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Human Polyclonal Antibodies Produced through DNA Vaccination of Transchromosomal Cattle Provide Mice with Post-Exposure Protection against Lethal Zaire and Sudan Ebolaviruses.

Bounds CE, Kwilas SA, Kuehne AI, Brannan JM, Bakken RR, Dye JM, Hooper JW, Dupuy LC, Ellefsen B, Hannaman D, Wu H, Jiao JA, Sullivan EJ, Schmaljohn CS - PLoS ONE (2015)

Bottom Line: Passive transfer by IP injection of the purified IgG (100 mg/kg) to groups of BALB/c mice one day after IP challenge with mouse adapted (ma) EBOV resulted in 80% protection while all mice treated with non-specific pAbs succumbed.Similarly, interferon receptor 1 knockout (IFNAR(-/-)) mice receiving the purified IgG (100 mg/kg) by IP injection one day after IP challenge with wild type SUDV resulted in 89% survival.These results are the first to demonstrate that filovirus GP DNA vaccines administered to TcBs by IM-EP can elicit neutralizing antibodies that provide post-exposure protection.

View Article: PubMed Central - PubMed

Affiliation: United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland, United States of America.

ABSTRACT
DNA vaccination of transchromosomal bovines (TcBs) with DNA vaccines expressing the codon-optimized (co) glycoprotein (GP) genes of Ebola virus (EBOV) and Sudan virus (SUDV) produce fully human polyclonal antibodies (pAbs) that recognize both viruses and demonstrate robust neutralizing activity. Each TcB was vaccinated by intramuscular electroporation (IM-EP) a total of four times and at each administration received 10 mg of the EBOV-GPco DNA vaccine and 10 mg of the SUDV-GPco DNA vaccine at two sites on the left and right sides, respectively. After two vaccinations, robust antibody responses (titers > 1000) were detected by ELISA against whole irradiated EBOV or SUDV and recombinant EBOV-GP or SUDV-GP (rGP) antigens, with higher titers observed for the rGP antigens. Strong, virus neutralizing antibody responses (titers >1000) were detected after three vaccinations when measured by vesicular stomatitis virus-based pseudovirion neutralization assay (PsVNA). Maximal neutralizing antibody responses were identified by traditional plaque reduction neutralization tests (PRNT) after four vaccinations. Neutralizing activity of human immunoglobulins (IgG) purified from TcB plasma collected after three vaccinations and injected intraperitoneally (IP) into mice at a 100 mg/kg dose was detected in the serum by PsVNA up to 14 days after administration. Passive transfer by IP injection of the purified IgG (100 mg/kg) to groups of BALB/c mice one day after IP challenge with mouse adapted (ma) EBOV resulted in 80% protection while all mice treated with non-specific pAbs succumbed. Similarly, interferon receptor 1 knockout (IFNAR(-/-)) mice receiving the purified IgG (100 mg/kg) by IP injection one day after IP challenge with wild type SUDV resulted in 89% survival. These results are the first to demonstrate that filovirus GP DNA vaccines administered to TcBs by IM-EP can elicit neutralizing antibodies that provide post-exposure protection. Additionally, these data describe production of fully human IgG in a large animal system, a system which is capable of producing large quantities of a clinical grade therapeutic product.

No MeSH data available.


