Viswanath Kurup - Academia.edu (original) (raw)

Papers by Viswanath Kurup

Research paper thumbnail of Aspergillus antigens associated with allergic bronchopulmonary aspergillosis

Frontiers in Bioscience, 2003

Research paper thumbnail of Characterization of a major latex allergen associated with hypersensitivity in spina bifida patients

The Journal of Immunology

Hypersensitivity to latex proteins present in health care products, particularly in latex gloves,... more Hypersensitivity to latex proteins present in health care products, particularly in latex gloves, has emerged as an important public health problem in recent years. Most of these latex allergens have not been purified or characterized. Here we report the purification and characterization of a 23-kDa latex polypeptide from nonammoniated natural rubber latex. This purified 23-kDa polypeptide reacted with IgE from 13 of 17 (76%) latex-allergic spina bifida patients, but from only 1 of 5 health care workers with latex allergy. Furthermore, all the sera of patients with anaphylaxis to latex showed reactivity with the 23-kDa polypeptide. This allergen also induced significant lymphocyte proliferation by PBMC from spina bifida patients, but not from health care workers. The 90 amino acid residues sequenced from the 23-kDa polypeptide showed a 45.6% homology with a previously reported latex protein, rubber elongation factor. By using a mAb LAM-1, produced against the 23-kDa allergen, we cou...

Research paper thumbnail of IgE from latex-allergic patients binds to cloned and expressed B cell epitopes of prohevein

The Journal of Immunology

Prohevein is one of the major allergens associated with latex allergy. In the present study, we i... more Prohevein is one of the major allergens associated with latex allergy. In the present study, we identified IgE binding regions of prohevein, and expressed multiple IgE binding epitopes by selective cloning. These truncated polypeptides were then used to demonstrate IgE in the sera of patients. Decapeptides of prohevein were synthesized on derivatized cellulose membrane with an offset of one amino acid. The IgE reactivity of these linear peptides was evaluated separately using pooled sera from latex-allergic health care workers (HCW) and spina bifida (SB) patients. A total of 10 IgE binding epitopes representing unique as well as shared epitopes from both the N- and C-domains of the prohevein were identified. Recombinant polypeptides were constructed based on the identified epitopes, and clones carrying DNA fragments were overexpressed. These recombinant peptides were evaluated for IgE binding with sera from HCW, SB, and normal individuals. Recombinant prohevein, hevein, and the C-do...

Research paper thumbnail of Immunochemical characterization of Aspergillus fumigatus antigens

Infection and Immunity, 1983

Culture filtrate antigens of Aspergillus fumigatus Ag 534 were purified by preparative isoelectri... more Culture filtrate antigens of Aspergillus fumigatus Ag 534 were purified by preparative isoelectric focusing and affinity chromatography. One of the pooled antigen fractions from the preparative isoelectric focusing step (pool 2) was passed through a concanavalin A column and yielded two components, designated antigens IIa and IIb. Antigen IIb reacted more strongly than antigen IIa with all of the aspergilloma and allergic bronchopulmonary aspergillosis sera tested by enzyme-linked immunosorbent assay. The glycoprotein nature of antigen IIb was shown by the concanavalin A binding properties and staining reactions of the components.

Research paper thumbnail of Immunoreactivity of antigen extracts of Aspergillus fumigatus isolated from different sources

Revista iberoamericana de micología, 1997

Allergic bronchopulmonary aspergillosis (ABPA) is the result of hypersensitivity to Aspergillus a... more Allergic bronchopulmonary aspergillosis (ABPA) is the result of hypersensitivity to Aspergillus antigens in patients with long-standing atopic asthma. In the present study mycelial and culture filtrate antigens from Aspergillus fumigatus cultures isolated from diverse sources were tested against sera of 10 ABPA patients and 10 control individuals by an ELISA methodology. The results indicate higher antibody reactivity against both antigens in the sera of ABPA patients, while culture filtrate antigens also gave non-specific reactivity with control sera. Mycelial extracts, in general, were useful in the diagnosis of ABPA.

