BACKGROUND AND OBJECTIVES: Since allergic fungal sinusitis (AFS) was first described in 1983, there have been occasional reports of patients that have typical allergic mucin without evidence of fungal disease. Clinically these patients were similar and undistinguishable from those of AFS. "Allergic mucin sinusitis without fungus" was first coined by Allphin in 1991 to describe this group of patients. The objective of this study was to evaluate the clinical features of chronic sinusitis with allergic mucin. MATERIAL AND METHOD: A review of medical records of twelve patients who had classic allergic mucin within their sinuses without documentation of the presence of fungi was undertaken. RESULTS: Eleven patients (92%) had nasal polyposis and eleven patients (92%) had one or more allergic conditions. However, only four patients (33%) had the evidence of type l hypersensitivity for fungal antigens. Postoperatively, eleven patients (92%) had recurrent symptoms and showed good response to systemic steroid therapy. In these patients, however, systemic steroid therapy was needed several times and additional surgeries were required in three patients. CONCLUSION: It remains unclear whether the chronic sinusitis of these patients is a new separate disease entity or a spectrum of the AFS. Further study will be needed to elucidate the disease entity of these patients.
Antigens, Fungal ; Fungi ; Humans ; Hypersensitivity ; Medical Records ; Mucins* ; Numismatics ; Sinusitis*

OBJECTIVETo establish an immunological method for detecting antibodies of Penicillium marneffei.

METHODSThe recombinant Mp1p protein of Penicillium marneffei was expressed in Pichia pastoris and labeled with HRP (Mp1p-HRP) with a modified sodium periodate method. A double-antigen sandwich enzyme-linked immunosorbant assay (ELISA) was established by determining the optimal coating concentration of Mp1p protein and the concentration of the detecting protein Mp1p-HRP. The sensitivity and specificity of the assay was evaluated by detecting Mp1p antibodies in 100 serum samples from healthy donors, 15 samples from culture-confirmed penicilliosis patients, and 21 samples from patients with culture-confirmed other fungal infections.

RESULTSA double-antigen sandwich ELISA was successfully established for detecting Mp1p-specific antibody. The specificity of the assay was 100% (121/121) for detecting Mp1p-specific antibody in the sera from healthy donors and patients with other fungal infection. The detection results of the 15 serum samples from patients with culture-confirmed penicilliosis showed positivity for Mp1p antibody in 2 samples and Mp1p antigen positivity in 12 samples; combining the detection results of Mp1p antigen and antibody obviously increased the diagnostic sensitivity to 93.3% (14/15).

CONCLUSIONThe double-antigen sandwich ELISA shows a high specificity in detecting Mp1p-specific antibody, and simultaneous detection of Mp1p antigen and antibody can increase the diagnostic sensitivity for penicilliosis.

Antibodies, Fungal ; blood ; immunology ; Antigens, Fungal ; blood ; immunology ; Enzyme-Linked Immunosorbent Assay ; methods ; Humans ; Mycoses ; blood ; diagnosis ; microbiology ; Penicillium ; immunology ; Pichia ; immunology ; Sensitivity and Specificity

We report a case of hypersensitivity pneumonitis in a 30-yr-old female housewife caused by Penicillium species found in her home environment. The patient was diagnosed according to history, chest radiograph, spirometry, high-resolution chest CT, and transbronchial lung biopsy. To identify the causative agent, cultured aeromolds were collected by the open-plate method. From the main fungi cultured, fungal antigens were prepared, and immunoblot analysis with the patient's serum and each fungal antigen was performed. A fungal colonies were isolated from the patient's home. Immunoblotting analysis with the patient's sera demonstrated a IgG-binding fractions to Penicillium species extract, while binding was not noted with control subject. This study indicates that the patient had hypersensitivity pneumonitis on exposure to Penicillium species in her home environment.
Adult ; Alveolitis, Extrinsic Allergic/*etiology/immunology/*microbiology ; Antibodies, Fungal/blood ; Antigens, Fungal ; Environmental Microbiology ; Female ; Housing ; Humans ; Immunoglobulin G/blood ; Korea ; Penicillium/*immunology/isolation and purification/*pathogenicity

OBJECTIVETo establish two double-antigen sandwich ELISA systems to detect anti-Afmp1cr and Afmp2cr antibodies of Aspergillus fumigatus.

