VETERINARY IMMUNOLOGY

Author: AYATS PÉREZ ARTURO.
Year: 2004.
University: COMPLUTENSE DE MADRID [www.ucm.es].
Place of defense: FACULTAD DE VETERINARIA.
Place of preparation: FACULTAD DE VETERINARIA.

Summary: The Keratitis Superfical Chronicle Inmunomediada (QSCI) is an inflammatory process, chronic, possibly inmunomediado, which affects the outer layer of the cornea and is presented in very specific breeds: German Shepherd, Collie, Mestizos, and so on. Traditionally, treatment has been based on the control of inflammation through administration of topical corticosteroids throughout the animal's life. A few years ago, with the emergence of a selective immunosuppressant cyclosporine as is the (CsA), opened new perspectives for the treatment of this disease. Recently there have proposed alternative therapies based on the association of two or more drugs with the intent to seek synergistic effects in treating processes eye inmunomediados. In this study we compared the efficacy of monotherapy with CsA oftámica 0.2% and the sequential therapy of corticosteroids (triamcinolone 4.0% and dexamethasone 0.1%) and CsA 0.2% in the treatment of QSCI We have determined that both CsA monotherapy as sequential therapy are effective in the treatment of QSCI both in the remission of clinical symptoms, their decline over time and the emergence of recurrence of such signs. But the sequential therapy has been shown statistically more effective, with a trend towards more rapid improvement and manifests itself in the eyes treated with the same at the 74% compared to monotherapy with a 34% CsA. Also, the computation of recurrence were similar in both treated groups. We propose, based on our results, the use of sequential therapy as an alternative and more effective as the treatment of choice for controlling the clinical signs of QSCI

Author: ORTEGA HERNÁNDEZ NIEVES.
Year: 2005.
University: MURCIA [www.um.es].
Place of defense: FACULTAD DE VETERINARIA.
Place of preparation: FACULTAD DE VETERINARIA.

Summary: Chlamydophila abortus is a parasitic intracellular bacterium, which causes abortion Enzootic sheep (AEO), a disease that causes significant economic losses in the livestock sector. The principal measure is to control the vaccination of animals. This paper evaluates the protection and analyzes the immune response induced different inactivated vaccines or commercial pilot and an attenuated against abortion clamidial. It used different lines of mice as the CBA / J SWISS OF1 or C57BL / 6 and the parameters were measured mortality, morbidity, insulation C.abortus in spleen, liver or uterus, the number of mice live offspring the presence of IFN-gamma serum, the production of cytokines (IFN-gamma, IL-4) from supernatant of cultured splenocytes posts antibody production and isotipos total serum IgG. First, it valued the effectiveness of vaccines marketed in our country over C.abortus. To do so, compared five vaccines business (four inactivated and an attenuated) with an inactivated vaccine is experimental and found that inactivated vaccines business confer protection very limited, unlike the vaccine, which performed well protected. Similarly, the inactivated vaccine designed by us showed some results of protection closest to the vaccine atenudad that the inactivated business. In second place were evaluated various methods of inactivation for C.abortus and found that the method that offered the best preservation of the antigenic structure was used by the Etilenimina binary (EIB). Subsequently, was assessed the effectiveness of experimental new inactivated vaccines, made with various adjuvants, compared with C.aboruts. It showed that the vaccine made with the saponin QS21 was the one that offered better protection, while the worst results were obtained with the vaccine formulated with aluminum hydroxide. He then was assessed the effectiveness of vaccines preselected adyuvadas with QS21 or Montanide ISA773 compared with a strain homologous or heterologous strains of C.abortus. To that end, we used a murine model pregnant and non-pregnant and another was observed that the two vaccines protected compared to both strains in both pregnant and non-pregnant mice. Thirdly, we explored the protective role of the cells involved in the innate immune response (NK cells and neutrophils), and the adaptive immune response (B lymphocytes, produced antibodies after vaccination and CD4 + T cells and CD8 +), in animals Immunized with vaccine candidate adyuvadas with QS21 or aluminum hydroxide and the vaccine atenudad 1B. To do so, after the vaccination of animals were deplecinados or using knockout mice (genetically deplecionados), depending on the population studied and were subsequently infected by C.abortus. It was observed that the influence of neutrophils, lymphocytes B and the protective power of the antibodies induced by vaccination, depended on the type of adjuvant used in the vaccine formulation. Thus, the influence of neutrophils was higher in mice immunized with the vaccine adjuvanted with aluminum hydroxide. While the influence of linfoctios B was higher in mice immunized with the vaccine adjuvanted with QS21. In addition, the antibodies productive with the latter vaccine, transferred to another mouse led to the best protection against infection by C.abortus. However, the absence of NK cells affected to a greater degree to animals immunized with the vaccine 1B, those inmun 8 ised c 757 on the inactivated vaccines. Paradoxically, T lymphocytes not played an essential role in any of the groups immunized, unlike d which occurs in a primary infection. Vaccinated animals can submit simultaneously pulmonary and gastrointestinal parasites, which could set immune responses type Th2. To do so, the effect was assessed immunosuppressive phase inducing dela vaccination and the efectora phase of the same. It was noted that the parasitación of animals before the vaccines (Phase inducing) did not alter the protection induced by the vaccine tested, while on the other hand if the parasitación was produced after immunization of mice (Phase efectora) and close in time with the infective dose C.abortus (statement of co-infection), the effectiveness d adjuvanted vaccine with aluminum hydroxide. The best results were obtained with the inactivated vaccine with QS21 and adjuvanted vaccine attenuated 1B. These results indicate that the selection of adjuvant is essential in securing an effective vaccine that can prevent the vaccine stems associated with co-interacciones natural.

