scholarly journals Protective and nonprotective monoclonal antibodies to Cryptococcus neoformans originating from one B cell.

1995 ◽  
Vol 181 (1) ◽  
pp. 405-409 ◽  
Author(s):  
J Mukherjee ◽  
G Nussbaum ◽  
M D Scharff ◽  
A Casadevall
Keyword(s):  
Monoclonal Antibodies ◽  
B Cell ◽  
Cryptococcus Neoformans ◽  
Somatic Mutations ◽  
Capsular Polysaccharide ◽  
Important Determinant ◽  
Protective Efficacy ◽  
Immunoglobulin M ◽  
The Other ◽  
Pathogenic Yeast

Two immunoglobulin M monoclonal antibodies (mAbs) derived from the same B cell recognize different epitopes on the capsular polysaccharide of the pathogenic yeast, Cryptococcus neoformans. Their respective epitopes are located in spatially distinct regions of the capsule. Passive administration of one mAb prolonged survival whereas the other mAb did not. The results indicate that specificity is an important determinant of antibody efficacy against C. neoformans and that somatic mutations occurring during the antibody response can affect the protective efficacy of antibodies to C. neoformans.

scholarly journals Protective and Nonprotective Human Immunoglobulin M Monoclonal Antibodies to Cryptococcus neoformans Glucuronoxylomannan Manifest Different Specificities and Gene Use Profiles

2004 ◽  
Vol 72 (8) ◽  
pp. 4810-4818 ◽  
Author(s):  
Robert W. Maitta ◽  
Kausik Datta ◽  
Qing Chang ◽  
Robin X. Luo ◽  
Bradley Witover ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Sequence Analysis ◽  
Cryptococcus Neoformans ◽  
Somatic Mutations ◽  
Capsular Polysaccharide ◽  
Germ Line ◽  
Immunoglobulin M ◽  
Human Immunoglobulin ◽  
Human Antibodies ◽  
Line Configuration

ABSTRACT The features of protective murine antibodies to the Cryptococcus neoformans capsular polysaccharide glucuronoxylomannan (GXM) have been rigorously investigated; however, the characteristics of protective human antibodies to GXM have not been defined. We produced monoclonal antibodies (MAbs) from XenoMouse mice (transgenic mice that express human immunoglobulin M [IgM], IgG2, and κ) which were immunized with a C. neoformans serotype D strain 24067 GXM-diphtheria toxoid conjugate. This study reports the specificity and efficacy of three human IgM MAbs, G14, G15, and G19, generated from these mice. Each MAb was specific for GXM, but G14 and G19 had different specificity based on their binding to serotype A strain H99 and SB4 GXMs, to which G15 did not bind. Nucleic acid sequence analysis revealed that G15 uses VH3-64 in the germ line configuration. G14 and G19 use VH6-1, which has somatic mutations. All of the MAbs use Vκ DPK22/A27. Studies of MAb efficacy in BALB/c mice showed that administration of 0.1 mg, but not 1 or 0.01 mg, of G15 prolonged survival against lethal C. neoformans strain 24067 challenge, whereas G14 and G19 were not protective at any dose. This panel of MAbs illustrates that serotype D GXM has epitopes that elicit human antibodies that can be either protective or nonprotective. Our findings suggest that VH gene use may influence GXM specificity and efficacy, and they provide insights into the possible contribution that VH gene use may have in resistance and susceptibility to cryptococcosis.

Comparative gene genealogical analyses of strains of serotype AD identify recombination in populations of serotypes A and D in the human pathogenic yeast Cryptococcus neoformans

Microbiology ◽  
2003 ◽  
Vol 149 (8) ◽  
pp. 2147-2154 ◽  
Author(s):  
Jianping Xu ◽  
Thomas G. Mitchell
Keyword(s):  
Monoclonal Antibodies ◽  
Cryptococcus Neoformans ◽  
Natural Populations ◽  
The Other ◽  
Pathogenic Yeast ◽  
Gene Genealogy ◽  
Serotype A ◽  
The World ◽  
Significant Divergence ◽  
Allelic Group

Cryptococcus neoformans is a major pathogen of humans throughout the world. Using commercial monoclonal antibodies to capsular epitopes, strains of C. neoformans manifest five serotypes: A, B, C, D and AD. Previous studies demonstrated significant divergence among serotypes A, B, C and D, which are typically haploid. In contrast, most strains of serotype AD are diploid or aneuploid and result from recent hybridization between strains of serotypes A and D. Whether serotypes A, B, C and D represent strictly asexual lineages is not known. Using comparative genealogical analyses of two genes, the authors investigated whether recombination occurred among strains within serotypes A and D. For each of 14 serotype AD strains, a portion (642 bp) of the orotidine monophosphate pyrophosphorylase (URA5) gene was cloned and sequenced. Each of these 14 strains contained two different alleles and sequences for both alleles were obtained. The URA5 gene genealogy was compared to that derived from the laccase (LAC) gene, which was reported recently for the same 14 strains. For both genes, each of the 14 serotype AD strains contained two phylogenetically distinct alleles: one allele was highly similar to those from serotype A strains and the other to alleles from serotype D strains. However, within both the serotype A allelic group and the serotype D allelic group, there was significant incongruence between genealogies derived from URA5 and LAC. The results suggest recombination in natural populations of both serotypes A and D.

