A vaccine using Anaplasma marginale subdominant type IV secretion system recombinant proteins was not protective against a virulent challenge
Anaplasma marginale is the most prevalent tick-borne livestock pathogen with worldwide distribution. Bovine anaplasmosis is a significant threat to cattle industry. Anaplasmosis outbreaks in endemic areas are prevented via vaccination with live A. centrale produced in splenectomized calves. Since A....
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Published in | PloS one Vol. 15; no. 2; p. e0229301 |
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Main Authors | , , , , , , |
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Language | English |
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21.02.2020
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Abstract | Anaplasma marginale is the most prevalent tick-borne livestock pathogen with worldwide distribution. Bovine anaplasmosis is a significant threat to cattle industry. Anaplasmosis outbreaks in endemic areas are prevented via vaccination with live A. centrale produced in splenectomized calves. Since A. centrale live vaccine can carry other pathogens and cause disease in adult cattle, research efforts are directed to develop safe recombinant subunit vaccines. Previous work found that the subdominant proteins of A. marginale type IV secretion system (T4SS) and the subdominant elongation factor-Tu (Ef-Tu) were involved in the protective immunity against the experimental challenge in cattle immunized with the A. marginale outer membrane (OM). This study evaluated the immunogenicity and protection conferred by recombinant VirB9.1, VirB9.2, VirB10, VirB11, and Ef-Tu proteins cloned and expressed in E. coli. Twenty steers were randomly clustered into four groups (G) of five animals each. Cattle from G1 and G2 were immunized with a mixture of 50 μg of each recombinant protein with Quil A® or Montanide™ adjuvants, respectively. Cattle from G3 and G4 (controls) were immunized with Quil A and Montanide adjuvants, respectively. Cattle received four immunizations at three-week intervals and were challenged with 107 A. marginale-parasitized erythrocytes 42 days after the fourth immunization. After challenge, all cattle showed clinical signs, with a significant drop of packed cell volume and a significant increase of parasitized erythrocytes (p<0.05), requiring treatment with oxytetracycline to prevent death. The levels of IgG2 induced in the immunized groups did not correlate with the observed lack of protection. Additional strategies are required to evaluate the role of these proteins and their potential utility in the development of effective vaccines. |
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AbstractList | Anaplasma marginale is the most prevalent tick-borne livestock pathogen with worldwide distribution. Bovine anaplasmosis is a significant threat to cattle industry. Anaplasmosis outbreaks in endemic areas are prevented via vaccination with live A. centrale produced in splenectomized calves. Since A. centrale live vaccine can carry other pathogens and cause disease in adult cattle, research efforts are directed to develop safe recombinant subunit vaccines. Previous work found that the subdominant proteins of A. marginale type IV secretion system (T4SS) and the subdominant elongation factor-Tu (Ef-Tu) were involved in the protective immunity against the experimental challenge in cattle immunized with the A. marginale outer membrane (OM). This study evaluated the immunogenicity and protection conferred by recombinant VirB9.1, VirB9.2, VirB10, VirB11, and Ef-Tu proteins cloned and expressed in E. coli. Twenty steers were randomly clustered into four groups (G) of five animals each. Cattle from G1 and G2 were immunized with a mixture of 50 [mu]g of each recombinant protein with Quil A.sup.® or Montanide.sup.[TM] adjuvants, respectively. Cattle from G3 and G4 (controls) were immunized with Quil A and Montanide adjuvants, respectively. Cattle received four immunizations at three-week intervals and were challenged with 10.sup.7 A. marginale-parasitized erythrocytes 42 days after the fourth immunization. After challenge, all cattle showed clinical signs, with a significant drop of packed cell volume and a significant increase of parasitized erythrocytes (p<0.05), requiring treatment with oxytetracycline to prevent death. The levels of IgG2 induced in the immunized groups did not correlate with the observed lack of protection. Additional strategies are required to evaluate the role of these proteins and their potential utility in the development of effective vaccines. Anaplasma marginale is the most prevalent tick-borne livestock pathogen with worldwide distribution. Bovine anaplasmosis is a significant threat to cattle industry. Anaplasmosis outbreaks in endemic areas are prevented via vaccination with live A. centrale produced in splenectomized calves. Since A. centrale live vaccine can carry other pathogens and cause disease in adult cattle, research efforts are directed to develop safe recombinant subunit vaccines. Previous work found that the subdominant proteins of A. marginale type IV secretion system (T4SS) and the subdominant elongation factor-Tu (Ef-Tu) were involved in the protective immunity against the experimental challenge in cattle immunized with the A. marginale outer membrane (OM). This study evaluated the immunogenicity and protection conferred by recombinant VirB9.1, VirB9.2, VirB10, VirB11, and Ef-Tu proteins cloned and expressed in E. coli. Twenty steers were randomly clustered into four groups (G) of five animals each. Cattle from G1 and G2 were immunized with a mixture of 50 μg of each recombinant protein with Quil A® or Montanide™ adjuvants, respectively. Cattle from G3 and G4 (controls) were immunized with Quil A and Montanide adjuvants, respectively. Cattle received four immunizations at three-week intervals and were challenged with 107 A. marginale-parasitized erythrocytes 42 days after the fourth immunization. After challenge, all cattle showed clinical signs, with a significant drop of packed cell volume and a significant increase