OBJECTIVE: To explore the relationship between GATA-3 and IL-12 in patients with allergic rhinitis. METHOD: The expression of GATA-3 was detected by reverse transcriptase polymerase chain reaction (RT-PCR) in 37 patients with allergic rhinitis and 12 control samples. IL-12 was detected by enzyme-linked immunosorbent assay. RESULT: The relative density ratio of GATA-3 to GAPDH in AR patients was 0.579 +/- 0.102, and in control group was 0.128 +/- 0.021. The concentrations of IL-12 were (53.7 +/- 12.3) ng/g and (121.9 +/- 20.4) ng/g in the two groups respectively. The expression of GATA-3 was negatively correlated to the expression of IL-12 in patients with allergic rhinitis. CONCLUSION The overexpression of GATA-3 in patients with allergic rhinitis was related to the decrease of IL-12, and to increasing the expression of IL-12 in topical tissue could suppress the expression of GATA-3 and may improve the therapeutic effectiveness of AR.
Case-Control Studies ; GATA3 Transcription Factor ; metabolism ; Humans ; Interleukin-12 ; metabolism ; Nasal Mucosa ; metabolism ; Rhinitis, Allergic, Perennial ; metabolism

BACKGROUNDInterleukin-13 (IL-13) is recognized to be a key modulator in the pathogenesis of Th2-induced allergic inflammation. Transcription factors GATA3 and NFAT1 have been both implicated in the regulation of Th2 cytokines. We previously demonstrated the GATA3-NFAT1 association during human T cell activation. However, the function of the GATA3-NFAT1 complex in Th2 cytokines regulation is still unknown. Small interference RNA (siRNA) was constructed to knock down GATA3 expression in Hut-78 cells to investigate the possible role of GATA3-NFAT1 complex in IL-13 transcription.

METHODSCells were stimulated with anti-CD3 plus anti-CD28 antibodies to mimic in vivo antigen-mediated co-stimulation; the expression of IL-13 mRNA was determined by real-time PCR; chromation immunoprecipitation (CHIP) assay was employed to investigate the NFAT1 binding to IL-13 promoter.

RESULTSGATA3 siRNA suppressed the expression of GATA3 both in mRNA and protein levels in Hut-78 cells. The binding of NFAT1 to IL-13 promoter was inhibited by GATA3 siRNA in activated T cells, which was followed by the reduction of IL-13 transcription.

CONCLUSIONGATA3-NFAT1 complex may play an important role in the regulation of IL-13 transcription in human T cells.

Cells, Cultured ; GATA3 Transcription Factor ; antagonists & inhibitors ; genetics ; Humans ; Interleukin-13 ; genetics ; NFATC Transcription Factors ; metabolism ; Promoter Regions, Genetic ; RNA, Small Interfering ; genetics ; T-Lymphocytes ; metabolism ; Transfection

