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Background:Exposure to cisplatin leads to cochlear cell death by apoptosis; these changes are most marked on the seventh day after exposure. Heat shock proteins are induced in inner ear cells in response to a variety of stimuli. This study examined the role of heat shock protein 70 in cisplatin-induced cochlear cell death.Methods:Fifty-six Sprague–Dawley rats were involved. Some were injected with cisplatin (5 mg/kg body weight), some with cisplatin plus the caspase inhibitor Z-Asp(OMe)-Glu(OMe)-Val-Asp(OME)-fluoromethylketone (5 mg/kg body weight) and others were left as controls (being injected only with saline). Seven days later, we examined the expression of heat shock protein 70 and several other apoptosis-related proteins within the rat cochlear cells; we also assessed total superoxide dismutase activity, auditory brainstem response and auditory steady state response.Results:Seven days after cisplatin injection, significantly increased expression of heat shock protein 70 was found within the rat cochleae. This correlated with increased executioner caspase levels, total superoxide dismutase activity and auditory brainstem response thresholds, and a significant elevation in auditory steady state response thresholds. Inhibition of caspase-3 activity significantly reduced cochlear heat shock protein 70 expression and total superoxide dismutase activity, and improved auditory brainstem response and auditory steady state response thresholds.Conclusions:Seven days after cisplatin exposure, we found disturbances of the cochlear cellular machinery involving heat shock protein 70, other apoptotic proteins and total superoxide dismutase.

The purpose of this study was to examine the expression of human leukocyte antigen-G (HLA-G) in patients with systemic lupus erythematosus (SLE) and its relation with interleukin-10 (IL-10) production. The study included 50 female SLE patients and 59 healthy female donors. HLA-G expression in peripheral blood and cutaneous biopsies was determined by flow cytometry and immunohistochemistry, respectively. Soluble HLA-G (sHLA-G) and IL-10 were quantified in serum samples by enzyme-linked immunosorbent assay. SLE patients presented with serum sHLA-G and IL-10 levels significantly higher than that observed in controls (median [interquartile range (IQR)] = 43.6 U/ml [23.2-150.2] vs 26.84 U/ml [6.0-45.2], p = 0.004; and 1.4 pg/ml [0-2.3] vs 0 pg/ml [0-1.5], p = 0.01, respectively). But no correlation was observed between sHLA-G and both IL-10 levels and the disease activity index for SLE patients. The expression of membrane HLA-G in peripheral lymphocytes from SLE patients was low, but higher than in controls (median [IQR] = 1.5% [0.6-1.8] and 0.3% [0.2-0.8], respectively; p = 0.02). Finally, these findings were in accordance with the weak expression of HLA-G in skin biopsies. Despite the fact that patients present higher levels of HLA-G than healthy controls, which suggests a possible relevance of this molecule in SLE, it seems not to be related to IL-10 production or disease activity.

Objective The development of common variable immunodeficiency (CVID) or hypogammaglobulinemia in systemic lupus erythematosus (SLE) is rare. The purpose of this article is to provide a detailed review of lupus-associated CVID and to identify clinical characteristics and laboratory features in patients with SLE-associated CVID. Methods We describe 2 patients with SLE and CVID and review the cases published in the English literature highlighting both the demographic and the clinical characteristics and the laboratory and therapeutic aspects of this disorder. Results Detailed descriptions of 18 patients were available; 89% were females with a mean age at the onset of SLE of 23.8 years. In 50% of patients CVID developed within the first 5 years after the diagnosis of SLE. All patients had been treated with corticosteroids and 72% had also received immunosuppressive therapy. Sinopulmonary infections were the most frequent symptom. SLE disease activity decreased after the development of CVID in 67% of patients. Most patients (89%) were treated with gammaglobulin therapy. The most notable immunological feature was a reduced number or percentage of B-cells in 60% of patients. Conclusions CVID should be suspected in any SLE patient with recurrent sinopulmonary infections in the absence of SLE activity and/or immunosuppressive treatment.

