RESUMO
BACKGROUND: Chlamydia abortus, an obligate intracellular pathogen with an affinity for placenta, causes reproductive failure. In non-pregnant animals, an initial latent infection is established until the next gestation, when the microorganism is reactivated, causing abortion. The precise mechanisms that trigger the awakening of C. abortus are still unknown. Sexual hormones such as estradiol and progesterone have been shown to affect the outcome of infection in other species of the family Chlamydiaceae, while estrogens increase chlamydial infection, progesterone has the opposite effect. To try to establish whether there is a relationship between these events and the latency/ reactivation of C. abortus in the reproductive tract of small ruminants, ovine endometrial (LE) and trophoblastic (AH-1) cells were treated with estradiol or progesterone prior to their infection with C. abortus. The results are compared with those obtained for treatment with penicillin prior to infection, which is a well-established model for studying persistent infection in other chlamydial species. Cells were examined by transmission electron microscopy, and an mRNA expression analysis of 16 genes related to the chlamydial developmental cycle was made. RESULTS: The changes observed in this study by the action of sex hormones seem to depend on the type of cell where the infection develops. In addition, while the changes are morphologically similar to those induced by treatment with penicillin, the patterns of gene expression are different. Gene expression patterns therefore, seem to depend on the persistence induced models of C. abortus used. Hormone treatments induced aberrant forms in infected endometrial cells but did not affect the chlamydial morphology in trophoblast cells. At the genetic level, hormones did not induce significant changes in the expression of the studied genes. CONCLUSIONS: The results suggest that penicillin induces a state of persistence in in vitro cultured C. abortus with characteristic morphological features and gene transcriptional patterns. However, the influence of hormones on the C. abortus developmental cycle is mediated by changes in the host cell environment. Furthermore, a persistent state in C. abortus cannot be characterised by a single profile of gene expression pattern, but may change depending on the model used to induce persistence.
Assuntos
Chlamydia/efeitos dos fármacos , Estradiol/farmacologia , Progesterona/farmacologia , Animais , Linhagem Celular , Chlamydia/crescimento & desenvolvimento , Chlamydia/ultraestrutura , Infecções por Chlamydia/veterinária , Feminino , Expressão Gênica , Microscopia Eletrônica de Transmissão/veterinária , Penicilinas/administração & dosagem , RNA Mensageiro , OvinosRESUMO
Chlamydia abortus produces ovine enzootic abortion (OEA). Symptoms are not observed until the organism colonises the placenta, eventually causing abortion. Infected animals become carriers and will shed the organism in the following oestruses. This process suggests that sex hormones might play an important role in the physiopathology of OEA, affecting the success of chlamydial clearance and also jeopardising the effectiveness of vaccination. However, the mechanisms through which sex hormones are involved in chlamydial pathogenicity remain unclear. The aim of this study, therefore, was to determine the effect of progesterone on the immune response against C. abortus and on the protection conferred by an experimental inactivated vaccine in sheep. Eighteen sheep were ovariectomised and divided into four groups: vaccinated and progesterone-treated (V-PG), vaccinated and non-treated (V-NT), non-vaccinated and non-treated (NV-NT) and non-vaccinated and progesterone-treated sheep (NV-PG). Animals from both PG groups were treated with commercial medroxyprogesterone acetate impregnated intravaginal sponges before and during the vaccination (V-PG) or just before challenge (NV-PG). The animals from both V groups were subcutaneously immunised with an experimental inactivated vaccine, which was seen to confer high protection in previous studies. All sheep were challenged intratracheally with C. abortus strain AB7 and were sacrificed on day 8 post-infection. Morbidity was measured as the variation in rectal temperature and samples of sera were collected for antibody and cytokine (IFN-γ and IL-10) analysis by commercial ELISA. In addition, lung and lymph node samples were collected for chlamydial detection by qPCR and for histopathological and immunohistochemical analyses. Sheep from the V-PG group showed less severe or no lesions and lower morbidity than the other groups. They also had the highest abundance of regulatory T-cells. The sheep from V-NT also manifested high antibody levels against C. abortus and less severe lesions than those observed in non-vaccinated sheep, which showed high morbidity, low antibody levels and severe lesions, especially in NV-NT. These results confirm the effectiveness of the experimental vaccine employed and suggest that progesterone could enhance the effect.
