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Stress in the regulation of the immune response in Leishmaniasis Felix J. Tapia Instituto de Biomedicina Facultad de Medic...
Neuroendocrine-immune axis Diffuse neuroendocrine system Peripheral Immune system Bioactive Polypeptides hormones  neurope...
Murine models of American cutaneous Leishmaniasis  C57BL/6 BALB/c Th1 IFN-  Th2 IL-4 L. mexicana MHOM/BZ/82/BEL21 Sánchez...
Sensory peptides:  Calcitonin gene-related peptide (CGRP) Substance (Sub P)    mediators of neurogenic inflammation Epide...
Background  <ul><li>Dendritic Langerhans cells are involved in the pathogenesis of cutaneous leishmaniasis.  </li></ul><ul...
Phases of Cutaneous Regulation Activation Inmunostimulation Effector Phase Challenging Antigen Capture Migration Antigen P...
epidermis dermis Mo ganglio linfático endotelio vascular C T T Mo ag .. . granuloma IL-4 mastocyte IFN-  NK M Φ IL-12 T m...
Objective <ul><li>Analyze the effect of acute stress (immobilization  and odorant) on Langerhans cells, Sub P, CGRP, and t...
Acute stressor (2, 8 hrs): Immobilization by placement in 50 ml polypropylene centrifuge tubes, before inoculation of  Lei...
BALB/c C57/BL6 Measurement footpad thickness every week for 12 weeks Mice stressed for 2 hr and infected with  L. mexicana...
Progression of  L. mexicana  infection  in experimental groups of BALB/c and C57BL/6 mice  0 1 2 3 4 5 6 7 8 9 10 11 12 13...
Langerhans cell density in experimental groups  of BALB/c and C57BL/6 at weeks O, 4 and 8 0.0 4.0 8.0 12.0 0 250 500 750 1...
Progression of infection with  L. mexicana  in  BALB/c mice subjected to acute stress
Langerhans cells Ia d +  in separated  epidermis of BALB/c mice infected with  L. mexicana Non stressed Stressed for 2 hr
Substance P in skin lesions of BALB/c  mice stressed and infected with  L. mexicana week 4 -  stressed for 8 hr Vascular e...
 
Conclusions <ul><li>Acute immobilization stress affects the numbers and function of Langerhans cells and Substance P and C...
Odorant inhalation stress in resistant and susceptible mouse models of cutaneous leishmaniasis
Acute stressor (48 hrs): Inhalation of citral for 48 hours.  At 24 hrs of stress, mice were infected  Leishmania  mexicana...
Odorant inhalation stress alters the immune response in resistant and susceptible mice infected with  L. mexicana
Odorant inhalation stress alters the immune response in resistant and susceptible mice infected with  L. mexicana
Odorant inhalation stress alters the immune response in resistant and susceptible mice infected with  L. mexicana
Conclusions Citral inhalation stress induces clinical and immunological alterations in the natural course of infection in ...
Macrophages treated by sera from mice sressed and infected with  L. mexicana 24 hours , Giemsa
1 3 5 9 11 0 100 200 BALB/c  stressed & infect BALB/c infect Weeks of infection Density of  DC-SIGN  positive cells in les...
1 3 5 9 11 0 1000 2000 3000 BALB/c infectado y estresado BALB/c infectado * * * * Density of  macrophages  MOMA-2   positi...
TLR-2   positive  cells in BALB/c mice stressed and infected with  L. mexicana 5 weeks  1 3 5 9 11 0 100 200 300 BALB/c  s...
1 3 5 9 11 0 100 200 BALB/c  stressed & infect BALB/c infect * Weeks of infection TLR- 4   positive  cells in BALB/c mice ...
CD14   positive  cells in BALB/c mice stressed and infected with  L. mexicana 5 weeks  1 3 5 9 11 0 100 200 300 BALB/c  st...
1 3 5 9 11 0 100 200 TLR- 9   positive  cells in BALB/c mice stressed and infected with  L. mexicana 5 weeks  Weeks of inf...
Laboratorio de Biología Molecular Instituto de Biomedicina Estudiantes: Ysamar Chirinos Alba Quiñones María del Rosario Ru...
Obrigado Thanks Gracias
American cutaneous leishmaniasis C57BL/6 IFN-  BALB/c IL-4 LCL Intermediate forms MCL CCL DCL C BA IFN   / IL-4
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Stress in the regulation of the immune response in Leishmaniasis

