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Immunopathology of bovine neosporosis
426
Immunity to parasites
25 June 1997 - Poster presentations
Treatment with chloroquine on 7w day after prknary infection, reduces significantly the cross protection to heterologous challenge. However, immune mice treated with chloroquine and challenged with homologous parasites still remain protected. Reinfection of immune mice (treated or not with chloroquine) with the mixtures of I? c. chabaudiand Ryoe/ii 17XLshowed very high parasftemias however after day 12 post challenge mice started to control the parasitemia in contrast to the controls. Conoiueion: These experiments confirm and extend earliest studies on the specificity of immunity induced in inbred mice to rodent malarias. They also demonstrate that cross protection against heterotogous challenge is lost when primary parasitemia is stopped at 7 day after infection using chloroquine. This work was supported by DGAPA F’oyect 7N.211195 and CONACyT @#P-M.
09:00-18:30/12:09-14:00
P.4.08
Forum lounges
Immunity to parasites
1P.4.08.01 ] Fish immune responses and disease resistance W.B. Van Muiswinkel, B.M.L. Verburg, Van Kemenade, G.F. Wiegertjes. Wageningen Institute of Animal Sciences (WIAS), Waganingen Agricultural University (WAU), RO. Box 3336700 AH Wageningen, The Netherlands introduction: During the last 20 years considerable progress has been made in describing and understanding the immune system of fish. Fish are the oldest animal group with an immune system showing clear similarities with the defence systems of mammals and birds. Both innate immunity and acquired immunity are important for the defence against invading pathogens. Methods and Results: Humoral and cellular responses show the expected characteristics of specificity and memory after injection of standard antigens in carp (Cyprinus carpio L.). There is a clear influence of external factors such as water temperature and stress conditions (e.g. handling, oral application of cortisol). Moreover, the genetic background of the fish can also play a rote in the immune response against DNP-KLH as well in disease resistance to a blood flagellate, Twpanoplasma bone/i. This aspect was investigated by developing high and low responding inbred lines of fish. Conclusion: It can be concluded that the present knowledge can be used for evaluation of the health status of fish in the wild or in culture, but also for preventive measures such as vaccination or selection for disease resistance.
P.4.08.02
Cellular immunity to Sc/t/sfosoma infection In Zlmbabween children
heemefobium
J.T. Spicer ‘, C.M.R. Turner’, M. Woolhouse *, P. Ndlova 3, P. Hagan 4. ’ Division of infection and Immunity IBLS, University of Glasgow. UK, ‘Depanment of Zoology University of Oxford, UK, 3Blair Research Institute, Hamre, Zimbabwe, 4Commission of the European Communities, Brussels introduction: This study aimed to elucidate the role of cytokines in mediating the immune response to S. haematobium infection in children from two areas in Zimbabwe. Matariaia and Methods: In Zimbabwe, blood was taken from children from two sites: one with a high and one with a low prevalence of infection. Levels of lnterleukin (IL)-2, IL-4, IL-5 IFNy, TNFar,and GM-CSF released into the culture supematants on stimulation with schistosome egg antigen (SEA), Whole worm homogenate (WWH) or Phytohaemaggluttnin (PHA) were measured one year afler drug treatment. Reaulta: We observed that children produce either IL-4 or IL-5 but not both in reswnse to antiaen and mitocten.On stimulation with PHA. children from the low intensity area produced m&e IL-4, IFNy and GM-CSF than children from the high intensity area. On stimulation with SEA, children from the high intensity area produced GM-CSF and IL-IO than children from the low intensitv area. Morechildren who were infected before treatment produced IL-5 on stimulation with PHA than uninfected. On stimulation with SEA, production of IL-4 was associated with lack on infection before treatment whereas production of IFNy on stimulation with WWH was associated with protection from re-infection. Conclusions: The results lend some support to the hypothesis that the Th2 helper cell population is partitioned into two sub-populations. Area was the dominant factor in determining cytokine profile. We would like to speculate that the cytokine production was of a ThOtype in the low prevalence area, compared to the high prevalence area, whereas in the high prevalence area, a pattern that could be described as either macrophage or Th2 type predominated.
