BACKGROUNDRecent studies have shown that T helper type-2 (Th2) cells can induce the apoptosis of CD4+CD25+ Treg cells or resist the immunosuppressive effect of Treg cells. We hypothesize that an imbalance of Th2/Treg is present in patients with allergic asthma.

METHODSTwenty-two patients with mild asthma, 17 patients with moderate to severe asthma, and 20 healthy donors were enrolled. All patients were allergic to house dust mites. The proportion of peripheral blood CD4+CD25+ Treg cells and Th2 cells were determined by flow cytometry. The concentration of interleukin (IL)-10, transforming growth factor (TGF)-β and IL-4 in plasma was determined by enzyme linked immunosorbent assay. In these subjects, peripheral blood mononuclear cells from 17 mild asthmatic patients, 13 moderate to severe asthmatic patients and 14 healthy donors were acquired and expression of forkhead box P3 (Foxp3) and GATA-3 mRNA was detected by reverse-transcriptase polymerase chain reaction.

RESULTSCompared with healthy donors and patients with mild asthma, the percent of CD4+CD25+ Treg cells and plasma IL-10 levels were decreased in patients with moderate to severe asthma. There were no significant differences in Foxp3 mRNA expression among three groups, but a downward trend seen among patients with asthma. However, the percent of Th2 cells, IL-4 levels and expression of GATA-3 mRNA was markedly higher in patients with mild and moderate to severe asthma than in the control group. The ratio of Th2/Treg and their cytokines was increased in allergic asthma, especially for moderate to severe asthma. The ratio of GATA-3/Foxp3 mRNA was also increased in allergic asthma. In patients with moderate to severe asthma, the percentage of peripheral blood Treg cells was negatively correlated to the percentage of Th2 cells and IL-4 levels.

CONCLUSIONSThe decline of CD4+CD25+ Treg cells in patients with moderate to severe asthma may play an important role in progress of the disease. Furthermore, the deficiency of CD4+CD25+ Treg cells was associated with the over-expression of Th2 response.

Asthma ; etiology ; immunology ; Cytokines ; blood ; Forkhead Transcription Factors ; genetics ; GATA3 Transcription Factor ; genetics ; Humans ; RNA, Messenger ; analysis ; T-Lymphocytes, Regulatory ; immunology ; Th2 Cells ; immunology

OBJECTIVETo investigate the role of CD4+CD25+high regulatory T cells in the pathogenesis of autoimmune hepatitis.

METHODSCD4+CD25+ high regulatory T cells and CD4+ T cells were measured by using flow cytometry in 16 patients with autoimmune hepatitis, 22 patients with chronic hepatitis B and 20 healthy blood donors. Foxp3 protein was detected by immunohistochemical assay in liver tissues from the patients with autoimmune hepatitis or chronic hepatitis B.

RESULTSThe percentage of CD4+CD25+high/CD4+ in patients with autoimmune hepatitis was significantly lower than that in healthy controls and patients with chronic hepatitis B. Meanwhile, the percentage of CD4+CD25+high/CD4+ highly increased in patients with chronic hepatitis B, compared with healthy controls; Foxp3 positive cells were mostly located in the hepatic lobular perisinusoidal spaces and the portal tract, and there was a significant difference in the quantity of Foxp3 positive cells between patients with autoimmune hepatitis and chronic hepatitis B.

CONCLUSIONPatients with autoimmune hepatitis harbor a decreased percentage of CD4+CD25+ high regulatory T cells, which may be associated with development of autoimmunity.

Adult ; Female ; Forkhead Transcription Factors ; analysis ; Hepatitis B, Chronic ; immunology ; Hepatitis, Autoimmune ; immunology ; Humans ; Immunohistochemistry ; Liver ; chemistry ; Male ; Middle Aged ; T-Lymphocytes, Regulatory ; immunology

OBJECTIVETo investigate the effect of heat shock factor 1 (HSF1) on airway hyperresponsiveness and airway inflammation in mice with asthma and possible mechanisms.