Research paper thumbnail of Immunochemical reactivity of Aspergillus fumigatus antigens from different sources

Revista iberoamericana de micología, 1997

We have studied the immunochemical and biochemical differences in 12 Aspergillus fumigatus strain... more We have studied the immunochemical and biochemical differences in 12 Aspergillus fumigatus strains isolated from different sources. The enzymatic activity of all these strains were studied by a rapid enzyme detection method (API-ZYM). One of 12 strains studied produced alkaline phosphatase, while two produced chymotrypsin, and three produced trypsin. By SDS-PAGE we studied proteins present in the antigen extracts from all 12 strains. Several of the protein bands were unique and may be used to differentiate the strains. One such protein is the 58 kDa band present in the mycelial extract and the 33 kDa in the culture filtrate. By crossed immunoelectrophoresis, differentiation of the strains isolated from cystic fibrosis patients can be made based on a few specific precipitin arcs developed against anti-Aspergillus rabbit serum.

Research paper thumbnail of Allergenic fungi and actinomycetes in smoking materials and their health implications

Mycopathologia, 1983

Street marijuana, commercial cigarettes and pipe tobaccos were studied for the presence of fungi ... more Street marijuana, commercial cigarettes and pipe tobaccos were studied for the presence of fungi and actinomycetes associated with hypersensitivity pneumonitis. Aspergillus species and thermophilic actinomycetes were isolated from the smoking materials. In addition, Aspergillusfumigatus spores were isolated from marijuana smoke, indicating the potential hazard involved in developing serious disease. Precipitin antibodies against fungi, particularly Aspergillus, showed a higher prevalence in marijuana smokers, whereas only very few cigarette smokers and nonsmokers demonstrated antibodies to fungi. Cigarette smokers and nonsmokers showed more or less similar prevelance of antibodies against thermophilic actinomycetes.

Research paper thumbnail of 852 C-terminal cysteine residues determine the IgE binding of Aspergillus fumigatus allergen 2 (Asp f 2)

Journal of Allergy and Clinical Immunology, 2000

Research paper thumbnail of Enhanced expression of CTLA-4 and concurrent downregulation of CD28 on lung cells of mice exposed to Aspergillus antigen

Journal of Allergy and Clinical Immunology, 2005

RATIONALE: Although IgE is implicated in viral immunity, its role in parvovirus B19 immunity has ... more RATIONALE: Although IgE is implicated in viral immunity, its role in parvovirus B19 immunity has not been studied. METHODS: Serum immmunoglobulins, IgG and IgE anti-Parvovirus B19, blood lymphocyte numbers, and epsilon and cytokine specific mRNA, were determined in pediatric patients with normal serum IgA levels (IgA+), and selective IgA deficiency (IgA-) on day 0 (diagnosis), 14 and 3 yrs after recovery (nephelometry, Western blot, flow cytometry, RT-PCR). RESULTS: All immunoglobulin levels of both patients were within normal ranges on days 0-3 years, excluding IgG 1 and IgA in the IgA-patient, which were elevated and negative, respectively. Both patients made IgE (and IgG) anti-parvovirus B19. Excluding CD8+CD60+ T cells, T, B and NK lymphocyte subset distributions were normal at all times. In both patients, CD8+CD60+ T cell numbers did not increase on day 0, but increased on day 14 (>5 fold). At 3 years, they returned to normal in the IgA+ patient, but remained high in the IgA-patient. On days 0-3 years, PBMC of both patients expressed epsilon and IFN-␣ specific mRNA.

