The purpose of this study was to separate acute and chronic effects of moderate exercise on the immune system by analyzing three sets of experimental and control groups; (1) 72 hours, (2) 1 week, (3) 2 weeks post exercise. Mice swam 5 days per week for 3 weeks accumulating a total of 125, 225, and 225 minutes of exercise in weeks 1, 2, and 3, respectively. Moderate swimming exercise did not result in a significant increase in SDH levels (p > 0.05). There was no change in tissue cell responses as measured by mitogen responsiveness, nor in splenic and thymic cell counts in response to the training regimen at any time point (p ≥ 0.05). Total, CD4, CD8, and T cell counts in the lymph nodes were significantly suppressed at 72 hours and 2 weeks post exercise (p ≤ 0.05). It appears that chronic exercise resulted in an increased trafficking of lymphatic cells, which could be interpreted as a sign of heightened immune reactivity.

Thesis (M. Clin. Sc.)--University of Adelaide, 1985. / Includes bibliographical references (leaves 99-106).

Thesis (Ph. D.)--University of Chicago, Department of Linguistics, December 2002. / Includes bibliographical references. Also available on the Internet.

Thesis (M.Sc.)--University of Alberta, 2010. / A thesis submitted to the Faculty of Graduate Studies and Research in partial fulfillment of the requirements for the degree of Master of Science in Experimental Medicine, Department of Medicine. Title from pdf file main screen (viewed on April 24 2010). Includes bibliographical references.

Thesis (Ph.D.)--University of Alberta, 2010. / A thesis submitted to the Faculty of Graduate Studies and Research in partial fulfillment of the requirements for the degree of Doctor of Philosophy, Department of Medical Microbiology and Immunology. Title from pdf file main screen (viewed on April 24, 2010). Includes bibliographical references.

Thesis (Ph. D.)--Illinois State University, 1994. / Title from title page screen, viewed March 21, 2006. Dissertation Committee: H. Tak Cheung (chair), Herman E. Brockman, Lynne A. Lucher, Philip D. Morse, Anthony J. Otsuka. Includes bibliographical references (leaves 101-111) and abstract. Also available in print.

