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Research Article

volume 53 issue 11 (november 2019) : 1509-1513,   Doi: 10.18805/ijar.B-3714

Cytokine response to killed Staphylococcus pseudintermedius antigen in dogs with skin diseases

H
Hridya Susan Varughese
M
M. Ananda Chitra
R
Ranjani Rajasekaran
S
S.Rajalakshmi
G
G. Dhinakar Raj
1Department of Veterinary Microbiology, Madras Veterinary College Tamil Nadu Veterinary and Animal Sciences University, Vepery, Chennai-600 007, Tamil Nadu, India.
Cite article:- Varughese Susan Hridya, Chitra Ananda M., Rajasekaran Ranjani, S.Rajalakshmi, Raj Dhinakar G. (2019). Cytokine response to killed Staphylococcus pseudintermedius antigen in dogs with skin diseases. Indian Journal of Animal Research. 53(11): 1509-1513. doi: 10.18805/ijar.B-3714.

ABSTRACT

The attempt was made to study the response of the dog having skin infection with various underlying causes to killed S.pseudintermedius antigen. PBMC were separated and sensitized with killed S. pseudintermedius for 24 hours. cDNA was made from extracted RNA and quantitative PCR were carried out for 8 cytokines with GAPDH as housekeeping gene. IL-6 was the cytokine which was expressed in a statistically significant high level in PBMC of healthy animals to killed antigen than PBMC of infected animals. Except for the IL-6, all other cytokines were expressed at high level in pyoderma animals than healthy control, demodicosis and allergy cases. In the present study, IL-1â, IL-8 and TNF-á were the cytokines that were up-regulated and, IL-6 and IFN-ã were down-regulated in demodicosis dogs with pyoderma at apparently significant level than the other tested cytokines. IL-4 was the only cytokine that was expressed in measurable quantity in allergic cases when compared to other cytokines. Thus, it was concluded that dogs with staphylococcal pyoderma infections developed a Th1/Th2 response to fight the infection. 

