To confirm local production of IgE, and evaluate role of immunoglobulins on eosinophil activation in nasal polyp (NP) tissue, we measured IgG, IgA, secretory IgA(sIgA), total (tIgE) and specific IgE (sIgE) to Dermatophagoides pteronyssinus(DP) by ELISA in NP tissue homogenates from 51 subjects. They were classified according to skin reactivity to DP: group I, 15 highly atopic subjects; group II, 18 weakly atopic subjects; and group III, 18 non-atopic subjects. Eosinophil cationic protein (ECP) level was measured by CAP system. Highest level of DP-sIgE was noted in group I, followed by group II and III (p<0.05). Nine (60%) of group I and 4 (22%) of group II subjects had detectable level of DP-sIgE with no significant differences in IgA, sIgA, and IgG. All of NP tissue had eosinophilic infiltration with no significant difference in activated eosinophil count or ECP level among 3 groups. A significant correlation was noted between EG2+ cell count and tIgE (r=0.55, p<0.05), and DP-sIgE level (r=0.60, p<0.05). A significant correlation was also noted between ECP and IgG (r=0.51, p<0.05) and DP-sIgE level (r=0.47, p<0.05) with no significant correlation with IgA or sIgA. These results suggest that DP-sIgE was detectable in NP tissue from weakly atopic subjects as well as highly atopic subjects. IgG and sIgE may have potential roles in eosinophil degranulation in NP tissue.
Blood Proteins/analysis ; Cell Degranulation/immunology ; Dermatophagoides pteronyssinus/immunology ; Eosinophil Granule Proteins ; Eosinophils/immunology ; Humans ; Immunoglobulin A/analysis/immunology ; Immunoglobulin E/analysis/immunology ; Immunoglobulin G/analysis/immunology ; Immunoglobulins/analysis/*immunology ; Nasal Polyps/*immunology/pathology ; *Ribonucleases

AIMTo explore the role of humoral immunity in the pathophysiological process of freezing injury and the possible immune interference in the preventation and treatment of frostbite.

METHODSSevere experimental freezing injury model was made in Wistar rats( n = 20). The concentration of three types of immunoglobulin (IgG, IgA and IgM), two types of complement components (C3 and C4), and circulating immune complex (CIC) were measured respectively before and at 4h, 1d, 3d, and 5d after frostbite. At the same time, the tissue immune complex (TIC) in skeletal muscle and the contents of the red blood cell immune complex (RBC-IC) were also observed and then was the red blood cell immune adherence activity (RCIA).

RESULTSSerum IgG concentration decreased rapidly to the lowest level at 4 h after frostbite IgA concentration dropped to the nadir on 1 day after freezing. Decreases of both immunoglobulins were maintained during the 5 days after frostbite. The fate of both C3 and C4 were the same as those immunoglobulins. Freezing had rather less effect on IgM level. CIC concentration in serum, expressed as the percent of prefreezing increased rapidly and to the zenith on the 3 days post-freezing. By immunofluorescence microscopy, thin continuous linear pattern (IgG) was demonstrated along the SM on the first day post-freezing. Granular and nodular deposits (IgG) appeared along the SM as the time proceeded after frostbite. RBC-IC contents, expressed as the erythrocyte IC rosette rate, increased significantly and to the zenith on the 3 d post-freezing, while RCIA depressed to the nadir at the same time.

CONCLUSIONThe freezing frostbite is an immune complex related disease which have not been reported by others before.

Animals ; Antigen-Antibody Complex ; analysis ; immunology ; Frostbite ; blood ; immunology ; Immunoglobulin A ; immunology ; Immunoglobulin G ; immunology ; Immunoglobulin M ; immunology ; Immunoglobulins ; immunology ; Male ; Rats ; Rats, Wistar

OBJECTIVETo research the maturation regulation of dendritic cells (DCs) pulsed with hepatocellular carcinoma (HCC) cell soluble antigens.