scholarly journals Monoclonal Antibodies Reactive with Immunorecessive Epitopes of Glucuronoxylomannan, the Major Capsular Polysaccharide of Cryptococcus neoformans

2003 ◽  
Vol 10 (5) ◽  
pp. 903-909 ◽  
Author(s):  
Suzanne Brandt ◽  
Peter Thorkildson ◽  
Thomas R. Kozel
Keyword(s):  
Monoclonal Antibody ◽  
Immune Response ◽  
Monoclonal Antibodies ◽  
Cryptococcus Neoformans ◽  
Capsular Polysaccharide ◽  
Passive Immunization ◽  
The Other ◽  
Serotype A ◽  
Humoral Immune ◽  
New Information

ABSTRACT Cryptococcus neoformans is surrounded by an antiphagocytic capsule whose primary constituent is glucuronoxylomannan (GXM). An epitope shared by GXM serotypes A, B, C, and D is immunodominant when mice are immunized with serotype A GXM. In contrast, an epitope shared only by serotypes A and D is immunodominant when mice are immunized with serotype D. Hybridomas secreting antibodies reactive with subdominant epitopes were identified through a positive-negative screening procedure in which antibody-secreting colonies were characterized by reactivity with both the immunizing polysaccharide and GXMs from each of the four major serotypes. In this manner, a monoclonal antibody (MAb) that was reactive with an epitope shared only by serotypes A and B was identified and designated F10F5. Such an epitope has not been described previously. Immunization of mice with de-O-acetylated serotype A GXM generated a hybridoma that secreted an antibody, designated F12D2, that was reactive with all four serotypes. Unlike previously described monoclonal and polyclonal panspecific antibodies, the reactivity of MAb F12D2 was not altered by de-O-acetylation of GXM. These results indicate that there are at least two panspecific GXM epitopes; one epitope is dependent on O acetylation for antibody reactivity, and the other is independent of O acetylation. This study identifies strategies for production of MAbs that are reactive with subdominant or cryptic GXM epitopes and provides new information regarding the antigenic makeup and the humoral immune response to GXM, an essential virulence factor that is a target for active and passive immunization.

scholarly journals Contribution of Epitope Specificity to the Binding of Monoclonal Antibodies to the Capsule of Cryptococcus neoformans and the Soluble Form of Its Major Polysaccharide, Glucuronoxylomannan

2003 ◽  
Vol 10 (2) ◽  
pp. 252-258 ◽  
Author(s):  
Raymond M. Duro ◽  
Dale Netski ◽  
Peter Thorkildson ◽  
Thomas R. Kozel
Keyword(s):  
Monoclonal Antibodies ◽  
Cryptococcus Neoformans ◽  
Capsular Polysaccharide ◽  
Protective Efficacy ◽  
Biological Activities ◽  
Qualitative Assessment ◽  
Soluble Form ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serological Assay ◽  
Epitope Specificity

ABSTRACT Incubation of encapsulated cryptococci with monoclonal antibodies (MAbs) specific for glucuronoxylomannan (GXM), the major capsular polysaccharide of Cryptococcus neoformans, produces two distinct capsular quellung-type reactions termed rim and puffy. The type of capsular reaction that occurs is determined by the epitope specificity of the MAb and the serotype of the yeast cell. Several biological activities, including opsonic activity, complement activation, and protective efficacy, are associated with the type of capsular reaction produced by a MAb. The goal of this study was to examine the reactivities of two families of anti-GXM MAbs with serotype A and D capsular polysaccharides in several immunochemical assays, including agglutination, immunofluorescence, quantitative precipitation, and enzyme-linked immunosorbent assay, in an effort to identify serological assays that are predictive of the capsular quellung reaction. The results showed that the type of capsular reaction (rim versus puffy) is a qualitative assessment of antibody-capsule interaction that cannot be predicted on the basis of a serological assay. The results further showed that antibody reactivity demonstrated in one serological assay is not necessarily predictive of results in another assay, particularly in cases where one assay examines antibody-capsule interactions, e.g., agglutination, and another assay examines interaction of antibody with soluble GXM. Taken together, the results suggest caution in interpretation of immunochemical assays for anti-GXM antibodies and recommend the use of multiple assays formats when studying anticryptococcal antibodies.