Presentacion en la Rede Leish vacina de CYTED, Salvador, Bahia, Brasil 21-09-09

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Stress in the regulation of the immune response in Leishmaniasis

  1. 1. Stress in the regulation of the immune response in Leishmaniasis Felix J. Tapia Instituto de Biomedicina Facultad de Medicina Universidad Central de Venezuela Reuniäo Cyted 2009 Projecto Mecanismo de controle e imunoprofilaxia na Leishmaniose Faculdade de Medicina da Bahia (FAMEB)
  2. 2. Neuroendocrine-immune axis Diffuse neuroendocrine system Peripheral Immune system Bioactive Polypeptides hormones neuropeptides cytokines Molecules of communication
  3. 3. Murine models of American cutaneous Leishmaniasis C57BL/6 BALB/c Th1 IFN-  Th2 IL-4 L. mexicana MHOM/BZ/82/BEL21 Sánchez et al. Acta Micros 1993, 2: 180-187 Aguilar-Torrentera et al. Am J Trop Med Hyg 2002, 66: 273–279 Díaz et al. Clin Exp Dermatol 2003, 28: 288-293
  4. 4. Sensory peptides: Calcitonin gene-related peptide (CGRP) Substance (Sub P)  mediators of neurogenic inflammation Epidermal dendritic (Langerhans) cells: Skin  take up and antigen processing Peripheral lymphoid organs  initiate primary T cell responses Leishmania infection of BALB/c and C57BL/6 mice : 10 3 amastigotes L. mexicana strain MHOM/BZ/82/BEL21 Course of infection 12 weeks Sites of evaluation
  5. 5. Background <ul><li>Dendritic Langerhans cells are involved in the pathogenesis of cutaneous leishmaniasis. </li></ul><ul><li>Epidermal cell signaling is essential to determine the type of cytokine-related immune response to be generated against Leishmania parasites. </li></ul><ul><li>Dendritic Langerhans cells, SubP and CGRP are affected by acute stress in contact hypersensitivity models. </li></ul>
  6. 6. Phases of Cutaneous Regulation Activation Inmunostimulation Effector Phase Challenging Antigen Capture Migration Antigen Presentation Clonal Expansion Recruitment Retention Proliferation Suppression
  7. 7. epidermis dermis Mo ganglio linfático endotelio vascular C T T Mo ag .. . granuloma IL-4 mastocyte IFN-  NK M Φ IL-12 T memory QC CL GM-CSF IL-1 TNF-  chemokines
  8. 8. Objective <ul><li>Analyze the effect of acute stress (immobilization and odorant) on Langerhans cells, Sub P, CGRP, and the natural course of infection in susceptible and resistant mouse models of cutaneous leishmaniasis </li></ul>
  9. 9. Acute stressor (2, 8 hrs): Immobilization by placement in 50 ml polypropylene centrifuge tubes, before inoculation of Leishmania parasites. Materials and methods Stress procedure
  10. 10. BALB/c C57/BL6 Measurement footpad thickness every week for 12 weeks Mice stressed for 2 hr and infected with L. mexicana Mice stressed for 8 hr and infected with L. mexicana Mice infected with L. mexicana Infection Control Mice stressed for 2 hr and non-infected with L. mexicana Stress Control (n=20) (n=20) (n=20) (n=20) (n=20) (n=20) (n=20) (n=20) Experimental design Footpad biopsies taken at weeks 0, 4 and 8 after infection Cryopreservation Immunodetection of CD205+ Langerhans cells, CGRP and Sub P
  11. 11. Progression of L. mexicana infection in experimental groups of BALB/c and C57BL/6 mice 0 1 2 3 4 5 6 7 8 9 10 11 12 13 1 2 3 4 5 6 Weeks of i infec tio n Size of lesi o n i n mm 0 1 2 3 4 5 6 7 8 9 10 11 12 13 1 2 3 4 5 6 2hr stress healthy 2hr stress infect ed 8hr stress infect ed Non-stressed infect ed
  12. 12. Langerhans cell density in experimental groups of BALB/c and C57BL/6 at weeks O, 4 and 8 0.0 4.0 8.0 12.0 0 250 500 750 1000 2hr stress healthy 2hr stress infect ed 8hr stress infec ed Non-stressed infect ed 0 1000 2000 0 sem 4 sem 8 sem Weeks of infec tion c ells /mm 2
  13. 13. Progression of infection with L. mexicana in BALB/c mice subjected to acute stress
  14. 14. Langerhans cells Ia d + in separated epidermis of BALB/c mice infected with L. mexicana Non stressed Stressed for 2 hr