P.4.08.03
lmmunopetholcgy
of bovine neosporosia
K. Taylor, D.J.L. Williams, J. McGarry, F.M. Guy, A.J. Trees. LiverpootSchco/of TropicalMedicine, Pembroke Place, Liverpool, UK introduotion: Neospom caninum is a recently described apicomplexan protozoan parasite closely related to Taxoplasma gondii. Neosporosis can result in neonatal mortality in dogs and is considered to be a major cause of abortion in cattle throughout the world. Unlike T:gondii, abortlon caused by N. caninum can reoccur in subsequent pregnancies with potentially devastating effects on the dairy cattle industry. We are investigating whether modulation of the immune response to N. caninum during pregnancy leads to reactivation of chronic infection with subsequent congenital infection. The subject of this presentation is to describe the dynamics of the immune response to N. caninum following experimental infection of caffle prior to the onset of pregnancy. Mat&ala and Mathoda: NC liverpooi isolate of N. caninum were grown in tissue culture by serial passage in Vero cells maintained in RPM1 1840 supplemented with glutamine (2 mM), foetal calf serum (2%) and penicillin (100 iu/ml)/streptomycin (50 &ml). 1 x IO’ tachyzoites were inoculated by IV injection. Proliferative peripheral blood lymphocyte responses and IFNy production was monitored following stimulation wtth parasite antigen (1 &ml). Antibody responses were analysed by ELISA and IFAT. Results:Strong proliferative responses were evident at day 8 post infection through to day 77 accompanied by the production of high levels of IFNy as measured by ELISA. Antibody levels rose sharply at day 10 and peaked around day 40. Conclusion: These results confirm that N. caninum induces strong cellmediated responses and will provide the base line for studies of the immune response during experimental infection of pregnant animals.
P.4.08.04
Lelshmanla infection: Generation of 2 (Th2) cells in susceptible and reslstant
Experimental
Mter
F. Sommer, M. Meixner, M. Rollinghoff, M. Lohoff. institute of Clinica/ Microbiology and Immunology Wassertunnstr.3, 91954 Edangan, Germany introduction: The fatal course of Leishmania (L.) major infection in susceptible BALB/c mice is thought to be causally linked to a Leishmaniaspecitic Th2 cell response. However, the existence of Th2 cells in the lymph nodes of infected, susceptible BALB/c mice has never been directly demonstrated. We were interested to investigate whether indeed Th2 cells exist in the lymph nodes of L. major-infected. susceptible BALB/c mice. Matariai and Mathoda: To approach this question we used the method of intracellular staining to determine the combined or exclusive production of 11-2, IL-4 and IFN-y by PMAAonomycin stimulated CD4+ T cells derived from lymph node cells of L. major-infected susceptible BALB/c and resistant C57BU8 mice. We operationally defined a Th2 cell as a CD4+ cell that has the capacity to produce 11-4,but neither IL-2 nor IFN-y after stimulation. Results: Th2 cells were not detected by thii method in lymph nodes of uninfected mice. After 7 days of infection with L. majo Th2 cells became detectable in susceptible and, surprisingly, to a similar extent also in resistant mice. However, in resistant mice Th2 cells disappeared between day 8 and 14 after infection, whereas they were still detectable in susceptible mice by week 2 and 3. There was no significant difference between susceptible and resistant mice in the percentage of CD4+ cells having the capacity to produce IFN-y by week 1 and 2. Conclusion: The data suggest that Th2 cells are induced by Leishmania infection in both susceptible and resistant mice during the early course of disease. Maintenance of Th2 cells in susceptible mice may be the consequence of rather than the reason for a non protective immune response against L. major.
P.4.08.05
Paraslte load during the acute phase of murine T. cruz/ infection da&rmlnes the actlvatlon pattern of the immune system in late chronic phase
C.R.F. Martnho, M.R. D’lmperio Lima, M.G. Grtsotto, J.M. Alvarez. Department of Immunology, Univemidade de Silo Paula, SLio Pauio, SF: Brasil introduction: The chronic phase of murtne T: cruzi infection is characterized by the presence of scanty parasites in the blood and tissues, which can only be detected using amplification methods. We have previously shown, however, that this low parasitaemia is directly influenced by the parasite load of the acute phase of the infection. In the present work, we have examined the effects of acute’s phase parasite load on the size and activation state of splenic lymphocyte compartments in long-term 5: cruziinfected mice. Wterlaia and Methods:Ten to twetve AIJ mice were infected i.p. with lo3 or IO5 T: cruzi blood trypomastigotes of the Y strain and treated at day 6 with a single oral dose of Benzonidazol (Rwhagan 1 gkg) in order to interrupt the parasitemia curves. One year later, the animals were sacrificed and their spleens screened for total and isotype specific Ig-secreting cells (ESA),
Immunity to parasites
25 June 1997 - Poster presentations
Treatment with chloroquine on 7w day after prknary infection, reduces significantly the cross protection to heterologous challenge. However, immune mice treated with chloroquine and challenged with homologous parasites still remain protected. Reinfection of immune mice (treated or not with chloroquine) with the mixtures of I? c. chabaudiand Ryoe/ii 17XLshowed very high parasftemias however after day 12 post challenge mice started to control the parasitemia in contrast to the controls. Conoiueion: These experiments confirm and extend earliest studies on the specificity of immunity induced in inbred mice to rodent malarias. They also demonstrate that cross protection against heterotogous challenge is lost when primary parasitemia is stopped at 7 day after infection using chloroquine. This work was supported by DGAPA F’oyect 7N.211195 and CONACyT @#P-M.
