OBJECTIVEInterleukin-4 plays a key role in the development of asthma. Overseas studies have shown that Q576R polymorphism in the interleukin-4 receptor (IL-4R) gene is related to asthma as well as increased serum IgE levels. This study was designed to investigate the association of Q576R polymorphism in IL-4R gene with childhood asthma and serum IgE levels.

METHODSThe polymorphism of IL-4R Q576R was determined by PCR/RFLP and serum total IgE level was measured using ELISA in 94 children with asthma. Sixty-eight healthy children served as controls.

RESULTSThe distribution frequency of heterozygous genotype Q576R (41%) and mutant allele R576 (26%) was significantly higher in children with asthma than that of controls (16% each) (P < 0.01; P < 0.05). The total serum IgE level between patients with genotype Q576R and Q576Q was not significantly different (225.78 +/- 51.43 IU/mL vs 163.24 +/- 31.32 IU/mL, P> 0.05).

CONCLUSIONSThe mutant R576 allele of IL-4R may be one of the candidate genes for susceptibility to asthma. Allele R576 of IL-4R is related to asthma but is irrelevant to the total serum IgE level in children with asthma.

Adolescent ; Asthma ; genetics ; immunology ; Child ; Child, Preschool ; Female ; Humans ; Immunoglobulin E ; blood ; Male ; Polymorphism, Genetic ; Receptors, Interleukin-4 ; genetics

Orai1 is the key subunit of the Ca2+-release-activated Ca2+ channel. Our previous report has demonstrated that Orai1 expression in the airway was upregulated in the ovalbumin (OVA)-induced allergic rhinitis (AR) mouse models. To observe whether inhibition of Orai1 expression in the airway could suppress symptoms in a murine model of AR and to assess the impacts of this inhibition on the responses of local and systemic immunocytes, we administered recombinant lentivirus vectors that encoded shRNA against ORAI1 (lenti-ORAI1) into the nostrils of OVA-sensitized mice before the challenges, and analyzed its effect on allergic responses, as compared with the unsensitized mice and untreated AR mice. Administration of lenti-ORAI1 into the nasal cavity successfully infected cells in the epithelial layer of the nasal mucosa, and significantly decreased the frequencies of sneezing and nasal rubbing of the mice. Protein levels of leukotriene C4, OVA-specific IgE, and IL-4 in the nasal lavage fluid and serum and eosinophil cation protein in the serum were also significantly reduced by lenti-ORAI1, as were the mRNA levels of these factors in the nasal mucosa and spleen. These data suggested that administration of lenti-ORAI1 into the nasal cavity effectively decreased Orai1 expression in the nasal mucosa, alleviated AR symptoms, and partially inhibited the hyperresponsiveness of the local and systemic immune cells including T cells, B cells, mast cells and eosinophils that are involved in the pathogenesis of AR.
Animals ; Calcium Channels/analysis/*genetics/immunology ; *Down-Regulation ; Eosinophil Cationic Protein/blood/genetics ; Glutathione Transferase/blood/genetics/immunology ; Immunoglobulin E/blood/genetics/immunology ; Interleukin-4/blood/genetics/immunology ; Lentivirus/genetics ; Mice ; Mice, Inbred BALB C ; Nasal Mucosa/immunology/metabolism ; Ovalbumin/immunology ; RNA, Messenger/genetics ; RNA, Small Interfering/*administration & dosage/genetics ; Rhinitis, Allergic, Perennial/*genetics/immunology ; Spleen/immunology/metabolism ; *Transfection

OBJECTIVETo investigate the relationship between the dendritic cell (DC) subsets and transcriptive factors, T-bet, GATA-3, and immune imbalance in acquired severe aplastic anemia (SAA).

METHODSThe DC1 (HLA-DR+Lin-CD11c+) and DC2 (HLA-DR+Lin-CD123+) in peripheral blood mononuclear cells (PBMNC) were measured with flow cytometry (FCM), the expressions of T-bet mRNA and GATA-3 mRNA in PBMNC with semiquantitative RT-PCR and the plasma level of IFN gamma and IL-4 with ELISA in 29 SAA patients and 16 healthy controls.

