transforming-growth-factor-beta and Anemia--Aplastic

Immune-mediated hematopoietic destruction is a key factor in idiopathic severe aplastic anemia (SAA). With great immunomodulatory functions, mesenchymal stem cells (MSCs) are important for bone marrow niche. While the underlying etiology of immunologic changes in SAA bone marrow remains unknown, dysfunctional MSCs are implicated as a major cause. To provide evidence for their defects in immunomodulation, alterations of SAA MSCs in regulating T cell differentiation were determined. During differentiation from CD4+ T cells into T helper 17 (Th17) cells under polarization conditions, impaired inhibition on IL-17 and IL-1β production was noted when cocultured with SAA MSCs compared to control MSCs (

Topics: Anemia, Aplastic; Cell Differentiation; Humans; Interleukin-17; Lymphocyte Activation; Mesenchymal Stem Cells; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

To study the expression of B lymphocyte-induced mature protein-1 (BLIMP-1) in regulatory T cells (Tregs) of children with aplastic anemia (AA), and analyze its correlation with the number of Tregs and the levels of inhibitory cytokines interleukin (IL)-10 and transforming growth factor (TGF)-β in plasma.. The peripheral blood samples of 10 newly diagnosed AA children and 10 healthy children were collected for experiment. qPCR was used to detect FOXP3 and PRDM1 mRNA expression levels. Flow cytometry was used to detect the proportion of Tregs, the expression of BLIMP-1 in Tregs, and the levels of cytokines such as IL-2, IL-17A, IL-6, interferon (IFN)-γ, IL-10 and TGF-β in plasma. Pearson correlation model was used to evaluate the relationship between the expression of BLIMP-1 in Treg and the number of Tregs, as well as the levels of IL-10 and TGF-β in plasma.. Compared with control group, the proportion of Tregs in peripheral blood of AA children was decreased significantly (P<0.001); The plasma levels of proinflammatory cytokines IL-2, IL-6 and IFN-γ in AA children were increased significantly (P=0.033, P=0.031, P=0.006), and IL-17A also was increased but the difference was not statistically significant (P=0.052), while anti-inflammatory cytokines IL-10 and TGF-β were significantly reduced (P=0.048, P=0.002). The relative expressions level of FOXP3 and PRDM1 mRNA in AA children were significantly lower than those in control group (P=0.037, P=0.016). The expression of BLIMP-1 protein in Tregs of AA children was significantly lower than that in control group (P<0.001). The expression level of BLIMP-1 protein in Tregs was positively correlated with the percentage of Tregs in lymphocytes (r=0.671, P=0.001), and was also positively correlated with the levels of IL-10 and TGF-β in plasma (r=0.500, P=0.029; r=0.486, P=0.030).. The expression of BLIMP-1 in Tregs of AA children is impaired, and the low expression of BLIMP-1 is related to the decrease of the number in Tregs and IL-10 and TGF-β expressions.. BLIMP-1在再生障碍性贫血患儿Treg细胞中的表达及意义.. 研究B淋巴细胞诱导成熟蛋白-1（BLIMP-1）在再生障碍性贫血（AA）患儿Treg细胞中的表达并分析其与Treg细胞数量、血浆中抑制性细胞因子白介素（IL）-10和转化生长因子（TGF）-β水平的相关性.. 收集10例初诊AA患儿和10例健康儿童的外周血标本。采用qPCR法检测FOXP3、PRDM1的mRNA表达水平。采用流式细胞术检测Treg细胞比例、Treg细胞中BLIMP-1表达以及血浆中IL-2、IL-17A、IL-6、γ干扰素、IL-10和TGF-β等细胞因子水平。采用Pearson相关模型分析Treg细胞中BLIMP-1表达与Treg细胞数量、血浆中IL-10和TGF-β水平的相关性.. 与对照组相比，AA患儿外周血Treg细胞比例显著下降（P＜0.001）；AA患儿血浆中促炎细胞因子IL-2、IL-6、IFN-γ水平显著升高（P=0.033，P=0.031，P=0.006），IL-17A水平亦升高但不显著（P=0.052），而抑炎细胞因子IL-10和TGF-β水平明显降低（P=0.048，P=0.002）；AA患儿FOXP3和PRDM1 mRNA的相对表达量显著低于对照组（P=0.037，P=0.016）；AA患儿Treg细胞的BLIMP-1蛋白表达显著低于对照组（P＜0.001）。Treg细胞中BLIMP-1蛋白的表达水平与Treg细胞占淋巴细胞百分比呈正相关（r=0.671，P=0.001)，与血浆中IL-10和TGF-β水平亦呈正相关（r=0.500，P=0.029；r=0.486，P=0.030).. AA患儿Treg细胞中BLIMP-1表达受损，且BLIMP-1表达低下与Treg细胞数量减少和IL-10、TGF-β水平减少具有相关性.