09:00-18:30/12:09-14:00
P.4.08
Forum lounges
Immunity to parasites
1P.4.08.01 ] Fish immune responses and disease resistance W.B. Van Muiswinkel, B.M.L. Verburg, Van Kemenade, G.F. Wiegertjes. Wageningen Institute of Animal Sciences (WIAS), Waganingen Agricultural University (WAU), RO. Box 3336700 AH Wageningen, The Netherlands introduction: During the last 20 years considerable progress has been made in describing and understanding the immune system of fish. Fish are the oldest animal group with an immune system showing clear similarities with the defence systems of mammals and birds. Both innate immunity and acquired immunity are important for the defence against invading pathogens. Methods and Results: Humoral and cellular responses show the expected characteristics of specificity and memory after injection of standard antigens in carp (Cyprinus carpio L.). There is a clear influence of external factors such as water temperature and stress conditions (e.g. handling, oral application of cortisol). Moreover, the genetic background of the fish can also play a rote in the immune response against DNP-KLH as well in disease resistance to a blood flagellate, Twpanoplasma bone/i. This aspect was investigated by developing high and low responding inbred lines of fish. Conclusion: It can be concluded that the present knowledge can be used for evaluation of the health status of fish in the wild or in culture, but also for preventive measures such as vaccination or selection for disease resistance.
P.4.08.02
Cellular immunity to Sc/t/sfosoma infection In Zlmbabween children
heemefobium
J.T. Spicer ‘, C.M.R. Turner’, M. Woolhouse *, P. Ndlova 3, P. Hagan 4. ’ Division of infection and Immunity IBLS, University of Glasgow. UK, ‘Depanment of Zoology University of Oxford, UK, 3Blair Research Institute, Hamre, Zimbabwe, 4Commission of the European Communities, Brussels introduction: This study aimed to elucidate the role of cytokines in mediating the immune response to S. haematobium infection in children from two areas in Zimbabwe. Matariaia and Methods: In Zimbabwe, blood was taken from children from two sites: one with a high and one with a low prevalence of infection. Levels of lnterleukin (IL)-2, IL-4, IL-5 IFNy, TNFar,and GM-CSF released into the culture supematants on stimulation with schistosome egg antigen (SEA), Whole worm homogenate (WWH) or Phytohaemaggluttnin (PHA) were measured one year afler drug treatment. Reaulta: We observed that children produce either IL-4 or IL-5 but not both in reswnse to antiaen and mitocten.On stimulation with PHA. children from the low intensity area produced m&e IL-4, IFNy and GM-CSF than children from the high intensity area. On stimulation with SEA, children from the high intensity area produced GM-CSF and IL-IO than children from the low intensitv area. Morechildren who were infected before treatment produced IL-5 on stimulation with PHA than uninfected. On stimulation with SEA, production of IL-4 was associated with lack on infection before treatment whereas production of IFNy on stimulation with WWH was associated with protection from re-infection. Conclusions: The results lend some support to the hypothesis that the Th2 helper cell population is partitioned into two sub-populations. Area was the dominant factor in determining cytokine profile. We would like to speculate that the cytokine production was of a ThOtype in the low prevalence area, compared to the high prevalence area, whereas in the high prevalence area, a pattern that could be described as either macrophage or Th2 type predominated.