RESULTSThe percentages of DC1 in PBMNC were (0.44 +/- 0.24)% and (0.73 +/- 0.30)% in untreated and recovered SAA patients respectively, both were higher than that in controls (0.29 +/- 0.10)% (P < 0.05). The percentage of DC2 in the untreated cases was lower than that of recovered ones or controls [(0.18 +/- 0.14)% vs (0.28 +/- 0.20)% and (0.29 +/- 0.13)%] (P < 0.05). DC1/DC2 ratios were 3.45 +/- 2.71 and 2.90 +/- 0.95 in untreated and recovered groups respectively, both were higher than that in controls (1.15 +/- 0.56) (P < 0.05). No statistic difference in DC1/DC2 ratio was found between untreated and recovered patients (P < 0.05). The relative mRNA expression levels of transcriptive factor T-bet were 0.37 +/- 0.07, 0.20 +/- 0.07 and 0.17 +/- 0.05 in the above 3 groups, respectively, untreated group being higher than that of recovered group or healthy controls (P < 0.05). There was no statistic difference of GATA-3 expression among the 3 groups (P > 0.05). T-bet/GATA-3 ratio was 0.72 +/- 0.13 in untreated group, being higher than that of recovered group (0.33 +/- 0.08) or controls (0.35 +/- 0.11). The plasma level of IFN gamma in the untreated group was (50.9 +/- 1.1) ng/L, which was higher than that of recovered group [(49.7 +/- 0.9) ng/L] or controls [(49.7 +/- 0.7) ng/L]. There was significant positive correlations between T-bet and DC1/DC2 ratio (r = 0.445, P < 0.01), as well as between T-bet and IFN gamma (r = 0.402, P < 0.01).

CONCLUSIONEither DC1/DC2 or T-bet/GATA-3 ratio might become an index to estimate immune imbalance. High-expressed T-bet was related to the progress of SAA. In patients with SAA, DC1/DC2 ratio returns to normal range later than that of routine blood test does, indicating that immunosuppressive therapy should not be withdrawn too earlier.

Adolescent ; Adult ; Anemia, Aplastic ; blood ; immunology ; Child ; Dendritic Cells ; immunology ; Female ; GATA3 Transcription Factor ; blood ; genetics ; Humans ; Interferon-gamma ; blood ; Interleukin-4 ; blood ; Male ; Middle Aged ; RNA, Messenger ; genetics ; T-Box Domain Proteins ; blood ; genetics ; Young Adult

Neospora caninum is the etiologic agent of bovine neosporosis, which affects the reproductive performance of cattle worldwide. The transmembrane protein, NcSRS2, and dense-granule protein, NcGRA7, were identified as protective antigens based on their ability to induce significant protective immune responses in murine neosporosis models. In the current study, NcSRS2 and NcGRA7 genes were spliced by overlap-extension PCR in a recombinant adenovirus termed Ad5-NcSRS2-NcGRA 7, expressing the NcSRS2-NcGRA7 gene, and the efficacy was evaluated in mice. The results showed that the titer of the recombinant adenovirus was 10(9)TCID50/ml. Three weeks post-boost immunization (w.p.b.i.), the IgG antibody titer in sera was as high as 1:4,096. IFN-gamma and IL-4 levels were significantly different from the control group (P<0.01). This research established a solid foundation for the development of a recombinant adenovirus vaccine against bovine N. caninum.
Adenoviridae/*genetics ; Animals ; Antibodies, Fungal/blood ; Antigens, Fungal/genetics/*immunology ; *Drug Carriers ; Fungal Proteins/genetics/*immunology ; Fungal Vaccines/administration & dosage/genetics/*immunology ; Immunoglobulin G/blood ; Interferon-gamma/blood ; Interleukin-4/blood ; Mice ; Mice, Inbred BALB C ; Neospora/genetics/*immunology ; Recombinant Fusion Proteins/genetics/immunology ; Vaccines, Synthetic/administration & dosage/genetics/immunology

OBJECTIVETo investigate the expressions of chemokine receptors and interleukin (IL) receptors on the peripheral blood mononuclear cells (PBMCs) from systemic lupus erythematosus (SLE) patients and their correlations with clinical features as well as SLE disease activity index (SLEDAI).

