Les patients admis en réanimation semblent pouvoir présenter des infections sévères à CMV, en dehors de tout traitement immunosuppresseur, mais l’incidence exacte de ces infections est difficile à évaluer. Nous avons réalisé un dépistage systématique du CMV chez 242 patients consécutifs, considérés non immunodéprimés avant leur admission en réanimation. Nous avons ainsi identifié que 16% des patients développaient au cours de leur séjour en réanimation une infection à CMV. La mortalité des patients ayant présenté une infection à CMV était supérieure à ceux n’ayant pas présenté l’infection. La physiopathologie des réactivations à CMV chez ces patients est incomprise. Nous avons sélectionné les patients de réanimation non-immunodéprimés et séropositif pour le CMV à l’admission, et nous avons comparé le statut des NK circulants de 15 patients ayant présenté une réactivation CMV dépistée par antigénémie circulante (les cas) à celui de 15 patients contrôles appariés aux premiers pour l’âge, le sexe, et la gravité à l’admission. Dans la période précédant la réactivation CMV, alors que les capacités de cytotoxicité sont comparables, nous mettons en évidence une déficience NK en termes de capacité de production d’interféron gamma chez les cas en comparaison des contrôles, et également de témoins sains. Le sepsis, cad le tableau clinique résultant de la réponse à toute infection, est dans sa forme la plus sévère, l’une des principales causes d’admission en service de réanimation. Nous avons réalisé un « immunomonitoring » complet des NK circulantes à la phase initiale d’états pro-inflammatoires chez 42 patients admis en réanimation pour des motifs infectieux (sepsis sévère ou choc septique) ou des tableaux de SIRS non infectieux. Les patients septiques présentaient une capacité réduite de dégranulation en comparaison des patients non-septique (SIRS). Ils présentaient également une réduction de la sécrétion d’interféron-, toujours en comparaison des patients SIRS. / Patients admitted to the intensive care unit (ICU) seem to have severe infections with CMV, without any immunosuppressive therapy, but the exact incidence of these infections is difficult to assess. We conducted a systematic screening of CMV in 242 consecutive patients non-immunosuppressed before ICU admission. We identified that 16% of patients developed CMV infection during their stay in ICU. The mortality of patients with CMV infection was higher than those without such infection. The pathophysiology of CMV reactivation occurring in critically ill patients with no previous immunosuppression is misunderstood. We selected non-immunocompromised ICU patients, CMV seropositive at admission, and we compared the status of circulating NK of 15 patients with CMV reactivation detected by antigenemia (cases) to 15 controls matched for age, gender, and severity on admission. In the period preceding the CMV reactivation, whereas capabilities cytotoxicity (degranulation) are comparable, we show impaired NK in terms of production of gamma interferon in cases compared to controls, and also healthy. Levels of IL-10 were significantly higher in cases with a strong correlation between the levels of this cytokine and severity of CMV reactivation as measured by the number of cells positive antigenemia. Sepsis, the clinical picture resulting from the response to any infection, in its severest form, is one of the main causes of admission in ICU. We conducted a complete immunomonitoring of circulating NK in the initial phase of pro-inflammatory state in 42 patients admitted to intensive care for reasons of infection (severe sepsis or septic shock) or of non-infectious SIRS (systemic inflammatory response syndrome). Septic patients showed a reduced ability of degranulation compared with non-septic patients (SIRS). They also showed a reduction in the secretion of gamma interferon, compared with patients with SIRS.

Mast cells respond to a variety of signals, are associated with both increased inflammation and regulation of the immune response, and are able to interact with a variety of hematopoietic and non-hematopoietic cells. The majority of the work that highlights mast cell pleiotropic abilities has been completed in murine models. Though these models have significantly advanced our understanding of what mast cells can do, they cannot inform us as to what mast cells actually do in human beings. The goal of this dissertation is to assess fully mature, primary human mast cell function beyond the well-defined type 1 hypersensitivity function and place mature human mast cells in the context of interactions with other immune cells. The first project addresses the ability of IFNγ, a historically Th1 associated cytokine, to dramatically alter mast cell phenotype. In particular, IFNγ stimulation allows mast cells to act as antigen presenting cells to CD4+ T cells. The second project describes and addresses the T cell suppressive function of myeloid derived suppressor cells in Mastocytosis, a disease of clonal mast cells.

