied subjects and associated with an increased frequency of TNF-α+ and IFN-γ+ CD4+ T cells. The blockade of IL-10R did not show further effect in T cells activation.ConclusionsThere is an association between the inflammatory state and a reduction of IL-10+ B cells that could contribute to the development of atherosclerosis.

Comparative investigation of percutaneous plating and intramedullary nailing effects on IL-6 production in patients with tibia shaft fracture

ObjectiveThe aim of this study was to analyze the effect of intramedullary nailing (IMN), open plating and percutaneous plating on the induction of IL-6 in patients with tibia fractures. MethodsA total of 30 patients with tibia shaft fracture underwent either intramedullary nailing (IMN, n = 15; 14 males and 1 female; mean age: 32.1 ± 15.6), ORIF plating (n = 8; 5 males and 3 females; mean age: 60.0 ± 17.8), or percutaneous plating (n = 7; 6 males and 1 female; mean age: 43.1 ± 21.4). Serum IL-6 cytokine levels were measured prior to, and 6 and 24 h after the surgery, using a special ELISA kit. ResultsThe IL-6 concentration increased to peak levels at 6 h in both IMN and percutaneous plating groups, and at 24 h in ORIF plating group (p < 0.001). The mean IL-6 concentration of percutaneous plating group was significantly lower than that of the IMN group at 6 h following the surgery (p = 0.022). In addition, the mean IL-6 concentration of ORIF plating group was significantly higher than that of the percutaneous plating group at 24 h post operation (p = 0.009). ConclusionOur results suggest that percutaneous plating compared to the IMN has lower effects on IL-6 production in patients with tibia fracture. Level of evidenceLevel III, therapeutic study

Cytokine Status of Serum in Ovarian Cancer Patients with Different Tumor Neoadjuvant Chemotherapy Response.

Polychemotherapy in ovarian cancer (OC) is the second major component of treatment. However, treatment with cytostatics is stopped in 25% of cases because of significant side effects. It is shown that concentration of certain cytokines and their balance is largely formed in accordance with a genetic polymorphism. The objective of the study was to evaluate the cytokine status of blood serum of patients with ovarian cancer with different tumor response to neoadjuvant chemotherapy (NACT). Patients received 2 courses NACT according to the scheme AP. The levels of IL-1β and IL-1Ra, IL-10, TNF-α in blood serum were determined by solid phase ELISA. For molecular genetic studies we selected polymorphic variants in the promoters of the gene represented in dbS`NP NCBI and SNP500 Cancer databases. The sharply declined in patients with ovarian cancer compared with the normal, level of IL-1β correlates with increased levels of IL-1RA. It is found that 75% of patients who had progression of the disease after NACT bear CT genotype of gene IL-1β associated with a low expression of the cytokine, while the TT genotype, providing a high level of the expression of IL-1β gene had met only 25% among patients in this group. At the same time 70% of patients with a complete response are the carriers of the T allele, while a complete response was associated with a higher level of IL-1β than in the progression group. Low secretion of TNF-α in all types of tumor response when testing TNF-α G-308A gene polymorphisms was associated with carriage of GA and AA genotypes, which are associated with low production of this cytokine. Increased compared to the control IL-10 production in patients with ovarian cancer associated with genotype replacement at position 1082 G/A IL-10 gene, which occured in 10% of patients with a complete response and 25% of patients with tumor progression after NACT. All the studied polymorphisms of IL-1β, IL-10 and TNF-α genes in patients with OC are associated with the level of these cytokines and tumor NACT response.

ProNGF-p75NTR axis plays a proinflammatory role in inflamed joints: a novel pathogenic mechanism in chronic arthritis

ObjectiveTo identify the role of mature nerve growth factor (mNGF), its immature form proNGF and their receptors in arthritis inflammation.MethodsReal-time PCR, western blot and ELISA were performed to evaluate NGF,...