METHODSRecombinant Afmp1cr and Afmp2cr proteins of A.fumigatus expressed in Pichia pastoris were obtained. Double-antigen sandwich ELISA systems for detecting specific anti-Afmp1cr and anti-Afmp2cr antibodies were developed after chessboard titrating to determine the appropriate concentrations of the recombinant proteins and HRP-labeled proteins. The sensitivity of the assay was evaluated using serum samples of rabbits immunized with Afmp1cr and Afmp2cr. The specificity of the assay was evaluated by detecting serum samples from healthy donors and patients with other pathogenic fungal and baterial infections. The performance of the two ELISA kits was furthered evaluated using serum samples from patients with suspected Aspergillus infection.

RESULTSThe established ELISA kits were capable of detecting anti-Afmp1cr and anti-Afmp2cr antibodies in immunized rabbit serum at the maximum dilutions of 800 and 3200, respectively. No cross-reactivity was observed in detecting serum from patients with other pathogenic fungal or bactetial infections. Both of the two kits yielded positive results in sera from two established Aspergillus-infected cases and a suspected case.

CONCLUSIONSTwo antibody-capture ELISA kits were developed for the laboratory diagnosis of A.fumigatus infection and can be potentially useful in the clinical diagnosis of Aspergillosis infections.

Animals ; Antibodies, Fungal ; isolation & purification ; Antigens, Fungal ; Aspergillosis ; diagnosis ; Aspergillus fumigatus ; Cross Reactions ; Enzyme-Linked Immunosorbent Assay ; Humans ; Pichia ; Rabbits ; Recombinant Proteins ; Sensitivity and Specificity

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

Adult ; Alternaria ; immunology ; Alveolitis, Extrinsic Allergic ; immunology ; Animals ; Antibodies ; immunology ; Antibodies, Bacterial ; immunology ; Antibodies, Fungal ; immunology ; Antigens ; immunology ; Antigens, Bacterial ; immunology ; Antigens, Fungal ; immunology ; Aspergillus fumigatus ; immunology ; Asymptomatic Diseases ; Candida albicans ; immunology ; Cladosporium ; immunology ; Columbidae ; immunology ; Female ; Healthy Volunteers ; Humans ; Immunoglobulin G ; immunology ; Male ; Melopsittacus ; immunology ; Middle Aged ; Mucor ; immunology ; Nocardia ; immunology ; Parrots ; immunology ; Penicillium chrysogenum ; immunology ; Stachybotrys ; immunology ; Thermoactinomyces ; immunology

Allergic fungal rhinosinusitis (AFRS) is a noninvasive fungal infection of the paranasal sinuses that are usually seen in young immunocompetent patients with atopy and/or asthma. Fungus balls can grow in moist cavities of the paranasal sinuses of a host with normal immunologic status. Cases of AFRS with concurrent fungus balls is very rare. We present a case of a patient who had AFRS on one side of the paranasal sinus and allergic fungal sinusitis on the other side. A 51-year-old female with atopy presented with a few-year history of nasal obstruction and rhinorrhea, as well as a history of high-dose systemic steroid therapy. The patient had nasal polyps and showed an elevated level of total IgE and positive MAST to fungal antigens. Endoscopic sinus surgery was performed. Allergic mucin from the right maxillary sinus contained sheets of eosinophils and Charcot-Leyden crystals. Also, a clay-like dark brown material from the left maxillary sinus was revealed to be a fungus ball.
Antigens, Fungal ; Aspergillosis ; Asthma ; Eosinophils ; Female ; Fungi* ; Humans ; Immunoglobulin E ; Maxillary Sinus ; Middle Aged ; Mucins ; Mycetoma ; Nasal Obstruction ; Nasal Polyps ; Paranasal Sinuses ; Sinusitis

OBJECTIVETo evaluate the diagnostic value of serum galactomannan antigen (GM) and (1→3)-β-D-glucan antigen (BG) assay in invasive fungal infections (IFI) in the patients with hematologic malignancies and the role in monitoring therapeutic response.

METHODSFifty one patients with hematological malignancies met the criteria for inclusion: (1) body temperature above 38°C for 48 hours, (2) failure to respond to broad-spectrum antibiotic treatment, or (3) temperature rose again after the responded drop. Blood samples were collected twice at the first week, then once a week in at least four weeks. The double antibody sandwich enzyme-linked immunosorbent assay (ELISA) and colorimetric assay were used for detecting GM and BG. The positive GM test is defined as two consecutive tests at different time GM value > 0.5 or > 0.8 and the positive G test is defined as BG value > 80 pg/ml. The patients were assigned into four groups as proven, probable, possible, and non-fungal infection respectively, and 21 normal volunteers were as controls.