Research paper thumbnail of Aspergillus antigens associated with allergic bronchopulmonary aspergillosis

Frontiers in Bioscience, 2003

Research paper thumbnail of Aspergillose bronchopulmonaire allergique chez l'enfant

Archives de Pédiatrie, 1999

Research paper thumbnail of Indoor environment and mold-related illness

Research paper thumbnail of Latex-induced early inflammatory responses in sensitized mice

Research paper thumbnail of Monoclonal antibodies bind identically to both spores and hyphae of Aspergillus fumigatus

Clinical & Experimental Allergy, May 1, 1992

Immunoelectron microscopy (IEM) was used to determine the binding of six monoclonal antibodies (M... more Immunoelectron microscopy (IEM) was used to determine the binding of six monoclonal antibodies (MoAbs) produced against Aspergillus fumigatus antigens present on or within the conidia and hyphae ofthe fungus. Antigen-antibody complexes were demonstrated in EM using labelled colloidal gold particles (15 nm). Three out of 6 MoAbs (C9. FI2 and HIO) reacted only with the cytoplasmic components of A. fumigatus while the remaining three (B12, F6G5 and D6E6) showed reactivity to both cytoplasm and cell wall of the conidia and hyphae. The results indicate that IEM is of considerable value in determining and seleeting monoclonal antibodies having specific reactivity with diverse antigenic components.

Research paper thumbnail of A murine model of allergic bronchopulmonary aspergillosis with elevated eosinophils and IgE

The Journal of Immunology

A model of allergic bronchopulmonary aspergillosis was developed by exposing BALB/c mice to Asper... more A model of allergic bronchopulmonary aspergillosis was developed by exposing BALB/c mice to Aspergillus fumigatus (AF) Ag. Animals immunized intranasally (i.n.) with soluble AF Ag produced low levels of serum IgE compared to animals given alum precipitated AF Ag i.p. The latter treatment also produced higher levels of serum IgG1 and AF-specific IgG1 than soluble AF given i.p. or i.n.. Blood and lung eosinophilia was detected in mice repeatedly exposed to AF by i.n. but not in the groups injected i.p. Particulate AF Ag-induced striking blood and lung eosinophilia and elevated levels of serum IgE in mice preexposed to AF Ag. The results indicate that route of inoculation and physical nature of Ag determine the immune response and can be manipulated to obtain enhanced IgE, eosinophils, or both in the animal model.

Research paper thumbnail of Aspergillus antigen-induced eosinophil differentiation in a murine model

Infection and Immunity, 1992

Eosinophilia is a prominent feature of the cellular response in allergic and parasitic diseases. ... more Eosinophilia is a prominent feature of the cellular response in allergic and parasitic diseases. Allergic bronchopulmonary aspergillosis due to colonization of the lungs of some asthmatics with Aspergillus fumigatus is characterized by high levels of serum immunoglobulin E and peripheral blood (PB) and lung eosinophilia. This study investigates the role of eosinophils in the pathogenesis of allergic bronchopulmonary aspergillosis by using a mouse model. BALB/c mice were immunized intranasally and intraperitoneally with A. fumigatus antigens (Ag), and the eosinophils in PB and bone marrow (BM) were enumerated. Eosinophilopoiesis in BM cultures was studied in the presence of murine recombinant interleukin-5 (mrIL-5) and supernatants from pokeweed mitogen-stimulated spleen cells as the source of eosinophil differentiation factors. Eosinophils were quantitated by direct counting and by estimating eosinophil peroxidase activity. The results indicate that the percentage of eosinophils in ...

Research paper thumbnail of Conformational and Linear B-Cell Epitopes of Asp f 2, a Major Allergen ofAspergillus fumigatus, Bind Differently to Immunoglobulin E Antibody in the Sera of Allergic Bronchopulmonary Aspergillosis Patients

Infection and Immunity, 1999

ABSTRACTAsp f 2 is a majorAspergillus fumigatusallergen involved in allergic bronchopulmonary asp... more ABSTRACTAsp f 2 is a majorAspergillus fumigatusallergen involved in allergic bronchopulmonary aspergillosis. Knowledge of the B-cell epitopes may contribute to the understanding of immunoregulation and immunodiagnosis. To elucidate the immunoglobulin E (IgE) binding epitopes in the linear sequence of Asp f 2, we synthesized decamer peptides spanning the whole molecule of Asp f 2 on derivatized cellulose membranes and evaluated IgE binding in ABPA patient and control sera. Peptides three to five amino acids long were synthesized based on amino acid sequences within the IgE binding regions and evaluated for the specificity of epitope antibody interactions. Nine IgE binding regions were recognized in this protein of 268 amino acid residues. Of the nine epitopes, seven (ATQRRQI, RKYFG, HWR, YTTRR, DHFAD, ALEAYA, and THEGGQ) are present in the hydrophilic regions of Asp f 2. Immunologic evaluation of the three recombinant fragments, Asp f 2A encompassing the N-terminal epitope region, As...

