We report an important observation that the surface conductivity of antibody layer immobilized on polylysine-coated glass substrate decreases upon the formation of complex with their specific antigens. This change in conductivity has been observed for both monoclonal and polyclonal antibodies. The conductance of monoclonal mouse IgG immobilized on polylysine-coated glass substrate changed from 1.02x10(-8) ohm(-1) to 1.41x10(-11) ohm(-1) at 10 V when complex is formed due to the specific biomolecular interactions with rabbit anti-mouse IgG F(ab')(2). Similar behavior was observed when the same set up was tested in two clinical assays: (1) anti-Leishmania antigen polyclonal antibodies taken from Kala Azar positive patient serum interacting with Leishmania promastigote antigen, and (2) anti-p21 polyclonal antibodies interacting with p21 antigen. The proposed concept can represent a new immunodiagnostic technique and may have wide ranging applications in biosensors and nanobiotechnology too.
Animals ; Antigen-Antibody Complex ; analysis ; Biosensing Techniques ; Electric Conductivity ; Humans ; Immunoassay ; methods ; Rabbits

To investigate leukocyte inhibitory factor(LlF) production and circulating immune complexes (CIC) in leprosy, peripheral blood mononuclear cells (PBMC) from 61 patients and sera from 6O patients were tested. The results indicate that there is a defect in LlF production in the lepromatous (LL) or borderline lepromatous (BL) types compared to the tubrculoid (TT) type (mean migration index=66.O +/- 16.O in LL 61.1 +/- 15.3 in BL, 51.9 +/- 11.2 in TT) (p < 0.05). The number of patients with positive CIC was higher among the LL patents (30%) than the TT patients (20%). There was also positive correlation between the bacterial index (Bl) and the CIC level (r=0.46, p < 0.05). The correlation between CIC and LIF in LL patients and the possibility (p=0.06) that the inuease m CIC may account for the decrease in LIF production in LL patients and vice versa are discussed.
Antigen-Antibody Complex/*analysis ; Human ; Leprosy/*immunology ; Leprosy, Lepromatous/immunology ; Leprosy, Tuberculoid/immunology ; Lymphokines/*analysis ; Support, Non-U.S. Gov't

BACKGROUNDSkin lesions are common manifestations in systemic lupus erythematosus (SLE). It is still unknown what the definite pathogenesis of skin involvement was and whether DNA participated in it. Our study was designed to explore the pathogenetic role and nature of nuclear antigen (DNA) deposited in the skin lesions of patients with SLE.

METHODSThirty skin samples from patients with SLE and 2 normal skin samples were studied. Extracellular DNA was evaluated by indirect immunofluorescence methods. The deposited immune complexes were extracted by cryoprecipitation, and DNA was then isolated with phenol and chloroform. DNA fragment sizes were detected by agarose gel electrophoresis. Finally, 8 different probes were used to analyze the origin of these DNA molecules using Dot hybridization.

RESULTSExtracellular DNA staining was found only in skin lesions, mainly those located in the basement membrane zone, vascular wall, and hair follicle wall. Normal skin and non-lesion SLE skin showed no fluorescence at locations outside the nuclei. There were no differences in the rate and intensity of extracellular DNA staining when comparing active phase to remission phase patients. No relationship was found between extracellular DNA and circulating anti-dsDNA antibodies. Deposited DNA fragments clustered into four bands of somewhat discrete sizes: 20 000 bp, 1300 bp, 800-900 bp, 100-200 bp. Small sized fragments (100-200 bp) were positively correlated with disease activity (P < 0.05, r = 0.407). Dot hybridization showed significant homology of the various extracellular DNA fragments examined with human genomic DNA, but not with DNA from the microorganisms and viruses we examined. There were also homologies between DNA samples from different individuals.

CONCLUSIONSDNA and its immune complexes may contribute to the pathogenesis of skin lesions in SLE. These DNA molecules range in size from 100 bp to 20 kb and may be endogenous in origin.

