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Protective mAbs and Cross-Reactive mAbs Raised by Immunization with Engineered Marburg Virus GPs.

Fusco ML, Hashiguchi T, Cassan R, Biggins JE, Murin CD, Warfield KL, Li S, Holtsberg FW, Shulenin S, Vu H, Olinger GG, Kim do H, Whaley KJ, Zeitlin L, Ward AB, Nykiforuk C, Aman MJ, Berry JD, Berry J, Saphire EO - PLoS Pathog. (2015)

Bottom Line: Antibodies against the filovirus surface glycoprotein (GP) have been shown to provide life-saving therapy in nonhuman primates, but such antibodies are generally virus-specific.Surprisingly, two of the mAbs raised against MARV GP also cross-react with the mucin-deleted GP cores of all tested ebolaviruses (Ebola, Sudan, Bundibugyo, Reston), but these epitopes are masked differently by the mucin-like domains themselves.The most efficacious mAbs in this panel were found to recognize a novel "wing" feature on the GP2 subunit that is unique to Marburg and does not exist in Ebola.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California, United States of America.

ABSTRACT
The filoviruses, which include the marburg- and ebolaviruses, have caused multiple outbreaks among humans this decade. Antibodies against the filovirus surface glycoprotein (GP) have been shown to provide life-saving therapy in nonhuman primates, but such antibodies are generally virus-specific. Many monoclonal antibodies (mAbs) have been described against Ebola virus. In contrast, relatively few have been described against Marburg virus. Here we present ten mAbs elicited by immunization of mice using recombinant mucin-deleted GPs from different Marburg virus (MARV) strains. Surprisingly, two of the mAbs raised against MARV GP also cross-react with the mucin-deleted GP cores of all tested ebolaviruses (Ebola, Sudan, Bundibugyo, Reston), but these epitopes are masked differently by the mucin-like domains themselves. The most efficacious mAbs in this panel were found to recognize a novel "wing" feature on the GP2 subunit that is unique to Marburg and does not exist in Ebola. Two of these anti-wing antibodies confer 90 and 100% protection, respectively, one hour post-exposure in mice challenged with MARV.

No MeSH data available.


Related in: MedlinePlus

Antibody characterization.(A) Overall summary of mAb generation, characterization, in vitro neutralization of MARV GP-pseudotyped VSV, and in vivo protection in mice. Antibodies were generated from a single immunogen with the exception of 54G1, 54G2 and 54G3 which were primed with Ravn GPΔmuc and boosted with GPΔmuc 30G4 Fab complex. (B) ELISA EC50 values against different forms of purified MARV Ravn GP or against VLPs in ng/ml. Relative potency is indicated by orange highlight (high, EC50 <20ng/ml), dark yellow (medium, EC50 20-200ng/ml), or light yellow (low, EC50 >200ng/ml). (C) Relative ELISA binding to purified mucin-containing GPs from other MARV strains. No binding at 10μg/ml maximum mAb concentration is represented by >.
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ppat.1005016.g001: Antibody characterization.(A) Overall summary of mAb generation, characterization, in vitro neutralization of MARV GP-pseudotyped VSV, and in vivo protection in mice. Antibodies were generated from a single immunogen with the exception of 54G1, 54G2 and 54G3 which were primed with Ravn GPΔmuc and boosted with GPΔmuc 30G4 Fab complex. (B) ELISA EC50 values against different forms of purified MARV Ravn GP or against VLPs in ng/ml. Relative potency is indicated by orange highlight (high, EC50 <20ng/ml), dark yellow (medium, EC50 20-200ng/ml), or light yellow (low, EC50 >200ng/ml). (C) Relative ELISA binding to purified mucin-containing GPs from other MARV strains. No binding at 10μg/ml maximum mAb concentration is represented by >.

Mentions: To generate MARV GP-specific mAbs, BALB/c mice were immunized with GPΔmuc antigens from either MARV strain Ci67, Musoke, Angola, or Ravn (Fig 1A). Mice for each subset were immunized and boosted with the same antigen with the exception of the 54 series (54G1, 54G2, 54G3). Eight of the ten mAbs in the panel are mouse IgG1. The remaining two mAbs, 9A11 and 2D8, are IgG2a (Fig 1A).


Protective mAbs and Cross-Reactive mAbs Raised by Immunization with Engineered Marburg Virus GPs.