of parasitized erythrocytes (p<0.05), requiring treatment with oxytetracycline to prevent death. The levels of IgG2 induced in the immunized groups did not correlate with the observed lack of protection. Additional strategies are required to evaluate the role of these proteins and their potential utility in the development of effective vaccines. Anaplasma marginale is the most prevalent tick-borne livestock pathogen with worldwide distribution. Bovine anaplasmosis is a significant threat to cattle industry. Anaplasmosis outbreaks in endemic areas are prevented via vaccination with live A . centrale produced in splenectomized calves. Since A . centrale live vaccine can carry other pathogens and cause disease in adult cattle, research efforts are directed to develop safe recombinant subunit vaccines. Previous work found that the subdominant proteins of A . marginale type IV secretion system (T4SS) and the subdominant elongation factor-Tu (Ef-Tu) were involved in the protective immunity against the experimental challenge in cattle immunized with the A . marginale outer membrane (OM). This study evaluated the immunogenicity and protection conferred by recombinant VirB9.1, VirB9.2, VirB10, VirB11, and Ef-Tu proteins cloned and expressed in E . coli . Twenty steers were randomly clustered into four groups (G) of five animals each. Cattle from G1 and G2 were immunized with a mixture of 50 μg of each recombinant protein with Quil A ® or Montanide ™ adjuvants, respectively. Cattle from G3 and G4 (controls) were immunized with Quil A and Montanide adjuvants, respectively. Cattle received four immunizations at three-week intervals and were challenged with 10 7 A . marginale -parasitized erythrocytes 42 days after the fourth immunization. After challenge, all cattle showed clinical signs, with a significant drop of packed cell volume and a significant increase of parasitized erythrocytes (p<0.05), requiring treatment with oxytetracycline to prevent death. The levels of IgG2 induced in the immunized groups did not correlate with the observed lack of protection. Additional strategies are required to evaluate the role of these proteins and their potential utility in the development of effective vaccines. |
Audience | Academic |
Author | Echaide, Ignacio E Mazzucco, Matilde N Primo, María E Sarli, Macarena de Echaide, Susana T Signorini, Marcelo L Novoa, María B |
AuthorAffiliation | 2 Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Rafaela, Santa Fe, Argentina 1 Instituto Nacional de Tecnología Agropecuaria, Estación Experimental Agropecuaria Rafaela, Rafaela, Santa Fe, Argentina Instituto Butantan, BRAZIL |
AuthorAffiliation_xml | – name: Instituto Butantan, BRAZIL – name: 2 Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Rafaela, Santa Fe, Argentina – name: 1 Instituto Nacional de Tecnología Agropecuaria, Estación Experimental Agropecuaria Rafaela, Rafaela, Santa Fe, Argentina |
Author_xml | – sequence: 1 givenname: Macarena surname: Sarli fullname: Sarli, Macarena organization: Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Rafaela, Santa Fe, Argentina – sequence: 2 givenname: María B orcidid: 0000-0001-6726-5239 surname: Novoa fullname: Novoa, María B organization: Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Rafaela, Santa Fe, Argentina – sequence: 3 givenname: Matilde N surname: Mazzucco fullname: Mazzucco, Matilde N organization: Instituto Nacional de Tecnología Agropecuaria, Estación Experimental Agropecuaria Rafaela, Rafaela, Santa Fe, Argentina – sequence: 4 givenname: Marcelo L surname: Signorini fullname: Signorini, Marcelo L organization: Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Rafaela, Santa Fe, Argentina – sequence: 5 givenname: Ignacio E orcidid: 0000-0002-0621-9885 surname: Echaide fullname: Echaide, Ignacio E organization: Instituto Nacional de Tecnología Agropecuaria, Estación Experimental Agropecuaria Rafaela, Rafaela, Santa Fe, Argentina – sequence: 6 givenname: Susana T surname: de Echaide fullname: de Echaide, Susana T organization: Instituto Nacional de Tecnología Agropecuaria, Estación Experimental Agropecuaria Rafaela, Rafaela, Santa Fe, Argentina – sequence: 7 givenname: María E orcidid: 0000-0002-6873-8867 surname: Primo fullname: Primo, María E organization: Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Rafaela, Santa Fe, Argentina |
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CitedBy_id | crossref_primary_10_1016_j_vetimm_2021_110285 crossref_primary_10_1016_j_vprsr_2020_100513 crossref_primary_10_3389_fvets_2022_946545 crossref_primary_10_3390_ijms21228813 |
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Snippet | Anaplasma marginale is the most prevalent tick-borne livestock pathogen with worldwide distribution. Bovine anaplasmosis is a significant threat to cattle... Anaplasma marginale is the most prevalent tick-borne livestock pathogen with worldwide distribution. Bovine anaplasmosis is a significant threat to cattle... |
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SubjectTerms | Adjuvants Anaplasma marginale Anaplasma marginale - immunology Anaplasma marginale - pathogenicity Anaplasmosis Animals Antibodies, Bacterial - immunology Bacterial Vaccines - genetics Bacterial Vaccines - immunology Biology and Life Sciences Calves Cattle Cattle industry Cell adhesion & migration Cell size Cloning Deoxyribonucleic acid Diseases DNA E coli EDTA Elongation Erythrocytes Escherichia coli Immunization Immunogenicity Immunoglobulin G Infections Livestock Medical research Medicine and Health Sciences Oxytetracycline Pathogenic microorganisms Pathogens Plasmids Protein expression Proteins Recombinant proteins Recombinant Proteins - genetics Recombinant Proteins - immunology Research and Analysis Methods Saponins Tick-borne diseases Type IV Secretion Systems - genetics Type IV Secretion Systems - immunology Vaccination Vaccines Virulence - immunology |
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Title | A vaccine using Anaplasma marginale subdominant type IV secretion system recombinant proteins was not protective against a virulent challenge |
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