Browsing by Author "Pais, ML"
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- Expression patterns of HLA-DR+ or HLA-DR- on CD4+/CD25++/CD127low regulatory T cells in patients with allergyPublication . Geraldes, L; Morgado, J; Almeida, A; Todo-Bom, A; Santos, P; Chieira, C; Pais, MLBACKGROUND: Allergic rhinoconjunctivitis induced by pollen is a highly prevalent chronic inflammatory disease in Europe. Parietariajudaica is a frequent trigger in the Mediterranean area. The function of regulatory T cells (Treg cells) in allergy has recently been investigated, but further data are necessary to better understand their role and to find new strategies to treat allergic diseases such as allergic rhinoconjunctivitis. OBJECTIVE: To characterize gene expression of HLA-DR+ or HLA-DR- on peripheral CD4+/CD25++/CD127low Treg cells in patients with allergy. METHODS: Peripheral Treg cells (CD4+/CD25++/CD127low) were quantified using flow cytometry and sorted according to HLA-DR expression during the pollen season in patients with allergic rhinoconjunctivitis caused by P. judaica. The results were compared with those of nonatopic controls. Expression of associated cytokines and their receptors was measured using quantitative reverse transcription-polymerase chain reaction after extraction of mRNA in sorted populations. RESULTS: During the pollen season, no significant differences were observed between allergic patients with rhinoconjunctivitis and healthy controls in terms of the absolute number or the percentage of Treg cells in peripheral blood. All patients had a higher number/percentage of HLA-DR- Treg cells than HLA-DR+ Treg cells. In both groups we found high levels of FOXP3 mRNA expression. Despite being lower in number, HLA-DR+ Treg cells presented higher expression of CD28, PRF1, GZMB, and FASL than HLA-DR-Treg cells. CONCLUSIONS: The most relevant results obtained suggest that HLA-DR+ Treg cells tend to present higher gene expression of molecules associated with contact-dependent cell activation and cytotoxicity.
- Functional characterization of peripheral blood dendritic cells and monocytes in systemic lupus erythematosusPublication . Henriques, A; Inês, L; Carvalheiro, T; Couto, M; Andrade, A; Pedreiro, S; Laranjeiro, P; Morgado, JM; Pais, ML; Pereira da Silva, JA; Paiva, AWith the purpose of contributing to a better knowledge of the APCs functional activity in SLE, we evaluated the distribution and functional ability to produce pro-inflammatory cytokines (TNF-α, IL-1β, IL-6 and IL-12) of peripheral blood (PB) monocytes and DC (tDC), particularly myeloid (mDC) and CD14(-/low)CD16(+) DC subpopulations comparing them with those obtained from healthy individuals. The study was performed in 34 SLE patients with diverse disease activity scores (SLEDAI) and 13 healthy age- and sex-matched controls (NC). Our results show an overall decrease in absolute number and relative frequency of tDC in SLE patients with active disease when compared to those with inactive disease and NC, although this decrease did not seem to have an effect on the distribution of PB DC subsets. The monocytes number in SLE patients was similar to those found in NC, whereas a higher frequency of monocytes producing cytokines as well as the amount of each cytokine per cell found without stimulation was particularly observed in those patients with active disease. After stimulation, we observed a higher frequency of IL-12-producing monocytes in active SLE patients. On the other hand, we found among DCs higher frequencies of cytokine-producing CD14(-/low)CD16(+) DCs and a higher amount of cytokines produced per cell, particularly in active disease. These findings support an increased production of inflammatory cytokines by APCs in active SLE, mostly associated with alterations in CD14(-/low)CD16(+) DC subset homeostasis that might contribute to explain the dynamic role of these cells in disease pathogenesis
- Perioperative tumor cell dissemination in patients with primary or metastatic colorectal cancerPublication . Tralhão, JG; Hoti, E; Serôdio, M; Laranjeiro, P; Paiva, A; Abrantes, AM; Pais, ML; Botelho, MF; Castro e Sousa, FINTRODUCTION: Although there is general correlation between the TNM stage of colorectal cancer (CRC) and its prognosis, there is often significant variability of tumor behaviour and individual patient outcome, which is unaccounted for by pathologic factors alone. Our aim was to estimate perioperative tumor cell dissemination in patients with primary or CRC liver metastases as a possible factor influencing the outcome. METHODS: Forty patients were prospectively enrolled in the study from the year 2007 to 2008. Eighteen patients had histologically proven CRC (50% rectal, 44% colonic, 6% colonic and rectal). Sixteen patients (47%) had CRC liver metastases only. The remaining six patients who underwent colon or liver resection for benign conditions, acted as the control group. All patients with malignant pathologies had R0 resections. Blood samples were taken before the surgical incision (T0), immediately after tumor resection (T1) and at the end of the surgical intervention (T2). Data acquisition was performed using a dual-laser FACSCalibur flow cytometer. Circulating malignant cells were identified as being CD45-/cytokeratin+. RESULTS: The analysis of patients overall (CRC resection subgroup and hepatectomy subgroup) revealed that there was no statistically significant difference of the tumoral cell count in the blood per million of hematopoietic cells at T0, T1 and T2. CONCLUSIONS: This study demonstrates no differences in the detected circulating numbers of tumor cells at different stages of surgical intervention.
