Preparation of monoclonal antibodies reactive with beta-1,2-linked oligomannosyl residues in the phosphomannan-protein complex of Candida albicans NIH B-792 strain.

1988 ◽  
Vol 34 (3) ◽  
pp. 539-543 ◽  
Author(s):  
M Tojo ◽  
N Shibata ◽  
M Kobayashi ◽  
T Mikami ◽  
M Suzuki ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Candida Albicans ◽  
Protein Complex ◽  
Protein Complexes ◽  
Myeloma Cell ◽  
Polyclonal Antiserum ◽  
Ascites Fluid ◽  
Serotype A ◽  
Mouse Myeloma Cell Line ◽  
Serotype B

Abstract Hybridomas obtained by fusing the spleen cells of BALB/c female mice hyperimmunized with heat-killed yeast-form cells of Candida albicans NIH B-792 strain and a mouse myeloma cell line, P3X63Ag8.653, produced antibodies to beta-1,2-linked oligomannosyl residues in the phosphomannan-protein complex of the parent cells. Most of these monoclonal antibodies were IgM, but about 10% of the hybridomas produced IgG1 immunoglobulins. Ascites fluid from BALB/c mice inoculated with an IgG1-producing hybridoma showed different precipitability with the phosphomannan-protein complexes of three representative C. albicans strains, with NIH B-792 (serotype B) greater than NIH A-207 (serotype A) greater than J-1012 (serotype A, formerly serotype C). In contrast, a rabbit polyclonal antiserum to C. albicans NIH B-792 cells was unable to distinguish these same complexes. This ascites fluid agglutinated the heat-killed cells of three Candida strains, but not those of three others or of Torulopsis glabrata IFO 0622. The other ascites fluids, containing antibodies of the IgM class, agglutinated cells from three C. albicans strains and also C. tropicalis IFO 0587 cells.

scholarly journals β-1,2-Mannosylation of Candida albicans Mannoproteins and Glycolipids Differs with Growth Temperature and Serotype

2002 ◽  
Vol 70 (9) ◽  
pp. 5274-5278 ◽  
Author(s):  
P. A. Trinel ◽  
T. Jouault ◽  
J. E. Cutler ◽  
D. Poulain
Keyword(s):  
Molecular Weight ◽  
Monoclonal Antibodies ◽  
Candida Albicans ◽  
Growth Temperature ◽  
Serotype A ◽  
Relative Molecular Weight ◽  
Serotype B

ABSTRACT Increasing the growth temperature from 28 to 37°C reduced the expression of β-1,2-oligomannoside epitopes on mannoproteins of Candida albicans serotypes A and B. In contrast, β-1,2-mannosylation of phospholipomannan (PLM) remained constant despite a slight decrease in the relative molecular weight (M r) of this compound. At all growth temperatures investigated, serotype A PLM displayed an M r and an antigenicity different from those of serotype B PLM when they were tested with a panel of monoclonal antibodies.

scholarly journals Production and Characterization of Monoclonal Antibodies to Psittacine Beak and Feather Disease Virus

1992 ◽  
Vol 4 (1) ◽  
pp. 13-18 ◽  
Author(s):  
Branson W. Ritchie ◽  
Frank D. Niagro ◽  
Kenneth S. Latimer ◽  
W. L. Steffens ◽  
Denise Pesti ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Myeloma Cell ◽  
Enzyme Linked Immunosorbent Assay ◽  
Spleen Cells ◽  
Myeloma Cell Line ◽  
Elisa System ◽  
Mouse Myeloma Cell Line ◽  
Feather Disease Virus ◽  
Mouse Myeloma

Monoclonal antibodies specific for the virus that causes psittacine beak and feather disease (PBFD) were produced by fusing spleen cells from mice immunized with purified concentrated PBFD virus with mouse myeloma cell line Sp2/0. The resulting hybridomas were tested for reactivity against whole purified virus by an enzyme-linked immunosorbent assay (ELISA) system. Four clones, designated 15H8, 8E3, 11G12, and 2C3, were subcloned by limiting dilution. Isotyping indicated that clone 15H8 was secreting IgG, whereas the remaining clones secreted IgM. The secreted immunoglobulins were characterized by reactivity against purified PBFD virus using immunoblotting procedures, by immunohistochemical staining of virus-induced lesions in infected tissues, and by inhibition of PBFD virus agglutination of cockatoo erythrocytes. Antibodies secreted by clones 15H8 and 8E3 had the strongest activity against purified whole virus. Only immunoglobulin secreted by the clone 15H8 could be used to detect viral antigen in infected tissues. None of the monoclonal antibodies had hemagglutination-inhibition activity.