OBJECTIVE: To investigate kinetic expression of genes T-bet and GATA-3 in blood mononuclear cells (BMCs) in allergic rhinitis rats model at different stages of development of allergic rhinitis. METHOD: Twenty SD rats (including 10 male and 10 female) were divided into 2 groups, experimental group and control group, randomly, 10 rats for each group. Ten rats in experimental group were sensitized and intranasally challenged by OVA, aluminium hydroxide hydrate gel and Bordelella pertussis inactive microorganism suspension (B. pertussis) adjuvants, as allergic rhinitis models, Ten rats in control group were investigated using physiological saline only. BMCs were separated from 2 ml blood which was extracted from rat heart at the end of sensitization, 10- hour after the first challenge and 10-hour after the final challenge, respectively. RT-PCR was utilized to detect the expression of T-bet and GATA-3. RESULT: At the end of sensitization, 10-hour after the first challenge and 10-hour after the final challenge, in experimental group, Relative quantitation of expression of T-bet was 0.404 +/- 0.187, 1.676 +/- 0.708, 0.503 +/- 0.514 and that of GATA-3 was 0.434 +/- 0.147, 0.600 +/- 0.480, 1.029 +/- 0.690, respectively. While, In control group, Relative quantitation of expression of T-bet was 0.487 +/- 0.212, 0.486 +/- 0.148, 0.495 +/- 0.103 and and its of GATA-3 was 0.596 +/- 0.249, 0.474 +/- 0.101, 0.550 +/- 0.119, respectively. At 10-hour after the first challenge, relative quantitation of expression of T-bet in experimental group was increased markedly and there was significant difference compared with contol group (t=4.18, P<0.01). In experimental group, The amount of expression of T-bet at 10-hour after first challenge was higher than it at 10-hour after the final challenge and there was markedly difference (t=5.14, P<0.01). The amount of expression of T-bet at 10-hour after first challenge was increased significantly compared with it at the end of sensitization (t= 5.27, P<0.01). while, the expression of GATA-3 at 10-hour after final challenge was markedly increased compared with it at the end of sensitization (t= 3.51, P<0.05) and was higher than it at 10-hour after first challenge (t=2.53, P<0.05). At 10-hour after final challenge, The amount of expression of GATA-3 in experimental group was significantly higher than it in control group (t=2.71, P<0.05). However, both the expression of GATA-3 and T-bet had not markedly changed in control group. CONCLUSION It is thought that the development of allergic rhinitis is a successive and sequencing kinetic course, and imbalance of expression of GATA-3 and T-bet may be genetic base on allergic rhinitis, both GATA-3 and T-bet were involved in allergic rhinitis only in different phase of development of allergic rhinitis.
Animals ; Asthma ; metabolism ; Disease Models, Animal ; Female ; GATA3 Transcription Factor ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley ; Rhinitis ; metabolism ; T-Box Domain Proteins ; metabolism

Adult ; Aged ; Female ; GATA3 Transcription Factor ; genetics ; metabolism ; Humans ; Leukocytes, Mononuclear ; metabolism ; Lymphoma, Large B-Cell, Diffuse ; blood ; genetics ; Male ; Middle Aged ; RNA, Messenger ; genetics

GATA-3 plays a central role in the Th2-mediated immunoreaction. This study was aimed to construct and select plasmid vectors of siRNA which can effectively and specifically suppress the gene expression of GATA-3. Plasmid including PSi338, PSi717 and PSi1232 were designed and constructed for GATA-3 regarded as target gene and transfected into murine P815 cells. Reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blot were used to detect the inhibition of GATA-3 mRNA as well as its protein in P815 cells. The results demonstrated that the expressions of mRNA and protein of GATA-3 in P815 cells were inhibited significantly by both of PSi338 and PSi717. It is concluded that PSi338 and PSi717 siRNA plasmid vectors have been successfully constructed, and both vectors are effective and specific siRNA plasmids for suppressing GATA-3 gene expression.
Animals ; Down-Regulation ; GATA3 Transcription Factor ; metabolism ; Genetic Vectors ; Mice ; Plasmids ; RNA Interference ; RNA, Messenger ; metabolism ; RNA, Small Interfering ; genetics ; Th2 Cells ; immunology ; Transfection

OBJECTIVETo observe the therapeutic effect and mechanism of new percutaneous absorption herbal patch for asthma of paracmasis, and to optimize the form of the patch.

METHODSOne hundred and twenty cases of paracmasis asthma were randomly divided into medicine patch group, medicinal vesiculation group and western medication group with 40 cases for each. The new percutaneous absorption herbal patch was applied on medicine patch group. Traditional medicinal herbs cake of the hospital were applied on medicinal vesiculation group. Feishu (BL 13), Fengmen (BL 12) and Dazhui (GV 14) were adopted for both groups. Each patch was applied for 6 hours and once every other day. Accuhaler was applied on the western medicine group with 2 inhalations a time and twice a day. Clinical symptom scores, number of attacks and asymtomatic days were observed right before, after and half a year after the treatment. Meanwhile, the expression level of IgE, IL-4, GATA-3 mRNA and T-bet mRNA were observed and compared before and after the treatment.

