Characterization of cytokine production in murine Trypanosoma cruzi infection by in situ immunocytochemistry: Lack of association between susceptibility and type 2 cytokine production

Abstract

Cytokine production in the spleens of mice infected with the protozoan parasite Trypanosoma cruzi was analyzed in three models which differ in the outcome of the infection. Using immunocytochemical techniques to detect cytokine-producing cells, the production of type 1 [interleukin-2 (IL-2) and interferon (IFN)-gamma], type 2 (IL-4, IL-5, IL-10), inflammatory [tumor necrosis factor (TNF)-alpha, IL-1 alpha, IL-6] and regulatory (transforming growth factor-beta) cytokines were examined. With the exception of IL-4 and IL-5, cells producing all of the cytokines assayed were detected in both the resistant and susceptible models of T. cruzi infection. Cells producing IL-4 and IL-5 were not detected until later in infection in the resistant mice (> 34 days), at about the time animals of the susceptible strain succumb to the infection. Mice of the susceptible model showed a slight delay in the appearance of cells producing the type 1 cytokines IL-2 and IFN-gamma and an earlier appearance of TNF-producing cells, in comparison to resistant mice. Cells producing IL-2 or IL-10 were transient in their appearance in the spleen while cells producing IL-1, IL-4, IL-5, IL-6, IFN-gamma, TNF, or TGF-beta were first detectable in either the acute or post-acute stage of the infection and persisted up to 700 days post infection in two different resistant models of the infection. Cells producing IFN-gamma, TNF-alpha and TGF-beta were particularly numerous even very late in the infection. Double-staining techniques were used to show that the vast majority of the IFN-gamma-producing cells in the spleen were CD4-, CD8- alpha/beta TCR+T cells. This study confirms the transience of IL-2 production in the acute stage of T. cruzi infection and the persistent and simultaneous production of type 1 and type 2 cytokines during the late-acute and chronic stages of the infection. Susceptibility or resistance to T. cruzi infection does not associate with a Th2 pattern of cytokine production in the three models examined in this study. The overlapping pattern of type 1 and type 2 cytokine-producing cells in both the acute and chronic stages of T. cruzi infection demonstrates that longterm infections do not necessarily lead to a dominance of either type 1 or type 2 cytokine production.

Participation of cytokines in the necrotic-inflammatory lesions in the heart and skeletal muscles of Calomys callosus infected with Trypanosoma cruzi

“…Pro-inflammatory cytokines as TNF-α and IFN-γ, as well as the regulatory cytokine TGF-β were identified during the course of infection in this study, and persisted until an advanced phase (70 days) (Zhang & Tarleton 1996). It has been demonstrated histochemically (Lima et al 2001), in highly susceptible inbred mice (C3H/ He), infected with a macrophagotropic strain (Biodeme Type I), the participation of TNF-α in the massive parasite destruction and spleen necrosis.…”

“…In situ demonstration, of the cytokines involved in the response to infection with T. cruzi, has been done by Zhang and Tarleton (1996), in the spleen of infected mice. Pro-inflammatory cytokines as TNF-α and IFN-γ, as well as the regulatory cytokine TGF-β were identified during the course of infection in this study, and persisted until an advanced phase (70 days) (Zhang & Tarleton 1996).…”

Immunological imbalance between IFN-³ and IL-10 levels in the sera of patients with the cardiac form of Chagas disease

“…Cytokine profiling has been demonstrated as fundamental to defining the immunopathological mechanisms involved in chronic chagasic cardiomyopathy and controlling the immune response during T. cruzi infection (Zhang & Tarleton 1996). The proinflammatory cytokines IL-12, IFN-γ and TNF-α (Th1 response) act in concert to activate macrophages to kill the parasites through the production of nitric oxide and NO-derived nitrogen free radicals.…”

Antigen-Specific Th1 But Not Th2 Cells Provide Protection from Lethal Trypanosoma cruzi Infection in Mice

“…Sections (5 m) from paraffin-embedded tissues were stained with hematoxylin and eosin for histopathological analysis. To detect OVA-specific Th1 and Th2 cells posttransfer, lymph nodes, spleens, and skeletal muscle tissues from recipient mice were frozen in liquid nitrogen and 5-to 10-m-thick sections were analyzed as previously described with some modifications (33). Briefly, acetone-fixed tissue sections were quenched with PBS containing 0.3% of H 2 O 2 and 0.1% of sodium azide and incubated with biotinylated KJ1-26 Ab in PBS at 4°C overnight.…”

“…The Th cell response to T. cruzi in both susceptible and resistant strains of mice normally exhibits a mixed Th1/Th2 cytokine production profile (33). Thus, it was of interest to determine whether the adoptive transfer of a combination of Th1 and Th2 cells or of naive DO.11.10 T cells (that could differentiate into Th1 and Th2 cells in vivo) could confer protection to T. cruzi infection equivalent to that of Th1 cells alone.…”