scholarly journals Biological Activities of Naturally Occurring Antibodies Reactive with Candida albicans Mannan

2004 ◽  
Vol 72 (1) ◽  
pp. 209-218 ◽  
Author(s):  
Thomas R. Kozel ◽  
Randall S. MacGill ◽  
Ann Percival ◽  
Qing Zhou
Keyword(s):  
Candida Albicans ◽  
B Cells ◽  
Complement System ◽  
Biological Activities ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serotype A ◽  
Naturally Occurring ◽  
Serotype B ◽  
Opsonophagocytic Killing ◽  
The Complement System

ABSTRACT Sera from normal adult humans may contain high levels of antibody reactive with Candida albicans mannan. This study examined selected biological activities of such antibodies, focusing on sera that were collected from 34 donors and analyzed individually. The results showed that antimannan titers were normally distributed. Reactivity as determined by enzyme-linked immunosorbent assay with serotype A mannan generally paralleled reactivity with serotype B. Analysis of the kinetics for activation of the complement system and deposition of complement component 3 (C3) onto serotype A and serotype B cells showed a decrease in the lag time that occurred before the onset of rapid accumulation of C3 that correlated with increasing antimannan titers. In contrast, there was a decrease in the overall rate of accumulation of C3 on serotype A cells that was strongly correlated with increasing antibody titers; serotype B cells showed no such decrease. An evaluation of the contribution of mannan antibody to opsonophagocytic killing showed that mannan antibody in individual sera and antimannan immunoglobulin G (IgG) affinity purified from human plasma contributed to killing by neutrophils in a dose-dependent fashion in the absence of a functional complement system. However, affinity-purified antibody in very high concentrations was inhibitory to both complement-dependent and complement-independent opsonophagocytosis, and this finding suggests a prozone-like effect. In contrast, if the complement system was functional, antimannan IgG was not needed for opsonophagocytic killing. These results suggest that naturally occurring mannan antibodies and the complement system are functionally redundant for opsonophagocytic killing by neutrophils.

Production of Monoclonal Antibodies Specific to Clostridium botulinum Type B Neurotoxin

1995 ◽  
Vol 78 (2) ◽  
pp. 381-385 ◽  
Author(s):  
Charles W Noah ◽  
Sherilyn S Poteet ◽  
Nora C Ramos ◽  
John C Perez ◽  
Shyi Y Huang
Keyword(s):  
Monoclonal Antibodies ◽  
Clostridium Botulinum ◽  
Specific Binding ◽  
Myeloma Cell ◽  
Type B ◽  
Immunoblot Assay ◽  
Botulinum Type ◽  
Mouse Myeloma Cell Line ◽  
Pure Cultures ◽  
Botulinum Neurotoxins

Abstract Four monoclonal antibodies were produced for use in a rapid method to detect Clostridium botulinum type B neurotoxin. Cells of mouse myeloma cell line SP2/0 were fused with splenocytes of immunized BALB/c mice. An immunoblot assay of semipurified commercial neurotoxins of C. botulinum types A, B, C, D, E, and F was used to show specificity. All the monoclonal antibodies reacted with type B neurotoxin but did not cross-react with the other types. The monoclonal antibodies, separately and combined, did not neutralize the toxin in mice, and all showed specificity to the whole neurotoxin molecule and the heavy-chain component by immunoblot. No evidence of specific binding to the hemagglutinin molecule was noted. When tested against concentrated cultured supernatants of C. botulinum types A, B, E, and F, the 4 monoclonal antibodies reacted only against type B strains. They will be incorporated into a rapid assay with other specific monoclonal antibodies to detect C. botulinum neurotoxins from pure cultures or suspect foods.

Structural studies of mannans from Candida albicans (serotypes A and B), Candida parapsilosis, Candida stellatoidea, and Candida tropicalis

1967 ◽  
Vol 45 (19) ◽  
pp. 2205-2211 ◽  
Author(s):  
R. J. Yu ◽  
C. T. Bishop ◽  
F. P. Cooper ◽  
H. F. Hasenclever ◽  
F. Blank
Keyword(s):  
Candida Albicans ◽  
Candida Tropicalis ◽  
Candida Species ◽  
Candida Parapsilosis ◽  
Cross Reactivity ◽  
Structural Studies ◽  
Serotype A ◽  
Serotype B ◽  
Hydrolysis Of