REFERENCES

  1. Akilov, O.E. and Mumcuoglu, K.Y. (2004). Immune response in demodicosis. European Academy of Dermatology and Venereology: JEADV :440–444 ,DOI: 10.1111/j.1468-3083.2004.00964.
  2. AnandaChitra, M., Jayanthy, C. and Nagarajan, B. (2015). Detection andsequence analysis of accessory gene regulator genes of Staphylococcus pseudintermedius isolates, Vet World: 8(7):902 -907.
  3. Bastien, B.C., Patil, A. and Satyaraj, E. (2015). The impact of weight loss on circulating cytokines in Beagle dogs, Vet Immunol Immunopathol: 163(3):174–182.
  4. Biancotto, A., Wank, A., Perl, S., Cook, W., Olnes, M.J., Dagur, P.K., Fuchs, J.C., Langweiler, M., Wang, E. and McCoy, J.P. (2013). Baseline levels and temporal stability of 27 multiplexed serum cytokine concentrations in healthy subjects, PLoS One:8(12): p.e76091 
  5. Böyum, A. (1968). Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g., Scand J Clin Lab Invest Suppl: 97:77-89.
  6. Camp, R.D.R., Fincham, N.J., Ross, J.S., Bacon, K.B. and Gearing, A.J.H.(1990). Leukocyte chemoattractant cytokines of the epidermis, J of Invest Derm: 95(6): 5108-5110.
  7. Dinarello, C.A. (2011). A clinical perspective of IL-1â as the gatekeeper of inflammation. Eur J Immunol: 41:1203–17.
  8. Felix, A.O.C., Guiot, E.G., Stein, M., Felix, S.R., Silva, E.F. and Nobre, M.O. (2013). Comparison of systemic interleukin 10 concen- trations in healthy dogs and those suffering from recurring and first time Demodex canis infestations, Vet Parasit:193: 312– 315.
  9. Ferrer, L., Ravera, I. and Silbermayr, K. (2014). Immunology and pathogenesis of canine demodicosis, Vet. Dermatol.:25: 427–465
  10. Fowler, V.J., Justice, A., Moore, C., Benjamin, D.K. Jr,, Woods, C.W., Campbell, S., Reller, L.B., et al (2005). Risk factors for haemat ogenous complications of intravascular catheter-associated Staphylococcus aureus bacteremia, Clin Infect Dis:40(5):695–703.
  11. Maina, E., Devriendt, B. and Cox, E. (2017). Changes in cytokine profiles following treatment with food allergen-specific sublingual immunotherapy in dogs with adverse food reactions. Vet Dermatol: 28: 612–e149. 
  12. Marzano, A.V., Fanoni, D., Antiga, E., Quaglino, P., Caproni, M., Crosti, C., Meroni, P. L. and. Cugno, M. (2014). Expression of cytokines, chemokines and other effector molecules intwo prototypic auto inflammatory skin diseases, pyodermagangrenosum and Sweet’s syndrome. Clin and Exp Immun.:178: 48–56.
  13. McNicholas, S., Talento, A.F., O’Gorman, J., Hannan, M.M., Lynch, M., Greene, C.M., Humphreys, H. and Fitzgerald-Hughes, D. (2014). Cytokine responses to Staphylococcus aureus bloodstream infection differ between patient cohorts that have different clinical courses of infection. BMC Infect Dis. 15: (14)580.
  14. Miller, W.H., Griffin, C.E. and Campbell, K.L. (2013). Parasitic diseases. In: Mullerand Kirk’s Small Animal Dermatology, 7th edition. Philadelphia PA: W.B. Saunders Co.; 284–342.
  15. Nuttall, T.J., Knight, P.A. and McAleese, S.M. (2002). Expression of Th1, Th2 and immunosuppressive cytokine gene transcripts in canine atopic dermatitis. Clin Exp Allergy; 32: 789–795.
  16. Olivry, T., Dean, G.A., Tompkins, M.B., Dow, J.L. and Moore, P.F. (1999). Toward a canine model of atopic dermatitis: amplification of cytokine gene transcripts in the skin of atopic dogs, Exp derm :8 ;204-211.
  17. Pfaffl, M.W. (2001). A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 29(9):e45
  18. Richter, K.R., Nasr1, A.N. and Mexas, AM. (2018). Cytokine concentrations measured by multiplex assays in canine peripheral blood samples, Vet Path: 55(1) :53-67.
  19. Santos, S.S., Brunialti, M.K., Rigato, O., Machado, F.R., Silva, E. and Salomao, R. (2012). Generation of nitric oxide and reactive oxygen species by neutrophils and monocytes from septic patients and association with outcomes, Shock: 38(1):18–23.
  20. Sauder, D.N. (1990). The role of epidermal cytokines in inflammatory skin diseases, J Invest Dermatol.:95: 27S-28S.
  21. Schlotter, Y.M., Rutten, V.P., Riemers, F.M., Knol, E.F. and Willemse, T. (2011). Lesional skin in atopic dogs shows a mixed Type-1 and Type-2 immune responsiveness, Vet immunol and immunopathol: 143:20-26.
  22. Takata, T., Miyazaki, M., Futo, M., Hara, S., Shiotsuka, S., Kamimura, H., Yoshimura, H., Matsunaga, A., et al (2013). Presence of both heterogeneous vancomycin-intermediate resistance and beta-lactam antibiotic-induced vancomycin resistance phenotypes is associated with the outcome in methicillin-resistant Staphylococcus aureus blood stream infection. Scand J Infect Dis.:    45(3):203–212.
  23. Tamura, Y., Ohta, H., Yokoyama, N., Lim, S.Y., Osuga, T., Morishita, K., Nakamura, K., Yamasaki, M. and Takiguchi, M. (2014). Evaluation of selected cytokine gene expression in colonic mucosa from dogs with idiopathic lymphocytic-plasmacytic colitis. J Vet Med Sci: 76 (10):1407-10. 
  24. Tani, K., Morimoto, M., Hayashi, T., Inokuma, H., Ohnishi, T., Hayashiya, S., Nomura, T., Une, S., Nakaichi, M. and Taura, Y. (2002). Evaluation of cytokine messenger RNA expression in peripheral blood mononuclear cells from dogs with canine demodicosis. J Vet Med Sci: 64(6):513-518,.
  25. vanDeuren, M., Dofferhoff, A.S.M. and van der Meer, J.W.M. (1992). Cytokines and the response to infection. J Path; 168: 349-56.
  26. vonPfeil, D.J., Cummings, B.P., Loftus, J.P., Levine, C.B., Mann, S., Downey, R.L., Griffitts, C. and Wakshlag, J.J. (2015). Evaluation of plasma inflammatory cytokine concentrations in racing sled dogs. Can Vet J: 56(12):1252.
  27. Wang, J.E., Jorgensen, P.F., Almlof, M., Thiemermann, C., Foster, S.J., Aasen, A.O. and Solberg, R. (2000). Peptidoglycan and lipoteichoic acid from Staphylococcus aureus induce tumor necrosis factor alpha, interleukin 6 (IL-6), and IL-10production in both T cells and monocytes in a human whole blood model. Infect Immun: 68(7):3965–3970. 
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Indian Journal of Animal Research
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    Research Article

    volume 53 issue 11 (november 2019) : 1509-1513,   Doi: 10.18805/ijar.B-3714

    Cytokine response to killed Staphylococcus pseudintermedius antigen in dogs with skin diseases