Topics: Anemia, Aplastic; Child; Cytokines; Flow Cytometry; Forkhead Transcription Factors; Humans; Positive Regulatory Domain I-Binding Factor 1; T-Lymphocytes, Regulatory; Transforming Growth Factor beta

While the incidence of paroxysmal nocturnal hemoglobinuria (PNH) is relatively high in Northern China, the exact mechanism of the disease remains unknown. Immunoregulatory cytokine polymorphisms can directly regulate the expression levels of cytokines, which play a crucial role in many diseases. The purpose of this study was to study cytokine gene single nucleotide polymorphisms (SNPs) and the correlated cytokine expression levels in relationship to the PNH pathogenesis.. Peripheral blood samples were collected from 30 PNH patients and 40 healthy donors; all of the samples were collected from the Han people of Northern China. Eight SNP loci in five cytokine genes, including tumor necrosis factor-alpha (TNF-α), interferon-gamma (IFN-γ), transforming growth factor-beta (TGF-β), interleukin-6 (IL-6), and IL-10, and aplastic anemia (AA) were assessed. TNF-a, TGF-b, IFN-g, IL-6, and IL-10 were analyzed by sequence-specific primer polymerase chain reaction (PCR-SSP). The plasma protein levels of TNF-a, TGF-b, and IFN-g were assessed by an ELISA.. The PNH patients had a lower frequency of the TC/GG genotype of the TGF-b gene (P < 0.01) and a higher frequency of the C allele in the TGF-b gene (+10) compared to the controls (P < 0.05). The predominant genotype of the +874 locus of the IFN-g gene was TA in the PNH patients, while that in the predominant genotype was AA in the control group and was statistically significant (P < 0.001). The frequency of the T allele in the IFN-g gene was dramatically higher in the PNH patients than in the controls (P < 0.05). The PNH patients had a reduced frequency of the GC and CC genotypes, as well as the C allele at locus -174 of the IL-6 gene compared to the controls (P < 0.01). In addition, the plasma concentrations of TNF-a, TGF-b, and IFN-g were significantly higher in the PNH group compared to the control group (P < 0.01).. Expression levels of the TNF-a, TGF-b, and IFN-g cytokines play an important role in PNH. The GC and CC genotypes, as well as the C allele of the IL-6 gene may protect the Han people of Northern China against PNH. Additionally, the TC/GG genotype of the TGF-b gene may be the protective allele. In contrast, the TA genotype and the T allele for the IFN-g gene, as well as the C allele of TGF-b may be susceptible to PNH. However, SNPs in the TNF-a and IL-10 genes did not correlate with PNH development. Alternatively, the increased plasma concentrations of TNF-a, TGF-b, and IFN-g in PNH patients may also be related to PNH development.