P.4.08.03
lmmunopetholcgy
of bovine neosporosia
K. Taylor, D.J.L. Williams, J. McGarry, F.M. Guy, A.J. Trees. LiverpootSchco/of TropicalMedicine, Pembroke Place, Liverpool, UK introduotion: Neospom caninum is a recently described apicomplexan protozoan parasite closely related to Taxoplasma gondii. Neosporosis can result in neonatal mortality in dogs and is considered to be a major cause of abortion in cattle throughout the world. Unlike T:gondii, abortlon caused by N. caninum can reoccur in subsequent pregnancies with potentially devastating effects on the dairy cattle industry. We are investigating whether modulation of the immune response to N. caninum during pregnancy leads to reactivation of chronic infection with subsequent congenital infection. The subject of this presentation is to describe the dynamics of the immune response to N. caninum following experimental infection of caffle prior to the onset of pregnancy. Mat&ala and Mathoda: NC liverpooi isolate of N. caninum were grown in tissue culture by serial passage in Vero cells maintained in RPM1 1840 supplemented with glutamine (2 mM), foetal calf serum (2%) and penicillin (100 iu/ml)/streptomycin (50 &ml). 1 x IO’ tachyzoites were inoculated by IV injection. Proliferative peripheral blood lymphocyte responses and IFNy production was monitored following stimulation wtth parasite antigen (1 &ml). Antibody responses were analysed by ELISA and IFAT. Results:Strong proliferative responses were evident at day 8 post infection through to day 77 accompanied by the production of high levels of IFNy as measured by ELISA. Antibody levels rose sharply at day 10 and peaked around day 40. Conclusion: These results confirm that N. caninum induces strong cellmediated responses and will provide the base line for studies of the immune response during experimental infection of pregnant animals.
P.4.08.04
Lelshmanla infection: Generation of 2 (Th2) cells in susceptible and reslstant
Experimental
Mter
F. Sommer, M. Meixner, M. Rollinghoff, M. Lohoff. institute of Clinica/ Microbiology and Immunology Wassertunnstr.3, 91954 Edangan, Germany introduction: The fatal course of Leishmania (L.) major infection in susceptible BALB/c mice is thought to be causally linked to a Leishmaniaspecitic Th2 cell response. However, the existence of Th2 cells in the lymph nodes of infected, susceptible BALB/c mice has never been directly demonstrated. We were interested to investigate whether indeed Th2 cells exist in the lymph nodes of L. major-infected. susceptible BALB/c mice. Matariai and Mathoda: To approach this question we used the method of intracellular staining to determine the combined or exclusive production of 11-2, IL-4 and IFN-y by PMAAonomycin stimulated CD4+ T cells derived from lymph node cells of L. major-infected susceptible BALB/c and resistant C57BU8 mice. We operationally defined a Th2 cell as a CD4+ cell that has the capacity to produce 11-4,but neither IL-2 nor IFN-y after stimulation. Results: Th2 cells were not detected by thii method in lymph nodes of uninfected mice. After 7 days of infection with L. majo Th2 cells became detectable in susceptible and, surprisingly, to a similar extent also in resistant mice. However, in resistant mice Th2 cells disappeared between day 8 and 14 after infection, whereas they were still detectable in susceptible mice by week 2 and 3. There was no significant difference between susceptible and resistant mice in the percentage of CD4+ cells having the capacity to produce IFN-y by week 1 and 2. Conclusion: The data suggest that Th2 cells are induced by Leishmania infection in both susceptible and resistant mice during the early course of disease. Maintenance of Th2 cells in susceptible mice may be the consequence of rather than the reason for a non protective immune response against L. major.
P.4.08.05
Paraslte load during the acute phase of murine T. cruz/ infection da&rmlnes the actlvatlon pattern of the immune system in late chronic phase
C.R.F. Martnho, M.R. D’lmperio Lima, M.G. Grtsotto, J.M. Alvarez. Department of Immunology, Univemidade de Silo Paula, SLio Pauio, SF: Brasil introduction: The chronic phase of murtne T: cruzi infection is characterized by the presence of scanty parasites in the blood and tissues, which can only be detected using amplification methods. We have previously shown, however, that this low parasitaemia is directly influenced by the parasite load of the acute phase of the infection. In the present work, we have examined the effects of acute’s phase parasite load on the size and activation state of splenic lymphocyte compartments in long-term 5: cruziinfected mice. Wterlaia and Methods:Ten to twetve AIJ mice were infected i.p. with lo3 or IO5 T: cruzi blood trypomastigotes of the Y strain and treated at day 6 with a single oral dose of Benzonidazol (Rwhagan 1 gkg) in order to interrupt the parasitemia curves. One year later, the animals were sacrificed and their spleens screened for total and isotype specific Ig-secreting cells (ESA),