    H
    Hridya Susan Varughese
    M
    M. Ananda Chitra
    R
    Ranjani Rajasekaran
    S
    S.Rajalakshmi
    G
    G. Dhinakar Raj
    1Department of Veterinary Microbiology, Madras Veterinary College Tamil Nadu Veterinary and Animal Sciences University, Vepery, Chennai-600 007, Tamil Nadu, India.
    Cite article:- Varughese Susan Hridya, Chitra Ananda M., Rajasekaran Ranjani, S.Rajalakshmi, Raj Dhinakar G. (2019). Cytokine response to killed Staphylococcus pseudintermedius antigen in dogs with skin diseases. Indian Journal of Animal Research. 53(11): 1509-1513. doi: 10.18805/ijar.B-3714.

    ABSTRACT

    The attempt was made to study the response of the dog having skin infection with various underlying causes to killed S.pseudintermedius antigen. PBMC were separated and sensitized with killed S. pseudintermedius for 24 hours. cDNA was made from extracted RNA and quantitative PCR were carried out for 8 cytokines with GAPDH as housekeeping gene. IL-6 was the cytokine which was expressed in a statistically significant high level in PBMC of healthy animals to killed antigen than PBMC of infected animals. Except for the IL-6, all other cytokines were expressed at high level in pyoderma animals than healthy control, demodicosis and allergy cases. In the present study, IL-1â, IL-8 and TNF-á were the cytokines that were up-regulated and, IL-6 and IFN-ã were down-regulated in demodicosis dogs with pyoderma at apparently significant level than the other tested cytokines. IL-4 was the only cytokine that was expressed in measurable quantity in allergic cases when compared to other cytokines. Thus, it was concluded that dogs with staphylococcal pyoderma infections developed a Th1/Th2 response to fight the infection. 