Related in: MedlinePlus

Weight loss and survival of BALB/c mice administered high, medium, or low doses of purified EBOV/SUDV human pAbs after maEBOV challenge.Groups of BALB/c mice (N = 10) received a single IP injection of 100mg/kg NS pAbs, 100 mg/kg EBOV/SUDV pAbs, 50 mg/kg EBOV/SUDV pAbs, or 10 mg/kg EBOV/SUDV pAbs one day after challenge or 50 mg/kg EBOV/SUDV pAbs, 25 mg/kg EBOV/SUDV pAbs, or 5 mg/kg EBOV/SUDV pAbs on day 1 and day 2 after challenge with 100 pfu maEBOV. (A) Mean weight was determined daily for each dosing group and graphed as the percent mean of the starting weight. All mice receiving the NS pabs succumb by day 8 post-challenge. (B) Kaplan-Meier survival curves indicating the percentage of surviving mice at each day of the 21-day post-challenge observation period are shown. The p-values for the following comparisons are as follows: 100 mg/kg NS pAbs vs. 100 mg/kg EBOV/SUDV pAbs +1 (p = 0.0003), 100 mg/kg NS pAbs vs. 50 mg/kg EBOV/SUDV pAbs +1 (p = 0.0059), NS pAbs vs. 10 mg/kg EBOV/SUDV pAbs +1 (p = 0.0630), NS pAbs vs. 50 mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.002), NS pAbs vs. 25 mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.0046), NS pAbs vs. 5mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.1082), 100mg/kg EBOV/SUDV pAbs +1 vs. 50 mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.1988), 50 mg/kg EBOV/SUDV +1 vs. 25 mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.9968), and 10 mg/kg EBOV/SUDV pAbs +1 vs. 5 mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.7275). Significant differences between survival curves are denoted by (*) where p < 0.05.
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pone.0137786.g005: Weight loss and survival of BALB/c mice administered high, medium, or low doses of purified EBOV/SUDV human pAbs after maEBOV challenge.Groups of BALB/c mice (N = 10) received a single IP injection of 100mg/kg NS pAbs, 100 mg/kg EBOV/SUDV pAbs, 50 mg/kg EBOV/SUDV pAbs, or 10 mg/kg EBOV/SUDV pAbs one day after challenge or 50 mg/kg EBOV/SUDV pAbs, 25 mg/kg EBOV/SUDV pAbs, or 5 mg/kg EBOV/SUDV pAbs on day 1 and day 2 after challenge with 100 pfu maEBOV. (A) Mean weight was determined daily for each dosing group and graphed as the percent mean of the starting weight. All mice receiving the NS pabs succumb by day 8 post-challenge. (B) Kaplan-Meier survival curves indicating the percentage of surviving mice at each day of the 21-day post-challenge observation period are shown. The p-values for the following comparisons are as follows: 100 mg/kg NS pAbs vs. 100 mg/kg EBOV/SUDV pAbs +1 (p = 0.0003), 100 mg/kg NS pAbs vs. 50 mg/kg EBOV/SUDV pAbs +1 (p = 0.0059), NS pAbs vs. 10 mg/kg EBOV/SUDV pAbs +1 (p = 0.0630), NS pAbs vs. 50 mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.002), NS pAbs vs. 25 mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.0046), NS pAbs vs. 5mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.1082), 100mg/kg EBOV/SUDV pAbs +1 vs. 50 mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.1988), 50 mg/kg EBOV/SUDV +1 vs. 25 mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.9968), and 10 mg/kg EBOV/SUDV pAbs +1 vs. 5 mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.7275). Significant differences between survival curves are denoted by (*) where p < 0.05.

Mentions: Clinical signs of EBOV infection, including weight loss, were observed beginning on days 4 or 5 in all groups of mice and by day 8 all mice receiving a 100 mg/kg dose of the NS pabs succumbed to disease (Fig 5A). Eight of the 10 mice that received a single 100 mg/kg dose of the V3 EBOV/SUDV pAbs one day after challenge survived (Fig 5B). Mice that received 50 mg/kg dose of the V3 EBOV/SUDV human pAbs on days 1 and 2 following challenge had a survival rate of 50%, which was significantly higher than controls (p = 0.0059), and was not significantly less than those receiving the 100 mg/kg dose (p = 0.199). Additionally, mice that received a single 50 mg/kg dose of the EBOV/SUDV pAbs one day post challenge or serial doses of 25 mg/kg on days one and two post challenge had a survival rate of 40%, which was also still significantly higher (p = 0.0059; p = 0.0046) than those receiving the NS pAbs. Mice receiving a single 10 mg/kg dose of the V3 EBOV/SUDV human pAbs one day post challenge and mice receiving serial doses at 5 mg/kg on days one and two after challenge did not have survival rates that were statistically significant when compared to mice receiving the NS pAbs.


