Protective efficacy of recombinant proteins adenylate kinase, nucleoside diphosphate kinase, and heat-shock protein 70 against Mycobacterium tuberculosis infection in mice

Seung Heon Lee, Eun Gae Lee, Su Yeon Kim, Sangnae Cho, Young Kil Park, Gill Han Bai

Research output: Contribution to journalArticle

Abstract

Background: Priming and boosting vaccination strategy has been widely explored for new vaccine development against tuberculosis. As an effort to identify other vaccine candidates, this study was initiated to evaluate protective efficacy of adenylate kinase (AK), nucleoside diphosphate kinase (NdK), and heat shock protein 70 (Hsp70) of Mycobacterium tuberculosis. Method: M. tuberculosis genes encoding AK, NdK, and Hsp70 proteins were amplified by PCR and cloned into E. coli expression vector, pQE30. Recombinant AK, NdK, and Hsp70 was purified through Ni-NTA resin. To evaluate immune responses, we performed enzyme-linked immunosorbent assay (ELISA) for IgG isotype and IFN-γ after mice were immunized subcutaneously with recombinant proteins delivered in dimethyl dioctadecylammonium bromide (DDA). Immunized- and control groups were challenged by aerosol with M. tuberculosis. The spleens and lungs of mice were removed aseptically and cultured for CFU of M. tuberculosis. Result: Vaccination with recombinant proteins AK, NdK, and Hsp70 delivered in DDA elicited significant level of antibody and IFN-γ responses to corresponding antigens but no protective immunity comparable to that achieved with Mycobacterium bovis BCG. Conclusion: Recombinant proteins AK, NdK, and Hsp70 do not effectively control growth of M. tuberculosis in mice when immunized with DDA as an adjuvant.

Original languageEnglish
Pages (from-to)142-151
Number of pages10
JournalTuberculosis and Respiratory Diseases
Volume58
Issue number2
Publication statusPublished - 2005 Feb 1

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Nucleoside-Diphosphate Kinase
Adenylate Kinase
Mycobacterium Infections
HSP70 Heat-Shock Proteins
Mycobacterium tuberculosis
Recombinant Proteins
Protein Kinases
Bromides
Mycobacterium bovis
Vaccination
Vaccines
Aerosols
Antibody Formation
Immunity
Tuberculosis
Spleen
Immunoglobulin G
Enzyme-Linked Immunosorbent Assay
Escherichia coli
Antigens

All Science Journal Classification (ASJC) codes

  • Pulmonary and Respiratory Medicine
  • Infectious Diseases

Cite this

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title = "Protective efficacy of recombinant proteins adenylate kinase, nucleoside diphosphate kinase, and heat-shock protein 70 against Mycobacterium tuberculosis infection in mice",
abstract = "Background: Priming and boosting vaccination strategy has been widely explored for new vaccine development against tuberculosis. As an effort to identify other vaccine candidates, this study was initiated to evaluate protective efficacy of adenylate kinase (AK), nucleoside diphosphate kinase (NdK), and heat shock protein 70 (Hsp70) of Mycobacterium tuberculosis. Method: M. tuberculosis genes encoding AK, NdK, and Hsp70 proteins were amplified by PCR and cloned into E. coli expression vector, pQE30. Recombinant AK, NdK, and Hsp70 was purified through Ni-NTA resin. To evaluate immune responses, we performed enzyme-linked immunosorbent assay (ELISA) for IgG isotype and IFN-γ after mice were immunized subcutaneously with recombinant proteins delivered in dimethyl dioctadecylammonium bromide (DDA). Immunized- and control groups were challenged by aerosol with M. tuberculosis. The spleens and lungs of mice were removed aseptically and cultured for CFU of M. tuberculosis. Result: Vaccination with recombinant proteins AK, NdK, and Hsp70 delivered in DDA elicited significant level of antibody and IFN-γ responses to corresponding antigens but no protective immunity comparable to that achieved with Mycobacterium bovis BCG. Conclusion: Recombinant proteins AK, NdK, and Hsp70 do not effectively control growth of M. tuberculosis in mice when immunized with DDA as an adjuvant.",
author = "Lee, {Seung Heon} and Lee, {Eun Gae} and Kim, {Su Yeon} and Sangnae Cho and Park, {Young Kil} and Bai, {Gill Han}",
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Protective efficacy of recombinant proteins adenylate kinase, nucleoside diphosphate kinase, and heat-shock protein 70 against Mycobacterium tuberculosis infection in mice. / Lee, Seung Heon; Lee, Eun Gae; Kim, Su Yeon; Cho, Sangnae; Park, Young Kil; Bai, Gill Han.

In: Tuberculosis and Respiratory Diseases, Vol. 58, No. 2, 01.02.2005, p. 142-151.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Protective efficacy of recombinant proteins adenylate kinase, nucleoside diphosphate kinase, and heat-shock protein 70 against Mycobacterium tuberculosis infection in mice

AU - Lee, Seung Heon

AU - Lee, Eun Gae

AU - Kim, Su Yeon

AU - Cho, Sangnae

AU - Park, Young Kil

AU - Bai, Gill Han

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