METHODSBCG HSP 70 was purified by SDS-PAGE electrophoresis and its biological activity was determined with ELISA. Phenotypes of DCs pulsed with antigens or with both antigens and BCG HSP 70 were analysed with flow cytometry. MTT assay was used to estimate the proliferation of self lymphocytes and the mixed lymphocyte reaction (MLR) of BCG HSP 70 primed DCs.

RESULTSThe characteristics of DCs had changed after loaded with soluble antigens of HCC. There were about 10% DCs which had lost their specific markers. The expression levels of CD54, CD83, CD86 molecules and the stimulatory ability in allogeneic MLR decreased. However, after being activated by BCG HSP 70, the DCs pulsed with antigens could keep their special markers and the expression levels of CD54, CD83, CD86 molecules increased too. The stimulatory abilities in allogeneic MLR and proliferation of self lymphocytes also improved.

CONCLUSIONThis study shows that BCG HSP 70 can induce DCs pulsed with antigens maturation and improve their antigen-presenting ability, which may be a useful maturation inducer for dendritic cells.

Antigen Presentation ; Antigens, CD ; analysis ; Antigens, Neoplasm ; immunology ; B7-2 Antigen ; Carcinoma, Hepatocellular ; immunology ; Dendritic Cells ; cytology ; immunology ; HSP70 Heat-Shock Proteins ; immunology ; Humans ; Immunoglobulins ; analysis ; Intercellular Adhesion Molecule-1 ; analysis ; Liver Neoplasms ; immunology ; Membrane Glycoproteins ; analysis ; Mycobacterium bovis ; immunology

Of the many theoretical causes of rheumatoid arthritis(RA), the most widely held theory is the autoimmune mechanism. In order to clarify the clinical significance of the immunological tests in RA, we studied immunoglobulin and complement levels in sera and synovial fluids of 118 RA patients and the following results were obtained. 1) The levels of immunoglobulins were elevated in both serum and synovial fluid and this was more prominent in the seropositive cases than the seronegative ones. 2) The levels of C3 component were decreased in both serum and synovial fluid, while those of C4 were decreased only in synovial fluid. Serum C3 and C4 component levels were more decreased in the seropositive cases than the seronegative ones. 3) The immunoglobulin levels in serum (IgG, IgM and IgA) and synovial fluid (IgG and IgA) and the levels of C3, C4 component in serum were well correlated with the clinical forms of rheumatoid arthritis. 4) The IgA level in serum and IgM level in synovial fluid were more increased in the exacerbated cases than the chronic ones. 5) Serum IgG level was decreased after steroid medication over one month.
Arthritis, Rheumatoid/blood/*immunology/metabolism ; Complement 3/analysis ; Complement 4/analysis ; Female ; Human ; Immunoglobulins/analysis ; Immunologic Tests ; Male

OBJECTIVETo prepare highly specific anti-ouabain polyclonal antibody for detecting endogenous ouabain in tissues.

METHODSOuabain-BSA compound was used to immunize hens, and the eggs were collected one week after the first immunization. The IgY antibodies in the egg yolk were separated and purified by PEG-6000 Method, and analyzed by 12% SDS-PAGE and enzyme-linked immunosorbent assay (ELISA) for titration. The IgY antibodies obtained were applied subsequently in ELISA and immunohistochemistry.

RESULTSThe IgY titer increased rapidly after the second immunization, with the highest titer of 1:10240 that lasted for at least 4 weeks. Competitive ELISA for IgY detection showed an average intraassay coefficient of variation (CV) of 2.03% and an inter-assay CV of 2.34%. Immunohistochemistry visualized the location of the endogenous ouabain mainly in the cytoplasm of the zona reticularis of rat adrenal cortex.

CONCLUSIONImmunization of hens allows efficient preparation of IgY antibody which can be used in routine immunoassays.