Assuntos
Vacinas Bacterianas/uso terapêutico , Infecções por Chlamydia/veterinária , Imunidade Humoral , Progesterona/administração & dosagem , Doenças dos Ovinos/imunologia , Aborto Animal/imunologia , Aborto Animal/prevenção & controle , Animais , Anticorpos Antibacterianos/sangue , Vacinas Bacterianas/imunologia , Chlamydia/imunologia , Infecções por Chlamydia/imunologia , Ensaio de Imunoadsorção Enzimática , Feminino , Ovinos , Doenças dos Ovinos/microbiologia , Vacinas de Produtos Inativados/imunologia , Vacinas de Produtos Inativados/uso terapêuticoRESUMO
Caprine tuberculosis is caused by bacteria of the Mycobacterium tuberculosis complex (Mycobacterium bovis and Mycobacterium caprae). Although typical tuberculoid granulomata are usually observed in the lungs and lymph nodes of infected goats, the presence of cavitary lesions with exuberant mycobacterial growth is also a common feature in this species. The aim of this study was to characterize the immunological mechanisms that lead to liquefaction and cavity formation by comparing granulomata and cavitary lesions. Samples from animals positive by skin testing were collected for microscopical and immunohistochemical examination. Samples were also collected for analysis of cytokine gene expression in the lesions by real time polymerase chain reaction. There were marked differences between granulomata and cavitary lesions. In cavitary lesions there was a substantial population of neutrophils and a significant decrease in the number of CD4(+) T cells, with concomitant increases in other T-cell populations (CD8(+) and cells expressing the γδ form of the T-cell receptor). The enzyme iNOS was strongly expressed by macrophages in the cavitary lesions. There was no difference in the balance of pro- and anti-inflammatory cytokine gene expression in the lesions. These findings suggest that cavitary lesions are reactivation sites, where conditions are optimal for Mycobacterium proliferation and that immunological mechanisms may underlie the severe destruction of lung tissue that characterizes the cavitary pathology.
Assuntos
Doenças das Cabras/imunologia , Doenças das Cabras/patologia , Tuberculose Pulmonar/imunologia , Tuberculose Pulmonar/patologia , Tuberculose Pulmonar/veterinária , Animais , Expressão Gênica , Perfilação da Expressão Gênica , Cabras , Imuno-Histoquímica , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
CBA/J mice were used in the present study to establish differences between the immune response to three chlamydial strains: AB7 (Chlamydia psittaci wild-type strain), 1B (C. psittaci vaccinal strain) and iB1 (C. pecorum). The evolution of chlamydial infection was evaluated in each strain by studying the clinical signs, the number of bacteria isolated from the spleen and the pathology of the liver. Three aspects of the immune response were then studied: the characterization of the infiltrate of leukocytes in the liver, the percentages of T- and B-cells, macrophages and neutrophils in the spleen, and the presence of cytokines in the serum. Infection followed a different course in the C. psittaci-infected mice; 1B-infected mice showed milder levels in all the parameters analysed than their AB7-infected counterparts. The resolution of infection was earlier in 1B-infected mice and, although the immune response to both strains was Th1-like, a more intense CD8+ T-cell response and an earlier presence of TNF-alpha in serum were observed in this group. C. pecorum infection was controlled mainly by a non-specific immune response, since these mice showed no signs of a systemic specific immune response. Neutrophil depletion experiments showed that these cells play a very limited role in the non-specific response against C. pecorum.
Assuntos
Infecções por Chlamydia/veterinária , Chlamydia/imunologia , Doenças dos Ovinos/imunologia , Animais , Linfócitos B/imunologia , Embrião de Galinha , Chlamydia/isolamento & purificação , Infecções por Chlamydia/imunologia , Infecções por Chlamydia/patologia , Chlamydophila psittaci/imunologia , Citocinas/sangue , Feminino , Fígado/imunologia , Fígado/microbiologia , Fígado/patologia , Macrófagos/imunologia , Camundongos , Camundongos Endogâmicos CBA , Neutrófilos/imunologia , Ovinos , Doenças dos Ovinos/microbiologia , Baço/microbiologia , Baço/patologia , Linfócitos T/imunologia , Fatores de TempoRESUMO
Monoclonal antibodies (mAbs) against bovine leukocyte antigens specific for T cells (CD2, CD4, CD8 and gammadelta receptor) and B cells (surface IgM) were used in samples from one week and one-, three- and seven-month-old goats to study the evolution of lymphocyte subsets by flow cytometry in peripheral blood and the lymphoid organs: thymus, jejunal (JPP) and ileal (IPP) Peyer's patches, spleen and mesenteric lymph nodes. An increase in the values of alpha/beta receptor T cells with age was recorded whereas the gammadelta receptor T cells fell in number. In peripheral blood and in all tissues, except IPP the values for B cells (sIgM+) were low. The CD4+ and CD8+ cells predominated in JPP while B cells were the most important subpopulation in IPP. In the spleen, as in JPP, the CD4/CD8 ratio was less than one and the gammadelta T cells values were high. In mesenteric lymph nodes, CD8+ and B(sIgM) cells predominated in the youngest animals.