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Current IssueEditorial BoardOnline First ArticlesArchive

volume 47 issue 3 (june 2013) : 240-243

DETECTION OF CD4/CD8 RATIO IN MICE IMMUNIZED WITH A BICISTRONIC PLASMID CONSTRUCT ENCODING A PPE GENE OF MYCOBACTERIUM AVIUM PARATUBERCULOSIS AND A CYTOKINE GENE OF MURINE GAMMA INTERFERON*

R
Rajib Deb
P
P. P. Goswami
1Division of Veterinary Biotechnology Indian Veterinary Research Institute, Izatnagar – 243 122, India

  • Submitted|

  • First Online |

  • doi

Cite article:- Deb Rajib, Goswami P. P. (2025). DETECTION OF CD4/CD8 RATIO IN MICE IMMUNIZED WITH A BICISTRONIC PLASMID CONSTRUCT ENCODING A PPE GENE OF MYCOBACTERIUM AVIUM PARATUBERCULOSIS AND A CYTOKINE GENE OF MURINE GAMMA INTERFERON*. Indian Journal of Animal Research. 47(3): 240-243. doi: .

ABSTRACT

In the present study  a gene encoding PPE protein of M.a.paratuberculosis (FJ032182) was cloned with murine IFN-³  in a mammalian  bicistronic vector pIRES 6.1 to elucidate  the role of gamma interferon on the PPE gene for determination of CD4/CD8 ratio  in post immunized mice. Flow cytometric analysis with mononuclear mice splenocytes on 42nd day post immunization revealed significant reduction of the CD4/CD8 ratio in the mice group immunized with pIR PPE/IFN (1.65±0.0001) compared to the  pIR PPE group (2.92±0.0003). These result suggested that co-expression of murine IFNg in conjunction with PPE protein significantly enhanced the CMI response.

REFERENCES

  1. Andersen, P. (2001) TB vaccines: progress and problems. Trends Immunol., 22(3):160–8.
  2. Barouch, D.H., Santra, S., Tenner-Racz, K., Racz, P., Kuroda, M.J., Schmitz, J.E., Jackson, S.s., Lufton, M.A., Freed, D.C., Perry, H.C., Davies, M.E., Shiver, J.W. and Letvin, N.L. (2002). Potent CD4+ T cell response elicited by a bicistronic HIV-1 DNA vaccine expressing gp120 and GM-CSF. J. of Immunol., 168: 562-568.
  3. Chambers, M.A., Williams, A., Hatch, G., Garier-Widen, D., Hall, G., Huygen, K., Lowrie, D., Marsh, P.D. and Hewinson, R.G. (2002). Vaccination of guinea pigs with DNA encoding the Mycobacterial antigen MPB83 influences pulmonary pathology but not hematogenous spread following aerogenic infection with Mycobacterium bovis. Infect. Imun., 70: 2159-2165.
  4. Coussens, P.M., Verman, N., Coussens, M.A., Elftman, M.D. and McNulty, A.M. (2004). Cytokine gene expression in peripheral blood mononuclear cells and tissue of cattle infected with Mycobacterium avium subsp. paratuberculosis : evidence for an inherent proinflammatory gene expression pattern. Infect. Immun., 73(3): 1409-1422.
  5. Goldsby, R.A., Kindt, T.J., Osborne, B.A. and Kuby, J. (2003). Immunology, Cytokine, 5th ed., W.H. Freeman and Company, New York p 288.
  6. Harris, N.B. and Barletta, R.G. (2001). Mycobacterium avium subsp. Paratuberculosis in veterinary medicine. Clin. Microbiol. Rev. 14: 489-512.
  7. Kaufmann, SH. (2001). How can immunology contribute to the control of tuberculosis? Nat Rev Immunol ., 1(1):20– 30.
  8. Koets, A., Hoek, A., Langelaar, M., Overdijk, M., Santema, W., Franken, P., Eden, W. and Rutten, V. (2006). Mycobacterial 70 kDa heat-shock protein is an effective subunit vaccine against bovine paratuberculosis. Vaccine., 24: 2550- 2559.
  9. Martin, E., Kamath, A.T., Tricass, J.A. and Brilton, W.J. (2000). Protection against virulent Mycobacterium avium infection following DNA vaccination with the 35 kDa antigen is accompanied by induction of gamma interferon secreting CD4+ T cells. Infect. Immun., 68: 3090-3096.
  10. Mullerad, J., Hovav, A.H., Nahary, R., Fishman, Y. and Bercovier, H. (2003). Immunogenecity of 16.7 kDa Mycobacterium paratuberculosis antigen. Microbiol. Pathogenesis, 34: 81-90.
  11. Mullerad, J., Hovav, Avi-H., Fishman, Y., Barletta, R.G. and Bercovier, H. (2002a). Antigenicity of Mycobacterium paratuberculosis superoxide dismutase in mice. FEMS Immunol. Med. Microbiol., 34: 81-88.
  12. Mullerad, J., Michal, I., Fishman, V., HOrar, A.H., Barletta, R.G. and Bercovier, H. (2002b). The imunogenecity of Mycobacterium paratuberculosis 85B antigen. Mol. Microbiol. Immunol., 190: 179-187.
  13. Okkels, L. M., I. Brock, F. Follmann, E. M. Agger, S. M. Arend, T. H. M. Ottenhoff, F. Oftung, I. Rosenkrands, and P. Andersen. (2003). PPE protein (Rv3873) from DNA segment RD1 of Mycobacterium tuberculosis: strong recognition of both specific T-cell epitopes and epitopes conserved within the PPE family. Infect. Immun. 71: 6116–6123.
  14. Olsen, I., G. Sigurgardottir, and Djonne B.. (2002). Paratuberculosis with special reference to cattle: a review. Vet. Q., 24:12–28
  15. Pardini, M., Giannoni, F., Palma, C., Iona, E., Cafaro, A., Brunori, L., Rinaldi, M., Fazio, V.M., Laguardia, M.E., Carbonella, D.C., Magnani, M., Ensoli, B., Fattorini, L. and Cassone, A. (2006). Immune response and protection by DNA vaccines expressing antigen 85B of Mycobacterium tuberculosis. FEMS Microbiol Lett., 262(2): 210-5.
  16. Portillo, P.D., Murillo, L.A. and Patarroyo, M.E. (1991). Am Mark, T. D. (1998). Genetically engineered vaccines: An overview (review). Plasmid, 39: 100-113.
  17. Sechi, L.A., Mara, L., Cappai, P., Frothingam, R., Ortu, S., Leoni, A., Ahmed, N. and Zanetti, S. (2006). Immunization with DNA vaccines encoding different mycobacterial antigens elicits a Th1 type immune response in lambs and protects against Mycobacterium avium subspecies paratuberculosis infection. Vaccine, 24: 229-235.
  18. Skeiky, Y. A. W., P. J. Ovendale, S. Jen, M. R. Alderson, D. C. Dillon, S. Smith, C. B. Wilson, I. M. Orme, S. G. Reed, and A. Campos-Neto. (2000). T cell expression cloning of a Mycobacterium tuberculosis gene encoding a protective antigen associated with the early control of infection. J. Immunol., 165: 7140–7149.
  19. Stabel, J.R. (1996). Production of ã-interferon by peripheral blood mononuclear cells: an important diagnostic tool for detection of subclinical paratuberculosis. J. Vet. Diagn. Investig., 8: 345-350.
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Indian Journal of Animal Research
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    volume 47 issue 3 (june 2013) : 240-243