METHODSA total of 36 mice were randomly divided into four groups: control, asthma, HSF1 small interfering RNA negative control (siHSF1-NC), and siHSF1 intervention (n=9 each). Ovalbumin (OVA) sensitization and challenge were performed to induce asthma in the latter three groups. The mice in the siHSF1-NC and siHSF1 groups were treated with siHSF1-NC and siHSF1, respectively. A spirometer was used to measure airway responsiveness at 24 hours after the last challenge. The direct count method was used to calculate the number of eosinophils. ELISA was used to measure the serum level of OVA-specific IgE and levels of interleukin-4 (IL-4), interleukin-5 (IL-5), interleukin-13 (IL-13), and interferon-γ (IFN-γ) in lung tissues and bronchoalveolar lavage fluid (BALF). Quantitative real-time PCR was used to measure the mRNA expression of HSF1 in asthmatic mice. Western blot was used to measure the protein expression of HSF1, high-mobility group box 1 (HMGB1), and phosphorylated c-Jun N-terminal kinase (p-JNK).

RESULTSThe asthma group had significant increases in the mRNA and protein expression of HSF1 compared with the control group (P<0.05). The siHSF1 group had significantly reduced mRNA and protein expression of HSF1 compared with the siHSF1-NC group (P<0.05). The knockdown of HSF1 increased airway wall thickness, airway hyperresponsiveness, OVA-specific IgE content, and the number of eosinophils (P<0.05). Compared with the siHSF1-NC group, the siHSF1 group had significantly increased levels of IL-4, IL-5, and IL-13 and significantly reduced expression of IFN-γ in lung tissues and BALF (P<0.05), as well as significantly increased expression of HMGB1 and p-JNK (P<0.05).

CONCLUSIONSKnockdown of HSF1 aggravates airway hyperresponsiveness and airway inflammation in asthmatic mice, and its possible mechanism may involve the negative regulation of HMGB1 and JNK.

Animals ; Asthma ; etiology ; Bronchial Hyperreactivity ; etiology ; immunology ; Cytokines ; biosynthesis ; DNA-Binding Proteins ; analysis ; physiology ; Eosinophils ; physiology ; Female ; HMGB1 Protein ; analysis ; Heat Shock Transcription Factors ; Immunoglobulin E ; blood ; Mice ; Mice, Inbred BALB C ; Transcription Factors ; analysis ; physiology

BACKGROUNDActive suppression by CD4+CD25+ regulatory T lymphocytes plays an important role in the down-regulation of T cell responses to foreign and self-antigens. This study was conducted to analyze whether the CD4+CD25+ regulatory T cells exist and function normally in tuberculous pleural effusion.

METHODSThe percentages of CD4+CD25+ T cells in pleural effusion and peripheral blood from patients with tuberculous pleurisy and peripheral blood from healthy control subjects were determined by flow cytometry. The expression of forkhead transcription factor Foxp3 was also examined. CD4+CD25+ and CD4+CD25(-) T cells from pleural effusion and blood were isolated, and were cultured to observe the effects of CD4+CD25+ T cells on proliferation response of CD4+CD25(-) T cells in vitro.

RESULTSThere were increased numbers of CD4+CD25+ T cells in tuberculous pleural effusion compared with peripheral blood from both patients with tuberculous pleurisy and normal subjects, and these cells demonstrated a constitutive high-level expression of Foxp3. Moreover, CD4+CD25+ T cells mediated potent inhibition of proliferation response of CD4+CD25(-) T cells.

CONCLUSIONThe increased CD4+CD25+ T cells in tuberculous pleural effusion express a high level of Foxp3 transcription factor, while potently suppressing the proliferation of CD4+CD25(-) T cells.

Adult ; Female ; Forkhead Transcription Factors ; analysis ; Humans ; Lymphocyte Activation ; Male ; Middle Aged ; Pleural Effusion ; etiology ; immunology ; T-Lymphocytes, Regulatory ; physiology ; Tuberculosis, Pleural ; etiology ; immunology

OBJECTIVETo study the roles of CD4(+)CD25(+)Foxp3(+) regulatory T cells (Treg) and IL-33 in the pathogenesis of asthma in children.