Despite a fludarabine-based treatment is the first choice of therapy in chronic lymphocytic leukemia (CLL), not all patients achieve a partial or complete response and some of them develop autoimmune manifestations. The aim of this study was to evaluate the influence of CD154 on these adverse effects because CD154 is involved in both B-cell survival and autoimmunity.Peripheral blood mononuclear cells (PBMC) from 36 patients with CLL were cultured in vitro with fludarabine or 2-chlorodeoxyadenosine for 24, 48, and 72 hours.Seven patients (19.4%) presented CD154 expression in PBMC cultured with purine analogues in vitro for 24 and/or 48 hours, while no expression was found when cultured in media alone. These seven patients showed a decreased apoptotic rate in vitro after purine analogues compared with those patients who did not express CD154 (p = 0.01 for fludarabine; p0.001 for 2-chlorodeoxyadenosine). CD154 expression was found to have prognostic value for response to fludarabine in vivo and was associated with the development of autoimmune manifestations (odds ratio = 25; 95% confidence interval = 3.5-166.7; p0.001).Our preliminary results suggest that CD154 expression in CLL patients, which may be induced by purine analogues, is associated with resistance to fludarabine and with development of autoimmune manifestations.

Objective To investigate the contribution of T lymphocytes and monocytes to cytokine production in systemic lupus erythematosus (SLE). Methods Forty-five SLE patients and 19 healthy volunteers were included. Serum levels of tumor necrosis factor alpha (TNFα), interferon gamma (IFNγ), interleukin (IL)-6, and IL10 were quantified by ELISA. The cytokine production capacities of peripheral blood mononuclear cells were assessed by culturing in vitro with PMA+Ionomycin or LPS. The intracellular cytokine expression was measured by flow cytometry in T lymphocytes and monocytes, respectively. The influence of the disease activity (measured as the SLE-disease activity index; SLEDAI) and the treatment the patients were receiving was evaluated. Results Serum IL10, IL6, and TNFα levels were increased in patients (P ≤ 0.01), and a higher spontaneous (without stimuli) intracellular expression of IL10 in CD4+ and CD8+ T lymphocytes (P < 0.05) and of IL6 in monocytes (P = 0.01) were found. After stimulation, patients presented a higher percentage of CD4+ and CD8+ T lymphocytes producing IL4 and IL10 (P ≤ 0.01), and of monocytes producing IL6 (P = 0.04) and IL10 (P = 0.008). The SLEDAI score was positively correlated with the percentage of CD4+IL10+ and CD8+IL10+ T lymphocytes (P < 0.01), and inversely correlated with CD8+TNFα+ (P= 0.02), CD4+IFNγ+ (P = 0.04) and CD8+ IFNγ+ (P = 0.002) T lymphocytes. Patients receiving high dose prednisone produced higher IL10, but they also were the patients with a more active disease. Conclusion Monocytes and T lymphocytes (CD4+ and CD8+) contribute to an overproduction of IL6 and IL10 in SLE; this correlates with the disease activity but is independent of the treatment the patients are receiving. © 2009 Clinical Cytometry Society

Background: Not all the patients with sarcoidosis need pharmacological therapy, and the decision to start therapy is based mainly on clinical conditions. The aim of this study was to evaluate the prognostic value of the leukocyte and lymphocyte subpopulations in the bronchoalveolar lavage fluid from these patients. Methods: Thirty-three nonsmoking patients with sarcoidosis were included and classified based on the presence of Lofgren's syndrome (n = 11), the radiological stage (12 at Stage I, 17 at Stage II, and 4 at Stage III), and their follow-up. Differential leukocyte subsets and the lymphocyte subpopulations were determined by flow cytometry. Results: The percentage of neutrophils was lower in patients with Lofgren's syndrome (P = 0.038) and in patients at Stage I (P = 0.002). Patients with a poor outcome had a higher percentage of neutrophils (P = 0.004) and NK cells (P = 0.023) than those with a stable disease. Finally, a higher percentage of NK cells was found in those patients who needed a steroid treatment (P = 0.012). Conclusions: Increased percentages of neutrophils and NK cells in the bronchoalveolar lavage fluid from patients with sarcoidosis are associated with a poor outcome and a higher probability to need steroids treatment. The percentage of neutrophils was also lower in patients with Lofgren's syndrome. © 2006 International Society for Analytical Cytology

Current evidence indicates that regulatory T cells (Tregs) actively suppress autoreactive lymphocytes that escape central tolerance.1,2 Tregs are characterised by a high intensity CD25 constitutive expression3 on the surface of CD4 T lymphocytes, which distinguishes them from the non-regulatory CD4+CD25+ T lymphocytes.4 Because of their important role in the maintenance of tolerance, it has been suggested that Tregs are decreased in patients with systemic lupus erythematosus (SLE).5 We have quantified Tregs in patients with SLE and controls and evaluated their association with disease activity and treatment. Thirty three patients with SLE (26 women, 7 men) and 14 healthy volunteers matched for age and sex (11 women, 3 men) with a mean age of 39.9 and 36 years, respectively, were studied. All patients fulfilled at least four of the American College of Rheumatology 1982 revised criteria for the classification of SLE.6 Disease activity at the time of blood sampling was assessed using the SLE Disease Activity Index (SLEDAI).7 …