BAFF induces IL-6 production by monocytes through a BAFF receptor and promotes IgG production by B cells in patients with primary Sjögren's syndrome. (CCR3P.210)

Abstract Backgrounds and Objectives: We have found that the elevated expression of BAFF receptor (BR3) on monocytes is involved in activation of monocytes to promote IL-6 production in patients with primary Sjögren's syndrome (pSS). In addition, it is well known that pSS is often accompanied by hypergammaglobulinemia (HγG). In our present study, we investigated the possible involvement of monocytes in the development of HγG. Methods: Peripheral monocytes were cultured in vitro with or without peripheral B cells and stimulated with soluble BAFF (sBAFF). The amounts of IL-6 and IgG in the culture supernatants were measured by ELISA. FACS analysis of whole blood samples was employed to analyze the expression of BR3. Results: The serum level of IgG and the proportion of BR3-positive monocytes (BR3+/CD14+) were elevated in pSS patients as compared to those in normal individuals. Remarkably, the BR3+/CD14+ ratio was positively and significantly correlated with the serum IgG level. In addition, the ratio was positively and significantly correlated with the amount of IL-6 produced by pSS monocytes stimulated with sBAFF. Stimulation of co-culture of B cells and monocytes with sBAFF drastically enhanced IgG production by B cells. Conclusions: Our data collectively suggest that the abnormal expression of BR3 on monocytes is responsible for the overproduction of IgG by B cells in pSS patients.

320 THE HEPATITIS C VIRUS NON-STRUCTURAL 3/4A PROTEASE MEDIATES AN INTRAHEPATIC T HELPER(Th)-1 TO Th2 SHIFT BY CLEAVING THE T CELL PROTEIN TYROSINE PHOSPHATASE

Concentrations of TNF-a and IL-6 in supernatants and patient sera were assayed by ELISA. We designed a 5-plex Phosflow FACS protocol to determine activation of key transcription factors in innate immune signalling pathways (NF-kB, ERK1/2, STAT1 and STAT3) in CD14+ monocytes following stimulation with LPS and flagellin. Results: We included 28 patients and 6 normal controls. Patients with compensated cirrhosis had a selective defect in flagellininduced IL-6 production (330±117pg/ml) compared to patients with non-cirrhotic ALD or NAFLD (896±146pg/ml; p = 0.01) or healthy controls (764±96pg/ml). There were no significant differences in flagellin-induced TNFa, nor any differences between cirrhotics and non-cirrhotics in LPS-induced cytokines. There were no differences in serum TNF-a, IL-6 and RANTES, nor in cytokine production when monocytes from healthy volunteers were incubated with patients’ sera before stimulation with LPS or flagellin. Flagellin-induced ERK1/2 phosphorylation in CD14+ monocytes is greater in non-cirrhotics than in cirrhotics (32.8% vs 10.5%, p = 0.056), but there were no differences in LPS-induced ERK1/2 phosphorylation and LPSand flagellin-induced NF-kB phosphorylation. Conclusions: We find selective impairment of TLR5-mediated IL-6 production in patients with compensated cirrhosis compared to non-cirrhotic patients that may be related to defects in ERK1/2 phosphorylation. These data identify potential signalling pathways that may be involved in liver disease progression or in susceptibility of cirrhotic patients to bacterial infections.

Inflammatory cytokine IL-10 polymorphism in patients of ulcerative colitis and irritable bowel syndrome