Research paper thumbnail of Immunological Characterization of Asp f 2, a Major Allergen fromAspergillus fumigatusAssociated with Allergic Bronchopulmonary Aspergillosis

Infection and Immunity, 1998

ABSTRACTThe 37-kDa recombinant protein Asp f 2, encoding an allergen ofAspergillus fumigatus, was... more ABSTRACTThe 37-kDa recombinant protein Asp f 2, encoding an allergen ofAspergillus fumigatus, was expressed in a prokaryotic expression system and immunologically evaluated for its functional and structural properties. The open reading frame for a 310-amino-acid-long protein was shown to encode a signal peptide of 31 amino acids. A native 37-kDa culture filtrate protein and a 55-kDa mycelial glycoprotein (gp55) exhibited complete N-terminal sequence homology to Asp f 2. A GenBank search for homologous proteins revealed 60 and 44% sequence homologies to the cytosolic protein ASPND1 fromAspergillus nidulansand fibrinogen binding protein fromCandida albicans, respectively. The glycosylation sites and cysteine molecules are conserved in all the three proteins. The extracellular matrix protein laminin showed a dose-dependent interaction with Asp f 2. This protein, expressed as a major cell-associated protein within 24 h of in vitro fungal culture, comprises 20 to 40% of total fungal prot...

Research paper thumbnail of Antibody response to low-molecular-weight antigens of Aspergillus fumigatus in allergic bronchopulmonary aspergillosis

Journal of Clinical Microbiology, 1989

Sera from patients with allergic bronchopulmonary aspergillosis (ABPA) or aspergilloma and normal... more Sera from patients with allergic bronchopulmonary aspergillosis (ABPA) or aspergilloma and normal sera were analyzed for specific antibodies by Western (immuno-) blotting with Aspergillus fumigatus antigens transferred electrophoretically onto polyvinylidene difluoride membranes. Western blot analysis demonstrated consistent reactivity of low-molecular-weight A. fumigatus antigens against ABPA sera but not against uncomplicated aspergilloma or normal sera. None of these low-molecular-weight components had any lectin-binding activity. Sera from patients with aspergilloma, however, frequently reacted with high-molecular-weight components of A. fumigatus. The majority of these high-molecular-weight antigenic components demonstrated concanavalin A-binding activity. The low-molecular-weight bands were discernible in Western blots with sera from all ABPA patients irrespective of disease activities, such as relapse, flare, or treatment. Antibodies detected by methods such as immunodiffusio...

Research paper thumbnail of IgE and eosinophil regulation in a murine model of allergic aspergillosis

Journal of Leukocyte Biology, 1994

Exposure of BALB/c mice to Aspergillus fumigatus (Af), the antigen responsible for causing allerg... more Exposure of BALB/c mice to Aspergillus fumigatus (Af), the antigen responsible for causing allergic bronchopulmonary aspergillosis in humans, caused elevated levels of serum immunoglobulin E (IgE) and peripheral blood and lung eosinophilia similar to that observed in the human disease. We have investigated the role of interleukin-4 (IL-4), IL-5 and interferon-γ in regulating IgE and eosinophilia in the mouse model. Animals were immunized by intraperitoneal injections of soluble Af antigens adsorbed to alum. These animals developed elevated IgE and Af specific IgG1 and were then treated with anticytokine monoclonal antibodies before the final exposure to particulate Af antigens by the intranasal route. The results showed that anti-IL-5 abrogated eosinophilia in mice, while those treated with anti-IL-4 retained the same or reduced IgE levels compared to pretreatment levels. All anti–IL-5, anti–IFN-γ, and control antibody–treated animals showed enhanced IgE levels. Anti–IFN-γ treatment...

