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Studies on recombinant glucokinase (r-glk) protein of Brucella abortus as a candidate vaccine molecule for brucellosis.
Vaccine 2014; 32(43):5600-6V

Abstract

Brucellosis is one of the most prevalent zoonotic diseases of worldwide distribution caused by the infection of genus Brucella. Live attenuated vaccines such as B. abortus S19, B. abortus RB51 and B. melitensis Rev1 are found most effective against brucellosis infection in animals, contriving a number of serious side effects and having chances to revert back into their active pathogenic form. In order to engineer a safe and effective vaccine candidate to be used in both animals and human, a recombinant subunit vaccine molecule comprising the truncated region of glucokinase (r-glk) gene from B. abortus S19 was cloned and expressed in Escherichia coli BL21DE3 host. Female BALB/c mice immunized with purified recombinant protein developed specific antibody titer of 1:64,000. The predominant IgG2a and IgG2b isotypes signified development of Th1 directed immune responses. In vitro cell cytotoxicity assay using anti-r-glk antibodies incubated with HeLa cells showed 81.20% and 78.5% cell viability against lethal challenge of B. abortus 544 and B. melitensis 16M, respectively. The lymphocyte proliferative assay indicated a higher splenic lymphocyte responses at 25μg/ml concentration of protein which implies the elevated development of memory immune responses. In contrast to control, the immunized group of mice intra-peritoneal (I.P.) challenged with B. abortus 544 were significantly protected with no signs of necrosis and vacuolization in their liver and spleen tissue. The elevated B-cell response associated with Th1 adopted immunity, significant in vitro cell viability as well as protection afforded in experimental animals after challenge, supplemented with histopathological analysis are suggestive of r-glk protein as a prospective candidate vaccine molecule against brucellosis.

Authors+Show Affiliations

Division of Microbiology, Defence Food Research Laboratory, Siddartha Nagar, Mysore 570011, Karnataka, India.Division of Microbiology, Defence Food Research Laboratory, Siddartha Nagar, Mysore 570011, Karnataka, India.Division of Microbiology, Defence Food Research Laboratory, Siddartha Nagar, Mysore 570011, Karnataka, India.Division of Microbiology, Defence Food Research Laboratory, Siddartha Nagar, Mysore 570011, Karnataka, India.Division of Microbiology, Defence Food Research Laboratory, Siddartha Nagar, Mysore 570011, Karnataka, India. Electronic address: drharshvardhanbatra@gmail.com.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

25131740

Citation

Vrushabhendrappa, , et al. "Studies On Recombinant Glucokinase (r-glk) Protein of Brucella Abortus as a Candidate Vaccine Molecule for Brucellosis." Vaccine, vol. 32, no. 43, 2014, pp. 5600-6.
Vrushabhendrappa , Singh AK, Balakrishna K, et al. Studies on recombinant glucokinase (r-glk) protein of Brucella abortus as a candidate vaccine molecule for brucellosis. Vaccine. 2014;32(43):5600-6.
Vrushabhendrappa, ., Singh, A. K., Balakrishna, K., Sripathy, M. H., & Batra, H. V. (2014). Studies on recombinant glucokinase (r-glk) protein of Brucella abortus as a candidate vaccine molecule for brucellosis. Vaccine, 32(43), pp. 5600-6. doi:10.1016/j.vaccine.2014.07.106.
Vrushabhendrappa , et al. Studies On Recombinant Glucokinase (r-glk) Protein of Brucella Abortus as a Candidate Vaccine Molecule for Brucellosis. Vaccine. 2014 Sep 29;32(43):5600-6. PubMed PMID: 25131740.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Studies on recombinant glucokinase (r-glk) protein of Brucella abortus as a candidate vaccine molecule for brucellosis. AU - Vrushabhendrappa,, AU - Singh,Amit Kumar, AU - Balakrishna,Konduru, AU - Sripathy,Murali Harishchandra, AU - Batra,Harsh Vardhan, Y1 - 2014/08/12/ PY - 2014/04/30/received PY - 2014/07/09/revised PY - 2014/07/31/accepted PY - 2014/8/19/entrez PY - 2014/8/19/pubmed PY - 2015/4/2/medline KW - B. abortus KW - B. melitensis KW - Brucellosis KW - In vitro protection KW - In vivo protection KW - r-glk subunit vaccines SP - 5600 EP - 6 JF - Vaccine JO - Vaccine VL - 32 IS - 43 N2 - Brucellosis is one of the most prevalent zoonotic diseases of worldwide distribution caused by the infection of genus Brucella. Live attenuated vaccines such as B. abortus S19, B. abortus RB51 and B. melitensis Rev1 are found most effective against brucellosis infection in animals, contriving a number of serious side effects and having chances to revert back into their active pathogenic form. In order to engineer a safe and effective vaccine candidate to be used in both animals and human, a recombinant subunit vaccine molecule comprising the truncated region of glucokinase (r-glk) gene from B. abortus S19 was cloned and expressed in Escherichia coli BL21DE3 host. Female BALB/c mice immunized with purified recombinant protein developed specific antibody titer of 1:64,000. The predominant IgG2a and IgG2b isotypes signified development of Th1 directed immune responses. In vitro cell cytotoxicity assay using anti-r-glk antibodies incubated with HeLa cells showed 81.20% and 78.5% cell viability against lethal challenge of B. abortus 544 and B. melitensis 16M, respectively. The lymphocyte proliferative assay indicated a higher splenic lymphocyte responses at 25μg/ml concentration of protein which implies the elevated development of memory immune responses. In contrast to control, the immunized group of mice intra-peritoneal (I.P.) challenged with B. abortus 544 were significantly protected with no signs of necrosis and vacuolization in their liver and spleen tissue. The elevated B-cell response associated with Th1 adopted immunity, significant in vitro cell viability as well as protection afforded in experimental animals after challenge, supplemented with histopathological analysis are suggestive of r-glk protein as a prospective candidate vaccine molecule against brucellosis. SN - 1873-2518 UR - https://www.unboundmedicine.com/medline/citation/25131740/Studies_on_recombinant_glucokinase__r_glk__protein_of_Brucella_abortus_as_a_candidate_vaccine_molecule_for_brucellosis_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0264-410X(14)01088-3 DB - PRIME DP - Unbound Medicine ER -