Η ανοσολογική απάντηση και κυρίως η κυτταρική ανοσία, κατέχει σημαντικό ρόλο στην παθογένεια της λοίμωξης τόσο από τον ιό της ηπατίτιδας Β όσο και από τον ιό της ηπατίτιδας C. Στην παρούσα μελέτη εκτιμήθηκε η κυτταρική ανοσία, όπως αυτή εκφράζεται από τους Τ λεμφοκυτταρικούς υποπληθυσμούς (CD3, CD4, CD8) στο περιφερικό αίμα και στον ηπατικό ιστό ασθενών με χρόνια ηπατίτιδα Β (ΧΗΒ) και χρόνια ηπατίτιδα C (XHC). Οι ασθενείς με ΧΗΒ χωρίσθηκαν σε δύο κατηγορίες, ανάλογα με την ενεργότητα της νόσου (ασθενείς με χρόνια ενεργό ηπατίτιδα και ασυμπτωματικοί φορείς της νόσου). Επίσης μετρήθηκαν οι κυτταροκίνες του περιφερικού αίματος ( IFN-γ, TNF-a, IL-10, IL-5, IL-12, IL-6, IL-1b, IL-4, IL-8, IL-2) στις παραπάνω ομάδες ασθενών. Μέτρηση των Τ λεμφοκυτταρικών υποπληθυσμών και των κυτταροκινών έγινε στους ασθενείς με ενεργό ΧΗΒ και XHC πριν και μετά την έναρξη της αντιϊκής αγωγής. Όσο αφορά τους Τ κυτταρικούς υποπληθυσμούς του περιφερικού αίματος δεν προέκυψε διαφορά στους ασθενείς με ενεργό ΧΗΒ και ασυμπτωματικών φορέων της νόσου. Αντίθετα, σημαντική αύξηση των Τ κυτταρικών υποπληθυσμών, ιδιαίτερα στο επίπεδο των CD8 κυττάρων διαπιστώθηκε στους ασθενείς με XHC τόσο στο περιφερικό αίμα όσο και στο ηπατικό ιστό σε σχέση με τους ασθενείς με ΧΗΒ. Όσο αφορά τον ηπατικό ιστό υπήρξε άμεση συσχέτιση των CD3 κυττάρων με τη πυλαία φλεγμονή και τη λοβιακή δραστηριότητα και στις δύο ομάδες ασθενών, ενώ άμεση συσχέτιση των CD8 κυττάρων με τη πυλαία φλεγμονή και τη λοβιακή δραστηριότητα παρατηρήθηκε μόνο στους ασθενείς με XHC. Από τη μέτρηση των κυτταροκινών του περιφερικού αίματος προκύπτει αύξηση της IFN-γ, TNF-a, IL-10, IL-5 στους ασυμπτωματικούς φορείς της ηπατίτιδας β σε σχέση με τους ασθενείς με ενεργότητα ενώ σχετικά με τους ασθενείς με XHC και ενεργό ηπατίτιδα β, σημαντική αύξηση του TNF-a και μείωση της IL-4 και IL-12 στους ασθενείς με XHC. Δεν παρατηρήθηκε διαφορά στους Τ κυτταρικούς πληθυσμούς και στις κυτταροκίνες του περιφερικού αίματος στους ασθενείς με ΧΗΒ και XHC πριν και μετά την έναρξη αντιϊκής αγωγής. Από τα αποτελέσματα της μελέτης προκύπτει ότι οι ασθενείς με ΧΗΒ και XHC επιδεικνύουν διαφορετικό ανοσολογικό προφίλ όσο αφορά τους Τ κυτταρικούς υποπληθυσμούς στο περιφερικό αίμα και στον ηπατικό ιστό. Φαίνεται ότι η ανάπτυξη ανεπαρκούς ανοσολογικής απάντησης είναι υπεύθυνη για τη μη αποτελεσματική ιϊκή κάθαρση και την εξέλιξη της ηπατικής βλάβης στους ασθενείς με ΧΗΒ, ενώ αντίθετα στους ασθενείς με XHC η νόσος εξελίσσεται παρά την παρουσία ανοσολογικής απάντησης που είναι πιο έντονη στο επίπεδο των CD8 κυττάρων. Παρόμοια αποτελέσματα προκύπτουν και από τη μέτρηση των κυτταροκινών του περιφερικού αίματος. Έτσι, όσο αφορά την XHC φαίνεται ότι η νόσος εξελίσσεται παρά την παρουσία μιας έντονης Th1 απάντησης, ενώ στους ασθενείς με ενεργό ΧΗΒ φαίνεται ότι υπερτερεί η Th2 απάντηση. Στους ασθενείς με χρόνια ηπατίτιδα β, φαίνεται ότι το υψηλό ιικό φορτίο μπορεί να οδηγήσει σε διαταραχή της λειτουργικότητας των Τ λεμφοκυττάρων, όσο αφορά την παραγωγή IFN-γ που οδηγεί σε εξέλιξη της νόσου και ενεργό ιϊκό πολλαπλασιασμο. / The aim of this study was to evaluate the immune response in peripheral blood and liver, through the measurement of T cell subsets and serum cytokines, in patients with chronic active hepatitis B, asymptomatic HBV carriers and chronic hepatitis C. Thirty four patients with chronic hepatitis B (21 with active hepatitis and 13 asymptomatic carriers) and twenty one patients with chronic hepatitis C were enrolled in the study. We also evaluated 21 biopsies from patients with active CHB and 21 biopsies from patients with CHC. We measured CD3, CD4, CD8 T cell lymphocytes in peripheral blood and liver tissue as well serum cytokines (IFN-γ, TNF-a, IL-10, IL-5, IL-12, IL-6, IL-1b, IL-4, IL-8, IL-2) in these patients. We also evaluated the T – cell subsets and serum cytokines in patients with active CHB and CHC before and after the onset of antiviral therapy. We found no differences in the numbers of all T cells lymphocytes in peripheral blood between patients with active CHB and asymptomatic carriers. In contrast, we found a significant increase in the numbers of T cell subsets in CHC compared to CHB patients. We also found a significant increase in the number of T cell subsets in the area of portal tracts and lobules in CHC patients compared to CHB patients. In both groups there was a direct correlation between CD3 cells in portal tracts and lobules and HAI score but a direct correlation between CD8 cells and HAI score was found only in CHC patients. Regarding cytokine profile, patients with chronic active disease presented significantly decreased levels of IFN-γ, TNF-alpha, IL-10 and IL-5 as compared to inactive carriers. Also, a significant negative correlation between serum hepatitis B viral load and IFN-γ levels was noted. No correlation was found between viral load and the other cytokines. Patients with CHC presented significantly increased levels of TNF-alpha and significantly decreased levels of IL-12 and IL-4 as compared to patients with active hepatitis b infection. We found no difference in T – cell subsets and serum cytokines before and after the onset of antiviral therapy. Insufficient cellular immune response is critical for the ineffective virus clearance and liver damage in active CHB, while in CHC, immune response is present in peripheral blood and liver. However, there is an immunological escape of HCV, which seems to survive in the presence of an adequate immune response. Also, patients with active hepatitis B and HBsAg inactive carriers seem to display different cytokine profile. Decreased Th1 response observed in patients with active hepatitis B could be implicated in the persistence of virus replication and ongoing progression of liver disease. Hepatitis B viral load seems to be an important factor for T cell dysfunction, as expressed through reduced IFN-γ production.