Research paper thumbnail of Aspergillus antigens associated with allergic bronchopulmonary aspergillosis

Frontiers in Bioscience, 2003

Research paper thumbnail of Characterization of a major latex allergen associated with hypersensitivity in spina bifida patients

The Journal of Immunology

Hypersensitivity to latex proteins present in health care products, particularly in latex gloves,... more Hypersensitivity to latex proteins present in health care products, particularly in latex gloves, has emerged as an important public health problem in recent years. Most of these latex allergens have not been purified or characterized. Here we report the purification and characterization of a 23-kDa latex polypeptide from nonammoniated natural rubber latex. This purified 23-kDa polypeptide reacted with IgE from 13 of 17 (76%) latex-allergic spina bifida patients, but from only 1 of 5 health care workers with latex allergy. Furthermore, all the sera of patients with anaphylaxis to latex showed reactivity with the 23-kDa polypeptide. This allergen also induced significant lymphocyte proliferation by PBMC from spina bifida patients, but not from health care workers. The 90 amino acid residues sequenced from the 23-kDa polypeptide showed a 45.6% homology with a previously reported latex protein, rubber elongation factor. By using a mAb LAM-1, produced against the 23-kDa allergen, we cou...

Research paper thumbnail of IgE from latex-allergic patients binds to cloned and expressed B cell epitopes of prohevein

The Journal of Immunology

Prohevein is one of the major allergens associated with latex allergy. In the present study, we i... more Prohevein is one of the major allergens associated with latex allergy. In the present study, we identified IgE binding regions of prohevein, and expressed multiple IgE binding epitopes by selective cloning. These truncated polypeptides were then used to demonstrate IgE in the sera of patients. Decapeptides of prohevein were synthesized on derivatized cellulose membrane with an offset of one amino acid. The IgE reactivity of these linear peptides was evaluated separately using pooled sera from latex-allergic health care workers (HCW) and spina bifida (SB) patients. A total of 10 IgE binding epitopes representing unique as well as shared epitopes from both the N- and C-domains of the prohevein were identified. Recombinant polypeptides were constructed based on the identified epitopes, and clones carrying DNA fragments were overexpressed. These recombinant peptides were evaluated for IgE binding with sera from HCW, SB, and normal individuals. Recombinant prohevein, hevein, and the C-do...

Research paper thumbnail of Immunochemical characterization of Aspergillus fumigatus antigens

Infection and Immunity, 1983

Culture filtrate antigens of Aspergillus fumigatus Ag 534 were purified by preparative isoelectri... more Culture filtrate antigens of Aspergillus fumigatus Ag 534 were purified by preparative isoelectric focusing and affinity chromatography. One of the pooled antigen fractions from the preparative isoelectric focusing step (pool 2) was passed through a concanavalin A column and yielded two components, designated antigens IIa and IIb. Antigen IIb reacted more strongly than antigen IIa with all of the aspergilloma and allergic bronchopulmonary aspergillosis sera tested by enzyme-linked immunosorbent assay. The glycoprotein nature of antigen IIb was shown by the concanavalin A binding properties and staining reactions of the components.

Research paper thumbnail of Immunoreactivity of antigen extracts of Aspergillus fumigatus isolated from different sources

Revista iberoamericana de micología, 1997

Allergic bronchopulmonary aspergillosis (ABPA) is the result of hypersensitivity to Aspergillus a... more Allergic bronchopulmonary aspergillosis (ABPA) is the result of hypersensitivity to Aspergillus antigens in patients with long-standing atopic asthma. In the present study mycelial and culture filtrate antigens from Aspergillus fumigatus cultures isolated from diverse sources were tested against sera of 10 ABPA patients and 10 control individuals by an ELISA methodology. The results indicate higher antibody reactivity against both antigens in the sera of ABPA patients, while culture filtrate antigens also gave non-specific reactivity with control sera. Mycelial extracts, in general, were useful in the diagnosis of ABPA.