Author: MARTÍNEZ CÁCERES CARLOS MANUEL.
Year: 2005.
University: MURCIA [www.um.es].
Place of defense: UNIVERSIDAD DE MURCIA.
Place of preparation: UNIVERSIDAD DE MURCIA.

Summary: Chlamydophila abortus is a parasitic intracellular gram-negative bacterium, which causes abortion Enzootic sheep, a disease that causes significant economic losses to livestock sector, and that carries a potential risk zoonósico. Although it is well established the importance of the immune response, both innate and cellular Th1, the resolution of infection clamidial, know little data on the role of NK cells and T-lymphocyte subpopulations in front of the immune response C. Abortus. The objective of this study is to determine the role of these cells in a mouse model of primary infection. Using a model of infection with mice C57BL/6J, we studied the role of NK cells through depletion and subsequent analysis of the course of infection with C. Abortus. The lack of NK cells increased the susceptibility of animals to infection clamidial, resulted in a high mortality and increased morbidity and liver bacterial load. The study inmunohistopatológico showed inflammatory foci large, composed essentially of PMNs, with numerous inclusions clamidiales associated with the outbreaks. The results obtained from the study of the concentration of g-IFN serum levels showed significantly reduced compared with the control group not deplecionado. Our results indicate, therefore, NK cells as a population essential in the early control of infection by C. Abortus. To establish the relationship between NK cells and other components of the innate immune response is deplecionó a group of mice C57BL6 / J PMNs. The infection caused a steep rise in mortality and bacterial load in the liver of animals deplecionados. The inmunohistopatología showed the absence of inflammatory foci, with the presence of rich inclusions clamidiales in parenchyma. The PMNs therefore appear to be critical to cell migration of NK cells into the inflammatory focus. To establish the role of each subpopulation of T lymphocytes (CD4 + and CD8 +) in primary infection by C. Abortus is deplecionaron mice C57BL/6J of CD4 + T cells and / or CD8 + T cells. It valued the mortality, morbidity, liver bacterial load, inmunohistopatología in liver and spleen, characterization inmunofenotípica of infiltrated inflammatory liver, levels of apoptosis in the liver and spleen, and production of cytokines from supernatants of cultured splenocytes posts. All animals deplecionados of CD8 + T cells died before day 12 pi, while mortality was not observed in the other groups. Animals deplecionados of CD4 + T cells showed a low morbidity compared with the control group of animals not deplecionados. Animals deplecionados of CD8 + T cells showed a high level of apoptotic cells, and an exacerbation of production g-IFN by splenocytes posts in culture and stimulated with antigen clamidial. The simultaneous depletion of both T lymphocyte subpopulations did not cause deaths, but resulted in a chronic infection. These results demonstrate that the subpopulation of CD8 + T cells is crucial for the survival of the animals against primary infection by C. Abortus, and may have a regulatory role in the response of CD4 + T cells against infection.