O ferro é um elemento chave em muitos processos metabólicos, como transporte de oxigênio, síntese de hormônios esteróides, respiração celular, transporte de elétrons, síntese de DNA, proliferação e diferenciação celular e regulação gênica. A deficiência de ferro é a desordem nutricional mais comum afetando aproximadamente um terço da população mundial. Pequenos déficits no compartimento funcional de ferro têm sérias conseqüências sobre o sistema imune, principalmente na imunidade mediada por células. A abordagem dos pais ou responsáveis, as exigências éticas e a aderência de crianças da mesma faixa etária e sem outros problemas que afetem o metabolismo do ferro e o sistema imune são as principais dificuldades enfrentadas no desenvolvimento de pesquisas com seres humanos, sendo necessário o estabelecimento de modelos experimentais. Este trabalho teve como objetivo estabelecer um modelo de indução e recuperação de deficiência de ferro em camundongos, visando a sua utilização em estudos sobre alterações do sistema imune induzidas por esta deficiência. A deficiência de ferro foi induzida por ingestão de uma ração com baixo teor de ferro (5 mg /kg de ração) por 4 e 8 semanas. No termino deste período foram determinados: concentração de hemoglobina (colorimetrico), hematócrito (microhematócrito), estoques de ferro hepático (espectrometria de absorção atômica) e fenotipagem (citometria de fluxo) dos linfócitos presentes no sangue periférico e em suspensão de células do baço dos animais dos grupos controle (C) e deficiente em ferro (DF), sendo avaliado a porcentagem de células T CD4+ e CD8+, bem como a expressão do receptor de transferrina (CD71+) nessas subpopulações. Não houve diferenças na concentração de hemoglobina e no valor do hematócrito entre os animais dos grupos DF e C, porém os estoques de ferro estavam significantemente reduzidos nos animais do grupo DF de quatro (p<0,05) e oito (p<0,01) semanas. Não houve diferenças na porcentagem de linfócitos T CD4+ e T CD8+ entre os animais dos grupos DF e C, porém os animais deficientes em ferro apresentaram maior porcentagem de linfócitos T CD8+ do baço expressando CD71+ (p< 0,001). Este trabalho sugere que a depleção nos estoques de ferro não altera a proporção dos subtipos de linfócitos, porem as células T CD8 + do baço são mais sensíveis à deficiência de ferro. / Iron have a crucial role in several metabolic pathways, such oxygen transport, steroid hormone synthesis, cellular respiration, electron transport, DNA synthesis, cellular proliferation and differentiation and genic regulation. The iron deficiency is most common disorder nutrition, affecting about 30% world population. Deficits in iron functional compartment have serious delays about immunity systems, especially in the cellular immunity. Because of environmental problems, age, deficiency of nutrients other than iron, prevalence of infection, which may make human studies difficult, we used an animal model. This work aimed established iron deficiency induction and recuperation in mouse, for study about immune systems alteration. Iron deficiency was induced by feeding mice a diet that contained only 5 mg Fe/Kg for 4 and 8 weeks. After this period were determined: hemoglobin (colorimetry), hematocrit (microhematocrit), liver iron stores (atomic absorption spectrophotometer) and we performed a flow cytometry analyses in peripheral blood and spleen lymphocytes in control (C) and iron deficient (ID) mouse. We defined the effects of iron deficiency on T-cell subset and expression of cell-surface transferrin receptor (CD71+) in these cells. Hemoglobin concentration and hematocrit of ID mice were not difference those of C mice, but iron stores of ID mice (4 and 8 weeks) were reduced (p< 0,05 and p< 0,01; respectively). Although T-cells subsets in peripheral blood and spleen were not altered, iron deficiency significantly increased the number of spleen T CD8+ cells that express CD 71+ (p< 0,001). Data suggest that depletion of iron storage not alter T-cells subsets and spleen T CD8+ is the most sensible subset in iron deficiency.