METHODSThe mRNA expressions of chemokine receptors and IL receptors on PBMCs of 93 SLE patients and 30 healthy controls were detected by reverse transcription-polymerase chain reaction, including CCR2, CCR3, CCR4, CCR5, CCR6, CCR8, CXCR3, CXCRS, CX3CR1, XCR1, IL-4R, and IL-10R. The clinical features of SLE patients were recorded. The correlations of chemokine receptors and IL receptors mRNA expressions with clinical features as well as SLEDAI were assayed using linear regression analysis.

RESULTSThe level of CCR5 mRNA in SLE patients (including active and inactive SLE) was significantly higher than that in healthy controls (P < 0.05), and there was no significant difference between active and inactive patients in this respect (P > 0.05). CX3CR1 mRNA expression significantly increased from healthy control to inactive SLE to active SLE in sequence. The others (except for CCR8, CXCR3, and IL-10R) in active SLE patients were significantly higher than those in both inactive SLE patients and healthy controls (all P < 0.05). There were positive correlations between SLEDAI and CCR2 (r = 0.424, t = 4.313, P < 0.001), CCR3 (r = 0.518, t = 5.410, P < 0.001), CCR4 (r = 0.376, t = 3.851, P < 0.001), CCR6 (r = 0.457, t = 4.513, P < 0.001), CXCR5 (r = 0.455, t = 4.629, P < 0.001), CX3CR1 (r = 0.445, t = 4.523, P < 0.001), as well as XCR1 (r = 0.540, t = 5.445, P < 0.001). And CCR5 mRNA expression level was positively correlated with IL-4R mRNA (r = 0.313, t = 2.353, P < 0.05). The patients with myositis and cutaneous vasculitis simultaneously showed lower levels of CCR5 and CX3CR1, and CCR5 expression was negatively correlated with the scores of SLEDAI in SLE cases accompanied by photosensitivity (r = 0.426, t = -2.155, P < 0.05).

CONCLUSIONIncreased expressions of CCR5 and CX3CR1 on PBMCs may be indicators in clinical survey for SLE.

Adolescent ; Adult ; CX3C Chemokine Receptor 1 ; Child ; Female ; Humans ; Leukocytes, Mononuclear ; immunology ; Lupus Erythematosus, Systemic ; etiology ; immunology ; Male ; Middle Aged ; RNA, Messenger ; blood ; Receptors, CCR5 ; genetics ; Receptors, Chemokine ; genetics ; Receptors, Interleukin-10 ; genetics ; Receptors, Interleukin-4 ; genetics

OBJECTIVETo study the expression of transcriptional factors T-bet and GATA3 mRNA and the levels of IFN-gamma and IL-4 in blood in children with acute idiopathic thrombocytopenic purpura (ITP) and investigate the tendency of polarization of Th1/Th2 in children with ITP.

METHODSBlood T-bet and GATA3 mRNA expression were examined using RT-PCR and plasma IFN-gamma and IL-4 levels were measured using EIASA in children with acute ITP in acute (n=30) and remission stages (n=28). Twenty healthy children served as the controls.

RESULTSBlood T-bet mRNA expression and IFN-gamma levels in children with ITP in the acute stage were markedly higher than those in the remission stage and controls (p<0.01). In contrast, blood GATA3 mRNA expression and IL-4 levels in children with ITP in the acute stage were significantly lower than those in the remmission stage and controls (p<0.01).

CONCLUSIONSThe high expression of T-bet and IFN-gamma and the low expression of GATA3 and IL-4 indicate the existence of Th1 polarization in children with acute ITP. This might be related to the regulation of T-bet and GATA3.

Child ; Child, Preschool ; Female ; GATA3 Transcription Factor ; genetics ; Humans ; Interferon-gamma ; blood ; Interleukin-4 ; blood ; Male ; Purpura, Thrombocytopenic, Idiopathic ; immunology ; RNA, Messenger ; blood ; T-Box Domain Proteins ; genetics ; Th1 Cells ; immunology