RESULTSClinical symptom scores of all the 3 groups were improved (all P < 0.01). The differences of the total effective rate, number of attacks and asymtomatic days of all the 3 groups are without statistical significance (all P > 0.05). However, the long-term therapeutic effect in half a year after the treatment showed that the total effective rate of the medicine patch group was 80.0% (32/40), and the medicinal vesiculation group was 70.0% (28/40). Both of the them were obviously higher than 47.5% (19/40) of the western medicine group (P < 0.01, P < 0.05). And the medicine patch group surpassed the other 2 groups in controlling the number of attacks and increasing the asymtomatic days (P < 0.05, P < 0.01). The level of IgE and IL-4 of all the 3 groups decreased sharply after the treatment (P < 0.05, P < 0.01). And there was no statistic significance in differences among groups (all P > 0.05). The level of GATA-3 mRNA was obviously decreased, while the level of T-bet mRNA was obviouly increased in the medicine patch and medicinal vesiculation groups (P < 0.05, P < 0.01). And the medicine patch group had obvious superiority on increasing the level of T-bet mRNA when compared with the medicinal vesiculation and western medicine groups (P < 0.01, P < 0.05).

CONCLUSIONIt is concluded that the new percutaneous absorption herbal patch has exact effect on asthma. The treatment may reverse the imbalance condition of Th1/Th2 through regulation on cell factor and its specific transcription factors.

Acupuncture Points ; Adolescent ; Adult ; Aged ; Asthma ; drug therapy ; genetics ; metabolism ; Child ; Drugs, Chinese Herbal ; therapeutic use ; Female ; GATA3 Transcription Factor ; genetics ; metabolism ; Humans ; Interleukin-4 ; genetics ; metabolism ; Male ; Middle Aged ; Transcription Factors ; genetics ; metabolism ; Young Adult

OBJECTIVETo study mRNA expression of immune-related genes (Foxp3, GATA3, CTLA4 and T-bet) in peripheral blood of the patients with allergic dermatitis induced by trichloroethylene (TCE).

METHODSThe peripheral blood samples were collected from 8 healthy workers (control group) and 8 patients with allergic dermatitis induced by TCE (case group). Real-time quantitative PCR was applied to detect mRNA expression of immune-related genes (Foxp3, GATA3, CTLA4, T-bet).

RESULTSThe mRNA expression levels of Foxp3, GATA3 and CTLA4 genes increased by 115%, 97% and 241% in case group, as compared with control group (P < 0.01). The mRNA expression level of T-bet gene decreased by 47% in case group, as compared with control group (P < 0.01).

CONCLUSIONThe mRNA expression levels of some immune-related genes changed in patients with allergic dermatitis induced by TCE, those genes may play an important role in TCE-induced allergy.

Adult ; CTLA-4 Antigen ; metabolism ; Case-Control Studies ; Dermatitis, Occupational ; genetics ; immunology ; Female ; Forkhead Transcription Factors ; metabolism ; GATA3 Transcription Factor ; metabolism ; Gene Expression ; Humans ; Male ; RNA, Messenger ; genetics ; T-Box Domain Proteins ; metabolism ; Trichloroethylene ; Young Adult

OBJECTIVETo study the expression levels of annexin A1 (ANXA1), GATA-3, and T-bet in T lymphocytes of peripheral blood in burned mice with sepsis at early stage, and to analyze their immune regulatory mechanisms.