    DETECTION OF CD4/CD8 RATIO IN MICE IMMUNIZED WITH A BICISTRONIC PLASMID CONSTRUCT ENCODING A PPE GENE OF MYCOBACTERIUM AVIUM PARATUBERCULOSIS AND A CYTOKINE GENE OF MURINE GAMMA INTERFERON*

    R
    Rajib Deb
    P
    P. P. Goswami
    1Division of Veterinary Biotechnology Indian Veterinary Research Institute, Izatnagar – 243 122, India

    • Submitted|

    • First Online |

    • doi

    Cite article:- Deb Rajib, Goswami P. P. (2025). DETECTION OF CD4/CD8 RATIO IN MICE IMMUNIZED WITH A BICISTRONIC PLASMID CONSTRUCT ENCODING A PPE GENE OF MYCOBACTERIUM AVIUM PARATUBERCULOSIS AND A CYTOKINE GENE OF MURINE GAMMA INTERFERON*. Indian Journal of Animal Research. 47(3): 240-243. doi: .

    ABSTRACT

    In the present study  a gene encoding PPE protein of M.a.paratuberculosis (FJ032182) was cloned with murine IFN-³  in a mammalian  bicistronic vector pIRES 6.1 to elucidate  the role of gamma interferon on the PPE gene for determination of CD4/CD8 ratio  in post immunized mice. Flow cytometric analysis with mononuclear mice splenocytes on 42nd day post immunization revealed significant reduction of the CD4/CD8 ratio in the mice group immunized with pIR PPE/IFN (1.65±0.0001) compared to the  pIR PPE group (2.92±0.0003). These result suggested that co-expression of murine IFNg in conjunction with PPE protein significantly enhanced the CMI response.

