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Epitope location in the Cryptococcus neoformans capsule is a determinant of antibody efficacy.

Nussbaum G, Cleare W, Casadevall A, Scharff MD, Valadon P - J. Exp. Med. (1997)

Bottom Line: The IgM mAbs 12A1 and 13F1 originated from the same B cell and differ only by somatic mutations in their variable regions; yet mAb 12A1 protects against serotype D infection, while mAb 13F1 does not.Immunofluorescence and immunoelectron microscopy studies revealed differences in antibody localization within the capsule of serotype D strain; mAb 12A1 bound to the outer rim of the capsule resulting in an annular pattern, whereas mAb 13F1 bound throughout the capsule and had a punctate appearance.Annular binding, but not punctate binding, was associated with increased opsonic efficacy for phagocytosis of C. neoformans by J774.16 macrophage-like cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.

ABSTRACT
Monoclonal antibodies (mAbs) to the polysaccharide capsule of Cryptococcus neoformans can prolong survival in mice. However, the properties of antibodies that mediate protection are not fully understood. The IgM mAbs 12A1 and 13F1 originated from the same B cell and differ only by somatic mutations in their variable regions; yet mAb 12A1 protects against serotype D infection, while mAb 13F1 does not. Phage peptide display libraries were used to analyze the fine specificity of these two mAbs. The selection of distinct peptide motifs from identical libraries confirmed that mAbs 12A1 and 13F1 bound to two distinct epitopes. Immunofluorescence and immunoelectron microscopy studies revealed differences in antibody localization within the capsule of serotype D strain; mAb 12A1 bound to the outer rim of the capsule resulting in an annular pattern, whereas mAb 13F1 bound throughout the capsule and had a punctate appearance. The difference in the binding pattern of mAb 12A1 and 13F1 was not observed on serotype A organisms, where both mAbs bound to the capsule with an annular fluorescence pattern. The fluorescence pattern of binding correlated with protective efficacy; mAb 13F1 prolonged survival of mice infected with the J11 serotype A strain (annular fluorescence), but not serotype D strains (punctate pattern). Annular binding, but not punctate binding, was associated with increased opsonic efficacy for phagocytosis of C. neoformans by J774.16 macrophage-like cells. The correlation between capsular binding pattern, opsonic activity, and ability to prolong survival suggests that the efficacy of anticryptococcal antibodies is dependent upon where they bind in the polysaccharide capsule.

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Survival of A/J mice after intraperitoneal treatment with the  IgM mAbs 12A1 and 13F1 preceding infection with C. neoformans. (A) serotype D strain 34873, (B) serotype A strain J11.
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Figure 3: Survival of A/J mice after intraperitoneal treatment with the IgM mAbs 12A1 and 13F1 preceding infection with C. neoformans. (A) serotype D strain 34873, (B) serotype A strain J11.

Mentions: We first examined the ability of mAbs 12A1 and 13F1 to modify the course of infection with serotype D strain 34873 which had not been tested previously. Intraperitoneal administration of mAb 13F1 before infection decreased survival relative to controls (P = 0.0149), whereas mAb 12A1 prolonged survival (P = 0.0396 compared to controls) (Fig. 3 A). In addition, the animals which received mAb 13F1 died within a short time of each other (SD 0.88), in contrast to the saline controls which died over a more extended period (SD 9.53). This was similar to the previous protection experiment with serotype D strain 24067 where the standard deviations were 0.37 and 2.53 d for 13F1 and saline controls, respectively (5). This may reflect enhanced infection in the presence of 13F1.


Epitope location in the Cryptococcus neoformans capsule is a determinant of antibody efficacy.

Nussbaum G, Cleare W, Casadevall A, Scharff MD, Valadon P - J. Exp. Med. (1997)

Survival of A/J mice after intraperitoneal treatment with the  IgM mAbs 12A1 and 13F1 preceding infection with C. neoformans. (A) serotype D strain 34873, (B) serotype A strain J11.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2196155&req=5

Figure 3: Survival of A/J mice after intraperitoneal treatment with the IgM mAbs 12A1 and 13F1 preceding infection with C. neoformans. (A) serotype D strain 34873, (B) serotype A strain J11.
Mentions: We first examined the ability of mAbs 12A1 and 13F1 to modify the course of infection with serotype D strain 34873 which had not been tested previously. Intraperitoneal administration of mAb 13F1 before infection decreased survival relative to controls (P = 0.0149), whereas mAb 12A1 prolonged survival (P = 0.0396 compared to controls) (Fig. 3 A). In addition, the animals which received mAb 13F1 died within a short time of each other (SD 0.88), in contrast to the saline controls which died over a more extended period (SD 9.53). This was similar to the previous protection experiment with serotype D strain 24067 where the standard deviations were 0.37 and 2.53 d for 13F1 and saline controls, respectively (5). This may reflect enhanced infection in the presence of 13F1.

Bottom Line: The IgM mAbs 12A1 and 13F1 originated from the same B cell and differ only by somatic mutations in their variable regions; yet mAb 12A1 protects against serotype D infection, while mAb 13F1 does not.Immunofluorescence and immunoelectron microscopy studies revealed differences in antibody localization within the capsule of serotype D strain; mAb 12A1 bound to the outer rim of the capsule resulting in an annular pattern, whereas mAb 13F1 bound throughout the capsule and had a punctate appearance.Annular binding, but not punctate binding, was associated with increased opsonic efficacy for phagocytosis of C. neoformans by J774.16 macrophage-like cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Cell Biology, Albert Einstein College of Medicine, Bronx, New York 10461, USA.

ABSTRACT
Monoclonal antibodies (mAbs) to the polysaccharide capsule of Cryptococcus neoformans can prolong survival in mice. However, the properties of antibodies that mediate protection are not fully understood. The IgM mAbs 12A1 and 13F1 originated from the same B cell and differ only by somatic mutations in their variable regions; yet mAb 12A1 protects against serotype D infection, while mAb 13F1 does not. Phage peptide display libraries were used to analyze the fine specificity of these two mAbs. The selection of distinct peptide motifs from identical libraries confirmed that mAbs 12A1 and 13F1 bound to two distinct epitopes. Immunofluorescence and immunoelectron microscopy studies revealed differences in antibody localization within the capsule of serotype D strain; mAb 12A1 bound to the outer rim of the capsule resulting in an annular pattern, whereas mAb 13F1 bound throughout the capsule and had a punctate appearance. The difference in the binding pattern of mAb 12A1 and 13F1 was not observed on serotype A organisms, where both mAbs bound to the capsule with an annular fluorescence pattern. The fluorescence pattern of binding correlated with protective efficacy; mAb 13F1 prolonged survival of mice infected with the J11 serotype A strain (annular fluorescence), but not serotype D strains (punctate pattern). Annular binding, but not punctate binding, was associated with increased opsonic efficacy for phagocytosis of C. neoformans by J774.16 macrophage-like cells. The correlation between capsular binding pattern, opsonic activity, and ability to prolong survival suggests that the efficacy of anticryptococcal antibodies is dependent upon where they bind in the polysaccharide capsule.

Show MeSH
Related in: MedlinePlus