scholarly journals Modulation of Polymorphonuclear Cell Interleukin-8 Secretion by Human Monoclonal Antibodies to Type 8 Pneumococcal Capsular Polysaccharide

2003 ◽  
Vol 71 (12) ◽  
pp. 6775-6783 ◽  
Author(s):  
Tamika Burns ◽  
Zhaojing Zhong ◽  
Michael Steinitz ◽  
Liise-anne Pirofski
Keyword(s):  
Monoclonal Antibodies ◽  
Capsular Polysaccharide ◽  
Immunoglobulin M ◽  
Interleukin 8 ◽  
Blue Staining ◽  
Enzyme Linked Immunosorbent Assay ◽  
Polymorphonuclear Cells ◽  
Human Monoclonal Antibodies ◽  
Classical Complement Pathway

ABSTRACT Pneumococcal capsular polysaccharide (PS) vaccines induce type-specific immunoglobulin M (IgM), IgG, and IgA. Type-specific IgG to the PS is sufficient to confer protection against the homologous serotype of the pneumococcus, but the efficacies of type-specific IgM and IgA are less well understood. We examined the in vitro activities and efficacies in mice of two human monoclonal antibodies (MAbs) to type 8 PS, NAD (IgA) and D11 (IgM). MAb-mediated opsonophagocytic killing was evaluated after coculture of type 8 pneumococci with human polymorphonuclear cells (PMNs), type-specific or control MAbs, and human complement sources. The effects of the MAbs on PMN interleukin-8 (IL-8) and IL-6 secretion were determined in supernatants from cocultures containing pneumococci and PMNs by enzyme-linked immunosorbent assay. MAb efficacy was determined in an intratracheal model of type 8 infection in mice with classical complement pathway deficiency. Both MAbs were protective in 100% of infected mice. Neither MAb promoted a significant amount of killing of type 8 pneumococci compared to its isotype control MAb. Both type-specific MAbs mediated complement-dependent modulation of PMN IL-8 secretion, with increased secretion at effector/target (E:T) ratios of 500:1 and 50:1 and reduced secretion at 1:5. Trypan blue staining revealed that PMNs cocultured with D11 were less viable at an E:T ratio of 1:5 than PMNs cocultured with the control MAb. PMN IL-6 secretion was increased by both type-specific and control MAbs. These results suggest that certain type-specific IgM and IgAs might contribute to host defense by modulation of the inflammatory response to pneumococci.

scholarly journals Immunoglobulin G3 blocking antibodies to the fungal pathogen Cryptococcus neoformans.

1996 ◽  
Vol 183 (4) ◽  
pp. 1905-1909 ◽  
Author(s):  
G Nussbaum ◽  
R Yuan ◽  
A Casadevall ◽  
M D Scharff
Keyword(s):  
Cryptococcus Neoformans ◽  
Host Defense ◽  
Fungal Pathogen ◽  
Capsular Polysaccharide ◽  
Protective Efficacy ◽  
Antibody Therapy ◽  
Antibody Isotype ◽  
Cryptococcal Infection ◽  
Epitope Specificity ◽  
Blocking Antibodies

Vaccination and infection can elicit protective and nonprotective antibodies to the fungus Cryptococcus neoformans in mice. The effect of nonprotective antibodies on host defense is unknown. In this study we used mixtures of protective and nonprotective monoclonal antibodies (mAbs) to determine if nonprotective mAbs blocked the activity of the protective mAbs. Antibody isotype and epitope specificity are important in determining the ability to prolong survival in mice given a lethal C. neoformans infection. Three different nonprotective immunoglobulin (Ig) G23 mAbs to cryptococcal capsular polysaccharide were used to study the interaction between the IgG3 isotype and protective IgG1 and IgG2a mAbs in murine cryptococcal infection. One IgG3 mAb reduced the protective efficacy of an IgG1 with identical epitope specificity. A second IgG3 mAb with different epitope specificity also reduced the protection provided by the IgG1 mAb. The protective efficacy of an IgG2a mAb was also dramatically decreased by still another IgG3 mAb. To our knowledge this is the first report of blocking antibodies to a fungal pathogen. The results have important implications for the development of vaccines and passive antibody therapy against C. neoformans.

scholarly journals Both Th1 and Th2 Cytokines Affect the Ability of Monoclonal Antibodies To Protect Mice against Cryptococcus neoformans