METHODSFlow cytometry was used to detect peripheral blood CD4(+)CD25(+)Foxp3(+)Treg proportion in CD4(+)T lymphocytes in.45 children with asthma, 50 children with wheezing caused by respiratory syncytial virus infection and 40 healthy children. Serum levels of IFN-γ, IL-4, IL-5 and IL-33 were measured using ELISA.

RESULTSThe level of peripheral blood CD4(+)CD25(+)Foxp3(+)Treg in the asthma group was significantly lower than in the wheezing and control groups (P<0.05). In contrast, serum levels of IL-33 in the asthma group was significantly higher than in the wheezing and control groups (P<0.05). Peripheral blood CD4(+)CD25(+)Foxp3(+)Treg level was negatively correlated with serum IL-33 level in the asthma group(r=-0.156, P<0.01).

CONCLUSIONSCD4(+)CD25(+)Foxp3(+)Treg may interact with IL-33 in the pathogenesis of childhood asthma.

Asthma ; etiology ; immunology ; Child ; Child, Preschool ; Female ; Forkhead Transcription Factors ; analysis ; Humans ; Infant ; Interleukin-2 Receptor alpha Subunit ; immunology ; Interleukin-33 ; Interleukins ; physiology ; Male ; T-Lymphocytes, Regulatory ; physiology

OBJECTIVETo observe the pathology of AIDS-related lymphadenopathy and its relationship to the expression and distribution of CD4 + CD25 + regulatory T cells in lymphoid node tissue.

METHODSTotally 22 biopsy and 13 autopsy lymphoid node tissues from HIV-positive patients were examined under microscopy and pathological staging was performed. Specific marker for CD4 + CD25 + regulatory T cells in lymphoid node tissue was detected with anti-Foxp3 monoclonal antibody by immunohistochemistry.

RESULTSAmong all the 35 specimens, 5, 4, 14, and 12 specimens were histopathologically staged from 1 to 4, respectively. FoxP3 were detected in all lymphoid node tissues. The distribution of FoxP3-positive lymphocytes were mainly in intermediate zone of follicle and cortical area in stages 1 and 2. The counts of FoxP3-positive lymphocytes remarkably decreased in stages 3 and 4, following depletion of lymphocytes.

CONCLUSIONSCD4 + CD25 + regulatory T cells exist in lymphoid node tissue of patients with HIV infection. Their amounts decrease or deplete along with the progression of AIDS-related lymphadenopathy.

Acquired Immunodeficiency Syndrome ; immunology ; pathology ; Adult ; CD4 Lymphocyte Count ; Female ; Forkhead Transcription Factors ; analysis ; Humans ; Immunohistochemistry ; Lymph Nodes ; immunology ; pathology ; Lymphatic Diseases ; immunology ; Male ; Middle Aged ; T-Lymphocytes, Regulatory ; metabolism

OBJECTIVETo investigate variation of FOXP3 and it's expression in male children presented with severe and early-onset enteropathy, rash with or without insulin-dependent diabetes mellitus (IDDM).

METHODSFour male children presented with severe and early-onset enteropathy, rash, with or without IDDM were subjected to detection of FOXP3 expression on the PBMC by flow cytometry and FOXP3 gene analysis. The maternal gene analysis was subsequently performed once the variant FOXP3 gene was found. All 11 exons and splice sites within FOXP3 gene were amplified by polymerase chain reaction (PCR) from genomic DNA. Reverse transcription polymerase chain reaction was used to amplify the FOXP3 transcripts. Sequence analysis was performed directly on the bulk PCR products forwardly and reversely. The candidate mutation site was compared with that of 100 healthy controls to exclude polymorphism. Flow cytometry was used to determine FOXP3 expression on CD4+CD25+ T cells and the frequency of Tregs in CD4+ T cells.