METHODSSeven-hundred and eighty male mice of clean grade were divided into sham injury group (n=60, sham injured on the back by immersing in 37 ℃ warm water for 10 s), burn group (n=240, inflicted with 20% TBSA deep partial- thickness burn on the back by immersing in 100 ℃ hot water for 10 s), sepsis group (n=240, intraperitoneally injected with 6 mg/kg lipopolysaccharide), and burn+ sepsis group (n=240) according to the random number table. Mice of burn+ sepsis group were treated as that in burn group at first, and then they were treated as that in sepsis group. (1) Immediately after injury, six mice in sham injury group were selected to collect lymphocyte suspension of peripheral blood (1 tube each mouse) according to the random number table. According to the random number table, 6 mice of each of the other three groups were respectively selected at post injury hour (PIH) 12, 24, 48, and 72 for the collection of lymphocyte suspension from peripheral blood (1 tube each mouse). Each tube of cell suspension was equally divided into two parts. Fluorescein isothiocyanate (FITC)-labeled human anti-mouse CD4 monoclonal antibody and phycoerythrin (PE)-labeled human anti-mouse interferon-γ monoclonal antibody were added to one part of cell suspension to mark helper T lymphocyte 1 (Th1). FITC-labeled human anti-mouse CD4 monoclonal antibody and PE-labeled human anti-mouse interleukin-4 (IL-4) monoclonal antibody were added to the other part of cell suspension to mark Th2. The percentages of Th1 and Th2 were determined with flow cytometer, and the ratio of Th1 to Th2 was calculated. (2) According to the random number table, 18 mice in sham injury group were selected immediately after injury for the collection of lymphocyte suspension of peripheral blood (1 tube each mouse), and 18 mice of each of the other 3 groups were respectively selected at PIH 12, 24, 48, and 72 to collect the lymphocyte suspension of peripheral blood (1 tube each mouse). The mRNA expression levels of ANXA1, GATA-3, and T-bet were determined by real-time fluorescent quantitative reverse transcription-PCR. (3) Immediately after injury, 36 mice in sham injury group were selected to collect lymphocyte suspension of peripheral blood (1 tube each mouse) according to the random number table, and then 36 tubes of cell suspension were divided into 6 batches (6 tubes each batch). Each one of 6 kinds of antibody combinations: antibodies for labeling Th1 and Th2 in combination with PE-anthocyanin 7 labeled human anti-mouse ANXA1 monoclonal antibody, PE-anthocyanin 7 labeled human anti-mouse GATA-3 monoclonal antibody, and PE-anthocyanin 7 labeled human anti-mouse T-bet monoclonal antibody was added to 1 tube of cell suspension at each batch. According to the random number table, 36 mice of each of the other 3 groups were respectively selected at PIH 12, 24, 48, and 72 for the collection of lymphocyte suspension of peripheral blood (1 tube each mouse), and then 36 tubes of cell suspension at each time point were divided into 6 batches for marking with 3 kinds of surface markers of Th1 and Th2 (6 tubes each batch). Each one of above-mentioned 6 kinds of antibodies was added to 1 tube of cell suspension at each time point for each batch. The percentages of ANXA1, GATA-3, and T-bet positive cells in Th1 and Th2 were determined with flow cytometer. Data were processed with one-way analysis of variance, analysis of variance of factorial design, and SNK test. The relationship between the percentages of ANXA1 positive cell and the percentages of GATA-3 positive cell in Th1 and Th2, and mRNA expression level of ANXA1 and mRNA expression level of GATA-3 in lymphocytes were assessed by linear correlation analysis.