Research paper thumbnail of Immunochemical reactivity of Aspergillus fumigatus antigens from different sources

Revista iberoamericana de micología, 1997

We have studied the immunochemical and biochemical differences in 12 Aspergillus fumigatus strain... more We have studied the immunochemical and biochemical differences in 12 Aspergillus fumigatus strains isolated from different sources. The enzymatic activity of all these strains were studied by a rapid enzyme detection method (API-ZYM). One of 12 strains studied produced alkaline phosphatase, while two produced chymotrypsin, and three produced trypsin. By SDS-PAGE we studied proteins present in the antigen extracts from all 12 strains. Several of the protein bands were unique and may be used to differentiate the strains. One such protein is the 58 kDa band present in the mycelial extract and the 33 kDa in the culture filtrate. By crossed immunoelectrophoresis, differentiation of the strains isolated from cystic fibrosis patients can be made based on a few specific precipitin arcs developed against anti-Aspergillus rabbit serum.

Research paper thumbnail of Allergenic fungi and actinomycetes in smoking materials and their health implications

Mycopathologia, 1983

Street marijuana, commercial cigarettes and pipe tobaccos were studied for the presence of fungi ... more Street marijuana, commercial cigarettes and pipe tobaccos were studied for the presence of fungi and actinomycetes associated with hypersensitivity pneumonitis. Aspergillus species and thermophilic actinomycetes were isolated from the smoking materials. In addition, Aspergillusfumigatus spores were isolated from marijuana smoke, indicating the potential hazard involved in developing serious disease. Precipitin antibodies against fungi, particularly Aspergillus, showed a higher prevalence in marijuana smokers, whereas only very few cigarette smokers and nonsmokers demonstrated antibodies to fungi. Cigarette smokers and nonsmokers showed more or less similar prevelance of antibodies against thermophilic actinomycetes.

Research paper thumbnail of 852 C-terminal cysteine residues determine the IgE binding of Aspergillus fumigatus allergen 2 (Asp f 2)

Journal of Allergy and Clinical Immunology, 2000

Research paper thumbnail of Enhanced expression of CTLA-4 and concurrent downregulation of CD28 on lung cells of mice exposed to Aspergillus antigen

Journal of Allergy and Clinical Immunology, 2005

RATIONALE: Although IgE is implicated in viral immunity, its role in parvovirus B19 immunity has ... more RATIONALE: Although IgE is implicated in viral immunity, its role in parvovirus B19 immunity has not been studied. METHODS: Serum immmunoglobulins, IgG and IgE anti-Parvovirus B19, blood lymphocyte numbers, and epsilon and cytokine specific mRNA, were determined in pediatric patients with normal serum IgA levels (IgA+), and selective IgA deficiency (IgA-) on day 0 (diagnosis), 14 and 3 yrs after recovery (nephelometry, Western blot, flow cytometry, RT-PCR). RESULTS: All immunoglobulin levels of both patients were within normal ranges on days 0-3 years, excluding IgG 1 and IgA in the IgA-patient, which were elevated and negative, respectively. Both patients made IgE (and IgG) anti-parvovirus B19. Excluding CD8+CD60+ T cells, T, B and NK lymphocyte subset distributions were normal at all times. In both patients, CD8+CD60+ T cell numbers did not increase on day 0, but increased on day 14 (>5 fold). At 3 years, they returned to normal in the IgA+ patient, but remained high in the IgA-patient. On days 0-3 years, PBMC of both patients expressed epsilon and IFN-␣ specific mRNA.

