No abstract available.
Chlamydia* ; Fluorescent Antibody Technique, Indirect*

No abstract available.
Diagnosis* ; Fluorescent Antibody Technique, Indirect* ; Herpes Simplex* ; Simplexvirus*

The pathophysiology of glandular dysfunction in Sjögren's syndrome (SS) has not been fully elucidated. Previously, we reported the presence of autoantibodies to AQP-5 in patients with SS, which was associated with a low resting salivary flow. The purpose of this study was to investigate the presence of anti-AQP1 autoantibodies. To detect anti-AQP1 autoantibodies, cell-based indirect immunofluorescence assay was developed using MDCK cells that overexpressed human AQP1. By screening 112 SS and 52 control sera, anti-AQP1 autoantibodies were detected in 27.7% of the SS but in none of the control sera. Interestingly, the sera that were positive for anti-AQP1 autoantibodies also contained anti-AQP5 autoantibodies in the previous study. Different from anti-AQP5 autoantibodies, the presence of anti-AQP1 autoantibodies was not associated with the salivary flow rate. Although anti-AQP1 autoantibodies are not useful as a diagnostic marker, the presence of autoantibodies to AQP1 may be an obstacle to AQP1 gene therapy for SS.
Aquaporin 1 ; Autoantibodies* ; Fluorescent Antibody Technique ; Fluorescent Antibody Technique, Indirect ; Genetic Therapy ; Humans ; Madin Darby Canine Kidney Cells ; Mass Screening

Type VII collagen is one of the major structural components of the corneal epithelial adhesion complex. Using the immunogold technique combined with indirect immunofluorescence analysis, the fine structural distribution of type VII collagen was studied in the corneas obtained from 5 enucleated hyman eyes (age range, 1-77 years) including one pathologic cornea from graft rejection. The findings on normal cornea corroborated the results from previous studies. In pathologic cornea from graft rejection, type VII collagen antibodies generated linear and irregular patchy fluorescence staining along the epithelial-stromal interface and immunogold binding to type VII collagen mainly occurred within the undulating lamina densa, more densealy distributed anchoring plaques and anchoring fibrils. The distribution of type VII collagen in pathologic human cornea from graft rejection is similar to normal human cornea. But, in pathologic cornea, type VII collagen is more densely distributed in superficial stroma and forms more extended anchoring network, which may be derived from the increased secretion of the type VII collagen due to the activated basal epithelial cell during healing process.
Antibodies ; Collagen Type VII* ; Cornea* ; Epithelial Cells ; Fluorescence ; Fluorescent Antibody Technique, Indirect ; Graft Rejection ; Humans ; Immunohistochemistry

BACKGROUND: IFN-gamma plays an important role in host response to intracellular organisms such as mycobacterium. Human infection with mycobacterium leads to a wide variety of outcomes, ranging from asymptomatic infection to widespread and rapidly fatal disease. Recent reports suggest that alteration of the function of IFN-gamma caused by a defective IFN-gamma receptor gene can explain different host response to mycobacterium. In this study, we investigated the role of IFN-gamma in the development of chronic refractory tuberculosis. METHODS: The LPS-induced TNF-alpha production with or without IFN-gamma priming was compared by using monocytes taken from recently diagnosed tuberculosis, chronic refractory tuberculosis patients and controls. And the IFN-gamma receptor was measured by indirect fluorescent antibody technique to know whether change in the priming effect of IFN-gamma is related to IFN-gamma receptor deficiency or not. RESULTS: The ratio of TNF-alpha produced in response to stimulation with INF-gamma and LPS to LPS alone was 13.5 +/- 7.6 in controls, 10.8 +/- 6.4 in recently diagnosed tuberculosis patients and 6.7 +/- 3.9 in chronic refractory tuberculosis patients. The priming effect of IFN-gamma significantly decreased in chronic refractory tuberculosis patients compared with that in controls (p=0.002). However, IFN-gamma receptor deficiency was detected in one of chronic refractory tuberculosis patients. CONCLUSION: The decrease of the priming effect of IFN-gamma may play an important role in the development of chronic refractory tuberculosis, and in some patients, this may be related to the IFN-gamma receptor deficiency.
Asymptomatic Infections ; Fluorescent Antibody Technique, Indirect ; Humans ; Monocytes ; Mycobacterium ; Tuberculosis* ; Tumor Necrosis Factor-alpha

OBJECTIVETo compare indirect immunofluorescence assay (IIFA) and enzyme-linked immunosorbent assay (ELISA) for detecting antinuclear antibodies (ANA) and anti-double-stranded DNA antibodies (anti-dsDNA).

METHODSA total of 125 serum samples were obtained from patients with established or suspected autoimmune disease, and 82 samples were used for ANA detection and 57 for anti-dsDNA detection using both IIFA and ELISA. Fourteen samples were examined for both ANA and anti-dsDNA. In cases where discrepancy occurred in the results by the two methods, extractable nuclear antigens were detected using immunoblotting.

RESULTSThe positivity rate of ANA detected by IIFA and ELISA was significantly different (87.8% and 73.17%, respectively, P<0.01), but the positivity rate of anti-dsDNA was similar between IIFA and ELISA (77.19% and 71.93%, respectively, P>0.05). The percent agreement between the two testing methods with different cutoff values of ANA and anti-dsDNA showed significant differences (P<0.01), and for some uncommon patterns, the percent agreement of the two methods was lowered in ANA detection but remained unchanged for anti-dsDNA with different ANA patterns. High percent agreements of the two methods were obtained with the cutoff ANA titer of 1:100 and the cutoff anti-dsDNA value of weak positivity, but they demonstrated a significant difference in testing low-titer ANA and anti-dsDNA.