Author: VIGLIANO FRANSICETTI FABRICIO ANDRÉS.
Year: 2005.
University: SANTIAGO DE COMPOSTELA [www.usc.es].
Place of defense: FACULTAD DE VETERINARIA.
Place of preparation: FACULTAD DE VETERINARIA.

Summary: In this paper we describe the morphology of the bodies hemolinfopoyéticos of silverside bonaerense (Odontesthes bonariensis) through its structural study, ultrastructural and immunohistochemical. According to the cellular composition of these bodies identified the primary functions of each of them. In the kidney skull of O.bonariensis takes place erythropoiesis and granulopoeisis, while the spleen is involved in the production of monocytes and thrombocytes, and to a lesser extent in the differentiation of peritrocito. The variations observed between bodies in the type, quantity and location of cells linked to the production of antibodies suggest that the spleen is the central site for the generation of specific humoral immune responses. Moreover, it is the principal organ hemocaterético due to the action of macrophages associated with sinus esplénicos, melanomacrófagos free and centers melanomacrófagos. The weather in this species seems to evolve at a very early age since it was not observed in macroscopic or microscopic animals analyzed. The immunohistochemical study found that the populations of reticular cells of the kidney and spleen show different immune. Alongside it possible to identify two kinds of cells in the citorretículo latter body. Some cells were S100 + and distributed throughout the body, while the others were vimentina + / uNOS + and is confined to the breasts splenic. Likewise, the expression of the antigen recognized by the antibody anti-células dendritic follicular in melanomacrófagos free and grouped suggests the existence of an evolutionary relationship between the centers melanomacrófagos of fish and germinal centers of higher vertebrates. The reticular cells of eplipsoides and sinuses splenic cells and muscle isas of renal arteries and arterioles expressed different isoforms of óxidonítrico synthase. This is the first identification of cells producing nitric oxide in blood vessels órgnaos hemolinfopoyéticos fish and indicates that nitric oxide is involved in the regulation of blood flow spleen and kidney of silverside.

Author: ARIAS VÁZQUEZ MARÍA SOL.
Year: 2006.
University: SANTIAGO DE COMPOSTELA [www.usc.es].
Place of defense: FACULTADE DE VETERINARIA DE LUGO.
Place of preparation: FACULTADE DE VETERINARIA DE LUGO.

