L'étude des populations de lymphocytes T CD4+ effecteurs (Teff) chez l'Homme présente un intérêt croissant dans les enjeux actuels que constitue l'élaboration de nouvelles immunothérapies. Ces travaux détaillent la caractérisation et la régulation d'une population de Teff exprimant l'ecto-nucléotidase CD73 ayant pour fonction de dégrader l'AMP extracellulaire en adénosine (Ado) immunosuppresseur. Cette population, enrichie en lymphocytes T helper de type Th1.17, est très polyfonctionelle et pro-inflammatoire. Les Teff CD73+ expriment peu de points de contrôles immunitaires inhibiteurs mais sont régulés par leur production autocrine d'Ado qui limite leurs fonctions effectrices et leurs capacités prolifératives. Les Teff CD73+ expriment par ailleurs fortement le transporteur ABC multidrug-resistance 1 (MDR1), responsable de l'exclusion de nombreuses drogues du cytoplasme des cellules. L'étude de cette population dans différents contextes pathologiques a permis de détailler le fonctionnement de cette population. J'ai pu mettre en évidence que l'expression de CD73 est en effet dynamique. Elle est notamment diminuée dans des pathologies arthritiques auto-immunitaires (la polyarthrite rhumatoïde et le rhumatisme psoriasique) dans lesquelles les Th17 et les Th1.17 sont fortement activés. La diminution d'expression de CD73 sur ces cellules constitue la levée d'un frein qui leur permet de contribuer pleinement à l'inflammation chronique caractérisant ces pathologies. Les Teff CD73+, présents dans les tumeurs solides humaines de sein et d'ovaire, pourraient en revanche présenter un avantage sélectif en contexte tumoral de par leur expression de MDR1 leur permettant de résister aux traitements de chimiothérapie. Ces traitements substrats de MDR1, combinés à des thérapies inhibant la fonction enzymatique de CD73, pourrait permettre la restauration d'une réponse immunitaire anti-tumorale efficace médiée par cette population pro-inflammatoire / Requirement for CD4+ effector T lymphocytes (Teff) comprehensive study in human is increasing since it can contribute to the emergence of new immunotherapy strategies. This work brings up important information concerning the characterization and regulation of a Teff population expressing the CD73 ecto-nucleotidase, which is able to degrade extracellular AMP into immunosuppressive adenosine (Ado). This population, highly polyfunctional and pro-inflammatory, is enriched in Th1.17 cells. CD73+ Teff express low levels of inhibitory immune checkpoints but are negatively regulated by the autocrine Ado production that limits their pro-inflammatory function and proliferative capacities. In addition, CD73+ Teff express high levels of the ABC transporter multi-drug resistance 1 (MDR1), responsible for the exclusion of cells’ cytoplasm of many drugs. The study of this population in different pathological contexts enabled to decipher its functions. I could evidence that CD73 expression is dynamic. CD73 is notably decreased in autoimmune arthritic pathologies (rheumatoid arthritis (RA) and psoriatic arthritis (PsA)) in which Th1.17 and Th17 are highly activated. CD73 decreased expression by these cells is a mechanism that alleviates self-inhibition by autocrine Ado production and enables them to fully contribute to chronic inflammation characterizing these pathologies. In tumor context, CD73+ Teff present in breast and ovarian tumors could on the contrary bear a selective advantage due to their high MDR1 expression enabling them to resist MDR1 substrates-based chemotherapy treatments. These chemotherapy treatments combined to therapies blocking CD73 enzymatic function could allow the restauration of an efficient anti-tumor immune response