S 21 ANNUAL CONFERENCE—2013 M isella n eo u s Corresponding author: Jitendra Km Choudhary. E-mail: jitendra709@gmail.com INFLAMMATORY CYTOKINE IL-10 POLYMORPHISM IN PATIENTS OF ULCERATIVE COLITIS AND IRRITABLE BOWEL SYNDROME Manish Kumar Tripathi, Ashok Kumar Jain, Vinod Kumar Dixit, Pankaj Kaushik, Jitendra Kumar Choudhary, Manjita Srivastava, Neha Singh, Arttrika Ranjan, Neha Gupta, Sangey Chopel Lamtha, Smita Verma Department of Gastroenterology, Institute of Medical Sciences, BHU, Varanasi Background: Ulcerative colitis (U.C.) is a complex multifactorial trait involving both environmental and genetic factors. UC is characterized by activation of macrophages and T lymphocyte; pro-inflammatory cytokine, chemokines and adhesion molecule expression and an inability to adequately down-regulate immune activation. Aim: The aim of the present study is to examine Interleukin 10 gene polymorphism in patients with UC and IBS. Methods: Age and sex matched 56 patients of IBS and 36 patients of ulcerative colitis were enrolled in the study. Blood samples were obtained after informed consent for DNA isolation by phenol-chloroform method. PCR for IL 10 -1082, -819, -592 was done followed by restriction digestion by site specific enzyme Mnl1, RsaI and MaeIII respectively, the results was analyzed on 2% agarose gel. Results: The percentage distribution of genotypic frequencies of IL-10 -1082 homozygotes and heterozygotes are given in the table 1. Table 1 Genotypic frequencies of IL10 -1082, -819, -592 between IBS and UC patients Cytokines Genotype Frequency IL-10 (1082) AA AG GG IBS 32(57.1) 16(28.6) 8(14.3) Ulcerative Colitis 22(61.1) 8(22.2) 6(16.7) c2=0.479, p=0.787 IL-10 (819) TT TC CC IBS 13(23.2) 21(37.5) 22(39.3) Ulcerative Colitis 4(11.1) 20(55.6) 12(33.3) c2=3.550, p=0.169 IL-10 (592) AA AC CC IBS 36(64.3) 10(17.9) 10(17.9) Ulcerative Colitis 16(44.4) 12(33.3) 8(22.2) c2=3.934, p=0.140 S124 Conclusion: Genotypic frequencies of IL-10 between UC and IBS patients showed that there is no statistically significant difference in IL-10 -1082, -819, -592 cytokine genes. Genotype ATA/ATA correlate with low level of IL-10 production Hence the results of this study indicate low production of anti-inflammatory cytokines IL-10 in patients with UC and IBS. Corresponding author: V.K.Dixit. E-mail: manish.imsbhu@gmail.com

PMO-141 Flagellin-induced IL-6 production is selectively impaired in patients with cirrhosis

IntroductionThe innate immune response is an important determinant of progression in chronic inflammatory liver diseases such as alcoholic (ALD) and non-alcoholic fatty liver disease (NAFLD). Sepsis is a frequent cause...

Abstract 5416: Myeloid-derived suppressor cells (MDSC) are increased and correlated with immune suppression and chronic inflammation in patients with cancer

Abstract Chronic inflammation plays a key role in the progress of malignant diseases and has been reported to relate with immune suppression seen in patients with advanced diseases. MDSC: myeloid-derived suppressor cells found as a new type of immune suppressor cells are enhanced by inflammation and have been reported to be present in blood circulation, lymphnodes and tumor tissues in patients with cancer. The suppression of immune function via depleting arginine and producing immune-suppressing cytokines including IL-10 has also been reported. In order to investigate MDSC in patients, peripheral blood mononuclear cells (PBMC) were collected from 18 normal healthy volunteers, 53 patients with gastrointestinal cancer, 65 with breast cancer, 8 with thyroid cancer, and 5 with ovarian cancer. These cells were used for the detection of MDSC (CD11b+CD14−CD33+) by flow cytometry. PBMC was also used for the PHA-blastogenesis of lymphocytes which is a marker of cell mediated immunity (stimulation indices: SI) and for the production assay of cytokines including IL-6 and IL-10. MDSC (%PBMC) of whole patients (5.68 + 5.92%), those of gastrointestinal cancer patients (3.76 + 4.90%) and those of breast cancer patients (2.92 + 4.13%) were significantly higher than those of normal volunteers (p&amp;lt;0.005, p=0.05, p&amp;lt;0.05, respectively, vs 1.59 + 1.08%). MDSC were inversely correlated with the SI (r=−0.271, p&amp;lt;0.05) in patients with gastrointestinal cancer and positively correlated with IL-6 production in whole patients(r=0.287, p=0.05) and with IL-10 production in patients with breast cancer (r=0.363, p&amp;lt;0.05). They were also significantly correlated to neutrophil counts (r=0.287, p&amp;lt;0.05) and neutrophil/lymphocyte ratios (NLR, r=0.623, p=0.05), and inversely correlated to lymphocyte counts (r=−0.251, p&amp;lt;0.01). Thus MDSC were successfully detected in peripheral blood and were significantly higher in patients with cancer. There were strong relationships among the inhibition of cell-mediated immunity, production of type 2 cytokines and inflammatory markers such as NLR. Therefore, MDSC seemed to work as a suppressor of immune reaction and may be an important marker of chronic inflammation in patients with cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5416. doi:1538-7445.AM2012-5416