Thesis (Ph. D.)--University of Washington, 2007. / Vita. Includes bibliographical references (leaves 117-142).

Thesis (Ph.D.)--University of Tennessee Health Science Center, 2009. / Title from title page screen (viewed on August 19, 2009). Research advisor: Edward Rosloniec, Ph.D. Document formatted into pages (xi, 126 p. : ill.). Vita. Abstract. Includes bibliographical references (p. 103-117).

When grown under hypothermic conditions, Chinese Hamster Ovary (CHO) cells become growth arrested in the G₀/G₁ phase of the cell cycle and also often exhibit increased recombinant protein production. In this study, we have validated this hypothesis by stimulating hypothermic growth using basic fibroblast growth factor and fetal bovine serum supplementation. This method led to 7.7- and 4.9-fold increase in total production compared to the 37°C and 32°C control cultures, respectively. This proof-of-concept study will motivate the creation of cell lines capable of growing at low temperatures for use in industrial processes. / Singapore-MIT Alliance (SMA)

Parkinson’s disease (PD) is characterized by the progressive loss of dopamine (DA) neurons in the substantia nigra pars compacta (SNc). Mechanisms regulating this neurodegeneration, however, are unclear. Evidence from PD pathology and models of PD, indicate mitochondrial disfunction triggers several death signalling pathways. Accordingly, in vivo and in vitro mitochondrial stress models of PD were employed to explore the role of two divergent molecular influences on dopaminergic neuronal survival. We examined neuroinflammatory and death signalling pathways arising from MPTP-induced mitochondrial stress. Interferon-gamma (IFN-ɣ) is a cytokine known to activate cellular components of inflammation, including microglia of the central nervous system (CNS). Results of a screen for cytokines in PD patient plasma revealed elevated levels of IFN-ɣ, suggesting a correlation between IFN-ɣ and PD associated DA cell death. In an MPTP mouse model of PD, germline deletion of IFN-ɣ improved survival of DA neurons and the nigrostriatal system, along with a reduction in microglia activation. Employing a survival co-culture system of neurons and microglia, it was found that neutralizing IFN-ɣ reduced DA cell loss induced by the mitochondrial complex I inhibitor, rotenone. DA cell death required localized microglia, activated through the IFN-ɣ-receptor (IFN-ɣ-R), with DA survival inversely proportional to IFN-ɣ expression, found to be up-regulated following rotenone. Investigation of the calpain-Cdk5-MEF2 signalling pathway in the MPTP model of DA cell death, motivated an examination of the nuclear orphan receptor, Nur77, following a review of potential MEF2 regulatory targets. MPTP induced a reduction in Nur77 mRNA from basal ii levels in SNc tissue, further regulated by ectopic Nur77 expression. These results strengthened our new model of MEF2 Nur77 regulation in DA neurons. In MPP+/MPTP DA survival experiments, loss in germline Nur77 expression presented an elevation in DA neuronal death both in vitro and in vivo, with a greater impairment in the nigrostriatal circuitry in comparison with normal expressing animals and cells. Dopaminergic supersensitivity related to Nur77 deficiency was attenuated with the ectopic expression of AV-Nur77 in vivo. These opposing mediators of survival yield new mechanisms by which DA neurons die, suggesting a mutitargeting approach to halt the progression of DA cell death.