Animals ; Calibration ; Cattle ; Cell Line ; Enzyme-Linked Immunosorbent Assay ; Immunization ; methods ; Immunoglobulins ; immunology ; Immunohistochemistry ; Ouabain ; analysis ; immunology ; Rats

AIMTo improve specificity and accuracy of endogenous ouabain measurement assay.

METHODSAnti-ouabain polyclonal antibody egg yolk (IgY) and anti-ouabain rabbit antibody (IgG) were prepared respectively. In the presence of two kinds of antibody, then the specificity and accuracy of enzyme-linked immunosorbent assay (ELISA) were compared.

RESULTSThe ELISA, in the presence of IgY, provided a sensitivity of the average intraassay coefficient of variation(CV) was 2.03%, and the inter-assay CV was 2.34% respectively. In contrast, IgG were 2.83% and 3.29%. No significant interferences were observed with hydrocortisone and dexamethasone. There was 3.45% vs. 5.95%, 3.20% vs. 5.20% of crossreaction with cedilanid and digoxin.

CONCLUSIONThe specificity and accuracy of ELISA, in which IgY was used, were more better than IgG.

Animals ; Antibody Specificity ; Chickens ; immunology ; Cross Reactions ; Egg Yolk ; immunology ; Enzyme-Linked Immunosorbent Assay ; methods ; Immunoglobulin G ; immunology ; Immunoglobulins ; immunology ; Male ; Ouabain ; analysis ; Rabbits

Adolescent ; Child ; Child, Preschool ; Complement System Proteins ; analysis ; Female ; Humans ; Immunoglobulins ; analysis ; Infant ; Male ; Pneumonia, Mycoplasma ; immunology

OBJECTIVETo investigate the effect of laparoscopic Roux-en-Y gastric bypass on the short-term immune function of type 2 diabetic patients and to explore the correlation between the immune regulatory effect and blood glucose control.

METHODSClinical data of 28 patients with type 2 diabetes mellitus who underwent laparoscopic Roux-en-Y gastric bypass surgery in our hospital during January 2014 to January 2015 were retrospectively collected. Fasting blood glucose (FBG), glycosylated hemoglobin (HbA1c), immunoglobulin (IgG, IgA, IgM), T lymphocytes CD3(+), killer T cells CD3(+)CD8(+), helper T cells CD3(+)CD4(+), natural killer(NK) cell subsets CD16(+),CD56(+) and B cell subsets CD19(+),CD45(+) levels were detected and compared between before and after surgery. Correlation of immunoglobulin and immune cell subsets with the level of FBG and HbA1c were examined.

RESULTSAfter operation, FBG and HbA1c decreased significantly from (5.78±1.15) mmol/L to (14.21±1.89) mmol/L, and (9.96±0.97)% to (6.87±0.69)%, respectively (all P<0.05), and immunoglobulin (IgG, IgA, IgM) increased significantly [IgG: from(9.41±1.23) g/L to (12.74±1.61) g/L, IgM: from (1.71±0.22) g/L to (2.43±0.39) g/L, IgA: from (1.25±0.26) g/L to (1.97±0.23) g/L, all P<0.05]. Besides, T lymphocytes CD3(+), killer T cells CD3(+)CD8(+) and B cell subsets CD19(+)CD45(+) elevated significantly as well [T lymphocytes CD3(+): from (55.3±3.8)% to (67.6±4.6)%, killer T cells CD3(+)CD8(+): from (15.6±5.3)% to (28.7±4.2)%, B cell subsets CD19(+)CD45(+): from (8.4±3.7)% to (18.1±4.1)%, respectively, all P <0.05]. There was no significant difference in the expression of helper T cells CD3(+)CD4(+) and natural killer cell subsets CD16(+)CD56(+) between pre-operation and post-operation(all P>0.05). Levels of immunoglobulin (IgG, IgA, IgM), CD4(+)/CD8(+) ratio and B cell subsets were negatively correlated with HbA1c and FBG levels (IgG with FBG: r=-0.865, IgA with FBG: r=-0.887, IgM with FBG: r=-0.902, CD4(+)/CD8(+) with FBG: r=-0.956, CD19(+)CD45(+) with FBG: r=-0.834; IgG with HbA1C: r=-0.859, IgA with HbA1C: r=-0.957, IgM with HbA1C: r=-0.843, CD4(+)/CD8(+) with HbA1C: r=-0.912, CD19(+)CD45(+) and HbA1C: r=-0.885, all P<0.05), but the proportion of NK cells was not significantly correlated with HbA1c and FBG (P>0.05).

