Thesis (Ph.D.)--University of Alabama at Birmingham, 2009. / Title from PDF title page (viewed on July 16, 2010). Includes bibliographical references.

Resumo: O objetivo do estudo foi avaliar a transferência de imunidade passiva (TIP) de vacas Canchim aos seus bezerros. Um grupo experimental foi constituído por 13 vacas primíparas e seus bezerros, e o outro por 13 vacas pluríparas e seus bezerros. Foram coletadas amostras de sangue venoso dos bezerros e secreções lácteas das vacas até 1 hora e 1, 2, 7, 15 e 30 dias após o nascimento/parto. No hemograma, notaram-se alterações características do período neonatal, como redução na contagem de hemácias, teor de hemoglobina e volume globular após a ingestão do colostro, além de aumento na contagem de linfócitos e redução da contagem de neutrófilos segmentados. Os exames bioquímicos séricos revelaram aumento nas atividades de GGT (até 3.746 U/L) e ALP (até 1.030 U/L) e nos teores de proteína total (até 7,77 g/dL), globulinas (até 6,01 g/dL), IgA (até 322 mg/dL) e IgG (até 2.918 mg/dL) após a ingestão do colostro, seguidos por redução gradual nestes parâmetros até os 30 dias de idade. As avaliações bioquímicas das secreções lácteas revelaram alta concentração de todos os componentes colostrais analisados, com redução gradual nos seus teores no decorrer do período experimental, à exceção dos minerais, cujas concentrações oscilaram pouco entre os momentos. A TIP foi eficiente em ambos os grupos de bezerros, e a qualidade das secreções lácteas, embora diferentes entre vacas primíparas e pluríparas, não interferiu na passagem de imunoglobulinas da vaca para o bezerro. / Abstract: The aim of this study was to evaluate the passive immunity transfer from Canchim cows to its calves. One experimental group comprised 13 primiparous cows and its calves and the other group, 13 multiparous cows and its calves. Samples of calves' venous blood and cow's lacteal secretions were collected until 1 hour and 1, 2, 7, 15 and 30 days after birth/parturition. In the hemogram, characteristic alterations of this period, such as reduction in the erytocyte count, hemoglobin concentration and corpuscular volume after colostrum intake and rise in the lymphocyte count and reduction in the neutrophil count were noticed. The serum biochemistry revealed augmentation in the GGT (up to 3,746 U/L) and ALP (up to 1,030 U/L) activities and in the total protein (up to 7.77 g/dL), globulin (up to 6.01 g/dL), IgA (up to 322 mg/dL) and IgG (up to 2,918 mg/dL) concentrations after colostrum intake, followed by a gradual reduction in these parameters until 30 days of age. The biochemical evaluation of the lacteal secretions revealed high concentrations of all the colostral components analysed, with gradual reduction in their concentrations along the experimental period, except for the minerals, which showed little oscillation between the moments. The passive immunity transfer was efficient in both groups of calves, and the quality of the lacteal secretions, although different between primiparous and multiparous cows, did not interfere in the transference of immunoglobulins from the cows to the calves. / Orientador: José Jurandir Fagliari / Coorientador: Alexandre Amstalden Moraes Sampaio / Banca: Eduardo Harry Birgel Júnior / Banca: Daniela Gomes da Silva / Mestre