Mechanisms of apoptotic cell-mediated IL-10 production in patients with systemic lupus erythematosus

Mechanisms of apoptotic cell-mediated IL-10 production in patients with systemic lupus erythematosus

Impact of antibiotic therapy on systemic cytokine expression in pneumococcal pneumonia

The aim of this study was to compare the evolution of systemic cytokine levels over time in patients with pneumococal pneumonia treated either with β-lactam monotherapy or with combination therapy (β-lactam plus fluoroquinolone). Prospective observational study of hospitalized non-immunocompromised adults with PP. Concentrations of IL-6, IL-8, IL-10, and TNF-α were determined on days 0, 1, 2, 3, 5, and 7. Patients on β-lactam monotherapy were compared with those receiving combination therapy. Fifty-two patients were enrolled in the study. Concentrations of IL-6, IL-8, and IL-10 decreased rapidly in the first days after admission, in accordance with the mean time to defervescence. High levels of IL-6 were found in patients with the worst outcomes, measured by the need for intensive care unit admission and mortality. No major differences in demographic or clinical characteristics or severity of disease were found between patients treated with β-lactam monotherapy and those treated with combination therapy. IL-6 levels fell more rapidly in patients with combination therapy in the first 48h (p = 0.016). Our data suggest that systemic expression of IL-6 production in patients with PP correlates with prognosis. Initial combination antibiotic therapy produces a faster decrease in this cytokine in the first 48h.

Abstract 2449: A novel humanized anti-IL-1beta antibody is highly effective at inhibiting IL-1 induced IL-6 production in myeloma patients in vitro

Abstract Background: IL-6 is a central myeloma growth factor and in vitro abnormal production of IL-1beta in the myeloma microenvironment stimulates the generation of paracrine IL-6. Mimicking in vitro observations, we have shown that responsive early stage myeloma patients at risk for progression to active myeloma who were treated with IL-1 inhibitors demonstrated decreases in the myeloma proliferative rate and C-reactive protein (IL-6 surrogate marker) leading to a chronic disease state with an improved progression free survival. In this study we investigated the effect of XOMA 052, a humanized anti-IL-1 antibody to inhibit stromal cell IL-6 production induced by either recombinant IL-1beta or supernatants generated from unsorted bone marrow cells from patients with smoldering multiple myeloma. Methods: Bone marrow stromal cells were incubated with varying concentrations of recombinant IL-1β (100, 10, 1, 0.1 pg/ml) in the absence or presence of anti-IL-1 antibody, anti-TNF antibody, or dexamethasone. In a similar fashion, supernatants from unsorted bone marrow cells from six patients with smoldering myeloma were co-cultured with bone marrow stromal cells and the effect of the humanized anti-IL-1β antibody was tested. Results: The anti-IL-1β antibody inhibited the IL-1 induced IL-6 production in vitro by &amp;gt; 85% at 100 pg/ml of IL-1 and &amp;gt; 90% at 10, 1, and 0.1 pg/ml of IL-1; specifically, using 10 pg/ml of IL-1, IL-6 production was inhibited from 149 down to 5.2 ng/ml in the presence of anti-IL-1 antibody. Anti-TNF antibody had minimal effect. At 10 pg/ml of IL-1 beta, a physiologically relevant amount of IL-1 in vitro in bone marrow samples from myeloma patients, the anti-IL-1 antibody was superior to dexamethasone at inhibition of IL-6 production. In in vitro testing of patient supernatants from unsorted bone marrow cells, three patients were high inducers and three patients low inducers of paracrine IL-6 production. The results demonstrate &amp;gt; 85% reduction of IL-6 production in all patients' samples tested. Importantly, the anti-IL-1β antibody was highly effective in samples from the three patients who were high inducers of paracrine IL-6 production (IL-6 production was inhibited from 191 down to 12 ng/ml; 157 to 13 ng/ml and 320 to 11 ng/ml). In comparison, the anti-TNF antibody had marginal effects. Conclusion: The humanized anti-IL-1beta antibody is highly effective in vitro at the inhibition of IL-6 generated by supernatant cultures from patients with early stage myeloma or by recombinant IL-1. Combination therapy with IL-1 inhibitors and apoptosis inducing agents may be useful in patients with active myeloma that have elevated IL-6 levels and a high growth rate at diagnosis. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 101st Annual Meeting of the American Association for Cancer Research; 2010 Apr 17-21; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2010;70(8 Suppl):Abstract nr 2449.