Fusco ML, Hashiguchi T, Cassan R, Biggins JE, Murin CD, Warfield KL, Li S, Holtsberg FW, Shulenin S, Vu H, Olinger GG, Kim do H, Whaley KJ, Zeitlin L, Ward AB, Nykiforuk C, Aman MJ, Berry JD, Berry J, Saphire EO - PLoS Pathog. (2015)

Antibody characterization.(A) Overall summary of mAb generation, characterization, in vitro neutralization of MARV GP-pseudotyped VSV, and in vivo protection in mice. Antibodies were generated from a single immunogen with the exception of 54G1, 54G2 and 54G3 which were primed with Ravn GPΔmuc and boosted with GPΔmuc 30G4 Fab complex. (B) ELISA EC50 values against different forms of purified MARV Ravn GP or against VLPs in ng/ml. Relative potency is indicated by orange highlight (high, EC50 <20ng/ml), dark yellow (medium, EC50 20-200ng/ml), or light yellow (low, EC50 >200ng/ml). (C) Relative ELISA binding to purified mucin-containing GPs from other MARV strains. No binding at 10μg/ml maximum mAb concentration is represented by >.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4482612&req=5

ppat.1005016.g001: Antibody characterization.(A) Overall summary of mAb generation, characterization, in vitro neutralization of MARV GP-pseudotyped VSV, and in vivo protection in mice. Antibodies were generated from a single immunogen with the exception of 54G1, 54G2 and 54G3 which were primed with Ravn GPΔmuc and boosted with GPΔmuc 30G4 Fab complex. (B) ELISA EC50 values against different forms of purified MARV Ravn GP or against VLPs in ng/ml. Relative potency is indicated by orange highlight (high, EC50 <20ng/ml), dark yellow (medium, EC50 20-200ng/ml), or light yellow (low, EC50 >200ng/ml). (C) Relative ELISA binding to purified mucin-containing GPs from other MARV strains. No binding at 10μg/ml maximum mAb concentration is represented by >.
Mentions: To generate MARV GP-specific mAbs, BALB/c mice were immunized with GPΔmuc antigens from either MARV strain Ci67, Musoke, Angola, or Ravn (Fig 1A). Mice for each subset were immunized and boosted with the same antigen with the exception of the 54 series (54G1, 54G2, 54G3). Eight of the ten mAbs in the panel are mouse IgG1. The remaining two mAbs, 9A11 and 2D8, are IgG2a (Fig 1A).

Bottom Line: Antibodies against the filovirus surface glycoprotein (GP) have been shown to provide life-saving therapy in nonhuman primates, but such antibodies are generally virus-specific.Surprisingly, two of the mAbs raised against MARV GP also cross-react with the mucin-deleted GP cores of all tested ebolaviruses (Ebola, Sudan, Bundibugyo, Reston), but these epitopes are masked differently by the mucin-like domains themselves.The most efficacious mAbs in this panel were found to recognize a novel "wing" feature on the GP2 subunit that is unique to Marburg and does not exist in Ebola.

View Article: PubMed Central - PubMed

Affiliation: Department of Immunology and Microbial Science, The Scripps Research Institute, La Jolla, California, United States of America.

ABSTRACT
The filoviruses, which include the marburg- and ebolaviruses, have caused multiple outbreaks among humans this decade. Antibodies against the filovirus surface glycoprotein (GP) have been shown to provide life-saving therapy in nonhuman primates, but such antibodies are generally virus-specific. Many monoclonal antibodies (mAbs) have been described against Ebola virus. In contrast, relatively few have been described against Marburg virus. Here we present ten mAbs elicited by immunization of mice using recombinant mucin-deleted GPs from different Marburg virus (MARV) strains. Surprisingly, two of the mAbs raised against MARV GP also cross-react with the mucin-deleted GP cores of all tested ebolaviruses (Ebola, Sudan, Bundibugyo, Reston), but these epitopes are masked differently by the mucin-like domains themselves. The most efficacious mAbs in this panel were found to recognize a novel "wing" feature on the GP2 subunit that is unique to Marburg and does not exist in Ebola. Two of these anti-wing antibodies confer 90 and 100% protection, respectively, one hour post-exposure in mice challenged with MARV.

No MeSH data available.


Related in: MedlinePlus