In multiple sclerosis (MS), pathogenic T effector cells (Teff) are believed to orchestrate immune-mediated destruction of the central nervous system (CNS) myelin sheath. In experimental autoimmune encephalomyelitis (EAE), a mouse model of MS, CNS infiltration by regulatory T cells (Treg), producing the anti-inflammatory cytokine IL-10, promotes the resolution of disease. Currently, little is understood about how Treg function within the inflamed CNS and on which cells they exert their suppressive function. There is a debate as to whether Treg in MS patients are capable of infiltrating the CNS and if they do, it is unclear whether they are functional. Understanding Treg function in EAE and MS could open up new possibilities for treatment, as Treg could be modulated for immunosuppressive therapy. A key step in the development of EAE (and presumably MS) is the ability of Teff cells to cross the blood brain barrier (BBB) and enter the CNS parenchyma. The hypothesis of this work was that Treg facilitate resolution of the inflamed CNS by preventing entry of the pathogenic T cells into the CNS parenchyma, thus preventing further damage. As such, it is important to understand with which immune cells and CNS resident cells Treg communicate to achieve resolution of disease. The presence of Treg in MS lesions was investigated with double immunohistochemistry (IHC) in frozen post-mortem MS brain tissue. CD4+Foxp3+ Treg were present in a subset of patients and their presence was associated with perivascular retention of CD4+Foxp3- and CD8+Foxp3- T cells. Foxp3+ cells in MS lesions predominantly expressed IL-10, indicating regulatory activity, although low-level production of IL-17, TNF-α, IFN-γ and GM-CSF was observed as well. Generally, analysis of total cytokine expression identified distinct patterns of cytokine production between lesions. Nonetheless, these could not be used to discriminate individual patients. These studies were repeated in C57BL/6 mice in which the Treg population was depleted before onset of EAE to mimic lesions with and without Treg presence, as found in MS patients. An immunofluorescent technique to study up to 5 fluorochromes simultaneously was developed to study antigen presenting cell (APC), Teff and Treg location, spatial relationship and function (as measured by cytokine expression) in the CNS of EAE mice at different stages of disease. Using this technique it was found that CD4+Foxp3- Teff and CD4+Foxp3+ Treg were located within 50-100μm of CD11c+ APC in the CNS of EAE affected mice. CNS Teff and Treg predominantly produced IFN-γ or IL-10, although low levels of IL-17 were detected in Teff and Treg as well. IL-17+ Treg were close to IL-17+ Teff, IFN-γ+ Treg were close to IFN-γ+ Teff, but IL-10+ Treg were not in close proximity to IL-10+ T cells in the CNS during EAE. In conclusion, there is evidence for functional Treg in EAE and MS lesions, supporting the concept of enhancing Treg activity as a clinical intervention. Treg seem to be capable of retaining pathogenic T cells at the blood brain barrier in MS lesions. In addition, studies of cytokine expression in MS lesions indicated that there is no sound basis for patient stratification based on peripheral blood cytokine profile. This thesis advances our understanding of Treg location, function and spatial relationship with other immune cells within the inflamed CNS.

Thesis (doctoral)--Humboldt-Universität, Berlin, 2001. / Includes bibliographical references (p. 152-189).

This research was conducted at the University of KwaZulu-Natal, Pietermaritzburg, South Africa. The research encompasses five different studies to assess nutrient cycling in intensive and extensive grazing systems with a view to optimising livestock production. The first study was designed to assess the effect of teff-lucerne mixtures on teff, lucerne and teff-lucerne mixture yields. Lucerne and teff-lucerne mixtures benefited from the association. The overall soil N content of the teff-lucerne mixture plots was greater than the teff alone plots. The second study focused on teff-leucaena association evaluation. It had two leucaena plant row spacings as treatments, 180cm and 120cm, respectively. Teff grown in mixture with leucaena produced a total teff dry matter (DM) of 7931.57 kg ha¯¹ for the 180cm row spacing and 8329.57 for the 120cm row spacing compared to the 3548.93 kg ha¯¹ of DM obtained from the teff alone treatment. The teff-Ieucaena stand also had a greater DM yield response to leucaena row spacing compared to the teff alone. In terms of nutritive quality, all stands from the teff-leucaena plots were better than the quality obtained from the teff alone plots. Total N content of teff from the l80cm row spacing was 21.83 g kg¯¹ and that from the 120cm 16.07 g kg¯¹ compared to the total nitrogen (N) content of 19.77 g kg¯¹ of the teff alone treatment. The total phosphorus (P) content was 2.73, 1.96 and 2.07 g kg¯¹ for the 180cm, 120cm and teff alone treatments respectively. However, the total soil N content was higher for the teff alone plot than for the teff-leucaena plots, which are 1.91, 1.48 and 100 g kg¯¹ for the teff alone, 180cm and 120cm treatments respectively. The third study was designed to assess the effects of different N fertilizer application rates on teff yield response. The rates applied were 0, 50, 100 and 150 kg N ha¯¹. There was significant difference in teff response of the three N fertilizer application rates compared to the control and teff DM yield response was lower for the 150 kg N ha¯¹ (838 kg ha¯¹) treatment compared to the control (553 kg ha¯¹). Both teff DM and nutritive value were higher in the plots treated with N fertilizer than in the plot which received no N fertilizer (control). The soil N content was also higher in those plots treated with N fertilizer. Study four was conducted on the Department of Grassland Science's grassland management techniques trial field at Ukulinga. The effects of nutrient cycling under different management techniques such as burning, mowing and grazing on grass yield response, plant quality and soil nutrients were assessed. However, the response of grass DM yield and P content was not significant but the three treatments had a significant effect on grass N content. Their effect on soil N content was also significant and the grazing plot had the greatest soil N levels. The last study was conducted in the rural areas of Okhombe and Zwelitsha to assess the effects of grazing intensity on grass yield response, plant quality and soil nutrient status at different distances from homesteads. Grass DM yield and nutritive value declined when distance from the homestead increased. The soil N content also was higher nearer to the homestead than further away. Most farmers, particularly in developing countries including those in Eritrea, often experience that their animals prefer forages from some plants such as lucerne, leucaena, and other indigenous leguminous plants. They also observe that they get greater yield from crops grown near leguminous plants or in rotation with legumes. They are also still using manure from their animals to fertilize their croplands. Therefore, it is still the duty of the researchers to demonstrate to farmers on farm studies to convince farmers that it is because leguminous plants have the ability to add quality and quantity to the feed of the animals and soil nutrients to the croplands. Hopefully, this study will convey to farmers the use of growing integrated grassllegume pastures and crops, and illustrate that livestock have their own role in transporting nutrients and hence use them as good means of distributors of soil nutrients. / Thesis (M.Sc.)-University of KwaZulu-Natal, Pietermaritzburg, 2004.