Research paper thumbnail of Aspergillus antigens associated with allergic bronchopulmonary aspergillosis

Frontiers in Bioscience, 2003

Research paper thumbnail of Aspergillose bronchopulmonaire allergique chez l'enfant

Archives de Pédiatrie, 1999

Research paper thumbnail of Indoor environment and mold-related illness

Research paper thumbnail of Latex-induced early inflammatory responses in sensitized mice

Research paper thumbnail of Monoclonal antibodies bind identically to both spores and hyphae of Aspergillus fumigatus

Clinical & Experimental Allergy, May 1, 1992

Immunoelectron microscopy (IEM) was used to determine the binding of six monoclonal antibodies (M... more Immunoelectron microscopy (IEM) was used to determine the binding of six monoclonal antibodies (MoAbs) produced against Aspergillus fumigatus antigens present on or within the conidia and hyphae ofthe fungus. Antigen-antibody complexes were demonstrated in EM using labelled colloidal gold particles (15 nm). Three out of 6 MoAbs (C9. FI2 and HIO) reacted only with the cytoplasmic components of A. fumigatus while the remaining three (B12, F6G5 and D6E6) showed reactivity to both cytoplasm and cell wall of the conidia and hyphae. The results indicate that IEM is of considerable value in determining and seleeting monoclonal antibodies having specific reactivity with diverse antigenic components.

Research paper thumbnail of A murine model of allergic bronchopulmonary aspergillosis with elevated eosinophils and IgE

The Journal of Immunology

A model of allergic bronchopulmonary aspergillosis was developed by exposing BALB/c mice to Asper... more A model of allergic bronchopulmonary aspergillosis was developed by exposing BALB/c mice to Aspergillus fumigatus (AF) Ag. Animals immunized intranasally (i.n.) with soluble AF Ag produced low levels of serum IgE compared to animals given alum precipitated AF Ag i.p. The latter treatment also produced higher levels of serum IgG1 and AF-specific IgG1 than soluble AF given i.p. or i.n.. Blood and lung eosinophilia was detected in mice repeatedly exposed to AF by i.n. but not in the groups injected i.p. Particulate AF Ag-induced striking blood and lung eosinophilia and elevated levels of serum IgE in mice preexposed to AF Ag. The results indicate that route of inoculation and physical nature of Ag determine the immune response and can be manipulated to obtain enhanced IgE, eosinophils, or both in the animal model.

Research paper thumbnail of Aspergillus antigen-induced eosinophil differentiation in a murine model

Infection and Immunity, 1992

Eosinophilia is a prominent feature of the cellular response in allergic and parasitic diseases. ... more Eosinophilia is a prominent feature of the cellular response in allergic and parasitic diseases. Allergic bronchopulmonary aspergillosis due to colonization of the lungs of some asthmatics with Aspergillus fumigatus is characterized by high levels of serum immunoglobulin E and peripheral blood (PB) and lung eosinophilia. This study investigates the role of eosinophils in the pathogenesis of allergic bronchopulmonary aspergillosis by using a mouse model. BALB/c mice were immunized intranasally and intraperitoneally with A. fumigatus antigens (Ag), and the eosinophils in PB and bone marrow (BM) were enumerated. Eosinophilopoiesis in BM cultures was studied in the presence of murine recombinant interleukin-5 (mrIL-5) and supernatants from pokeweed mitogen-stimulated spleen cells as the source of eosinophil differentiation factors. Eosinophils were quantitated by direct counting and by estimating eosinophil peroxidase activity. The results indicate that the percentage of eosinophils in ...

Research paper thumbnail of Conformational and Linear B-Cell Epitopes of Asp f 2, a Major Allergen ofAspergillus fumigatus, Bind Differently to Immunoglobulin E Antibody in the Sera of Allergic Bronchopulmonary Aspergillosis Patients

Infection and Immunity, 1999

ABSTRACTAsp f 2 is a majorAspergillus fumigatusallergen involved in allergic bronchopulmonary asp... more ABSTRACTAsp f 2 is a majorAspergillus fumigatusallergen involved in allergic bronchopulmonary aspergillosis. Knowledge of the B-cell epitopes may contribute to the understanding of immunoregulation and immunodiagnosis. To elucidate the immunoglobulin E (IgE) binding epitopes in the linear sequence of Asp f 2, we synthesized decamer peptides spanning the whole molecule of Asp f 2 on derivatized cellulose membranes and evaluated IgE binding in ABPA patient and control sera. Peptides three to five amino acids long were synthesized based on amino acid sequences within the IgE binding regions and evaluated for the specificity of epitope antibody interactions. Nine IgE binding regions were recognized in this protein of 268 amino acid residues. Of the nine epitopes, seven (ATQRRQI, RKYFG, HWR, YTTRR, DHFAD, ALEAYA, and THEGGQ) are present in the hydrophilic regions of Asp f 2. Immunologic evaluation of the three recombinant fragments, Asp f 2A encompassing the N-terminal epitope region, As...