    REFERENCES

    1. Andersen, P. (2001) TB vaccines: progress and problems. Trends Immunol., 22(3):160–8.
    2. Barouch, D.H., Santra, S., Tenner-Racz, K., Racz, P., Kuroda, M.J., Schmitz, J.E., Jackson, S.s., Lufton, M.A., Freed, D.C., Perry, H.C., Davies, M.E., Shiver, J.W. and Letvin, N.L. (2002). Potent CD4+ T cell response elicited by a bicistronic HIV-1 DNA vaccine expressing gp120 and GM-CSF. J. of Immunol., 168: 562-568.
    3. Chambers, M.A., Williams, A., Hatch, G., Garier-Widen, D., Hall, G., Huygen, K., Lowrie, D., Marsh, P.D. and Hewinson, R.G. (2002). Vaccination of guinea pigs with DNA encoding the Mycobacterial antigen MPB83 influences pulmonary pathology but not hematogenous spread following aerogenic infection with Mycobacterium bovis. Infect. Imun., 70: 2159-2165.
    4. Coussens, P.M., Verman, N., Coussens, M.A., Elftman, M.D. and McNulty, A.M. (2004). Cytokine gene expression in peripheral blood mononuclear cells and tissue of cattle infected with Mycobacterium avium subsp. paratuberculosis : evidence for an inherent proinflammatory gene expression pattern. Infect. Immun., 73(3): 1409-1422.
    5. Goldsby, R.A., Kindt, T.J., Osborne, B.A. and Kuby, J. (2003). Immunology, Cytokine, 5th ed., W.H. Freeman and Company, New York p 288.
    6. Harris, N.B. and Barletta, R.G. (2001). Mycobacterium avium subsp. Paratuberculosis in veterinary medicine. Clin. Microbiol. Rev. 14: 489-512.
    7. Kaufmann, SH. (2001). How can immunology contribute to the control of tuberculosis? Nat Rev Immunol ., 1(1):20– 30.
    8. Koets, A., Hoek, A., Langelaar, M., Overdijk, M., Santema, W., Franken, P., Eden, W. and Rutten, V. (2006). Mycobacterial 70 kDa heat-shock protein is an effective subunit vaccine against bovine paratuberculosis. Vaccine., 24: 2550- 2559.
    9. Martin, E., Kamath, A.T., Tricass, J.A. and Brilton, W.J. (2000). Protection against virulent Mycobacterium avium infection following DNA vaccination with the 35 kDa antigen is accompanied by induction of gamma interferon secreting CD4+ T cells. Infect. Immun., 68: 3090-3096.
    10. Mullerad, J., Hovav, A.H., Nahary, R., Fishman, Y. and Bercovier, H. (2003). Immunogenecity of 16.7 kDa Mycobacterium paratuberculosis antigen. Microbiol. Pathogenesis, 34: 81-90.
    11. Mullerad, J., Hovav, Avi-H., Fishman, Y., Barletta, R.G. and Bercovier, H. (2002a). Antigenicity of Mycobacterium paratuberculosis superoxide dismutase in mice. FEMS Immunol. Med. Microbiol., 34: 81-88.
    12. Mullerad, J., Michal, I., Fishman, V., HOrar, A.H., Barletta, R.G. and Bercovier, H. (2002b). The imunogenecity of Mycobacterium paratuberculosis 85B antigen. Mol. Microbiol. Immunol., 190: 179-187.
    13. Okkels, L. M., I. Brock, F. Follmann, E. M. Agger, S. M. Arend, T. H. M. Ottenhoff, F. Oftung, I. Rosenkrands, and P. Andersen. (2003). PPE protein (Rv3873) from DNA segment RD1 of Mycobacterium tuberculosis: strong recognition of both specific T-cell epitopes and epitopes conserved within the PPE family. Infect. Immun. 71: 6116–6123.
    14. Olsen, I., G. Sigurgardottir, and Djonne B.. (2002). Paratuberculosis with special reference to cattle: a review. Vet. Q., 24:12–28
    15. Pardini, M., Giannoni, F., Palma, C., Iona, E., Cafaro, A., Brunori, L., Rinaldi, M., Fazio, V.M., Laguardia, M.E., Carbonella, D.C., Magnani, M., Ensoli, B., Fattorini, L. and Cassone, A. (2006). Immune response and protection by DNA vaccines expressing antigen 85B of Mycobacterium tuberculosis. FEMS Microbiol Lett., 262(2): 210-5.
    16. Portillo, P.D., Murillo, L.A. and Patarroyo, M.E. (1991). Am Mark, T. D. (1998). Genetically engineered vaccines: An overview (review). Plasmid, 39: 100-113.
    17. Sechi, L.A., Mara, L., Cappai, P., Frothingam, R., Ortu, S., Leoni, A., Ahmed, N. and Zanetti, S. (2006). Immunization with DNA vaccines encoding different mycobacterial antigens elicits a Th1 type immune response in lambs and protects against Mycobacterium avium subspecies paratuberculosis infection. Vaccine, 24: 229-235.
    18. Skeiky, Y. A. W., P. J. Ovendale, S. Jen, M. R. Alderson, D. C. Dillon, S. Smith, C. B. Wilson, I. M. Orme, S. G. Reed, and A. Campos-Neto. (2000). T cell expression cloning of a Mycobacterium tuberculosis gene encoding a protective antigen associated with the early control of infection. J. Immunol., 165: 7140–7149.
    19. Stabel, J.R. (1996). Production of ã-interferon by peripheral blood mononuclear cells: an important diagnostic tool for detection of subclinical paratuberculosis. J. Vet. Diagn. Investig., 8: 345-350.
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