Antibodies, Viral ; blood ; Betacoronavirus ; immunology ; Coronavirus Infections ; diagnosis ; Gold Colloid ; chemistry ; Humans ; Immunoassay ; methods ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Pandemics ; Pneumonia, Viral ; diagnosis ; Reagent Kits, Diagnostic

Diagnosis of Mycoplasma pneumoniae infection is important due to its variable clinical manifestations and absence of response to beta-lactams. Introduction of enzyme immunoassays (EIAs) for serologic diagnosis of M. pneumoniae has made it possible to separate the analyses of specific IgG and IgM antibodies. We compared four different commercial EIAs, ImmunoWELL IgG, IgM (GenBio), Medac IgG, IgA, IgM (Medac), Platelia IgG, IgM (Sanofi Pasteur), and Ridascreen IgG, IgA, IgM (r-Biopharm) with indirect particle agglutination assay (PA), Serodia-MycoII (Fujirebio). We tested 91 specimens from 73 pediatric patients (2-17 yr) hospitalized at a tertiary-care hospital between December 2005 and January 2006. The measurements of IgM EIAs were correlated with PA titers (Spearman's correlation coefficient, from 0.89 to 0.92) with high concordance rates, ranging from 82.4% to 92.3%. However, some negative IgM-EIA results in PA-positive specimens indicated that serial samplings with convalescent sera would be necessary to confirm M. pneumoniae infection.
Adolescent ; Antibodies, Bacterial/chemistry ; Child ; Child, Preschool ; Female ; Humans ; Immunoenzyme Techniques/*methods ; Immunoglobulin G/*chemistry ; Immunoglobulin M/*chemistry ; Infant ; Male ; Microbiology ; Mycoplasma pneumoniae/chemistry/*immunology/metabolism ; Pneumonia, Mycoplasma/*diagnosis/*immunology ; Polymerase Chain Reaction ; Serologic Tests

OBJECTIVES: Purpose of this study was to investigate physiological and neurocognitive effects of total sleep deprivation by using laboratory blood tests and the computerized neurocognitive function test in healthy subjects. METHODS: Sixteen healthy volunteers participated in this study. Subjects were recommended to remain awake for 48 hours under continuous surveillance. Lab tests of cortisol, prolactin, thyroid hormone, growth hormone, and immunoglobulin (IgG, IgA, IgM, IgD, IgE), CBC, BC and the Vienna test were performed before and after 48 hours of total sleep deprivation. RESULTS: Concentration of T3 and T4 significantly increased after deprivation. In the reaction test, distribution reaction time significantly increased, and correct reaction significantly decreased. In the vigilance test, amount of correct reaction significantly decreased, and the mean value of correct reaction time was significantly delayed. Level of fasting blood sugar, total protein, albumin, alkaline phosphatase and potassium significantly increased, respectively, except for the level of total bilirubin which was decreased. After total sleep deprivation, WBC counts significantly increased. Regarding immunoglobulin level, IgG, IgA and Ag M concentrations appeared to decrease, but the results were not statistically significant. CONCLUSION: The effect of total sleep deprivation on physiological function was significant in the level of thyroid hormone. Although statistically not significant, the results of growth hormone and the immune system showed a trend in relations to the effect of total sleep deprivation. Results of blood chemistry suggest that sleep deprivation may influence metabolism of hepatobiliary system. Cognitive impairment was also seen in reactive and vigilant functions after total sleep deprivation.
Alkaline Phosphatase ; Bilirubin ; Blood Glucose ; Chemistry ; Fasting ; Growth Hormone ; Healthy Volunteers ; Hematologic Tests ; Hydrocortisone ; Immune System ; Immunoglobulin A ; Immunoglobulin D ; Immunoglobulin G ; Immunoglobulin M ; Immunoglobulins ; Metabolism ; Potassium ; Prolactin ; Reaction Time ; Sleep Deprivation* ; Thyroid Gland

The antigen location of Cryptosporidium parvum, which stimulates antibody formation in humans and animals, was investigated using infected human sera. Immuno-electron microscopy revealed that antigenicity-inducing humoral immunity was located at various developmental stages of parasites, including asexual, sexual stages, and oocysts. The amount of antigen-stimulating IgG antibodies was particularly high on the oocyst wall. The sporozoite surface was shown to give stimulation on IgG and IgM antibody formation. Trophozoites implicated the lowest antigenicity to humoral immunity, both IgG and IgM, by showing the least amount of gold labeling. Immunogold labeling also provided clues that antigens were presented to the host-cell cytoplasm via feeder organelles and host-parasite junctions.
Animals ; Antibodies, Protozoan/*immunology ; Antigens, Protozoan/*analysis ; Cryptosporidium parvum/*chemistry/*immunology/ultrastructure ; Female ; Humans ; Immunoglobulin G/immunology ; Immunoglobulin M/immunology ; Mice ; Microscopy, Immunoelectron ; Sporozoites/chemistry/immunology/ultrastructure ; Staining and Labeling/methods ; Trophozoites/chemistry/immunology/ultrastructure