RESULTSOne of the 4 patients showed a G13128A genetic variation in exon 11, which resulted in a Met370Ile substitution. No sequence variations were found at the same site in any of 100 healthy controls, indicating that the Met370Ile substitution is not a polymorphism but a novel missense mutation. The patient's mother was identified as a carrier for this mutation. There was no reduced frequency of Tregs in the peripheral blood of the patient and FOXP3 protein expression is normal as compared with controls.

CONCLUSIONA novel missense mutation of FOXP3 which causes IPEX phenotype was identified in a Chinese child according to immunologic screening and gene sequencing. Infants with early-onset IDDM and persistent diarrhea should be suspected as IPEX, FOXP3 gene analysis will be a reliable diagnostic approach to IPEX.

Child, Preschool ; DNA Mutational Analysis ; Diabetes Mellitus, Type 1 ; genetics ; immunology ; Forkhead Transcription Factors ; genetics ; Genes, X-Linked ; Genetic Diseases, X-Linked ; genetics ; immunology ; metabolism ; Humans ; Infant ; Intestinal Diseases ; genetics ; metabolism ; Male ; Mutation, Missense ; Syndrome ; T-Lymphocytes, Regulatory ; immunology

OBJECTIVETo study the clinicopathologic features and immunophenotype of renal cell carcinomas, and to discuss their diagnostic value.

METHODSThe clinicopathologic features of 114 cases of renal cell carcinoma were reviewed and categorized on the basis of 2004 WHO classification. Immunohistochemical study for a panel of antibodies (including CK, CD10, vimentin, CD117, AMACR, CK7 and TFE3) was carried out. The follow-up data, if available, were also analyzed.

RESULTSThe cases were reclassified into 5 subtypes, including 77 cases (67.5%) of clear cell carcinoma (CCRCC), 11 cases (9.6%) of papillary renal cell carcinoma (PRCC), 14 cases (12.3%) of chromophobe renal cell carcinoma (chrRCC), 10 cases (8.8%) of renal carcinoma associated with Xp11.2 translocations/TFE3 gene fusions (Xp11.2RCC) and 2 cases (1.8%) of unclassified renal cell carcinoma (unRCC). Immunohistochemical study showed that the expression rates of CK, CD10 and vimentin in CCRCC were 93.5% (72/77), 93.5% (72/77) and 75.3% (58/77), respectively. On the other hand, all the 11 cases of PRCC studied were positive for AMACR. The expression rate of CD117 in chrRCC was 78.5% (11/14). In the 10 cases of Xp11.2 RCC studied, the expression rates of TFE3, AMACR, CD10 and CK were 100% (10/10), 100% (10/10), 90% (9/10) and 70% (7/10), respectively.

CONCLUSIONSThe various subtypes of renal cell carcinomas are heterogeneous in histologic appearance and demonstrate distinctive immunophenotype. The expressions of CD10, vimentin, CD117, AMACR, CK7 and TFE3 are helpful in the differential diagnosis.

Adenocarcinoma, Clear Cell ; pathology ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Basic Helix-Loop-Helix Leucine Zipper Transcription Factors ; genetics ; immunology ; metabolism ; Biomarkers, Tumor ; analysis ; genetics ; Carcinoma, Papillary ; immunology ; pathology ; Carcinoma, Renal Cell ; immunology ; metabolism ; pathology ; Female ; Gene Fusion ; Humans ; Immunophenotyping ; Kidney Neoplasms ; immunology ; metabolism ; pathology ; Male ; Middle Aged ; Neprilysin ; analysis ; Racemases and Epimerases ; analysis ; genetics ; Translocation, Genetic ; Vimentin ; analysis ; World Health Organization ; Young Adult