RESULTS(1) Compared with those in sham injury group immediately after injury, the percentages of Th1 and Th2 and the ratio of Th1 to Th2 of mice in burn group were significantly decreased from PIH 24 on, with P values below 0.05; the percentages of Th1 and Th2 and the ratios of Th1 to Th2 of mice in sepsis group and burn+ sepsis group were significantly decreased from PIH 12 on, with P values below 0.05. (2) Compared with those in sham injury group immediately after injury, the mRNA expression levels of ANXA1 and GATA-3 in lymphocyte of mice in burn group were significantly decreased from PIH 24 on, with P values below 0.05; the mRNA expression level of T-bet was significantly decreased at PIH 24 but significantly increased at PIH 48 and 72, with P values below 0.05. Compared with those in sham injury group immediately after injury, the mRNA expression levels of ANXA1 and GATA-3 in lymphocytes of mice in sepsis group were significantly decreased from PIH 12 on, and the mRNA expression level of T-bet was increased significantly from PIH 12 on, with P values below 0.05; the mRNA expression levels of ANXA1, GATA-3, and T-bet in lymphocytes of mice in burn+ sepsis group were significantly decreased from PIH 12 on, with P values below 0.05, reaching the nadir at PIH 72 (0.50±0.04, 0.45±0.03, 0.21±0.05, respectively). (3) A significant positive correlation was observed between ANXA1 mRNA expression level and GATA-3 mRNA expression level in lymphocytes of peripheral blood (r=0.862, P<0.05). (4) Compared with those in sham injury group immediately after injury, the percentages of ANXA1 and GATA-3 positive cellsin Th1 and Th2 of mice in burn group were significantly lowered from PIH 24 on, and the percentage of T-bet positive cells was significantly decreased at PIH 24, but it was increased from PIH 48 on, with P values below 0.05. The percentages of ANXA1 and GATA-3 positive cells in Th1 and Th2 of mice in sepsis group were continuously decreased from PIH 12 on, which were lower at most time points than those in sham injury group immediately after injury, with P values below 0.05. The percentages of T-bet positive cells in Th1 and Th2 of mice in sepsis group were significantly increased since PIH 12 as compared with those in sham injury group immediately after injury, with P values below 0.05. The percentages of ANXA1, GATA-3, and T-bet positive cells in Th1 and Th2 of mice in burn+ sepsis group were continuously lowered from PIH 12, with significantly statistical differences at most time points as compared with those in sham injury group immediately after injury, with P values below 0.05. (5) The percentages of GATA-3 positive cells in Th1 and Th2 were significantly positively correlated with those of ANXA1 (with r values respectively 0.747 and 0.787, P values below 0.05).

CONCLUSIONSThe expression levels of ANXA1, GATA-3, and T-bet were continuously lowered in burned mice with sepsis, and it may play an important role in Th1/Th2 balance switching to Th2 bias and immunosuppressive process.

Animals ; Biomarkers ; Burns ; immunology ; metabolism ; GATA3 Transcription Factor ; biosynthesis ; genetics ; Humans ; Interferon-gamma ; biosynthesis ; genetics ; Interleukin-4 ; metabolism ; Male ; Mice ; RNA, Messenger ; Real-Time Polymerase Chain Reaction ; Sepsis ; blood ; T-Lymphocytes ; metabolism ; Transcription Factors ; biosynthesis ; genetics

The study was aimed to explore the effects of T-bet (T-box expressed in T cell), GATA-3(GATA binding protein 3) and relevant signal transduction pathways on the immune-related pathogenesis of chronic aplastic anemia (CAA), and to investigate the immunological regulation mechanism in the treatment of CAA by using cyclosporine A (CsA) at the level of Th cell imbalance, transcriptional factors, and relevant signal pathways. The real-time fluorescent quantitative polymerase chain reaction (real-time FQ-PCR) was used to determine the mRNA expression of T-bet, GATA-3, signal transducers and activators of transcription 4 (STAT4) and signal transducers and activators of transcription 6 (STAT6) in peripheral blood mononuclear cell (PBMNC) of CAA patients before and after treatment with CsA; the flow cytometry (FCM) and enzyme linked immunosorbent assay (ELISA) were used to determine the Th1/Th2 proportion in peripheral blood, and levels of IFN-γ, IL-12 and IL-4 in PBMNC-cultured supernatant. Healthy people were included to test the above indexes. The results showed that the mRNA expression of PBMNC T-bet, STAT4, T-bet/GATA-3 ratio, Th1 proportion, Th1/Th2 ratio and levels of IFN-γ and IL-12 in PBMNC-cultured supernatant of CAA patients were significantly higher than those of healthy people (p < 0.01). After treating with CsA for 6 months of CsA treatment, expression of T-bet, STAT4, T-bet/GATA-3 ratio, Th1 proportion, IFN-γ and IL-12 levels were lower than before, however, the expression of T-bet, STAT4, T-bet/GATA-3 ratio, Th1 proportion and IFN-γ had not been reduced to normal state. Compared to healthy people, no significant difference existed in the mRNA expression of GATA-3, STAT6, Th2 proportion, as well as level of IL-4 before and after treatment (p>0.05). It is concluded that the abnormal activation of IFN-γ/T-bet and IL-12/STAT4 pathways, as well as Th balance deviating to Th1 excursion play vital roles in the immunological pathogenesis of AA. CsA lowers the abnormal activation of IFN-γ/T-bet and IL-12/STAT4 pathways to correct Th1 hyperpolarization, which may reduce the abnormally activated cell-mediated immunity and relax hematopoietic depression of AA patients.
Adolescent ; Adult ; Aged ; Anemia, Aplastic ; drug therapy ; immunology ; metabolism ; Case-Control Studies ; Cyclosporine ; pharmacology ; Female ; GATA3 Transcription Factor ; metabolism ; Humans ; Interferon-gamma ; metabolism ; Interleukin-12 ; metabolism ; Interleukin-4 ; metabolism ; Male ; Middle Aged ; STAT4 Transcription Factor ; metabolism ; STAT6 Transcription Factor ; metabolism ; Signal Transduction ; T-Box Domain Proteins ; metabolism ; Th1 Cells ; drug effects ; Th2 Cells ; drug effects ; Young Adult