The prevalent ages at onset for Kawasaki Disease (KD) and Epstein-Barr virus (EBV) infection are known to be similar in Korea and Japan. We evaluated the correlation between EBV infection and KD. The antibodies to EBV such as anti-viral capsid antigen (VCA) IgG and IgM, anti-diffuse and restricted early antigen IgG (anti-EADR IgG), and the anti-EBV determined nuclear antigen IgG (anti-EBNA IgG) were examined in 29KD patients at five separate times sequentially during a period of one year, and also in 14 other children with a past history of KD. The results of each group were compared with those of age-matched controls. The positive rates of anti-VCA IgG and IgM at presentation in the KD patients were 41.4% (12/29) and 0% (0/29), respectively. Only one patient was found to be anti-VCA IgM-positive within two months. There were no cases of anti-VCA IgG except one, anti-EADR IgG and anti-EBNA IgG positive to negative seroconversion during the year. The children with a past history of KD showed higher anti-EBNA IgG-positive rates than the controls (p=0.04). There was no difference in the seropositive rates of the antibodies to EBV, cytomegalovirus, herpes simplex virus and herpes zoster virus. In conclusion, children with KD were noted to have normal immune responses to EBV infection. Children with a past history of KD seemed to be infected with EBV at a later age than children with no history of KD.
Mucocutaneous Lymph Node Syndrome/*virology ; Male ; Korea ; Infant ; Immunoglobulins/metabolism ; Immunoglobulin M/chemistry ; Immunoglobulin G/chemistry ; Humans ; Herpesvirus 4, Human/*metabolism ; Female ; Epstein-Barr Virus Infections/*complications ; Child, Preschool ; Antibodies, Viral/*chemistry ; Age of Onset

OBJECTIVETo develop and evaluate a multiplex detection of IgM antibodies to pathogens caused viral hemorrhagic fever.

METHODSThe nucleocapsid proteins (NP) of HTN, SEO, Puu MBV, Lassa, RFV and HPS viruses expressed in prokaryotic cells and purified were coupled to 7 different xMAP fluorescent microbeads. The assay was evaluated and optimized when screened against a panel of reference sera collected from HFRS patients, and compared to commonly used MacELISA Kits.

RESULTSFor detection of anti-NP antibodies, the sensitivity and specificity of the assay were comparable to a commonly used MacELISA kit, but it could detect different antigen specific antibodies in one reaction simultaneously.

CONCLUSIONA robust, rapid and multiplex assay based on NPs could be developed via Luminex xMAP platform for laboratory diagnosis of viral hemorrhagic fever and seroepidemiological investigation.

Antibodies, Viral ; blood ; Fluorescence ; Hemorrhagic Fevers, Viral ; blood ; immunology ; virology ; Humans ; Immunoassay ; methods ; Immunoglobulin M ; blood ; Microspheres ; Nucleocapsid Proteins ; chemistry ; immunology ; Viruses ; immunology

OBJECTIVETo evaluate the feasibility and stability of chemically conjugating IgM on collagen films.

METHODSIgM was labeled with 125I using the chloramine-T method. Six collagen films were randomly divided into two groups. In chemical coupling group 125I-labeled IgM was chemically coupled with the films through N-succinmiclyl-3- (2-pyridyl-dithio) propionate reaction. In control group 125I-labeled IgM was absorbed onto collagen films. The amount of IgM on the collagen films and the amount of IgM remained on the films after extensive rinsing with phosphate buffered saline were monitored by counting the radioactivity of 125I.

RESULTSThe amount of antibodies loaded onto collagen films in the chemical coupling group was 15 times higher than that on the control films, showing significant statistical difference (P < 0.01). And the stability of conjugation antibodies on collagen films was significantly better than the control films.

CONCLUSIONChemical coupling is an effective approach to immobilize antibodies on collagen for further plasmid DNA tethering.

Angioplasty, Balloon, Coronary ; instrumentation ; Animals ; Antibodies, Antinuclear ; metabolism ; Cattle ; Coated Materials, Biocompatible ; chemistry ; metabolism ; Collagen ; chemistry ; metabolism ; Genetic Vectors ; Immunoglobulin M ; metabolism ; In Vitro Techniques ; Mice ; Protein Binding ; Stents ; Surface Properties

OBJECTIVETo study the effects and mechanism of Huchang Qingfei pellets on immune function in rats infected with mycoplasma pneumoniae.