  15. 15. Substance P in skin lesions of BALB/c mice stressed and infected with L. mexicana week 4 - stressed for 8 hr Vascular edothelium infiltrate
  16. 17. Conclusions <ul><li>Acute immobilization stress affects the numbers and function of Langerhans cells and Substance P and CGRP innervations. </li></ul><ul><li>The skew of the skin immune response after stress and infection may be diverted by epidermal accessory signals to distinct Th1 or Th2 responses in resistant or susceptible mice, respectively. </li></ul><ul><li>The genetic background of the mouse strain determined the response to acute stress in murine leishmaniasis. </li></ul>
  17. 18. Odorant inhalation stress in resistant and susceptible mouse models of cutaneous leishmaniasis
  18. 19. Acute stressor (48 hrs): Inhalation of citral for 48 hours. At 24 hrs of stress, mice were infected Leishmania mexicana Materials and methods Stress procedure
  19. 20. Odorant inhalation stress alters the immune response in resistant and susceptible mice infected with L. mexicana
  20. 21. Odorant inhalation stress alters the immune response in resistant and susceptible mice infected with L. mexicana
  21. 22. Odorant inhalation stress alters the immune response in resistant and susceptible mice infected with L. mexicana
  22. 23. Conclusions Citral inhalation stress induces clinical and immunological alterations in the natural course of infection in mice inoculated with L. mexicana. These alterations include decreased numbers of dendritic Langerhans cells and their morphological, and downregulation of Th1 and Th2 cytokines in both resistant and susceptible mice. Only IL-12 and iNOS remained in normal values. The results indicate that citral inhalation suppress the immune response against the Leishmania parasite regardless the genetic background of the host.
  23. 24. Macrophages treated by sera from mice sressed and infected with L. mexicana 24 hours , Giemsa
  24. 25. 1 3 5 9 11 0 100 200 BALB/c stressed & infect BALB/c infect Weeks of infection Density of DC-SIGN positive cells in lesions of stressed and L.mexicana -infected BALB/c mice c ells /mm 2
  25. 26. 1 3 5 9 11 0 1000 2000 3000 BALB/c infectado y estresado BALB/c infectado * * * * Density of macrophages MOMA-2 positive cells in lesions of stressed and L.mexicana -infected BALB/c mice * p  0.05 Weeks of infection c ells /mm 2
  26. 27. TLR-2 positive cells in BALB/c mice stressed and infected with L. mexicana 5 weeks 1 3 5 9 11 0 100 200 300 BALB/c stressed & infect BALB/c infect Weeks of infection c ells /mm 2
  27. 28. 1 3 5 9 11 0 100 200 BALB/c stressed & infect BALB/c infect * Weeks of infection TLR- 4 positive cells in BALB/c mice stressed and infected with L. mexicana 5 weeks * p  0.05 c ells /mm 2
  28. 29. CD14 positive cells in BALB/c mice stressed and infected with L. mexicana 5 weeks 1 3 5 9 11 0 100 200 300 BALB/c stressed & infect BALB/c infect Weeks of infec tion c ells /mm 2
  29. 30. 1 3 5 9 11 0 100 200 TLR- 9 positive cells in BALB/c mice stressed and infected with L. mexicana 5 weeks Weeks of infection BALB/c stressed & infect BALB/c infect c ells /mm 2
  30. 31. Laboratorio de Biología Molecular Instituto de Biomedicina Estudiantes: Ysamar Chirinos Alba Quiñones María del Rosario Ruiz Fabiola Cabrera María Alejandra Sarabia Eliana Figueira Mónica Fernández Adriana Arbeláez Celsy Hernández Alejandra Da Almeida Pedro Salazar María García Izaskun Urdanibia Laboratorio: Nilka L. Díaz Iraima B. Monsalve Mónica Suárez Miguel Marzal Luis González Colaboradores: Martín A. Sánchez Zelandia Fermín Ladys Sarmiento
  31. 32. Obrigado Thanks Gracias
  32. 33. American cutaneous leishmaniasis C57BL/6 IFN-  BALB/c IL-4 LCL Intermediate forms MCL CCL DCL C BA IFN  / IL-4

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