Antibodies, Antinuclear ; blood ; Antigen-Antibody Complex ; analysis ; DNA ; analysis ; immunology ; Humans ; Lupus Erythematosus, Systemic ; immunology ; Skin ; immunology ; Staining and Labeling

The occurrence of immune complexes in the serum from rats infected with M. leprae-murium and 38 patients with leprosy were studied by the polyethylene glycol precipitation complement consumption (PEG-CC) test and the results were compared in the various forms of the disease. Circulating immune complexes (CIC) were significantly increased in the sera from rats infected with M. lepraemurium compared to normal control rats (P < 0.005). There were no significant differences between the the level of CIC in the sera from lepromatous leprosy patients and that from tuberculoid leprosy patients, but in the sera from patients with erythema nodosum leprosum (ENL) the level of CIC was significantly increased (P < 0.005). And although we couldn't find a corre1ation between the level of CIC and bacterial indices in lepromatous leprosy patients, CIC tends to de-crease after negative conversion of their bacterial indices. These findings suggested that the detection of CIC can be of some practical interest in the early diagnosis of ENL and can be a valuable assessment in following the therapy after negative conversion of their bacterial indices.
Animal ; Antigen-Antibody Complex/analysis* ; Disease Models, Animal ; Human ; Leprosy/immunology* ; Male ; Rats ; Reference Values ; Time Factors

Higher levels of IgE-containing immune complexes (IC) have been reported in sera from patients with allergic diseases than in sera from controls. To evaluate the possibility of an IC-mediated mechanism in the pathogenesis of bronchial asthma, we measured circulating C3-containing IgE IC (C3-IgE IC) using anti-C3 ELISA from 20 house dust mite (HDM)-sensitive asthmatics, 20 non-atopic asthmatics, and 14 non-atopic controls. C3-IgE IC levels were significantly higher in HDM-sensitive asthmatics (mean +/- S.D.: 12.2 +/- 7.8 AU/ml) than in non-atopic asthmatics (6.5 +/- 7.5 AU/ml) or controls (5.8 +/- 4.4 AU/ml). C3-IgE IC levels were significantly correlated with HDM-specific IgE levels (r = 0.50, p<0.05), but not with total IgE levels (r = 0.36, p< 0.05) in HDM-sensitive atopic asthmatics. C3-IgE IC levels in sera did not significantly change during HDM-bronchoprovocation test in six HDM-sensitive asthmatics who showed positive reaction. Part of C3-IgE IC could be precipitated by protein G coupled beads. In conclusion, C3-IgE IC levels were elevated in sera from HDM-sensitive asthmatics; moreover IgG antibodies might be a component of C3-IgE IC. Our results suggest that an IgE IC-mediated mechanism could be involved in the pathogenesis of atopic asthma.
Adult ; Animal ; Antigen-Antibody Complex/*blood ; Asthma/*immunology ; Complement 3/*analysis ; Dust ; Human ; Immunoglobulin E/*blood ; Mites/immunology

Animals ; Antigen-Antibody Complex ; analysis ; DNA ; immunology ; DNA, Protozoan ; immunology ; Female ; In Situ Hybridization ; Lupus Nephritis ; etiology ; Mice ; Mice, Inbred BALB C ; Trypanosoma ; genetics

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

OBJECTIVEThis study investigated the clinical manifestations and renal pathological findings of 95 children with Henoch-Schonlein purpura nephritis (HSPN) in order to explore the relationship between clinical manifestations and renal pathology in HSPN.

METHODSAccording to clinical manifestations, 95 HSP patients were classified into six clinical groups: 1) normal urine analysis; 2) isolated hematuria or proteinuria; 3) proteinuria with hematuria; 4) acute nephritis; 5) nephrotic syndrome; 6) acute nephritis with over 50 mg/(kg.d) of proteinuria. The severity of the renal pathological findings was determined based on the classification of the International Study of Kidney Disease (ISKDC), including grades I-VI. The relationship between clinical manifestations and the severity of renal pathological findings was studied.

RESULTSNephrotic syndrome was the most common clinical diagnosis (26 cases), followed by proteinuria with hematuria (23 cases), normal urine analysis (20 cases), isolated hematuria or proteinuria (15 cases), acute nephritis with over 50 mg/(kg.d) of proteinuria (7 cases) and acute nephritis (4 cases). Twenty-five out of 26 patients with nephrotic syndrome had an ISKDC classification of grade III-IV. All of the four patients with acute nephrits had a classification of grade IIIb. The 7 cases of acute nephritis with over 50 mg/(kg.d) of proteinuria had a classification of grade IIIa-V. The 20 patients with normal urine analysis had a classification of grade Iia- IIIb. There were no significant differences in ISKDC classification among the patients with normal urine analysis, isolated hematuria or proteinuria, and hematuria plus proteinuria. As the course progressed, the degree of renal pathological changes in patients with isolated hematuria or proteinuria and hematuria plus proteinuria became more serious. Of all the 95 patients, 58% had co-deposition of immunoglobulins A, G and M. The percentage of co-deposition of immunoglobulins A, G and M was related to the disease course and the severity of renal pathological findings.