METHODA rat model of mycoplasmal pneumonia (MP) was developed in repeated intranasal infectious routes and then the humoral and cellular immunocompetences were detected by radioimmunoassay, immune-turbidimetry and flow cytometry.

RESULTThe levels of serum IgG,IgM and IL-2, IL-6 were enhinced obviously, the complement C3 and TNF-alpha were decreased and the ratio of CD4+ /CD8+ was improved significantly in the Huchang groups as compared with MP model group.

CONCLUSIONHuchang Qingfei pellets can reinforce immune function via preventing both cellular and humoral immunity from depression in the rats with MP.

Animals ; CD4-CD8 Ratio ; Complement C3 ; metabolism ; Drug Combinations ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Female ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Interleukin-2 ; blood ; Interleukin-6 ; blood ; Male ; Plants, Medicinal ; chemistry ; Platycodon ; chemistry ; Pneumonia, Mycoplasma ; blood ; immunology ; Rats ; Rats, Wistar ; Scutellaria baicalensis ; chemistry ; Tumor Necrosis Factor-alpha ; metabolism

We investigated the characteristic features of cervical lymph node B cells to determine whether their behavior differs from that of B cells located elsewhere, because cervical lymph nodes may be exposed to continual antigenic stimulation from the naso- and/or oropharynx. B cells were isolated from cervical lymph nodes, spleen and peritoneal fluid of mice, cultured in medium, and exposed to various stimuli. The expression of various surface molecules characteristic of lymphoid B cells was assayed by flow cytometry, and immunoglobulin secreted into the culture supernatants was evaluated by enzyme-linked immunosorbent assay. B220+ cells were cultured in medium alone or with lipopolysaccharide, and their entrance into S phase in response to stimuli was measured by proliferative assays. Phenotypic characteristics of cervical lymph node B cells included CD5low, CD23high, CD43low, B7.1low, B7.2low, and Syndecan-1low. Unstimulated lymphoid B cells did not secrete immunoglobulin, but, upon stimulation, secretion of IgM was increased more than secretion of IgA and IgG. B cells actively entered S phase after 48 hr stimulation. These results show that B cells in cervical lymph nodes are conventional B2 cells, like splenic B cells.
Spleen/cytology ; Phenotype ; Mice, Inbred BALB C ; Mice ; Male ; Lymph Nodes/*cytology ; Immunoglobulin M/chemistry ; Flow Cytometry ; Enzyme-Linked Immunosorbent Assay ; Culture Media/pharmacology ; Cell Proliferation ; Cell Culture Techniques/methods ; B-Lymphocytes/*cytology ; Antigens/metabolism ; Animals

OBJECTIVETo study the therapeutic effect and possible mechanism of total panax notoginseng saponins (PNS) for treatment of rheumatoid arthritis (RA), and to observe its safety and influence on RA immune related inner environment.

METHODSEighty-four patients were randomly assigned to two groups. All were treated with the routine therapy with diclofenac sodium, Leflunomide and prednisone, but for the 43 patients in the treatment group PNS was given additionally. The therapeutic course was 28 days for both groups. Clinical efficacy and change of indexes including platelet counts, immnuoglobulins (IgG, IgA, IgM), complement (C)3, rheumatoid factor (RF), C-reactive protein (CRP), ceruloplasmin (CER), haptoglobin (HPT), and alpha1-acid glycoprotein (AAG) were observed.

RESULTSSignificant improvement of clinical symptoms, including the joint swelling index, joint tenderness index, joint pain index, time of morning stiffness and VAS revealed in both groups after treatment, and the effect in the treatment group was better (P<0.05 or P<0.01). PLT, CER, AAG, HPT, CRP, IgG, IgA, IgM, C3 and RF were lowered in both groups (P<0.01), but the lowering in PLT, CER, AAG and CRP in the treatment group was more significant than that in the control group respectively (P < 0.05 or P < 0.01).

CONCLUSIONPNS can significantly improve the condition of patients, enhance the therapeutic effect in treating RA, through regulating the disordered immunity and improving the effect of anti-inflammatory and analgesia.

Adolescent ; Adult ; Aged ; Anti-Inflammatory Agents ; therapeutic use ; Anti-Inflammatory Agents, Non-Steroidal ; therapeutic use ; Arthritis, Rheumatoid ; drug therapy ; immunology ; Diclofenac ; therapeutic use ; Drug Therapy, Combination ; Female ; Humans ; Immunoglobulin A ; blood ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Isoxazoles ; therapeutic use ; Male ; Middle Aged ; Panax notoginseng ; chemistry ; Phytotherapy ; Prednisone ; therapeutic use ; Rheumatoid Factor ; blood ; Saponins ; therapeutic use ; Treatment Outcome ; Young Adult