This first detailed description of the pathology of tuberculosis, caused by Mycobacterium tuberculosis in springbok is reported. The springbok were part of a semi free-ranging herd kept on the grounds of iThemba Laboratory for Accelerator Based Science (LABS) in the Kuils River district of the Western Cape Province, South Africa. Of the 33 animals sampled, two animals had tuberculosis lesions. Mycobacterium tuberculosis was isolated from these two animals, as well as an animal that did not show tuberculosis pathology. The index case was an adult ewe that was presented for necropsy in a severely weakened condition. The ewe showed advanced miliary tuberculosis with marked macroscopic lesions in the lungs, pleura and respiratory lymph nodes. Limited sampling was done but microscopic tuberculosis lesions were found in almost all the organs sampled, and acid-fast bacilli were generally numerous. Six healthy rams were culled nine months later and a pilot study indicated miliary tuberculosis lesions in one ram, which again were macroscopically most prominent in the lungs, pleura and respiratory lymph nodes. Macroscopic lesions were also noted in the sternal, iliac, prefemoral and retropharyngeal lymph nodes. Microscopy in this animal revealed lesions in the macroscopically affected organs as well as numerous other lymph nodes, and suspected lesions occurred in the testicle and colon. Acid-fast bacilli were scarce to moderate in affected organs. Because of the miliary nature of the lesions in both affected animals, the route of infection could not be established conclusively. The lesions in most affected organs of both animals resembled classical tuberculous granulomas, viz. central caseous necrosis, with various degrees of calcification, surrounded by various numbers macrophages, epithelioid cells, multinucleated giant cells and lymphoplasmacells, and mild to moderate fibrous encapsulation. Necrotic lesions in the spleen, liver and kidney of the ewe were more disseminate and coagulative. A main study conducted on healthy culled animals 19 months after the pilot study failed to find any animal with tuberculosis lesions in the group of 25 sampled. These animals were all negative for mycobacteria via mycobacterial culture. The Interferon-gamma (INFg) assay was performed on all the animals of the pilot and main study but failed to identify the culture-positive animals and showed one false-positive reaction. / Dissertation (MMedVet (Pathology))--University of Pretoria, 2007. / Paraclinical Sciences / unrestricted

Please read the abstract in the 00 front of this document. / Thesis (PhD)--University of Pretoria, 2009. / Veterinary Tropical Diseases / unrestricted

Indiana University-Purdue University Indianapolis (IUPUI) / Chlamydia trachomatis is an obligate intracellular bacterial pathogen that can infect the eyes, genital tract, and disseminate to lymph nodes in humans. Many C. trachomatis infections are clinically asymptomatic and can become chronic if left untreated. When humans are infected with C. trachomatis, a cytokine that is produced is interferon-gamma (IFN-γ). In vitro, IFN-γ stimulates expression of the host enzyme indoleamine 2,3-dioxygenase. This enzyme converts free intracellular tryptophan to N-formylkynurenine. Tryptophan starvation induces C. trachomatis to enter a viable-but-nonculturable state termed persistence, which has been proposed to play a key role in chronic Chlamydial disease. To circumvent host induced tryptophan depletion, urogenital strains of C. trachomatis encode a functional tryptophan synthase (TS). TS synthesizes tryptophan from indole and serine, allowing Chlamydia to reactivate from persistence. Transcriptomic analysis revealed C. trachomatis differentially regulates hundreds of genes in response to tryptophan starvation. However, genes that mediate entry, survival, and reactivation from persistence remain largely unknown. Using a forward genetic screen, we identified six Susceptible to IFN-γ mediated Persistence (Sip) mutants that have diminished capacities to reactivate from persistence with indole. Mapping the deleterious persistence alleles in three of the Sip mutants revealed that only one of the mutants had a mutation in TS. The two other Sip mutants mapped had mutations in CTL0225, a putative integral membrane protein, and CTL0694, a putative oxidoreductase. Neither of these genes plays a known role in tryptophan synthesis. However, amino acid (AA) competitive inhibition assays suggest that CTL0225 may be involved in the transport of leucine, isoleucine, valine, cysteine, alanine, and serine. Additionally, metabolomics analysis indicates that all free amino acids are depleted in response to IFN-γ, making this amino acid transporter essential during persistence. Taken together we have identified two new chlamydial persistence genes that may play a role in chronic chlamydial disease.

Cyclosporine is a potent immunosuppressive agent used to treat a wide range of canine inflammatory diseases. Unfortunately, optimal dosing protocols for achieving immunosuppression with cyclosporine in dogs remain unclear, and standard methods that objectively monitor effectiveness of immunosuppression have not been established. We evaluated an already established panel of biomarkers of immunosuppression in vivo with two oral dosages of cyclosporine in seven normal dogs, a high dosage known to induce immunosuppression and a lower dosage used to treat atopy, with a washout period between the two dosages. The biomarker panel included the flow cytometric evaluation of T-lymphocyte cytokine expression (IL-2, IL-4, and IFN-gamma). High dosage cyclosporine resulted in significant decreases in IL-2 and INF-gamma expression, but not IL-4 expression. Low dosage cyclosporine was associated with a significant decrease in INF-gamma expression, while IL-2 expression was not affected. The results demonstrated suppression of biomarkers in a dose-dependent manner.