CONCLUSIONSHSPN children with nephrotic syndrome or acute nephritis with or without proteinuria had relatively severe renal pathological changes. The clinical manifestations were not always in parallel with the severity of renal pathological findings in HSPN children. With the course progressing, the renal pathological changes tended to be serious. The severe renal pathological manifestations came with co-deposition of immunogolobulins A, G and M in the glomerulin.

Adolescent ; Antigen-Antibody Complex ; analysis ; Child ; Child, Preschool ; Female ; Humans ; Kidney ; pathology ; Male ; Nephritis ; complications ; immunology ; pathology ; Purpura, Schoenlein-Henoch ; complications ; immunology ; pathology

Human immunodeficiency virus type 1 (HIV-1) phenotype plays an important role in the pathogenesis of AIDS. The presence of syncytium-inducing (SI) HIV-1 isolates in infected persons is associated with a rapid decline of CD4+T cells (CD4+), rapid disease progression, and reduced survival time after AIDS diagnosis. We have reported the effects of Korean red ginseng (KRG) on HIV-1 infected patients. To investigate whether KRG affects HIV-1 at gene level and there is a correlation between genotype and decline of CD4+, the C2-V3 region of env gene from 65 HIV-1 isolates were cloned and sequenced. Distributions of subtype were subtype B 57 (88%), subtype A 4 (6%), subtype C 2 (3%), subtype G 1 (2%), and subtype H 1 (2%). The prevalences of SI according to the number of CD4+ are as follows; 40% (6/15) in CD4+ <100/ ul, 14% (1/7) in 100-200/ul, and 2% (1/43) in >200/ul. Seventy-five percent (6/8) of SI were detected in rapid progressor with the decline of CD4+ over 60/ul per year. The correlation between SI genotype and the detection of immune complex dissociated (ICD) p24 antigen was significant (p<0.001). In the 40 patients followed-up over 60 months by CD4+, there was significant correlation between annual decrease of CD4+ and duration of KRG intake (R=-0.380, p<0.01), whereas no correlation between CD4+ and zidovudine (ZDV) was observed. The intrapatient variation of amino acid level showed significant inverse correlation with the months of KRG intake (R=-0.47, p<0.01). These results suggest that the determination of genotype by C2- V3 sequencing may be used for the evaluation of prognosis of AIDS patient, and long-term intake of KRG may prevent or delay the progression from NSI to SI.
Antigen-Antibody Complex ; Clone Cells ; Diagnosis ; Disease Progression ; Genes, env* ; Genotype ; HIV* ; HIV-1* ; Humans* ; Panax* ; Phenotype ; Prevalence ; Prognosis ; Sequence Analysis* ; Zidovudine

OBJECTIVETo study the correlation of erythrocyte immune function between normal neonates and their mothers and the influence of various obstetric factors on neonatal erythrocyte immune function.

METHODSThe adherent rate of complement 3b-receptor on the surface of red blood cells (RBC-C3bRR) and the immune complex adherent rate of red blood cells (RBC-ICR) were detected using the erythrocyte saccharomyces rosette test in 104 normal neonates and their mothers. The correlation of erythrocyte immune function between neonates and their mothers was evaluated by the maternal-infant paired test.

RESULTSThe levels of RBC-C3bRR (16.80 +/- 1.56% vs 16.23 +/- 1.63%; P < 0.05) and RBC-ICR (5.72 +/- 1.63% vs 5.02 +/- 1.38%; P < 0.01) in neonates were significantly higher than those in their mothers. There was a significantly positive correlation in RBC-ICR levels between neonates and their mothers (r = 0.28, P < 0.05). No correlation was found in RBC-C3bRR levels between the two groups. Neither RBC-C3bRR nor RBC-ICR levels of neonates were associated with various obstetric factors such as amniotic fluid, placenta, umbilical cord, parturient patterns, and puerperal anemia and pregnancy-induced hypertension syndrome.

CONCLUSIONSThe erythrocyte immune function in neonates has a relatively mature level and correlates with their mothers' erythrocyte immune function. Various obstetric factors have no influences on neonatal erythrocyte immune function.

Antigen-Antibody Complex ; immunology ; Erythrocytes ; immunology ; Female ; Fetal Blood ; immunology ; Humans ; Infant, Newborn ; immunology ; Linear Models ; Male ; Pregnancy ; Receptors, Complement 3b ; analysis ; Rosette Formation