Increased expression of interleukin‐7 in labial salivary glands of patients with primary Sjögren's syndrome correlates with increased inflammation

To study the expression levels and immunostimulatory capacities of interleukin-7 (IL-7) in primary Sjögren's syndrome. Labial salivary gland (LSG) IL-7 expression was determined by immunohistochemistry, using a quantitative scoring system, in 30 patients with sicca syndrome: 15 patients with primary Sjögren's syndrome (SS) and 15 patients with non-SS sicca syndrome. The correlation of IL-7 expression in LSGs with parameters of local and peripheral disease was studied, and serum and salivary IL-7 levels were determined. Additionally, the effects of IL-7 on cytokine production by peripheral blood mononuclear cells (PBMCs) from patients with primary SS were determined in vitro by Luminex multicytokine assay and compared with the effects in control subjects. The expression of IL-7 in LSGs was higher in patients with primary SS compared with that in patients with non-SS sicca syndrome. IL-7 was observed primarily in the vicinity of lymphocytic infiltrates. Salivary IL-7 levels in patients with primary SS were higher than those in control subjects. In all 30 patients with sicca syndrome, IL-7 expression in LSGs correlated with parameters of both local and peripheral disease. Furthermore, IL-7 stimulated T cell-attracting and T cell-differentiating cytokines (monokine induced by interferon-gamma [IFNgamma], IFNgamma-inducible 10-kd protein, IL-12, and IL-15), as well as Th1 (IFNgamma), Th2 (IL-4), Th17 (IL-17A), proinflammatory (tumor necrosis factor alpha and IL-1alpha), and regulatory (IL-10 and IL-13) cytokine production by PBMCs. All of these cytokines were previously shown to be associated with primary SS. The IL-7-induced increase in IL-10 production in patients with primary SS was reduced compared with that in control subjects. The correlation between LSG IL-7 expression and (local) disease parameters in primary SS as well as the IL-7-mediated induction of inflammatory cytokines indicate that IL-7 might contribute to the immunopathology of primary SS.

Toll-like receptor 2 expression on monocytes from patients with BCG vaccine-associated suppurative lymphadenitis

† Cheju National University Jeju Korea Purpose : Toll-like receptor 2 (TLR2 is critical in the immune response to mycobacterial infections. The purpose of this study was to analyze TLR2 surface expressions and TLR2-mediated tumor necrosis factor-alpha (TNF-α and interleukin-6 (IL-6 production in patients with BCG vaccine-associated suppurative lymphadenitis. Methods : Peripheral monocytes were separated from 16 patients with BCG vaccine-associated suppurative lymphadenitis and 10 healthy controls using a magnet cell isolation kit. Monocytes (1×10 6 cells/well were incubated with a constant amount of Pam3CSK4 (100 μg/mL for 24 hours. TLR2 surface expression on monocytes was analyzed by FACS analysis and TLR-2 mRNA expression was determined by RT-PCR. TLR2-mediated TNF-α and IL-6 production were measured by ELISA. Results : In patients with BCG vaccine-associated suppurative lymphadenitis low TLR2 expression on monocytes (3.39±1.2% versus 4.64±2.6% together with significantly lower TLR2 mRNA expression than in the healthy controls was seen after Pam3CSK4 stimulation. TLR2-mediated TNF-α and IL-6 production in patients with BCG vaccine-associated suppurative lymphadenitis (TNF-α 775.5±60.8 pg/mL; IL-6 4645.8±583.9 pg/mL were also lesser than that in healthy controls (TNF-α 1098.5±94.3 pg/mL; IL-6 6696.3±544.3 pg/mL. Conclusion : These findings suggest that low TLR2 expression on monocytes might be associated with increased susceptibility to BCG vaccine-associated suppurative lymphadenitis. (Korean J Pediatr 2009;52:667-673

