OBJECTIVETo characterize the specific monoclonal antibodies to Aspergillus conidia.

METHODSFlow cytometry was used to examine the reactivity of the specific monoclonal antibodies to Aspergillus conidia.

RESULTSBoth the monoclonal antibodies MA3 and Con2 showed specific reactivity to Aspergillus conidia suspensions. MA3 was capable of binding to the conidia of A.fumigatus, A.flavus, A.niger and A.terreus, while Con2 was reactive only to the conidia of A.fumigatus.

CONCLUSIONTwo specific monoclonal antibodies (MA3 and Con2) to Aspergillus conidia have been obtained.

Antibodies, Fungal ; immunology ; isolation & purification ; Antibodies, Monoclonal ; immunology ; isolation & purification ; Antibody Specificity ; Aspergillus ; immunology ; Flow Cytometry ; Spores, Fungal ; immunology

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 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

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

Encephalitozoon cuniculi is a microsporidian parasite commonly found in rabbits that can infect humans, causing encephalitozoonosis. The prevalence of encephalitozoonosis is not well documented, even when many clinics suspect pet rabbits as being highly infected. This study investigated the seropositivity of E. cuniculi using ELISA. The examination of 186 rabbits using ELISA showed that 22.6% (42/186) were seropositive against E. cuniculi. In analysis with healthy status, all 42 seropositive sera were collected from clinically normal rabbits. Moreover, the gender and age of pet rabbits did not have anysignificant effect on E. cuniculi infection. To the best of our knowledge, this is the first report to describe the seroprevalence of E. cuniculi in pet rabbits and suggests that pet rabbits could act as an important reservoir of encephalitozoonosis for both pet animals and humans in Korea.
Animals ; Antibodies, Fungal/*blood ; Encephalitozoon cuniculi/*immunology ; Encephalitozoonosis/epidemiology/*veterinary ; Enzyme-Linked Immunosorbent Assay ; Female ; Korea/epidemiology ; Male ; *Pets ; Rabbits ; Seroepidemiologic Studies

OBJECTIVETo study the respiratory system injury in fur processing environment.

METHODSEnvironmental fungal survey was conducted in the fur processing procedures. Investigation of respiratory symptoms and chest X-ray examination were also carried out in 138 fur processing workers and 40 control workers. At the same time, the serum antibodies to fungi were analyzed by ELISA.

RESULTSFungal number(629-3,681 cfu/m3) in fur processing procedures was much higher than those in the control environment. Cladosporium and Alternaria were the leading strains of fungi in fur processing procedures. The rates of respiratory symptoms(cough, sputum, chest tightness, dyspnea, and fever) in fur processing workers were higher than those in the control workers. The rates of the symptoms in female workers were 37.9%, 28.4%, 10.5%, 22.1%, 4.2%, respectively. Abnormalities of chest X ray were found in 7 workers. The serum antibodies to Cladosporium and Alternaria(A450 nm 0.631, 0.724, respectively) in fur workers were significantly higher than those in the control workers(P < 0.05). The positive rates of the antibodies to Cladosporium and Alternaria(44.2%, 42.8%) were significantly higher than those in the control workers(P < 0.01).

CONCLUSIONCladosporium and Alternaria may be the pathogens of occupational respiratory diseases in fur processing workers.

Alternaria ; isolation & purification ; Antibodies, Fungal ; blood ; Cladosporium ; isolation & purification ; Environmental Microbiology ; Female ; Hair ; Humans ; Occupational Diseases ; etiology ; Radiography, Thoracic ; Respiratory Tract Diseases ; etiology

This article reports two cases of children with B-cell acute lymphoblastic leukemia (B-ALL) complicated by invasive fungal disease (IFD) who received bridging treatment using blinatumomab. Case 1 was a 4-month-old female infant who experienced recurrent high fever and limb weakness during chemotherapy. Blood culture was negative, and next-generation sequencing (NGS) of peripheral blood, bronchoalveolar lavage fluid, and cerebrospinal fluid were all negative. Chest CT and cranial MRI revealed obvious infection foci. Case 2 was a 2-year-old male patient who experienced recurrent high fever with multiple inflammatory masses during chemotherapy. Candida tropicalis was detected in peripheral blood and abscess fluid using NGS, while blood culture and imaging examinations showed no obvious abnormalities. After antifungal and blinatumomab therapy, both cases showed significant improvement in symptoms, signs, and imaging, and B-ALL remained in continuous remission. The report indicates that bridging treatment with blinatumomab in children with B-ALL complicated by IFD can rebuild the immune system and control the underlying disease in the presence of immunosuppression and severe fungal infection.
Child, Preschool ; Female ; Humans ; Infant ; Male ; Antibodies, Bispecific/therapeutic use* ; Invasive Fungal Infections/drug therapy* ; Precursor B-Cell Lymphoblastic Leukemia-Lymphoma/drug therapy* ; Remission Induction

Behcet's disease (BD) is a multisystemic chronic inflammatory disease. It is characterized by recurrent oral and genital ulcers, uveitis, skin lesions and other manifestations, including neurologic, vascular, joint, and gastrointestinal ulcers of variable severity. Recurrent aphthous ulcer (RAU) represents a very common, but poorly understood, mucosal disorder. If a patient of RAU without any other typical symptoms of BD has gastrointestinal symptoms, it is difficult to distinguish this RAU from true BD with gastrointestinal involvement. Because pathognomonic clinical features and tools are absent, the differential diagnosis of these two diseases relies on the characteristic clinical features and the judgement of an experienced physician. Sixty-five out of a total 960 RAU patients and forty-four of 556 BD patients with gastrointestinal symptoms between January 1996 and December 2003 participated in this study. All were evaluated with esophagogastroduodenoscopy and colonoscopy. Clinical, endoscopic and histopathologic findings were analyzed and ELISA tests were conducted to detect serum levels of ASCA and pANCA. No significant difference was found between the two groups. Differential diagnosis between RAU with gastrointestinal symptoms and BD with gastrointestinal involvement requires further prospective, large-scale study.
Adolescent ; Adult ; Aged ; Antibodies, Antineutrophil Cytoplasmic/blood ; Antibodies, Fungal/blood ; Behcet Syndrome/*diagnosis/immunology/pathology ; Comparative Study ; Diagnosis, Differential ; Endoscopy ; Female ; Gastrointestinal Diseases/*diagnosis/immunology/pathology ; Humans ; Male ; Middle Aged ; Saccharomyces cerevisiae/immunology ; Serologic Tests ; Stomatitis, Aphthous/*diagnosis/immunology/pathology