The identification of primary location of a metastatic tumor is a difficult diagnostic problem and sometimes can be facilitated by the use of immunohistochemical markers. Thyroid transcription factor-1 (TTF-1) is a 38-kDa nuclear homeodomain transcription factor that is expressed specifically in lung or thyroid neoplasms. Cytokeratin 20 (CK20) is a 46-kDa low-molecular-weight cytokeratin that shows restricted expression in adenocarcinomas of the gastrointestinal tract (GIT) and transitional cell carcinomas of the urinary tract. We studied the immunohistochemical expression of TTF-1 and CK20 in 68 metastatic carcinomas in cervical lymph nodes. The primary sites were the lung in 29 cases, stomach in 13, colorectum in 3, and other sites in 23. TTF-1 expression was detected in 69.0% of metastatic lung carcinomas and none in metastatic GIT carcinomas, whereas CK20 expression was detected in 68.8% of metastatic GIT carcinomas and none of metastatic lung carcinomas. TTF-1 had a specificity of 0.95 and a sensitivity of 0.69 for metastatic lung carcinoma, whereas CK20 had a specificity of 1.00 and a sensitivity of 0.69 for metastatic GIT carcinoma. These results indicate that TTF-1 and CK20 should be the first choice as a component of antibody panel to prove or to exclude the lung and GIT origin, respectively, especially in patients presenting with metastatic carcinomas of unknown primary site.
Adenocarcinoma/chemistry/pathology/secondary ; Carcinoma/chemistry/pathology/*secondary ; Gastrointestinal Neoplasms/chemistry/pathology ; Homeodomain Proteins/analysis ; Humans ; Intermediate Filament Proteins/*analysis/immunology ; Keratin-20 ; Lung Neoplasms/chemistry/pathology ; Lymph Nodes/chemistry/pathology ; Lymphatic Metastasis/*diagnosis/pathology ; Neck ; Neoplasms, Unknown Primary/chemistry/pathology ; Nuclear Proteins/*analysis/immunology ; Sensitivity and Specificity ; Transcription Factors/*analysis/immunology ; Tumor Markers, Biological/analysis

OBJECTIVETo investigate the molecular mechanism of action of BET bromodomain inhibitor JQ1 in treating airway remodeling in asthmatic mice.

METHODSA total of 24 mice were randomly divided into control group, ovalbumin (OVA)-induced asthma group (OVA group), and JQ1 intervention group (JQ1+OVA group), with 8 mice in each group. OVA sensitization/challenge was performed to establish a mouse model of asthma. At 1 hour before challenge, the mice in the JQ1+OVA group were given intraperitoneal injection of JQ1 solution (50 μg/g). Bronchoalveolar lavage fluid (BALF) and lung tissue samples were collected at 24 hours after the last challenge, and the total number of cells and percentage of eosinophils in BALF were calculated. Pathological staining was performed to observe histopathological changes in lung tissue. RT-PCR and Western blot were used to measure the mRNA and protein expression of E-cadherin and vimentin during epithelial-mesenchymal transition (EMT).

RESULTSCompared with the control group, the OVA group had marked infiltration of inflammatory cells in the airway, thickening of the airway wall, increased secretion of mucus, and increases in the total number of cells and percentage of eosinophils in BALF (P<0.01). Compared with the OVA group, the JQ1+OVA group had significantly alleviated airway inflammatory response and significant reductions in the total number of cells and percentage of eosinophils in BALF (P<0.01). Compared with the control group, the OVA group had significant reductions in the mRNA and protein expression of E-cadherin and significant increases in the mRNA and protein expression of vimentin (P<0.01); compared with the OVA group, the JQ1+OVA group had significant increases in the mRNA and protein expression of E-cadherin and significant reductions in the mRNA and protein expression of vimentin (P<0.01); there were no significant differences in these indices between the JQ1+OVA group and the control group (P>0.05).

CONCLUSIONSMice with OVA-induced asthma have airway remodeling during EMT. BET bromodomain inhibitor JQ1 can reduce airway inflammation, inhibit EMT, and alleviate airway remodeling, which provides a new direction for the treatment of asthma.

Airway Remodeling ; drug effects ; Animals ; Asthma ; drug therapy ; Azepines ; pharmacology ; Cadherins ; analysis ; genetics ; Epithelial-Mesenchymal Transition ; Female ; Mice ; Nuclear Proteins ; antagonists & inhibitors ; Ovalbumin ; immunology ; RNA, Messenger ; analysis ; Transcription Factors ; antagonists & inhibitors ; Triazoles ; pharmacology ; Vimentin ; analysis ; genetics