Heavy chain isotype switch recombination is preceded by the appearance of RNA initiating 5' of the specific switch region which will undergo recombination. In an effort to understand the potential function of germline transcripts in switch recombination and the degree to which the regulation of germline transcripts correlates with the regulation of switching, we studied this process in the murine B-lymphoma cell line I.29μ, which in the presence of bacterial lipopolysaccharide (LPS) switches primarily to IgA and less frequently to IgE. Levels of α-germline transcripts initiating upstream of α switch (Sα) sequences are elevated in clones of this line which switch well as compared to clones which switch less frequently. TGFβ1 has been shown to increase α-germline transcripts and switching to IgA expression in LPS-stimulated murine splenic B-cells. We now demonstrate in I.29μ cells that TGFβ also increases switching to IgA and increases the level of α-germline transcripts 5 to 9 fold. Nuclear run-on analysis shows that this increase is at the level of transcription. Thus, TGFβ appears to direct switching to IgA by inducing transcription from the unrearranged Sα- CαDNA segment. Germline α RNA is quite stable in I.29μ cells, having a half life of about 3 to 5 hours, and we find only slight stabilization in the presence of TGFβ. Levels of ε-germline transcripts are not increased by TGFβ . IL-4, which modestly increases switching to IgA in I.29μ cells, slightly increases trancription of α-germline RNA. However, we present evidence suggesting that endogenously produced IL-4 may also act at additional levels to increase switching to IgA. IFNγ, which reduces IgA expression in these cells, also reduces the level of α-germline transcripts. IFNγ also reduces the level of ε-germline transcripts induced by IL-4. Our results support the hypothesis that the regulation of transcription of particular switch sequences by cytokines in turn regulates the specificity of recombination. In studies aimed at identifying other signalling pathways that promote class switching, we discovered that inhibitors of the nuclear enzyme poly(ADP-ribose) polymerase (PARP) increase lipopolysaccharide (LPS)-induced switching to IgA in the B cell lymphoma I.29μ and to IgG1 in LPS + IL-4-treated splenic B cells. PARP, which binds to and is activated by DNA strand breaks, catalyzes the removal of ADP-ribose from NAD+ and poly(ADP-ribosylation) of chromatin-associated acceptor proteins. This enzyme is believed to function in cellular processes involving DNA strand breaks as well as in modulating chromatin structure. In I.29μ cells, PARP inhibitors increase IgA switching by day 2 and cause a 5-fold average increase in switching on day 3 as assayed by immunofluorescence microscopy. The PARP inhibitor, nicotinamide, also causes a reduced intensity of hybridization of Cμ and Cα specific probes to genomic DNA fragments containing the expressed VDJ-Cμ and the unrearranged Sα - Cα segments, respectively, indicating that PARP inhibition increases rearrangment of these fragments. Induction of switching by PARP inhibitors is not mimicked by treatment with cAMP analogs or reduced by inhibitors of protein kinase A (PKA). Induction of switching by PARP inhibitors does not appear to involve increased levels of transcription of the unrearranged Cα gene, although TGFβ is required for optimal induction by PARP inhibitors, consistent with a requirement for transcription of the unrearranged CH gene. PARP inhibitors do not overcome the requirement for endogenously produced IL-4.

While a decrease of wild felid population has led to disruption of conservation programme, recent studies have shown the importance of immune regulation for determining health outcomes and co-infection. Immunoglobulin G is important for detecting and evaluating responses to infectious diseases and vaccination. But, there is limited information on felid immunoglobulins and their role for functional immunity. This study aimed at isolating and characterizing lion’s immunoglobulin G. Lions’ sera (n = 68) were processed using the MagReSyn® magnetic beads and the final protein concentration was determined using the Xpose™ Trinean Spectrophotometer. The cross-reactivity of goat anti-cat immunoglobulin with sera of lions and other species was analysed using ELISA. High cross-reactivity was observed in lions ranging from 87.7 to 100%, and low reactivity with rhino (22.4%) followed by chicken (0.01%). The protein concentration from purified sera yielded 39.09 mg/ml. Molecular weight of lion IgG 150-160 kDa was detected with both chains at 54-56 kDa and 24-26 kDa on SDS PAGE. These results indicate a potential aid in developing serological tools to monitor exposure to micro-organisms of lions. / Agriculture and  Animal Health / M. Sc. (Agriculture)

No description available.