Research paper thumbnail of Immunological Characterization of Asp f 2, a Major Allergen fromAspergillus fumigatusAssociated with Allergic Bronchopulmonary Aspergillosis

Infection and Immunity, 1998

ABSTRACTThe 37-kDa recombinant protein Asp f 2, encoding an allergen ofAspergillus fumigatus, was... more ABSTRACTThe 37-kDa recombinant protein Asp f 2, encoding an allergen ofAspergillus fumigatus, was expressed in a prokaryotic expression system and immunologically evaluated for its functional and structural properties. The open reading frame for a 310-amino-acid-long protein was shown to encode a signal peptide of 31 amino acids. A native 37-kDa culture filtrate protein and a 55-kDa mycelial glycoprotein (gp55) exhibited complete N-terminal sequence homology to Asp f 2. A GenBank search for homologous proteins revealed 60 and 44% sequence homologies to the cytosolic protein ASPND1 fromAspergillus nidulansand fibrinogen binding protein fromCandida albicans, respectively. The glycosylation sites and cysteine molecules are conserved in all the three proteins. The extracellular matrix protein laminin showed a dose-dependent interaction with Asp f 2. This protein, expressed as a major cell-associated protein within 24 h of in vitro fungal culture, comprises 20 to 40% of total fungal prot...

Research paper thumbnail of Antibody response to low-molecular-weight antigens of Aspergillus fumigatus in allergic bronchopulmonary aspergillosis

Journal of Clinical Microbiology, 1989

Sera from patients with allergic bronchopulmonary aspergillosis (ABPA) or aspergilloma and normal... more Sera from patients with allergic bronchopulmonary aspergillosis (ABPA) or aspergilloma and normal sera were analyzed for specific antibodies by Western (immuno-) blotting with Aspergillus fumigatus antigens transferred electrophoretically onto polyvinylidene difluoride membranes. Western blot analysis demonstrated consistent reactivity of low-molecular-weight A. fumigatus antigens against ABPA sera but not against uncomplicated aspergilloma or normal sera. None of these low-molecular-weight components had any lectin-binding activity. Sera from patients with aspergilloma, however, frequently reacted with high-molecular-weight components of A. fumigatus. The majority of these high-molecular-weight antigenic components demonstrated concanavalin A-binding activity. The low-molecular-weight bands were discernible in Western blots with sera from all ABPA patients irrespective of disease activities, such as relapse, flare, or treatment. Antibodies detected by methods such as immunodiffusio...

Research paper thumbnail of IgE and eosinophil regulation in a murine model of allergic aspergillosis

Journal of Leukocyte Biology, 1994

Exposure of BALB/c mice to Aspergillus fumigatus (Af), the antigen responsible for causing allerg... more Exposure of BALB/c mice to Aspergillus fumigatus (Af), the antigen responsible for causing allergic bronchopulmonary aspergillosis in humans, caused elevated levels of serum immunoglobulin E (IgE) and peripheral blood and lung eosinophilia similar to that observed in the human disease. We have investigated the role of interleukin-4 (IL-4), IL-5 and interferon-γ in regulating IgE and eosinophilia in the mouse model. Animals were immunized by intraperitoneal injections of soluble Af antigens adsorbed to alum. These animals developed elevated IgE and Af specific IgG1 and were then treated with anticytokine monoclonal antibodies before the final exposure to particulate Af antigens by the intranasal route. The results showed that anti-IL-5 abrogated eosinophilia in mice, while those treated with anti-IL-4 retained the same or reduced IgE levels compared to pretreatment levels. All anti–IL-5, anti–IFN-γ, and control antibody–treated animals showed enhanced IgE levels. Anti–IFN-γ treatment...