Please read the abstract in the 01chapter1 part of this document / Thesis (DPhil (Food Science))--University of Pretoria, 2006. / Food Science / unrestricted

Rhizoctonia solani and Pythium spp. are aggressive soil-borne fungal pathogens responsible for seed rot and seedling damping-off of many crops. With increased environmental and public concern over the use of chemicals, biological control of these diseases has been attracting more attention. However, success with this strategy depends on the development of effective antagonists, which requires repeated in vitro and in vivo tests. Bacillus spp. were isolated from a soil sample obtained from a field where sorghum and tef had been grown for at least two years. Potential Bacillus isolates were screened for their ability to inhibit in vitro growth of R. solani and Pythium sp. Among 80 isolates tested, endospore forming Bacillus spp. H44 and H51 gave highest antifungal activity against the two test-pathogens in three consecutive tests. Results demonstrated that both H44 and H51 have potential as biocontrol agents against diseases caused by these two pathogenic fungi. The interaction between three isolates of Trichoderma (T. harzianum Eco-T, Trichoderma spp. SY3 and SY4) and Pythium sp. were investigated using in vitro bioassays together with environmental scanning electron microscopy (ESEM). Visual observation on the dual culture tests revealed that hyphal growth of Pythium was inhibited by these antagonists soon after contact between the two organisms within 3-4 days of incubation. The ESEM investigations showed that all three isolates of Trichoderma grew toward the pathogen, attached firmly, coiled around and penetrated the hyphae of the pathogen, leading to the collapse and disintegration of the host's cell wall. Degradation of the host cell wall was postulated as being due to the production of lytic enzymes. Based on these observations, antibiosis (only by Eco-T) and mycoparasitism (by all three isolates) were the mechanisms of action by which in vitro growth of Pythium sp. was suppressed by these Trichoderma isolates. The reduction of seedling diseases caused by R. solani and a pythium sp. were evaluated by applying the antagonists as seed coating and drenching antagonistic Bacillus spp. (B81, H44 and H51) and Trichoderma (T. harzianum Eco-T and Trichoderma spp. SY3 and SY4). On both crops, R. solani and Pythium sp. affected stand and growth of seedlings severely. With the exceptions of H51, applications all of isoltes to seeds reduced damping-off caused by R. solani in both crops. Application of Eco-T, H44 and SY3 to sorghum controlled R. solani and Pythium sp. effectively by yielding similar results to that of Previcur®. On tef, biological treatments with Eco-T and SY4 reduced seedling damping-off caused by R. solani and Pythium sp., respectively, by providing seedling results similar to the standard fungicides, Benlate® and Previcur®. Most other treatments gave substantial control of the two pathogens on tef. Overall, Bacillus sp. H44 and T harzianum Eco-T were the best biocontrol agents from their respective groups in reducing damping-off by the two pathogens. In all instances, effects of application method on performance of biocontrol agents and adhesive on emergence and growth of seedlings were not significant. A field trial was conducted at Ukulinga Research Farm at the University of Natal, Pietermaritzburg, South Africa, to determine efficacy of biological and chemical treatments on growth promotion and reduction of damping-off incited by R. solani and Pythium sp., and to evaluate the effects of a seed coating material, carboxymethyl cellulose (CMC), on seedling emergence and disease incidence. Seeds of sorghum and tef were treated with suspensions of antagonistic Bacillus H44 or T harzianum Eco-T, or sprayed with fungicides, Benlate® or Previcur®. Application of Benlate® and Previcur® during planting significantly increased the final stand and growth of sorghum seedlings. Seed treatments with both H44 and Eco-T substantially controlled damping-off caused by Pythium, resulting in greater dry weights of seedlings than the standard fungicide. However, they had negative effects when they were tested for their growth stimulation and control of R. solani. The CMC had no significant effect on germination and disease levels. These results showed that these antagonists can be used as biocontrol agents against Pythium sp. However, repeated trials and better understanding of the interactions among the antagonists, the pathogens, the crop and their environment are needed to enhance control efficiency and growth promotion of these antagonists. Some of these biocontrol agents used in this study have the potential to diseases caused by R. solani and Pythium sp. However, a thorough understanding of the host, pathogen, the antagonist and the environment and the interactions among each other is needed for successful disease control using these antagonists. / Thesis (M.Sc.)-University of Natal, Pietermaritzburg, 2003.