Background IL10 is a pleiotrophic cytokine that inhibits the antigen specific activation of Th1 clones by reducing both the antigen presenting capacity and the production of pro-inflammatory cytokines by monocytes. It is also a potent activation and differentiation factor for B lymphocytes. SLE patients show increased IL10 serum levels associated with the pathogenesis of the disease. Peripheral blood mononuclear cells from SLE patients, and their relatives, constitutively produce high levels of the cytokine, suggesting an hereditary component in IL10 production. Anti-IL10 antibodies administration to SLE patients may be beneficial in the management of refractory cases. The IL10 gene promoter contains two dinucleotide repeats (microsatellites): IL10G and IL10R, close to transcription factor binding sites, probably playing an important role in the transcription regulation. Objectives To evaluate the association of the IL10G microsatellite with SLE in Spanish patients. Methods 80 unrelated SLE patients from Canary Islands (Spain) fulfilling at least four of the ACR criteria and 79 sex-matched healthy donors were included. A segment of the IL10 gene containing the microsatellite was amplified by PCR and the resulting products were electrophoresed in denaturing polyacrilamide gels. Chi-square test was used for statistical analysis. Results No particular IL10G allele was found to be significantly associated with SLE. Neverthaless, we found an elevated risk of serositis in carriers of the allele 19 (p = 0.007), a significant association of the 19/23 genotype with seizures (p = 0.018), and a reduced risk of renal disorder (p = 0.047) and thrombocytopenia (p = 0.049) in carriers of the allele 23. Conclusion Although IL10G alleles seem not to influence susceptibility to SLE in Spaniards, some of them may be related to the presence of several clinical manifestations of the disease.

Background PARP is a DNA-binding protein that catalyses the covalent addition of ADP-ribose to many nuclear proteins. It is involved in DNA repair and apoptosis, processes that have been implicated as causative factors for SLE. It has been shown that patients suffering SLE have decreased PARP mRNA levels and a low enzime activity. The PARP gene is located at 1q41-q42 and contains a microsatellite (CA)n polymorphism in the promoter region that probably affects the transcription and mRNA levels. A recent study found preferential transmission of the allele containing 12 CA repeats to affected offspring in Caucasian SLE families. Objectives To investigate the association between PARP polymorphism and SLE susceptibility in Spaniards. Methods 76 unrelated SLE patients from Canary Islands (Spain) fulfilling at least four of the ACR criteria and 79 sex-matched healthy donors were included. A segment of the PARP gene containing the microsatellite was amplified by PCR and the resulting products were electrophoresed in denaturing polyacrilamide gels. Chi-square test was used for statistical analysis. Results Allelic distributions of PARP polymorphism showed no statistical differences between SLE patients and controls. But considering particular clinical features we found several associations with specific alleles: a) homozygotes for 11rep. allele suffered more lymphopenia (p = 0.04) but less hypocomplementemia (p = 0.03), b) autoantibodies anti-Ro/La were more frequent (p = 0.02) in carriers of 15rep., c) carriers of 16rep. allele presented more malar rash (p = 0.03) and seizures (p = 0.04), and finally d) carriers of 17rep. allele showed a reduced risk of production of several autoantibodies as anti-Sm (p = 0.007) and anti-Ro/La (p = 0.04). Conclusion Although PARP alleles seem not to influence susceptibility to SLE in Spaniards, some of them may be related to the presence of several clinical and analytical manifestations of the disease.

Cardiac myxoma cells have been demonstrated to produce interleukin (IL)-6, but its role in the systemic and immunologic manifestations of patients with this tumor is controversial. There is no evidence of any other source of such cytokine, but here we report a case of myxoma with systemic manifestations at diagnosis in which we evaluated the IL-6 production by peripheral blood cells before and after surgical tumor resection. Peripheral blood mononuclear cells were isolated by density gradient centrifugation, and the IL-6 expression in monocytes and lymphocytes was evaluated by flow cytometry. Before surgery, 74.4% of monocytes produced IL-6, but 1 month after surgery, a decrease in both the number of monocytes and the percentage of these cells expressing IL-6 was found along with an improvement of systemic and immunologic manifestations. Here, we demonstrated for the first time that monocytes contribute to the elevated production of IL-6 in patients with myxoma.