Mannans have been isolated from cells of the following Candida species: C. albicans (serotype A), C. albicans (serotype B), C. parapsilosis, C. stellatoidea, and C. tropicalis. Methylation and hydrolysis of each mannan yielded the following methyl ethers of d-mannose (with only small variations in the relative amounts): 2,3,4,6-tetra-O-methyl-d-mannose, 3,4,6-tri-O-methyl-d-mannose (major), 2,3,4-tri-O-methyl-d-mannose (minor), 2,4,6-tri-O-methyl-d-mannose (minor), 3,4-di-O-methyl-d-mannose, and 3,5-di-O-methyl-d-mannose. The mannans therefore contained a predominance of 1 → 2 linkages in the linear portions, with smaller amounts of 1 → 6 and 1 → 3 linkages. Branching occurred through C-2 and C-6 of d-mannopyranose and d-mannofuranose units, and branches were terminated by d-mannopyranose units. The specific rotations of the mannans indicated that most of the glycosidic linkages were in the α configuration. The structural studies support the observation that the mannans are very similar serologically and show cross-reactivity in antisera to any of the parent organisms.

Structural analysis of phospho-d-mannan-protein complexes isolated from yeast and mold form cells of Candida albicans NIH A-207 serotype a strain

1989 ◽  
Vol 187 (2) ◽  
pp. 239-253 ◽  
Author(s):  
Nobuyuki Shibata ◽  
Shigeyuki Fukasawa ◽  
Hidemitsu Kobayashi ◽  
Minehiro Tojo ◽  
Toshio Yonezu ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Structural Analysis ◽  
Protein Complexes ◽  
Serotype A

scholarly journals Macrophage phagocytosis of Candida albicans. An in vitro study

1999 ◽  
Vol 13 (3) ◽  
pp. 233-238 ◽  
Author(s):  
Ilan WEINFELD ◽  
Esther Goldenberg BIRMAN ◽  
Claudete Rodrigues PAULA
Keyword(s):  
Candida Albicans ◽  
In Vitro Study ◽  
Neutral Red ◽  
Serotype A ◽  
Macrophage Phagocytosis ◽  
Serotype B ◽  
Vitro Model ◽  
Different Strains

Considering the role of macrophages in relation to fungi and the various utilized methodologies, the authors established an in vitro model to evaluate macrophage phagocytosis of Candida albicans. Activated macrophages were obtained from the peritoneal cavity of isogenic mice (A/Sn). Two different strains of Candida albicans serotype A and serotype B with different levels of pathogenicity in vivo and other similar characteristics were utilized in the study. Several microscopic fields containing about 200 macrophages were counted. The percentage of macrophages phagocytizing at least one viable or nonviable yeast cell determined an average number of phagocytized yeasts. Neutral red and fluorescein diacetate plus ethidium bromide were used for staining. It is possible to conclude that this is an efficient model related to the used methodology. The average number of yeasts in both strains were similar when inside macrophages, and there was a higher percentage of C. albicans serotype A phagocytosis, which was not experimentally pathogenic in vivo.

Use of monoclonal antibodies for radioimmunoassay of water buffalo milk progesterone

1987 ◽  
Vol 54 (4) ◽  
pp. 471-477 ◽  
Author(s):  
Rosanna Capparelli ◽  
Domenico Iannelli ◽  
Aldo Bordi
Keyword(s):  
Monoclonal Antibodies ◽  
Water Buffalo ◽  
Myeloma Cell ◽  
Buffalo Milk ◽  
Spleen Cells ◽  
Myeloma Cell Line ◽  
Mouse Myeloma Cell ◽  
Mouse Myeloma Cell Line ◽  
The Mean ◽  
Mouse Myeloma

SummaryIn order to standardize a radioimmunoassay of milk progesterone as a routine method for confirmation of oestrus and diagnosis of pregnancy in water buffalo, monoclonal antibodies against progesterone were produced. Hybridomas were prepared by fusing spleen cells from a Balb/c mouse immunized with progesterone 11α-hemisuccinate–bovine serum albumin conjugate with the mouse myeloma cell line NS-1. Thirty wells out of 94 secreted anti-progesterone antibodies. Of the ten independent hybridomas derived, one (AF65) was suitable for the quantification of milk progesterone by radioimmunoassay. The tracer used in the assay was progesterone-11α-hemisuccinate ([2-125I]iodohistamine). The sensitivity of the assay was 50 pg/tube. The mean progesterone concentration at oestrus was 0·8±0·2 ng/ml increasing to 8·5±0·8 ng/ml 24 d later in pregnant animals.

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