- Th17 cells in systemic lupus erythematosus share functional features with Th17 cells from normal bone marrow and peripheral tissuesPublication . Henriques, A; Inês, L; Pais, ML; Pereira da Silva, JA; Paiva, AAThis study was designed to investigate the functional heterogeneity of human Th17 and how their plasticity shapes the nature of immune cell responses to inflammation and autoimmune diseases, such as systemic lupus erythematosus (SLE). We evaluated functional Th17 cell subsets based on the profile of cytokine production in peripheral blood (PB), bone marrow aspirates (BM) and lymph node biopsies (LN) from healthy individuals (n = 35) and PB from SLE patients (n = 34). Data were analysed by an automated method for merging and calculation of flow cytometric data, allowing us to identify eight Th17 subpopulations. Normal BM presented lower frequencies of Th17 (p = 0.006 and p = 0.05) and lower amount of IL-17 per cell (p = 0.03 and p = 0.02), compared to normal PB and LN biopsies. In the latter tissues were found increased proportions of Th17 producing TNF-α or TNF-α/IL-2 or IFN-γ/TNF-α/IL-2, while in BM, Th17 producing other cytokines than IL-17 was clearly decreased. In SLE patients, the frequency of Th17 was higher than in control, but the levels of IL-17 per cell were significantly reduced (p < 0.05). Among the eight generated subpopulations, despite the great functional heterogeneity of Th17 in SLE, a significant low proportion of Th17 producing TNF-α was found in inactive SLE, while active SLE showed a high proportion producing only IL-17. Our findings support the idea that the functional heterogeneity of Th17 cells could depend on the cytokine microenvironment, which is distinct in normal BM as well as in active SLE, probably due to a Th1/Th2 imbalance previously reported by our group.
- Tolerogenic versus Inflammatory Activity of Peripheral Blood Monocytes and Dendritic Cells Subpopulations in Systemic Lupus ErythematosusPublication . Carvalheiro, T; Rodrigues, A; Lopes, A; Velada, I; Ribeiro, A; Martinho, A; Inês, L; Pereira da Silva, JA; Pais, ML; Paiva, AAbnormalities in monocytes and in peripheral blood dendritic cells (DC) subsets have been reported in systemic lupus erythematosus (SLE). We aim to clarify the tolerogenic or inflammatory role of these cells based on ICOSL or IFN-α and chemokine mRNA expression, respectively, after cell purification. The study included 18 SLE patients with active disease (ASLE), 25 with inactive disease (ISLE), and 30 healthy controls (HG). In purified plasmacytoid DC (pDC) was observed a lower ICOSL mRNA expression in ASLE and an increase in ISLE; similarly, a lower ICOSL mRNA expression in monocytes of ALSE patients was found. However, a higher ICOSL mRNA expression was observed in ASLE compared to HG in myeloid DCs. Interestingly, clinical parameters seem to be related with ICOSL mRNA expression. Regarding the inflammatory activity it was observed in purified monocytes and CD14(-/low) CD16(+) DCs an increase of CCL2, CXCL9, and CXCL10 mRNA expression in ASLE compared to HG. In myeloid DC no differences were observed regarding chemokines, and IFN-α mRNA expression. In pDC, a higher IFN-α mRNA expression was observed in ASLE. Deviations in ICOSL, chemokine, and IFN-α mRNA expression in peripheral blood monocytes and dendritic cells subpopulations in SLE appear to be related to disease activity.