CONCLUSIONIIFA is more sensitive than ELISA in detecting the total ANA and anti-dsDNA. ELISA prescreening combined with IIFA can obtain the information of the nuclear pattern and allow the observation of the titer alterations. The combination of two or more testing methods can greatly enhance the accuracy of the results.

The distribution of pemphigus subclass autoantibodies in a patient with pemphigus vulgaris (PV) has been investigated by semiquantitative indirect immunofluorescence (IIF), using the HP series monoclonal antibodies specific for four human IgG subclasses on human foreskins. IgG1 and IgG4 intercellular substance-specific autoantibodies were detected in the serum of the patient, whereas IgG2 and IgG3 autoantibodies were absent. In addition to foreskins, human tonsillar epithelia were used as substrates of IIF for detecting the PV autoantibodies and it was one of satisfactory substitutes for monkey esophagus.
Antibodies, Monoclonal ; Autoantibodies* ; Esophagus ; Fluorescent Antibody Technique, Indirect ; Foreskin ; Haplorhini ; Humans ; Immunoglobulin G* ; Pemphigus*

OBJECTIVES: The purpose of this study is to compare newly developed assay for identification of ENA antibody, Phadia EliA ENA with Euroimmun line immunoassay by analyzing the degree of agreement and the individual antibodies between two methods. METHODS: A total of 82 patient samples were used. Indirect immunofluorescence assay using Hep-2 cell was performed to screen the antinuclear antibody (ANA). Euroimmun line immunoassay (LIA) and Phadia EliA ENA assay were tested to identify the antibodies against extractable nuclear antigens (ENAs). Kappa statistics was used to evaluate the degree of agreement. RESULTS: Mean age of patients was 41.0 (8-79), and the M:F ratio was 21:61. ANA was positive in 74 samples, and negative were 8 samples. Kappa analysis of the 82 tested samples showed a moderate strength of agreement (kappa = 0.521, P = 0.000). There were differences in the order of identified individual antibodies between two methods (Ro > La = RNP > Centromere > Sm > Scl-70 in Phadia EliA ENA, Ro > RNP > Sm>La > Scl-70 > Centromere=Jo-1 in Euroimmun LIA). Ro antibody was most frequently identified in Phadia EliA ENA negative-Euroimmun LIA positive specimens (Ro > RNP = Jo-1 > La = Sm = Centromere > Scl-70). CONCLUSIONS: A moderate strength of agreement was observed between the Phadia EliA ENA and the Euroimmun LIA. There seemed to be a significant difference in the ratio of individual antibodies, especially in the anti-Ro and Sm antibodies.
Antibodies ; Antibodies, Antinuclear ; Antigens, Nuclear ; Centromere ; Fluorescent Antibody Technique, Indirect ; Humans ; Immunoassay

The authors investigated the incidence of stratum corneum and upper epidermal cytoplasmic antibodies with 30 untreated and 20 treated psoriasis sera, and normal human sera using normal human skins of 5 different sites and psoriatic lesion by the method of indirect immunofluorescence in order to evaluate immunologic responses i n psoreasis. The results are summarized as follows . I) The positivity of stratum corneum antibodies in untreated psoriasis sera(78.7%) was si, nificantly higher than that in normal sera(64.0%). The incidence of stratum corneum antibodies in untreated psoriasis sera was found to be the highest in the arm, followed by the scalp, leg, abdomen, and face as substrate. 2) The positivity of stratum corneum antibodies in psoriasis and normal human sera was significantly higher when tested with the psoriatic lesion as substrate than norma! skin as substrate, 3) The titer of stratum corneum antibodies in 5 sera using human skin obtained from 5 different sites on the body as substrate are the highest in the arm, and leg, and(in decreasing order of frequency) the scalp, abdomen, and face. 4) The positivity of upper epidermal cytoplhsmic antibodies in normal human sera (40.7%) was significa.ntly higher than that in untreated psoriasis sera(21.3%). 5) Ir.. the majority of cases, upper epidermal cytoplasmic antibodies coexisted with straturn corneum antibodies in the sera of patients with psoriasis and in the sera of normal humans.
Abdomen ; Antibodies* ; Arm ; Cytoplasm* ; Fluorescent Antibody Technique, Indirect ; Humans ; Humans* ; Incidence* ; Leg ; Psoriasis* ; Scalp ; Skin

In the present study, we have tried to isolate and identify herpes simplex virus type 2(HSV 2) from clinical specirnens, which were inoculated into Vero cell line and grown. Eight strains of viruses were isolated from 20 suspected cases diagnosed from the pr ivate clinics in Seoul. Viruses isolated from 4 rnale and 1 female cases with active lesion were identified to the HSV 2 by indirect immunofluorescence using monoclonal antibody to HSV-2. In addition, morphology of the isolated viruses were observed under electron microscope.
Female ; Fluorescent Antibody Technique, Indirect ; Herpes Simplex* ; Herpesvirus 2, Human* ; Humans ; Seoul ; Simplexvirus* ; Vero Cells