OBJECTIVE: To explore the role of combined heart-thymus transplantation for heart allograft in rats. METHODS: Vascularized heart-thymus combined transplantation was performed with microsurgical technique. Graft survival, histopathology, infiltration of CD4+, CD8+ T cells, level and mRNA expressions of IL-2 and IL-4 in the serum and cardiac grafts were investigated. RESULTS: Heart allograft in the controls had a survival time of (6.0+/-0.76) d. heart-thymus combined transplantation in non-thymectomized rats had a survival time of (6.88+/-0.64)d (P<0.05). Heart-thymus combined transplantation in thymectomized rats led to an evident survival time of (14.13+/-5.82)d (P<0.01) for cardiac graft, which further obtained long term survival after short course of treatment with cyclosporine. Pathologic lesion and infiltration of CD4+ and CD8+ T cells in cardiac grafts showed mitigated in the long term survival group. IL-2 level in the serum and cardiac grafts maintained low level in the long term survival group, whereas IL-4 maintained high level. CONCLUSION Whether thymectomized or not in recipient rats, heart-thymus combined transplantation has a positive effect to protect cardiac graft. Furthermore, in thymectomized rats heart-thymus combined transplantation may lead to evident survival prolongation of the heart grafts, which induces immune tolerance in short course of treatment with cyclosporine.
Animals ; CD4-Positive T-Lymphocytes ; immunology ; CD8-Positive T-Lymphocytes ; immunology ; Cyclosporine ; therapeutic use ; Graft Survival ; drug effects ; immunology ; Heart Transplantation ; Immune Tolerance ; drug effects ; immunology ; Immunosuppressive Agents ; therapeutic use ; Interleukin-2 ; blood ; genetics ; Interleukin-4 ; blood ; genetics ; Male ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; Reverse Transcriptase Polymerase Chain Reaction ; Thymectomy ; Thymus Gland ; transplantation ; Time Factors ; Transplantation Immunology ; immunology ; Transplantation, Homologous

OBJECTIVEIt is well known that vitamin A can improve mucosal immunity and anti-infection immunity. But the mechanisms thereof remain to be clarified. Previous studies on the role of vitamin A in immune regulation focused on lymphocytes, whereas little had been done about dendritic cells, which play very important roles in immune response. The objective of this study was to understand the effects of retinoic acid (RA), the metabolic product of vitamin A in vivo,on the differentiation, maturation and functions of dendritic cells from cord blood.

METHODSCord blood samples were collected from nine well-nourished full-term neonates. Mononuclear cells were isolated by Ficoll-Hypaque gradient centrifugation and cultured in the presence of 1000 u/ml GM-CSF, 500 u/ml IL-4 for 6 days, then TNF-alpha 20 ng/ml was added into the medium and cultured for another 3 days. The cells were incubated with or without 1 x 10(-6) MRA. Expression of surface molecules, CD1a, CD83, HLA-DR on DC was measured by flow cytometry. The ability of DC derived from the culture to induce proliferation of T cells in the mixed lymphocyte reaction (allo-MLR) was used for the evaluation of their function. IL-12, IFN-gamma, IL-4 and IL-10 were detected at mRNA levels by RT-PCR to understand the roles of DC treated with RA in regulation of Th1/Th2 balance.

RESULTSOn the sixth day of cell culture, the percentage of DC incubated with RA (57.28 +/- 9.22) was much lower than that without RA (79.57 +/- 11.85) (P < 0.001), but on the ninth day, there were no differences between the presence or absence of RA (76.18 +/- 10.27 vs. 73.72 +/- 15.58). When RA was added to the medium and the culture was continued for nine days, the percent of immature DC (CD1a + HLA-DR+) was much higher than that of the control (absence of RA) (58.93 +/- 4.70 vs. 45.80 +/- 7.88, t = 6.575, P < 0.001); whereas, mature DC (CD83 + HLA-DR+) percentage was markedly lower than that of the control (17.25 +/- 8.49 vs. 27.92 +/- 13.94, t = 4.435, P = 0.002). The T lymphocytes proliferation induced by the DC treated with RA was reduced from 16 857 +/- 3 643 to 11 924 +/- 2 576 cpm (t = 5.598, P < 0.001) in allo-MLR. Expression of mRNA for IL-12p35, IL-12p40, IFN-gamma in the cells that had been incubated with RA declined, but IL-10, IL-4 increased significantly.

CONCLUSIONVitamin A inhibited the differentiation and maturation of cord blood DC, reduced it's ability to stimulate allo-T lymphocytes proliferation, and down-regulated Th1 cytokines, up-regulated Th2 cytokines, consequently made immune response inclined to Th2.

