Temperature-mediated recombinant anthrax protective antigen aggregate development: Implications for toxin formation and immunogenicity
| Autor: | Juan Arciniega, Esther D. Valerdi-Madrigal, Lev Sirota, Rocío I. Domínguez-Castillo, Juan C. Amador-Molina |
|---|---|
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Hot Temperature Bacterial Toxins Enzyme-Linked Immunosorbent Assay Anthrax Vaccines complex mixtures Neutralization law.invention 03 medical and health sciences Mice Protein Aggregates Immune system Antigen law Neutralization Tests Animals Antigens Bacterial Anthrax vaccines 030102 biochemistry & molecular biology General Veterinary General Immunology and Microbiology biology Protein Stability Immunogenicity Macrophages Public Health Environmental and Occupational Health Antibody titer Molecular biology Antibodies Bacterial Antibodies Neutralizing Recombinant Proteins enzymes and coenzymes (carbohydrates) 030104 developmental biology Infectious Diseases RAW 264.7 Cells biology.protein Recombinant DNA Molecular Medicine Female Antitoxins Antibody |
| Zdroj: | Vaccine. 34(35) |
| ISSN: | 1873-2518 |
| Popis: | Anthrax vaccines containing recombinant PA (rPA) as the only antigen face a stability issue: rPA forms aggregates in solution after exposure to temperatures ⩾40°C, thus losing its ability to form lethal toxin (LeTx) with Lethal Factor. To study rPA aggregation's impact on immune response, we subjected rPA to several time and temperature combinations. rPA treated at 50°C for 30min formed high mass aggregates when analyzed by gel electrophoresis and failed to form LeTx as measured by a macrophage lysis assay (MLA). Aggregated rPA-formed LeTx was about 30 times less active than LeTx containing native rPA. Mice immunized with heat-treated rPA combined with Al(OH)3 developed antibody titers about 49 times lower than mice immunized with native rPA, as measured by a Toxicity Neutralization Assay (TNA). Enzyme Linked Immunosorbent Assay (ELISA) of the same immune sera showed anti-rPA titers only 2-7 times lower than titers elicited by native rPA. Thus, rPA's ability to form LeTx correlates with its production of neutralizing antibodies, and aggregation significantly impairs the protein's antibody response. However, while these findings suggest MLA has some value as an in-process quality test for rPA in new anthrax vaccines, they also confirm the superiority of TNA for use in vaccine potency. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Full Text from ScienceDirect