Octapharma tillverkar Octagam, en lösning innehållande humant immunglobulin för intravenös administrering, vars huvudkomponent är immunglobulin G (IgG). Aggregerade IgG förknippas med aktivering av komplementsystemet i frånvaro av antigen. Denna ospecifika aktivering av komplement har tidigare rapporterats orsaka biverkningar hos patienter. För att undvika detta använder Octapharmas laboratorium för kvalitetskontroll en metod som kallas test av antikomplementär aktivitet, för kontroll av batcher av Octagam innan de släpps ut på marknaden. Denna metod har flera problem, t.ex. låg tillgänglighet av viktiga reagens. Syftet med detta projekt var därför att undersöka om en alternativ metod, CIC-C1q ELISA, kunde mäta aktiveringen av komplementsystemet på immunglobuliner genom att testa olika batcher av IgG och värmebehandlade IgG-prover för att bedöma om det fanns en korrelation mellan resultaten från de två metoderna. Resultaten visade att det inte fanns någon korrelation mellan ACA-testet och CIC-C1q ELISA. Varken normala IgG-batcher eller värmebehandlade prover uppvisade någon tydlig korrelation. Den enda fördelen som C1q kan ha är att den gav ett högt resultat för en batch som tidigare har rapporterats orsaka biverkningar hos patienter, medan svaret i ACA-testet var relativt normalt. CIC-C1q ELISA metoden har dock visat sig ha sina egna problem eftersom standardprover ger inkonsekventa värden och mer tester behöver utföras för att hitta spädnings-linjäritet. / Octapharma manufactures Octagam, a liquid preparation of highly purified human immunoglobulin for intravenous administration, the main component of which is immunoglobulin G (IgG). Aggregated IgG is associated with activation of the complement system in the absence of antigen. This non-specific activation of complement has been reported to cause adverse reactions in patients. To avoid this, Octapharma's quality control laboratory uses a method called the determination of anti-complementary activity (ACA) in Immunoglobulin, for control of all batches of Octagam before release. This method has several problems, such as low availability of critical reagents. Therefore, the aim of this project was to investigate whether an alternative method, CIC-C1q ELISA, could measure the activation of the complement system on immunoglobulins by testing different batches of IgG and heat-treated IgG samples to assess whether there is a correlation between the results of the two methods. The results showed that there was no correlation between the ACA test and the CIC-C1q ELISA. Neither normal IgG batches nor heat-treated samples showed a clear correlation. The only advantage the C1q may have is that it gives a high response for a batch that has been reported to cause adverse reactions in patients, while the response in the ACA test was relatively normal. If this can be demonstrated by testing similar batches, it may be beneficial to continue testing the C1q ELISA. However, this method has also been shown to have its own problems, such as the standard sample giving inconsistent values, and more work is needed to find the linearity of the dilution.

No description available.

Diabetes mellitus Typ 1 und Allergien vom Soforttyp gehören zu den häufigsten chronischen Erkrankungen des Kindes- und Jugendalters. Diabetes mellitus Typ 1 wird verursacht durch eine autoimmune Zerstörung der Beta-Zellen des Pankreas. Aus immunologischer Sicht wird dieser Prozess durch TH1-Zellen dominiert. Im Gegensatz dazu wird vermutet, dass Allergien vom Soforttyp, wie die allergische Rhinitis, das allergische Asthma und die allergische Urtikaria mit TH2-Zellen assoziiert seien. Die Hypothese, dass TH1- und TH2-Zellen sich gegenseitig in ihrer Aktivität hemmen, ist immer noch gültig. Ziel unserer Fall-Kontroll-Studie war es, die Assoziation zwischen Typ 1 Diabetes und IgE-vermittelten Allergien zu untersuchen. Zur Prüfung unserer Forschungshypothese wurden ein standardisierter, evaluierter Fragebogen sowie verschiedene Laboranalysen herangezogen. Es konnte gezeigt werden, dass Diabetes mellitus Typ 1 mit einem erhöhten Risiko für das gleichzeitige anamnestische Vorliegen IgE-vermittelter allergischer Symptome assoziiert sein könnte. Somit konnten wir bestätigen, dass die noch heute weit verbreitete TH1/TH2-Hypothese eine Vereinfachung tatsächlich viel komplizierterer immunologischer Vorgänge darstellt. Um diese Assoziation im Detail zu prüfen, bedarf es jedoch weiteren populationsbasierten epidemiologischen Studien.

published_or_final_version / abstract / Pathology / Master / Master of Philosophy

<p>Analysis of M-components is an important part of the diagnosis of monoclonal gammopathies and for the evaluation of disease response during treatment. In this project, two widely used electrophoresis methods and their corresponding immunotyping method were compared to evaluate the sensitivity of each method for the detection of small M-components. The project included 30 plasma samples from patients with identified M-components; 10 samples containing each IgG, IgA and IgM, respectively. All samples were diluted with normal EDTA plasma to achieve M-components of 5,00g/L. The samples were then serially diluted to achieve M-component concentrations of; 5,00, 2,50, 1,25, 0,63, 0,31 and 0,16g/L. All 180 samples were analysed with agarose gel electrophoresis and capillary electrophoresis. The dilutions above and below the detection level of each method were then analysed with immunofixation and immunosubtraction. The results showed good agreement between agarose gel electrophoresis and capillary electrophoresis in the highest concentrations of IgG and IgM. With agarose gel electrophoresis, IgA was detected in the same location as transferrin and the lowest concentration detected were therefore 1,25g/L. Besides the samples containing IgG, immunofixation was the most sensitive method.</p>