Antigens, CD ; Antigens, CD1 ; analysis ; Cell Differentiation ; drug effects ; genetics ; immunology ; Cytokines ; genetics ; Dendritic Cells ; drug effects ; immunology ; metabolism ; Fetal Blood ; cytology ; immunology ; Flow Cytometry ; Gene Expression ; drug effects ; HLA-DR Antigens ; analysis ; Humans ; Immunoglobulins ; analysis ; Interferon-gamma ; genetics ; Interleukin-10 ; genetics ; Interleukin-12 ; genetics ; Interleukin-4 ; genetics ; Membrane Glycoproteins ; analysis ; Reverse Transcriptase Polymerase Chain Reaction ; Vitamin A ; pharmacology

In fascioliasis, T-helper 2 (Th2) responses predominate, while little is known regarding early immune phenomenon. We herein analyzed early immunophenotype changes of BALB/c, C57BL/6, and C3H/He mice experimentally infected with 5 Fasciola hepatica metacercariae. A remarkable expansion of CD19+ B cells was observed as early as week 1 post-infection while CD4+/CD8+ T cells were down-regulated. Accumulation of Mac1+ cells with time after infection correlated well with splenomegaly of all mice strains tested. The expression of tumor necrosis factor (TNF)-alpha mRNA in splenocytes significantly decreased while that of IL-4 up-regulated. IL-1beta expression was down-modulated in BALB/c and C57BL/6 mice, but not in C3H/He. Serum levels of transforming growth factor (TGF)-beta were considerably elevated in all mice during 3 weeks of infection period. These collective results suggest that experimental murine fascioliasis might derive immune suppression with elevated levels of TGF-beta and IL-4 during the early stages of infection.
Animals ; B-Lymphocytes/immunology ; CD4-Positive T-Lymphocytes/immunology ; CD8-Positive T-Lymphocytes/immunology ; Down-Regulation ; Fasciola hepatica/*immunology ; Fascioliasis/*immunology ; Immunophenotyping ; Immunosuppression ; Interleukin-4/blood/genetics/*immunology ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C3H ; Mice, Inbred C57BL ; Spleen/immunology ; Transforming Growth Factor beta/blood/genetics/*immunology

OBJECTIVETo explore the effect of moxibustion pretreatment on the immune function of athletes during the courses of heavy load training.

METHODSTwelve male middle-distance race athletes were randomly divided into an observation group (n = 6) and a control group (n= 6). In observation group, mild moxibustion was applied at Guanyuan (CV 4), Zusanli (ST 36) or Guanyuan (CV 4), Sanyinjiao (SP 6) alternatively once daily, for 4 weeks. In control group, there was no any intervention. The expressions of gamma-interferon (IFN-gamma) and interleukin-4 (IL-4) were detected by realtime fluorescent quantitative PCR method before heavy load training, after 3 weeks of treatment, after 1 week adjusting training.

RESULTS(1) The ratios of Testosterone/Cortisol (T/C) of (0.017 +/- 0.003) in the control group and of (0.021 +/- 0. 007) in the observation group after heavy load training for three weeks were significant lower than 0.036 +/- 0.009 and 0.037 +/- 0.013 before heavy load training in the same group, respectively (both P < 0.01) (2) IFN-gamma mRNA of (2.76 +/- 1.28) and ratio of IFN-gamma mRNA/IL-4 mRNA of (3.28 +/- 1.76) in the control group after heavy load training for three weeks were significant higher than 1.75 +/- 0.44 and 1.52 +/- 0.76 before heavy load training, respectively (both P < 0.05), and ratio of IFN-gamma mRNA/IL-4 mRNA of (0.86 +/- 0.66) after adjusting training for 1 week reduced significantly as compared with that after heavy load training for three weeks (P < 0.05). (3) In the observation group, IFN-gamma mRNA, IL-4 mRNA and ratio of IFN-gamma mRNA/IL-4 mRNA increased firstly and then restore with the time, but these changes had no significant differences (all P > 0.05).

CONCLUSIONMoxibustion pretreatment can prevent Th1/Th2 imbalance of male middle-distance race athlete during the course of heavy load training.

Adolescent ; Adult ; Athletes ; Humans ; Hydrocortisone ; blood ; Interferon-gamma ; genetics ; Interleukin-4 ; genetics ; Male ; Moxibustion ; methods ; Physical Education and Training ; RNA, Messenger ; analysis ; Testosterone ; blood ; Th1 Cells ; immunology ; Th2 Cells ; immunology