2001 ◽  
Vol 69 (10) ◽  
pp. 6445-6455 ◽  
Author(s):  
David O. Beenhouwer ◽  
Scott Shapiro ◽  
Marta Feldmesser ◽  
Arturo Casadevall ◽  
Matthew D. Scharff
Keyword(s):  
T Cells ◽  
Monoclonal Antibodies ◽  
Cryptococcus Neoformans ◽  
Capsular Polysaccharide ◽  
Variable Region ◽  
Interleukin 12 ◽  
Mouse Strains ◽  
Natural Immunity ◽  
Cryptococcal Infection ◽  
Th1 And Th2

ABSTRACT Variable-region-identical mouse immunoglobulin G1 (IgG1), IgG2b, and IgG2a monoclonal antibodies to the capsular polysaccharide ofCryptococcus neoformans prolong the lives of mice infected with this fungus, while IgG3 is either not protective or enhances infection. CD4+ T cells are required for IgG1-mediated protection, and CD8+ T cells are required for IgG3-mediated enhancement. Gamma interferon is required for both effects. These findings revealed that T cells and cytokines play a role in the modulation of cryptococcal infection by antibodies and suggested that it was important to more fully define the cytokine requirements of each of the antibody isotypes. We therefore investigated the efficacy of passively administered variable-region-identical IgG1, IgG2a, IgG2b, and IgG3 monoclonal antibodies against intravenous infection withC. neoformans in mice genetically deficient in interleukin-12 (IL-12), IL-6, IL-4, or IL-10, as well as in the parental C57BL/6J strain. The relative inherent susceptibilities of these mouse strains to C. neoformans were as follows: IL-12−/− > IL-6−/− > C57BL/6J ≈ IL-4−/− ≫ IL-10−/−. This is consistent with the notion that a Th1 response is necessary for natural immunity against cryptococcal infection. However, none of the IgG isotypes prolonged survival in IL-12−/−, IL-6−/−, or IL-4−/− mice, and all isotypes significantly enhanced infection in IL-10−/− mice. These results indicate that passive antibody-mediated protection againstC. neoformans requires both Th1- and Th2-associated cytokines and reveal the complexity of the mechanisms through which antibodies modulate infection with this organism.

scholarly journals Capsular Reactions of Cryptococcus neoformans with Polyspecific and Oligospecific Polyclonal Anticapsular Antibodies

2001 ◽  
Vol 69 (2) ◽  
pp. 1189-1191 ◽  
Author(s):  
Tracy C. MacGill ◽  
Randall S. MacGill ◽  
Thomas R. Kozel
Keyword(s):  
Monoclonal Antibodies ◽  
Cryptococcus Neoformans ◽  
Capsular Polysaccharide ◽  
Biological Activities ◽  
Polyclonal Rabbit

ABSTRACT Monoclonal antibodies to the capsular polysaccharide ofCryptococcus neoformans produce distinct capsular reactions and have biological activities that are determined by serotype specificity. In the present study, polyclonal rabbit anticapsular antibodies were cross-absorbed to produce serotype specificities similar to those of monoclonal antibodies. The results showed that polyclonal and monoclonal antibodies with similar serotype specificities have similar capsular reactions and biological activities.

scholarly journals Avidity as a Determinant of the Protective Efficacy of Human Antibodies to Pneumococcal Capsular Polysaccharides

1999 ◽  
Vol 67 (5) ◽  
pp. 2366-2370 ◽  
Author(s):  
William R. Usinger ◽  
Alexander H. Lucas
Keyword(s):  
Capsular Polysaccharide ◽  
Important Determinant ◽  
Protective Efficacy ◽  
Inverse Correlation ◽  
Capsular Polysaccharides ◽  
Human Antibodies ◽  
Invasive Diseases ◽  
20 Nm ◽  
The Relationship

ABSTRACT Antibodies reactive with capsular polysaccharides are considered the principal mediators of immunity against invasive diseases caused byStreptococcus pneumoniae. In this study, we tested the hypothesis that anti-pneumococcal capsular polysaccharide (PPS) antibody avidity can influence protective efficacy. We measured the avidities of individual adult postvaccination immunoglobulin G2 (IgG2) antibodies to PPS serotypes 6B and 23F and examined the relationship between avidity and opsonophagocytic and mouse-protective activities. The avidities of PPS 6B- and PPS 23F-specific IgG2 antibodies ranged from 6 to 31 nM−1 and from 3 to 20 nM−1, respectively. We observed an inverse correlation between the magnitude of avidity and the amount of antibody required to protect mice against lethal bacteremia caused by serotype 6B pneumococci. Similarly, higher-avidity antibodies were more effective than lower-avidity antibodies in vitro in mediating complement-dependent opsonophagocytosis of both 6B and 23F pneumococci. These data suggest that in adults, PPS antibodies are sufficiently polymorphic to possess biologically significant variations in avidity. We conclude that avidity functions as an important determinant of anticapsular antibody protective efficacy against pneumococci.

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