    REFERENCES

    1. Akilov, O.E. and Mumcuoglu, K.Y. (2004). Immune response in demodicosis. European Academy of Dermatology and Venereology: JEADV :440–444 ,DOI: 10.1111/j.1468-3083.2004.00964.
    2. AnandaChitra, M., Jayanthy, C. and Nagarajan, B. (2015). Detection andsequence analysis of accessory gene regulator genes of Staphylococcus pseudintermedius isolates, Vet World: 8(7):902 -907.
    3. Bastien, B.C., Patil, A. and Satyaraj, E. (2015). The impact of weight loss on circulating cytokines in Beagle dogs, Vet Immunol Immunopathol: 163(3):174–182.
    4. Biancotto, A., Wank, A., Perl, S., Cook, W., Olnes, M.J., Dagur, P.K., Fuchs, J.C., Langweiler, M., Wang, E. and McCoy, J.P. (2013). Baseline levels and temporal stability of 27 multiplexed serum cytokine concentrations in healthy subjects, PLoS One:8(12): p.e76091 
    5. Böyum, A. (1968). Isolation of mononuclear cells and granulocytes from human blood. Isolation of monuclear cells by one centrifugation, and of granulocytes by combining centrifugation and sedimentation at 1 g., Scand J Clin Lab Invest Suppl: 97:77-89.
    6. Camp, R.D.R., Fincham, N.J., Ross, J.S., Bacon, K.B. and Gearing, A.J.H.(1990). Leukocyte chemoattractant cytokines of the epidermis, J of Invest Derm: 95(6): 5108-5110.
    7. Dinarello, C.A. (2011). A clinical perspective of IL-1â as the gatekeeper of inflammation. Eur J Immunol: 41:1203–17.
    8. Felix, A.O.C., Guiot, E.G., Stein, M., Felix, S.R., Silva, E.F. and Nobre, M.O. (2013). Comparison of systemic interleukin 10 concen- trations in healthy dogs and those suffering from recurring and first time Demodex canis infestations, Vet Parasit:193: 312– 315.
    9. Ferrer, L., Ravera, I. and Silbermayr, K. (2014). Immunology and pathogenesis of canine demodicosis, Vet. Dermatol.:25: 427–465
    10. Fowler, V.J., Justice, A., Moore, C., Benjamin, D.K. Jr,, Woods, C.W., Campbell, S., Reller, L.B., et al (2005). Risk factors for haemat ogenous complications of intravascular catheter-associated Staphylococcus aureus bacteremia, Clin Infect Dis:40(5):695–703.
    11. Maina, E., Devriendt, B. and Cox, E. (2017). Changes in cytokine profiles following treatment with food allergen-specific sublingual immunotherapy in dogs with adverse food reactions. Vet Dermatol: 28: 612–e149. 
    12. Marzano, A.V., Fanoni, D., Antiga, E., Quaglino, P., Caproni, M., Crosti, C., Meroni, P. L. and. Cugno, M. (2014). Expression of cytokines, chemokines and other effector molecules intwo prototypic auto inflammatory skin diseases, pyodermagangrenosum and Sweet’s syndrome. Clin and Exp Immun.:178: 48–56.
    13. McNicholas, S., Talento, A.F., O’Gorman, J., Hannan, M.M., Lynch, M., Greene, C.M., Humphreys, H. and Fitzgerald-Hughes, D. (2014). Cytokine responses to Staphylococcus aureus bloodstream infection differ between patient cohorts that have different clinical courses of infection. BMC Infect Dis. 15: (14)580.
    14. Miller, W.H., Griffin, C.E. and Campbell, K.L. (2013). Parasitic diseases. In: Mullerand Kirk’s Small Animal Dermatology, 7th edition. Philadelphia PA: W.B. Saunders Co.; 284–342.
    15. Nuttall, T.J., Knight, P.A. and McAleese, S.M. (2002). Expression of Th1, Th2 and immunosuppressive cytokine gene transcripts in canine atopic dermatitis. Clin Exp Allergy; 32: 789–795.
    16. Olivry, T., Dean, G.A., Tompkins, M.B., Dow, J.L. and Moore, P.F. (1999). Toward a canine model of atopic dermatitis: amplification of cytokine gene transcripts in the skin of atopic dogs, Exp derm :8 ;204-211.
    17. Pfaffl, M.W. (2001). A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 29(9):e45
    18. Richter, K.R., Nasr1, A.N. and Mexas, AM. (2018). Cytokine concentrations measured by multiplex assays in canine peripheral blood samples, Vet Path: 55(1) :53-67.
    19. Santos, S.S., Brunialti, M.K., Rigato, O., Machado, F.R., Silva, E. and Salomao, R. (2012). Generation of nitric oxide and reactive oxygen species by neutrophils and monocytes from septic patients and association with outcomes, Shock: 38(1):18–23.
    20. Sauder, D.N. (1990). The role of epidermal cytokines in inflammatory skin diseases, J Invest Dermatol.:95: 27S-28S.
    21. Schlotter, Y.M., Rutten, V.P., Riemers, F.M., Knol, E.F. and Willemse, T. (2011). Lesional skin in atopic dogs shows a mixed Type-1 and Type-2 immune responsiveness, Vet immunol and immunopathol: 143:20-26.
    22. Takata, T., Miyazaki, M., Futo, M., Hara, S., Shiotsuka, S., Kamimura, H., Yoshimura, H., Matsunaga, A., et al (2013). Presence of both heterogeneous vancomycin-intermediate resistance and beta-lactam antibiotic-induced vancomycin resistance phenotypes is associated with the outcome in methicillin-resistant Staphylococcus aureus blood stream infection. Scand J Infect Dis.:    45(3):203–212.
    23. Tamura, Y., Ohta, H., Yokoyama, N., Lim, S.Y., Osuga, T., Morishita, K., Nakamura, K., Yamasaki, M. and Takiguchi, M. (2014). Evaluation of selected cytokine gene expression in colonic mucosa from dogs with idiopathic lymphocytic-plasmacytic colitis. J Vet Med Sci: 76 (10):1407-10. 
    24. Tani, K., Morimoto, M., Hayashi, T., Inokuma, H., Ohnishi, T., Hayashiya, S., Nomura, T., Une, S., Nakaichi, M. and Taura, Y. (2002). Evaluation of cytokine messenger RNA expression in peripheral blood mononuclear cells from dogs with canine demodicosis. J Vet Med Sci: 64(6):513-518,.
    25. vanDeuren, M., Dofferhoff, A.S.M. and van der Meer, J.W.M. (1992). Cytokines and the response to infection. J Path; 168: 349-56.
    26. vonPfeil, D.J., Cummings, B.P., Loftus, J.P., Levine, C.B., Mann, S., Downey, R.L., Griffitts, C. and Wakshlag, J.J. (2015). Evaluation of plasma inflammatory cytokine concentrations in racing sled dogs. Can Vet J: 56(12):1252.
    27. Wang, J.E., Jorgensen, P.F., Almlof, M., Thiemermann, C., Foster, S.J., Aasen, A.O. and Solberg, R. (2000). Peptidoglycan and lipoteichoic acid from Staphylococcus aureus induce tumor necrosis factor alpha, interleukin 6 (IL-6), and IL-10production in both T cells and monocytes in a human whole blood model. Infect Immun: 68(7):3965–3970. 
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