RESULTSTwo hundred and forty serum samples were collected from 51 patients including 2 of proven IFI, 26 probable IFI, 17 possible IFI and 6 non-fungal infection. The true-positive group including the proven and probable groups, and true negative group was the non-fungal infection group. GM tests were positive in 21 of 28 cases in true positive group, and only one of 6 cases in non-fungal infection. The sensitivity, specificity, positive predictive value and negative predictive value were 75%, 83.3%, 95.5% and 41.7%, respectively. G tests were positive in all 28 cases of the true positive group, and 4 in 6 non-fungal infection cases. The sensitivity, specificity, positive predictive value and negative predictive value were 100%, 33.3%, 87.5% and 100%, respectively. G test is more sensitive than GM test (P = 0.015), but there was no significant difference in specificity of the two tests (P = 0.242). In 19 of 21 patients with GM test positive, anti-fungal treatment was effective, and GM value gradually decreased to negative, two invalid patients were persistent with GM test positive. After two weeks treatment, the average GM value was significantly lower in the effective group than in the ineffective group (P < 0.05). BG values in the responded patients showed a gradual decline similar to that of GM values, but not to negative. The changes of BG value in ineffective group varied with a trend upward. The changes in BG value had no relation with treatment effectiveness.

CONCLUSIONSSerum GM and BG antigens detection provides strong evidence for early diagnosis of IFI. Combination of GM and G tests can improve the diagnostic specificity and reduce the false positive GM test seems superior to G test for monitoring GM and BG values during treatment.

Adult ; Aged ; Aged, 80 and over ; Antigens, Fungal ; blood ; immunology ; Female ; Hematologic Neoplasms ; immunology ; microbiology ; Humans ; Male ; Mannans ; immunology ; Middle Aged ; Mycoses ; blood ; immunology ; Young Adult ; beta-Glucans ; immunology

BACKGROUNDTo explore the feasibility of cloning of the hepatocyte receptor interacting with the Pre S1 protein of HBV by two hybrid system.

METHODSYeast expression plasmids encoding fusion proteins of full length or portions of Pre S1 of HBV and DNA binding domain of yeast protein GAL4 were constructed and used to transform yeast reporter strain SFY526. Reporter gene product ?galactosidase activity was assayed as a measure of transcription activation in yeast. Mammalian expression plasmid encoding fusion proteins of full length Pre S1 and DNA binding domain of GAL4 was constructed and used to cotransfect hepatoma cell line Huh?7 together with CAT reporter plasmid. Cell extracts were assayed for CAT activity by thin?layer chromatography.

RESULTSThe fusion proteins of full length Pre S1 protein and GAL4 DNA binding domain present transcriptional activation function in yeast. The transcription activating sequence is localized to the 21 to 47 amino acids of Pre S1 protein Fusion proteins of full length Pre S 1 and GAL 4 DNA binding domain do not show transcriptional activation function in mammalian cells.

CONCLUSIONThe transcriptional activating sequence of HBVPre S1 protein in yeast overlaps the hepatocyte receptor binding site. The transcriptional activation function of HBV Pre S1 protein in yeast may prevent researchers?from using yeast two hybrid system to clone HBV receptor interacting with Pre S1 protein. However, the Pre S1 protein does not show transcriptional activation function in mammalian cells. Mammalian two?hybrid system may be a practical method to clone the HBV hepatocyte receptor interacting with Pre S1 protein.

DNA-Binding Proteins ; genetics ; Fungal Proteins ; genetics ; Hepatitis B Surface Antigens ; genetics ; Humans ; Protein Precursors ; genetics ; Recombinant Fusion Proteins ; genetics ; Transcriptional Activation ; Tumor Cells, Cultured ; Yeasts

Since the initial description of allergic fungal sinusitis by Millar et al. in 1981, reported cases have been increasing. However, one case of allergic fungal sinusitis has been reported in Korea. The diagnosis can be established by demonstrating type I hypersensitivity reaction, nasal polyposis, characteristic CT scan, eosinophilic mucus without evidence of fungal invasion into sinus tissue, and a positive fungal stain or culture of sinus contents. The authors present a case which met the diagnostic criteria of allergic fungal sinusitis. This patient had nasal polyp and showed elevated level of total IgE, peripheral eosinophilia, positive MAST and strongly positive immediate skin reactivity to fungal antigens. On histologic examination, typical allergic mucin containing sheets of eosinophils, Charcot-Leyden crystals was found. Fungal hyphae were also found in the mucin content.
Antigens, Fungal ; Diagnosis ; Eosinophilia ; Eosinophils ; Humans ; Hypersensitivity, Immediate ; Hyphae ; Immunoglobulin E ; Korea ; Mucins ; Mucus ; Nasal Polyps ; Sinusitis* ; Skin ; Tomography, X-Ray Computed