Topics: Adult; Aged; Alleles; Anemia, Aplastic; Asian People; China; Cytokines; Enzyme-Linked Immunosorbent Assay; Female; Genotype; Hemoglobinuria, Paroxysmal; Humans; Interferon-gamma; Interleukin-10; Interleukin-6; Male; Middle Aged; Polymorphism, Genetic; Polymorphism, Single Nucleotide; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha; Young Adult

Glycosylphosphatidylinositol-anchored protein-deficient (GPI-AP(-) ) T cells can be detected in some patients with bone marrow failure (BMF), but the link between these cells and BMF pathophysiology remains to be elucidated. To clarify the significance of GPI-AP(-) T cells in BMF, peripheral blood from 562 patients was examined for the presence of CD48(-) CD59(-) CD3(+) cells using high-resolution flow cytometry (FCM), and the GPI-AP(-) T cells were characterized with regard to their phenotype and sensitivity to inhibitory molecules, including herpesvirus entry mediator (HVEM) and a myelosuppressive cytokine, TGF-β. A multi-lineage FCM analysis detected CD48(-) CD59(-) CD3(+) T cells in 72 (12.8%) of the patients, together with GPI-AP(-) myeloid cells. Unexpectedly, 12 patients (10 with aplastic anemia and 2 with myelodysplastic syndrome-refractory anemia, 2.1%), who showed clinical features similar to those of other BMF patients with GPI-AP(-) myeloid cells, such as a good response to immunosuppressive therapy, displayed 0.01-0.3% GPI-AP(-) cells exclusively in T cells. The CD48(-) CD59(-) T cells consisted of predominantly effector memory (EM) and terminal effector cells, while CD48(-) CD59(-) T cells from non-BMF patients who had received anti-CD52 antibody only showed EM and central memory phenotypes. TGF-β and HVEM capable of inhibiting T-cell proliferation via its GPI-AP CD160 ligation suppressed the in vitro proliferation of GPI-AP(+) T cells more potently than that of GPI-AP(-) T cells from the same patients. The presence of GPI-AP(-) T cells, as well as GPI-AP(-) myeloid cells, may therefore reflect the immunopathophysiology of BMF in which cytokine-mediated suppression of hematopoietic stem cells via GPI-AP-type receptors takes place.

Topics: Adolescent; Adult; Aged; Aged, 80 and over; Anemia, Aplastic; Bone Marrow Diseases; Bone Marrow Failure Disorders; Case-Control Studies; Child; Child, Preschool; Female; GPI-Linked Proteins; Hematopoietic Stem Cells; Hemoglobinuria, Paroxysmal; Humans; Immunophenotyping; In Vitro Techniques; Infant; Male; Membrane Proteins; Middle Aged; Myelodysplastic Syndromes; Myeloid Cells; Receptors, Tumor Necrosis Factor, Member 14; T-Lymphocytes; Transforming Growth Factor beta; Young Adult