OBJECTIVETo explore the correlation between the expression of GATA-3 and the level of local cytokines (IL-5, IL-6 and IL-8).

METHODSThe levels of IL-5, IL-6 and IL-8 in ethmoid sinus mucosa were titrated in 45 patients with chronic rhinosinusitis and 11 normal subjects by ELISA. Patients were divided into AR group (with allergic rhinitis) and NAR group (without allergic rhinitis) . Semi-quantitative RT-PCR and immunohistochemical staining were used to examine the GATA-3 expression in nasal mucosa. The correlation between the expression of GATA-3 and the levels of cytokines was evaluated.

RESULTSIL-5, IL-6 and IL-8 levels in both AR and NAR groups were significantly elevated compared with normal group (all P < 0.01 for AR group; P < 0.05, 0.05, 0.01 for NAR group, respectively), and they were much higher in AR group in comparison with NAR group (P < 0.01, 0.05, 0.01, respectively). Semi-quantitative RT-PCR showed that AR and NAR groups had markedly greater level of GATA-3 mRNA than that in control group (P < 0.01, respectively), and the level of GATA-3 mRNA in AR group was further higher than that in NAR group (P < 0.01). Immunohistochemical staining illustrated that GATA-3 was primarily presented in cytoplasma and the GATA-3 positive cells were mainly infiltrating inflammatory cells in submucosa. The mean GATA-3 positive-staining rate was (27. 90 +/- 16.75)% and (10.22 +/- 0.05)% in AR and NAR group, which were markedly higher than (1.30 +/- 1.78)% in control group (P < 0.01, respectively). Pearson correlation analysis demonstrated that GATA-3 positive-staining rate was closely correlated with IL-5 level, but not IL-6 and IL-8. The correlation coefficient was 0. 712 for GATA-3 and IL-5 (P < 0.01), 0.200 for GATA-3 and IL-6 (P > 0.05), 0.089 for GATA-3 and IL-8 (P > 0.05).

CONCLUSIONSActivation of GATA-3 might be one of the mechanisms for induction of IL-5 expression in chronic rhinosinusitis . Concomitance of allergic rhinitis with chronic rhinosinusitis further increased expression of GATA-3, and subsequently enhanced IL-5 expression. Chronic sinusitis may be related to allergy, and GATA-3 may play a key role in the pathogenesis of chronic sinusitis.

Adolescent ; Adult ; Aged ; Chronic Disease ; Female ; GATA3 Transcription Factor ; metabolism ; Humans ; Interleukin-5 ; metabolism ; Interleukin-6 ; metabolism ; Interleukin-8 ; metabolism ; Male ; Middle Aged ; Nasal Mucosa ; metabolism ; Sinusitis ; metabolism ; Young Adult