Summary: The infection F. Hepatica not affected equally to the different species of animals, sheep are very sensitive, which implies greater direct economic losses (death of the animals, especially in the acute fasciolosis) and indirect (confiscation of livers, decreased production costs and treatment of animals affected). In paragraph 1 through screening of a cDNA library with sera from infected sheep on an experimental (9Â th spi) were selected 19 clones; after confronting these clones to sera from rats with experimental fasciolosis obtained at 3 spinnaker, we see only reaction with 3 clones (5, 10 and 17), so we decided sequencing and confirm that contained DNA trematodo. The sequence of clones 5, 10 and 17 revealed the presence of complete reading frames (open reading frame, ORF), with a start codon which codified for the methionine (ATG), and a termination codon. With the expression of these clones were obtained 3 proteins 12'3 kDa (rFhRSA), 16 kDa (rFhMAV) and 2.9 kDa (rFhAPS). By ELISA and serums sheep and rats with experimental fasciolosis, that the best results were obtained with the lower molecular weight protein (rFhAPS), and the determination of its functional activity revealed that it was a serín-proteasa. In paragraph 2, in order to assess the usefulness of the protein rFhAPS were employed various diagnostic techniques in sheep infected experimentally. We had 49 sheep from a farm qualified sanitariamente who had never been in the grass. The infection of animals was conducted administrándoles oral metacercarias F. Hepatica. This study used the pattern of infection known as trickle infection, and that is to manage a small number of metacercarias repeatedly, specifically 25 metacercarias daily for 7 consecutive days. Lambs were distributed at 4 homogeneous groups, consisting of 14 lambs each, except for the control group which was composed of 7 animals remained without infect, or deal well before the group was infected (with 175 metacercarias / animal), the re-infected (animals primoinfectados and reinfected to 18 spi, with the same dose and schedule used in primary infection), and finally the group triclabendazol (animals primoinfectados and treaties to 4 spi, orally at a dose of 10 mg kg-1 of this drug thereafter; to 14 weeks of treatment, namely the 18 spi, the lambs were reinfectaron with the same dose and schedule used prior to treatment). At the end of the study, all animals were slaughtered to determine the number of copies of F. Hepatica housing and the injuries had resulted. Weekly stool samples were taken straight from each of the animals and remained 4Â ° C until analysis. In order to know how many eggs per gram of feces (hpg), each sample was analyzed in duplicate, using the sedimentation technique. To study the kinetics of antibodies and antigens compared to F. Hepatica, took blood samples from each animal, from 2 weeks before the infection to their sacrifice. The blood collection was made using vacuum tubes closed type Vacutainer without anticoagulant. We used the ELISA (ELISA-AgC) to determine the kinetics of circulating antigens of F. Hepatica, IgG using a polyclonal anti-F. Hepatica obtained from rabbits immunized with FhES. The kinetics obtained by ELISA antibody was similar when used antigen FhES or protein rFhAPS, demonstrating that protein is valid for inmunodiagnóstico of fascioliasis experimental sheep. The reinfection of the lambs led to a further increase in antigenemia and 8 of 12bb IgG versus rFhAPS. As had happened in animals primoinfectados, peak values of antigens appreciated at 3-4 spr, period of migration of youth flukes by the parenchyma, and were negative at 12 spr, which coincides with the arrival of flukes reinfection of the bile ducts. The fact that the levels of IgG detected by ELISA-iy protein rFhAPS not decrease can be related to an increase in the elimination of eggs in the feces as a result of reinfection. In infected animals and treated with triclabendazol from 1Â th spt The values of antigens and antibodies decreased markedly to be negative to 4Â th spt, demonstrating that the protein rFhAPS is very useful for evaluating the effectiveness of treatment fasciolicida by ELISA, and thus exceeds one of the major drawbacks presenting this technique for the assessment of anthelmintic treatments. In paragraph 3 was determined the prevalence of infection by F. Hepatica in sheep exploited regime semi-extensivo in the province of Lugo, took stool samples and blood of 340 sheep belonging to 22 farms, in order to carry out the studies coprológicos and serological (kinetics of antigens and antibodies by ELISA with FhES and rFhAPS). We believe that the animals had fasciolosis active when detected antigens F. Hepatica, removing eggs, or both, while not considered infected if not eliminated eggs and is not detected antigens F. Hepatica serum. Of the 340 animals studied, 198 had active fasciolosis and 67'7% of the sheep were also positive by ELISA and protein rFhAPS while only were the 52'5% when using the FhES. Thus, of the 142 animals had no fasciolosis, found that the 85'9% (122/142) were also negative rFhAPS, and the FhES only what was 47'8%. Again using the protein rFhAPS, that the percentage of false negatives (32'3%) was lower than that found with the FhES (47'5%). With protein rFhAPS the rate of false positives (14%) was lower than that found with antigen FhES (52'1%). From these results it appears that the use of ELISA with rFhAPS provides more reliable results. Finally, we assessed the effectiveness of triclabendazol in sheep naturally infected by F. Hepatica, using a group of 18 sheep that were grazing semiextensivo in an area which was previously found infection F. Hepatica. Prior to divide animals into 2 groups were conducted 2 analysis coprológicos with an interval of 2 weeks, to make sure that all were infected naturally. Separated 2 groups were homogeneous with respect to the elimination of hpg; the sheep of a first group failed to receive any worming and acted as witnesses for the infection, and the second was treated with triclabendazol (10 mg kg-1 pv) . During the 16 weeks of treatment, samples of blood and feces of all animals. The sheep out to pasture followed, and therefore could be reinfectar natural way to continue eating the metacercarias he had in the area. By ELISA-AgC see that, since the 2Â meetings until 6Â th spt, the values of OJ were negative, coinciding with the absence of removing eggs. Also, although the absorbance values obtained with ELISA and rFhAPS failed to be negative, a week to administer the triclabendazol rates antibodies decreased significantly until the 6Â th spt, which were found near the figures absorbance cutoff point . Animals continued grazing in the same plots that before treatment, which could reinfection with metacercarias F. Hepatica, which explains why, from 8Â th spt And, coinciding with the beginning of a new removing eggs were detected an increase in the values of OJ, both ELISA-AgC with rFhAPS.

Author: RODRIGUEZ CORTES ALHELÍ.
Year: 2006.
University: AUTÓNOMA DE BARCELONA [www.uab.es].
Place of defense: FACULTAT DE VETERINARIA.
Place of preparation: UNIVERSIDAD AUTÓNOMA DE BARCELONA.