To explore whether predisposition to bone marrow failure syndromes (BMF), such aplastic anemia (AA), paroxysmal nocturnal hemoglobinuria (PNH) and myelosysplastic syndromes (MDS), is found in killer cell immunoglobulin-like receptor (KIR) and human leukocyte antigen (HLA) ligand (KIR-L) gene variations or cytokine polymorphisms.. We studied a cohort of 77 patients with AA, 129 with MDS and 285 healthy controls for the frequencies of KIR-L and KIR genotypes and 22 selected single nucleotide polymorphisms (SNPs) located within 10 cytokine (IL-1α, IL-1β, IL-2, IL-4, IL-6, IL-10, IL12, IFN- γ, TNF- α, TGF- β) and 3 cytokine receptor (IL-1R, IL-1RA, IL-4Rα) genes.. In AA we found a decreased frequency of inhibitory KIR-2DL3 genes. In MDS, no difference in the frequency of KIR genotype was identified; however, a decreased frequency of 2DL3 was found in hypocellular MDS. Analysis of the KIR genotype in correlation with the corresponding KIR-L profile, revealed a decreased frequency of stimulatory 2DS1/C2 mismatch both in AA and MDS. In AA and MDS cohorts, compared to controls, we found a higher frequency of TT codon 10 variant and of GG codon 25 variant of TGF- β gene, consistent with a high secretory phenotype. This relationship was even more pronounced in PNH and hypocellular MDS. We confirm that the hypersecretory genotype T/T at position -874 of INF-γ gene was overrepresented only in AA and correlates with presence of a PNH clone. Instead in MDS patients, the frequency of G/A polymorphism at position -308 on the TNF- α gene promoter, which correlates with higher TNF- α production, was found significantly higher. Moreover, hypocellular MDS was characterized by a higher prevalence of IL-10 GCC/GCC haplotype, which is functionally associated with a low secretor phenotype.. Our findings suggest that alterations in KIR/KIR-L matching, such as increased 3DL2 and decreased 2DS1 mismatch, and in the polymorphisms of TGFβ1, IFN-γ, TNF- α and IL-10 may account for the propensity to immunemediated killing of hematopoietic stem cells and/or ineffective hematopoiesis characteristic of AA and MDS. Further studies are needed to elucidate whether these immunogenetic traits may be involved in increased risk of developing immune-mediated BMF.

Topics: Adolescent; Adult; Aged; Anemia, Aplastic; Bone Marrow Diseases; Bone Marrow Failure Disorders; Child; Child, Preschool; Cohort Studies; Cytokines; Gene Frequency; Genotype; Hemoglobinuria, Paroxysmal; Humans; Interferon-gamma; Interleukin-10; Middle Aged; Myelodysplastic Syndromes; Polymorphism, Single Nucleotide; Receptors, Cytokine; Receptors, KIR; Transforming Growth Factor beta

Transforming growth factor beta (TGF-beta) 1 is a ubiquitous bifunctional cytokine implicated in the regulation of haemopoietic stem cells and bone marrow stromal cells. We analysed sera from 63 patients with aplastic anaemia and describe a significant reduction of TGF-beta1 that was directly related to their treatment status. Untreated patients (n = 35), patients who did not respond (n = 15) and those with a partial response (n = 23) to treatment had significantly lower TGF-beta1 than the normal control group (n = 55), P < 0.0001, P < 0.0001 and P = 0.002 respectively. Patients in complete remission (n = 15) exhibited TGF-beta1 serum levels comparable to the control group. In addition, there was a correlation (r = 0.83, P < 0.0001) between serum TGF-beta1 and platelet count at time of sample. We have demonstrated that the primary source of TGF-beta1 in peripheral blood mononuclear cell (PBMC) cultures was not CD3-positive cells. These data indicate aplastic anaemia is associated with a decreased TGF-beta1 expression in peripheral blood circulation, which may be a direct consequence of thrombocytopenia. In vitro stromal layers grown from aplastic patient bone marrow (n = 14) produced significantly lower levels of TGF-beta1 (P = 0.02) when compared to normal stroma (n = 15). In the aplastic anaemia bone marrow compartment we postulate that accessory cells down-regulate TGF-beta1 expression to allow stem cell cycling to counteract hypoplasia. As TGF-beta1 is important in the regulation of haemopoiesis, dysregulation of this cytokine in combination with previously described abnormal cytokine expression may contribute significantly to the pathophysiology of aplastic anaemia by exacerbating primary stem cell defects.