Sublingual immunotherapy-induced IL-10 production is associated with changed response to the decongestion test: Preliminary results

Sublingual immunotherapy (SLIT) has been shown to be capable of inducing a regulatory T-cell response as evidenced by IL-10 production. Decongestion testing consists of spraying an intranasal vasoconstrictor drug to evaluate the recovery of nasal airflow limitation. The aim of this study was to assess the association of SLIT-induced IL-10 production with nasal airflow recovery after decongestion testing. Nineteen patients with perennial allergic rhinitis (PAR) were studied: 9 patients successfully assumed SLIT for 3 years and 10 patients were considered as control. In vitro IL-10 production was evaluated after SLIT. Rhinomanometry and decongestion testing were performed in all subjects before and after 3 years. After 3 years, SLIT patients showed a significant decrease of nasal resistances (0.048) and reduced response to decongestion testing (p = 0.044 for absolute values and p = 0.0051 for delta). The comparison with nontreated allergic patients shows significant differences concerning both pretest values (p = 0.02) and delta percentages (p = 0.00466). In addition, the decrease of nasal airflow resistance values and the percentages of reversibility were significantly associated with IL-10 levels (p = 0.0016 and p = 0.00294, respectively). This preliminary study provides the first evidence of a changed response to decongestion testing after SLIT that is associated with IL-10 production in patients with PAR.

Interleukin-6 production and deregulation of the hypothalamic-pituitary-adrenal axis in patients with major depressive disorders

The present study was designed to determine whether an association exists between HPA activity and cytokine production in major depression (MD). In 9 patients with MD and 11 control subjects of both sexes, all drug-free, activity of the HPA axis was evaluated by circadian rhythm of plasma cortisol, 24-h free urinary cortisol, an overnight 1 mg dexamethasone suppression test, and an oCRF stimulation test. Spontaneous and LPS-stimulated production of IL-1beta, IL-6, and TNFalpha by peripheral blood mononuclear cells were also determined. We found a significantly elevated spontaneous production of IL-6 in patients with MD (3541.2 +/- 726.8 vs 380.4 +/- 77.5 pg/mL in controls, p < 0.05), while LPS-stimulated production was significantly lower in patients than in control subjects (19,867.7 +/- 3649.2 vs 33,142.2 +/- 15,47.2 pg/mL, p < 0.05). The adrenocorticotropic hormone response to oCRF, evaluated as the area under the curve (AUCACTH) was significantly lower in patients than in control subjects (p = 0.02). A positive correlation between AUCACTH and LPS-stimulated IL-6 secretion was observed in patients with MD (r = 0.75, p < 0.05) but not in controls. These findings suggest that the activation of the inflammatory response described in depression might be associated with long-term hyperactivity of the HPA axis.

Therapy with human monoclonal anti-CTLA-4 antibody, CP-675,206, reduces regulatory T cells and IL-10 production in patients with advanced malignant melanoma (MM)