CONCLUSIONSLaparoscopic Roux-en-Y gastric bypass has good effect on the immune function of patients with type 2 diabetes mellitus. Improvement of immune function is correlated with the control of FBG and HbA1c.

Blood Glucose ; analysis ; Diabetes Mellitus, Type 2 ; immunology ; surgery ; Gastric Bypass ; Glycated Hemoglobin A ; analysis ; Humans ; Immunoglobulins ; blood ; Laparoscopy ; Lymphocyte Subsets ; immunology ; Postoperative Period ; Retrospective Studies

The serum level of Ig G, Ig M, Ig A, Ig D and C3 were determined by a Hyland Immunodiffusion plate in 60 patients and 29 normal healthy controls. Immunoglobulin G level was increased only in patient's with herniated nucleus pulposus. We also could demonstrate high levels of Ig G in group I, the ruptured nucleus cases, but there were no differences in the levels of other immunoglobulins and C3 in other groups. In the Ochterlony gel diffusion test for the detection of humoral antibody for the nucleus pulposus, only 4 cases gave precipitation lines using antigen excised from nucleus pulposus. In 2 cases high levels of Ig G were observed. All 89 patients gave negative precipitation lines using proteoglycan as antigen.
Adolescent ; Adult ; Autoimmune Diseases/complications* ; Back Pain/etiology ; Back Pain/immunology* ; Child ; Female ; Human ; Immunoglobulins/analysis ; Intervertebral Disk* ; Male ; Middle Age ; Spinal Diseases/immunology*

OBJECTIVETo observe the effects of Huaiqihuang granules on the immune function in children with severe Mycoplasma pneumoniae pneumonia.

METHODSPediatric inpatients with severe Mycoplasma pneumoniae pneumonia were randomly divided into Huaiqihuang granule treatment group (n=51) and conventional treatment group (n=47). The Huaiqihuang granule treatment group was orally administered Huaiqihuang granules in addition to the conventional treatment, while the conventional treatment group received conventional treatment only. Levels of serum IgA, IgG, and IgM, percentages of CD4and CD8 T lymphocyte subsets, and CD4/CD8ratio were examined in the two groups. The incidence rate of respiratory tract re-infection within three months following treatment was compared between the two groups.

RESULTSThe levels of serum IgA, IgG, and IgM, the percentage of CD4 T lymphocytes, and the CD4/CD8ratio were significantly higher in the Huaiqihuang granule treatment group than in the conventional treatment group three months after treatment (P<0.05). In contrast, the percentage of CD8T lymphocytes was significantly lower in the Huaiqihuang granule treatment group than in the conventional treatment group (P<0.05). In addition, the incidence rate of respiratory tract re-infection within three months following treatment was significantly lower in the Huaiqihuang granule treatment group than in the conventional treatment group (P<0.05).

CONCLUSIONSHuaiqihuang granules can regulate immune functions and reduce the incidence of short-term respiratory tract re-infection in children with severe Mycoplasma pneumoniae pneumonia.

Adolescent ; CD4-CD8 Ratio ; Child ; Child, Preschool ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Immunoglobulins ; analysis ; Male ; Pneumonia, Mycoplasma ; drug therapy ; immunology ; T-Lymphocyte Subsets ; drug effects ; immunology