Human Polyclonal Antibodies Produced through DNA Vaccination of Transchromosomal Cattle Provide Mice with Post-Exposure Protection against Lethal Zaire and Sudan Ebolaviruses.

Bounds CE, Kwilas SA, Kuehne AI, Brannan JM, Bakken RR, Dye JM, Hooper JW, Dupuy LC, Ellefsen B, Hannaman D, Wu H, Jiao JA, Sullivan EJ, Schmaljohn CS - PLoS ONE (2015)

Weight loss and survival of BALB/c mice administered high, medium, or low doses of purified EBOV/SUDV human pAbs after maEBOV challenge.Groups of BALB/c mice (N = 10) received a single IP injection of 100mg/kg NS pAbs, 100 mg/kg EBOV/SUDV pAbs, 50 mg/kg EBOV/SUDV pAbs, or 10 mg/kg EBOV/SUDV pAbs one day after challenge or 50 mg/kg EBOV/SUDV pAbs, 25 mg/kg EBOV/SUDV pAbs, or 5 mg/kg EBOV/SUDV pAbs on day 1 and day 2 after challenge with 100 pfu maEBOV. (A) Mean weight was determined daily for each dosing group and graphed as the percent mean of the starting weight. All mice receiving the NS pabs succumb by day 8 post-challenge. (B) Kaplan-Meier survival curves indicating the percentage of surviving mice at each day of the 21-day post-challenge observation period are shown. The p-values for the following comparisons are as follows: 100 mg/kg NS pAbs vs. 100 mg/kg EBOV/SUDV pAbs +1 (p = 0.0003), 100 mg/kg NS pAbs vs. 50 mg/kg EBOV/SUDV pAbs +1 (p = 0.0059), NS pAbs vs. 10 mg/kg EBOV/SUDV pAbs +1 (p = 0.0630), NS pAbs vs. 50 mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.002), NS pAbs vs. 25 mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.0046), NS pAbs vs. 5mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.1082), 100mg/kg EBOV/SUDV pAbs +1 vs. 50 mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.1988), 50 mg/kg EBOV/SUDV +1 vs. 25 mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.9968), and 10 mg/kg EBOV/SUDV pAbs +1 vs. 5 mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.7275). Significant differences between survival curves are denoted by (*) where p < 0.05.
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getmorefigures.php?uid=PMC4589376&req=5

pone.0137786.g005: Weight loss and survival of BALB/c mice administered high, medium, or low doses of purified EBOV/SUDV human pAbs after maEBOV challenge.Groups of BALB/c mice (N = 10) received a single IP injection of 100mg/kg NS pAbs, 100 mg/kg EBOV/SUDV pAbs, 50 mg/kg EBOV/SUDV pAbs, or 10 mg/kg EBOV/SUDV pAbs one day after challenge or 50 mg/kg EBOV/SUDV pAbs, 25 mg/kg EBOV/SUDV pAbs, or 5 mg/kg EBOV/SUDV pAbs on day 1 and day 2 after challenge with 100 pfu maEBOV. (A) Mean weight was determined daily for each dosing group and graphed as the percent mean of the starting weight. All mice receiving the NS pabs succumb by day 8 post-challenge. (B) Kaplan-Meier survival curves indicating the percentage of surviving mice at each day of the 21-day post-challenge observation period are shown. The p-values for the following comparisons are as follows: 100 mg/kg NS pAbs vs. 100 mg/kg EBOV/SUDV pAbs +1 (p = 0.0003), 100 mg/kg NS pAbs vs. 50 mg/kg EBOV/SUDV pAbs +1 (p = 0.0059), NS pAbs vs. 10 mg/kg EBOV/SUDV pAbs +1 (p = 0.0630), NS pAbs vs. 50 mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.002), NS pAbs vs. 25 mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.0046), NS pAbs vs. 5mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.1082), 100mg/kg EBOV/SUDV pAbs +1 vs. 50 mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.1988), 50 mg/kg EBOV/SUDV +1 vs. 25 mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.9968), and 10 mg/kg EBOV/SUDV pAbs +1 vs. 5 mg/kg EBOV/SUDV pAbs +1 and +2 (p = 0.7275). Significant differences between survival curves are denoted by (*) where p < 0.05.
Mentions: Clinical signs of EBOV infection, including weight loss, were observed beginning on days 4 or 5 in all groups of mice and by day 8 all mice receiving a 100 mg/kg dose of the NS pabs succumbed to disease (Fig 5A). Eight of the 10 mice that received a single 100 mg/kg dose of the V3 EBOV/SUDV pAbs one day after challenge survived (Fig 5B). Mice that received 50 mg/kg dose of the V3 EBOV/SUDV human pAbs on days 1 and 2 following challenge had a survival rate of 50%, which was significantly higher than controls (p = 0.0059), and was not significantly less than those receiving the 100 mg/kg dose (p = 0.199). Additionally, mice that received a single 50 mg/kg dose of the EBOV/SUDV pAbs one day post challenge or serial doses of 25 mg/kg on days one and two post challenge had a survival rate of 40%, which was also still significantly higher (p = 0.0059; p = 0.0046) than those receiving the NS pAbs. Mice receiving a single 10 mg/kg dose of the V3 EBOV/SUDV human pAbs one day post challenge and mice receiving serial doses at 5 mg/kg on days one and two after challenge did not have survival rates that were statistically significant when compared to mice receiving the NS pAbs.