Topics: Adolescent; Adult; Aged; Anemia, Aplastic; Cells, Cultured; Female; Humans; Leukocytes, Mononuclear; Male; Middle Aged; Platelet Count; Stromal Cells; Transforming Growth Factor beta

The aim of this study was to measure the level of cytokines produced by peripheral blood mononuclear cells (PBMNC) in patients with aplastic anemia (AA) and determine their effect on normal bone marrow (BM) colony growth. Thirty-five patients with AA and 21 normal controls were enrolled in the study. Medium conditioned by PBMNC of AA patients in the presence of phytohemagglutinin (PHA) was found to be suppressive to the clonal growth of normal BM cells. Thus, we further determined the presence in the PBMNC conditioned medium (CM) of inhibitory cytokines (macrophage inflammatory protein-1 alpha [MIP-1 alpha], transforming growth factor-beta 2 [TGF-beta 2], interferon-gamma [IFN-gamma], and tumor necrosis factor-alpha [TNF-alpha]) and stimulatory cytokines (granulocyte-macrophage colony-stimulatory factor [GM-CSF], interleukin-3 [IL-3], and stem cell factor [SCF]). The results show no significant difference between AA patients and normal controls in the spontaneous production of all cytokines by PBMNC. After PHA stimulation, the production of MIP-1 alpha, IFN-gamma, TNF-alpha, and GM-CSF significantly increased in the cultures of AA patients (p = 0.0009, 0.0002, 0.0022, and 0.0156, respectively). However, both TGF-beta 2 and SCF were undetectable in most of the tested samples. IL-3 was measured in the conditioned medium only after PHA stimulation, but without significant difference between the two groups (p = 0.67). Furthermore, the myelopoietic suppressing effect of AA-PBMNC CM could be significantly blocked by pretreatment with specific antibodies to the corresponding inhibitory cytokines (MIP-1 alpha, IFN-gamma, and TNF-alpha). After antibody neutralization, an apparent change occurred in the clonal growth of normal BM cells incubated with AA-PBMNC CM, resulting in colony enhancement of 205, 131, and 237% by anti-MIP-1 alpha, anti-IFN-gamma, and anti-TNF-alpha, respectively. These results suggest that overproduction of inhibitory cytokines, rather than underproduction of stimulating cytokines, may play a role in the progression of at least some patients with AA.

Topics: Anemia, Aplastic; Antibodies; Chemokine CCL4; Culture Media, Conditioned; Cytokines; Female; Granulocyte-Macrophage Colony-Stimulating Factor; Hematopoiesis; Humans; Interferon-gamma; Interleukin-3; Leukocytes, Mononuclear; Macrophage Inflammatory Proteins; Male; Middle Aged; Monokines; Stem Cell Factor; Transforming Growth Factor beta; Tumor Necrosis Factor-alpha

Levels of transforming growth factor-beta (TGF-beta) activity in the conditioned medium of blood lymphocytes of twelve patients with aplastic anemia (AA), nine patients with myelodysplastic syndromes (MDS) and five normal volunteers were investigated. We were able to observe growth inhibitory activity on porcine endothelial cells only after acidification of the materials. The growth inhibitory activity is neutralized by anti-TGF-beta antibody. It indicates that TGF-beta exists as a latent form in the conditioned medium. On the basis of growth inhibition assay, the mean level of TGF-beta production of MDS patients was estimated to be 188 +/- 199 pg/1 x 10(7) cells and that of normal volunteers was 668 +/- 314 pg/1 x 10(7) cells. In contrast, the lymphocytes of almost all of the AA patients failed to produce detectable amounts of TGF-beta. No correlation between TGF-beta levels and peripheral blood parameters could be detected. Stimulation of lymphocytes by phytohemagglutinin is known to increase the production of TGF-beta. Induction of TGF-beta production was also observed in AA (45% of normal controls). Possible roles of the decreased production of TGF-beta in the pathogenesis of AA were discussed.

Topics: Adult; Anemia, Aplastic; Cells, Cultured; Female; Humans; Lymphocytes; Male; Middle Aged; Myelodysplastic Syndromes; Transforming Growth Factor beta

Topics: Anemia, Aplastic; Animals; Glomerulonephritis; Humans; Liver Regeneration; Myocardial Infarction; Pulmonary Fibrosis; Receptors, Cell Surface; Receptors, Transforming Growth Factor beta; Signal Transduction; Transforming Growth Factor beta