7505 Background: T-regulatory (T-reg) cells are CD3+CD4+CD25+ and produce IL-10. Engagement of CTLA-4 on activated T cells with CD80/CD86 on APC diminishes T-cell activation and proliferation via inhibition of IL-2 transcription and production. To block signaling of this repressive pathway, 26 patients (pts) with MM were treated with CP-675,206 in a Multi-dose Phase I/II Trial with the intent of augmenting host T-cell mediated responses against the tumor. Methods: Sixteen pts received monthly infusions of CP-675,206 at 10 mg/kg, and another 10 pts at 15 mg/kg every 3 months; all patients were assessed for objective tumor responses (OR) by WHO criteria. Peripheral blood (PB) was obtained from pts at baseline and day 14 of each course of treatment to determine PB T cells (CD3+) and subsets (CD4, CD8, and CD25) by FACS, IL-2, IL-10, and interferon-gamma production, and proliferation of T cells in response to activation of PB mononuclear cells (PBMC) with anti-CD3. Results: Of the 18 evaluable pts, 5 (3 at the 10 mg/kg and 2 at the 15 mg/kg doses) attained OR, 13 progressed (PD); and 8 were stable (SD) or too early for tumor evaluation. After two months of treatment, OR and SD pts had significant increases in %CD3+CD4+ T cells (48.4% vs 57.6%; p=0.018) and a concomitant decrease in %CD3+CD4+CD25+ T-reg cells (38.9% vs 26.7%; p=0.025) whereas PD pts had a significant increase in %CD3+CD4+ T cells (50.4% vs 55.1%; p<0.05) without a reduction in T-reg cells. PBMC from OR, and not PD, had a significant reduction in the constitutive IL-10 production (11.3 pg/mL vs 2.9 pg/mL; p=0.032). Moreover, the ratio of IL-2/IL-10 production by anti-CD3 activated PBMC of OR, and not PD, pts was significantly higher after 1 cycle of treatment (14.2 vs 29.4; p=0.028), compared to baseline. Conclusions: Multiple doses of CP-675,206 can induce OR in pts with MM. CP-675,206 can also reduce the %CD3+CD4+CD25+ T-reg cells and constitutive IL-10 production, and increase the ratio of IL-2/IL-10 production by anti-CD3 activated PBMC. Collectively, these data suggest that CP-675,206 is biologically active in MM and may improve T-cell mediated immunity in clinical responders. Author Disclosure Employment or Leadership Consultant or Advisory Role Stock Ownership Honoraria Research Funding Expert Testimony Other Remuneration Pfizer Pfizer Pfizer Pfizer Pfizer

Nucleotide-Binding Oligomerization Domain-2 Modulates Specific TLR Pathways for the Induction of Cytokine Release

The recognition of peptidoglycan by cells of the innate immune system has been controversial; both TLR2 and nucleotide-binding oligomerization domain-2 (NOD2) have been implicated in this process. In the present study we demonstrate that although NOD2 is required for recognition of peptidoglycan, this leads to strong synergistic effects on TLR2-mediated production of both pro- and anti-inflammatory cytokines. Defective IL-10 production in patients with Crohn's disease bearing loss of function mutations of NOD2 may lead to overwhelming inflammation due to a subsequent Th1 bias. In addition to the potentiation of TLR2 effects, NOD2 is a modulator of signals transmitted through TLR4 and TLR3, but not through TLR5, TLR9, or TLR7. Thus, interaction between NOD2 and specific TLR pathways may represent an important modulatory mechanism of innate immune responses.

Evaluation of In Vitro Production of IFN-gamma, IL-10, IL-12 and IL-13 by Blood Cells in Patients with Cutaneous Leishmaniasis Lesions.

This study investigated the in vitro production of interferon-gamma, interleukin (IL)-10, IL-12, and IL-13, after antigenic stimulation of the cells (with Leishmania antigen and lipopolysaccharide) using whole blood from patients with cutaneous leishmaniasis lesions caused by Leishmania tropica and in normal volunteers with history of cutaneous leishmaniasis.ELISA results showed that the mean production of interferon-gamma by cells of whole blood in patients with lesions in response to Leishmania antigen was significantly lower than corresponding values in volunteers with history of cutaneous leishmaniasis (P< 0.05) and significantly higher levels of IL-10 production in patients with lesions were observed compared with cured volunteers of the disease (P<0.01). A similar level of IL-12, including p40 subunit of IL-12, was detected in both groups tested in this study in response to stimulation of parasite antigen. The levels of the IL-13 after stimulation with Leishmania antigen were significantly more in patients compared with volunteers with history of cutaneous leishmaniasis (P< 0.01). There was no significant difference in the mean production of IFN-gamma, IL-10, IL-12 and IL-13 by PHA or LPS stimulated cells from patients with lesions and volunteers with history of the disease, indicating that there was no qualitative defect in cytokine production in these patients.In this study, we have detected the decreased production of interferon- gamma by cells of patients with lesions of cutaneous leishmaniasis in response to parasite antigen and unbalanced production of regulatory cytokines such as IL-10 and IL-13 using the whole-blood stimulation assay technique. The required small volume of blood and the rapid set up time are the advantages in this assay technique. Using this assay for further immunodetection of cytokines may confirm its value for clinical investigation.