Bottom Line: Passive transfer by IP injection of the purified IgG (100 mg/kg) to groups of BALB/c mice one day after IP challenge with mouse adapted (ma) EBOV resulted in 80% protection while all mice treated with non-specific pAbs succumbed.Similarly, interferon receptor 1 knockout (IFNAR(-/-)) mice receiving the purified IgG (100 mg/kg) by IP injection one day after IP challenge with wild type SUDV resulted in 89% survival.These results are the first to demonstrate that filovirus GP DNA vaccines administered to TcBs by IM-EP can elicit neutralizing antibodies that provide post-exposure protection.

View Article: PubMed Central - PubMed

Affiliation: United States Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland, United States of America.

ABSTRACT
DNA vaccination of transchromosomal bovines (TcBs) with DNA vaccines expressing the codon-optimized (co) glycoprotein (GP) genes of Ebola virus (EBOV) and Sudan virus (SUDV) produce fully human polyclonal antibodies (pAbs) that recognize both viruses and demonstrate robust neutralizing activity. Each TcB was vaccinated by intramuscular electroporation (IM-EP) a total of four times and at each administration received 10 mg of the EBOV-GPco DNA vaccine and 10 mg of the SUDV-GPco DNA vaccine at two sites on the left and right sides, respectively. After two vaccinations, robust antibody responses (titers > 1000) were detected by ELISA against whole irradiated EBOV or SUDV and recombinant EBOV-GP or SUDV-GP (rGP) antigens, with higher titers observed for the rGP antigens. Strong, virus neutralizing antibody responses (titers >1000) were detected after three vaccinations when measured by vesicular stomatitis virus-based pseudovirion neutralization assay (PsVNA). Maximal neutralizing antibody responses were identified by traditional plaque reduction neutralization tests (PRNT) after four vaccinations. Neutralizing activity of human immunoglobulins (IgG) purified from TcB plasma collected after three vaccinations and injected intraperitoneally (IP) into mice at a 100 mg/kg dose was detected in the serum by PsVNA up to 14 days after administration. Passive transfer by IP injection of the purified IgG (100 mg/kg) to groups of BALB/c mice one day after IP challenge with mouse adapted (ma) EBOV resulted in 80% protection while all mice treated with non-specific pAbs succumbed. Similarly, interferon receptor 1 knockout (IFNAR(-/-)) mice receiving the purified IgG (100 mg/kg) by IP injection one day after IP challenge with wild type SUDV resulted in 89% survival. These results are the first to demonstrate that filovirus GP DNA vaccines administered to TcBs by IM-EP can elicit neutralizing antibodies that provide post-exposure protection. Additionally, these data describe production of fully human IgG in a large animal system, a system which is capable of producing large quantities of a clinical grade therapeutic product.

No MeSH data available.


Related in: MedlinePlus