Tuberculosis (TB) is a global health problem, which is proving extremely difficult to control in the absence of an effective vaccine. Bacille Calmette-Gu&eacute;rin (BCG), the only vaccine currently licensed against TB, demonstrates variable efficacy in humans and cattle. A greater understanding of what constitutes a protective host immune response is required in order to aid the development of improved vaccines. Tissue-resident memory T cells (T_RM) are a recently-identified subset of T cells, which may represent an important aspect of protective immunity to TB. This thesis aims to characterise the role of lung T_RM in BCG-induced protection against TB. In a mouse model, intravascular staining allowed discrimination between lung-vascular and lung-parenchymal T cells. Experiments demonstrated that BCG vaccination induced a population of antigen-specific lung-parenchymal CD4⁺ T cells, a putative tissue-resident population. This lung-parenchymal population was significantly increased in frequency following mucosal BCG vaccination, compared to systemic BCG vaccination. This correlated with enhanced protection against Mycobacterium tuberculosis (M.tb) infection in the lungs of mice receiving mucosal BCG, compared to those receiving systemic BCG. Mucosal BCG induced lung-parenchymal CD4⁺ T cells with enhanced proliferative capacity and a PD1⁺KLRG1^- cell-surface phenotype, a memory-like phenotype associated with improved protection against M.tb infection. These cells may represent a BCG-induced lung T_RM population responsible for the enhanced protection observed following mucosal BCG. Overall, this thesis highlights the potential of mucosal vaccination to elicit lung T_RM and identifies this as a possible immunological mechanism underlying enhanced protection against M.tb infection. These cells may constitute an important target for future vaccination strategies.

Submitted by Ana Maria Fiscina Sampaio (fiscina@bahia.fiocruz.br) on 2012-11-27T17:22:07Z No. of bitstreams: 1 Andréa Mendes Pereira Ativação de celulas... 2006.pdf: 40276494 bytes, checksum: 9f4656b98e4c74bae9c81e7797e85988 (MD5) / Made available in DSpace on 2012-11-27T17:22:07Z (GMT). No. of bitstreams: 1 Andréa Mendes Pereira Ativação de celulas... 2006.pdf: 40276494 bytes, checksum: 9f4656b98e4c74bae9c81e7797e85988 (MD5) Previous issue date: 2006 / Fundação Oswaldo Cruz. Centro de Pesquisas Gonçalo Moniz. Salvador, Bahia, Brasil / Visando avaliar futuramente o potencial de citocinas na indução de resposta imune celular específica do tipo ThI quando associadas a antígeno(s) recombinante(s) de Leishmania chagasi/infantum no cão, a combinação de IL-2 e IL-12 caninas recombinantes é analisada no presente trabalho. Aqui, descrevemos a clonagem do DNA complementar (cDNA) e, pela primeira vez, a expressão de IL-2 canina recombinante biologicamente ativa em Escherichia coli e por células de mamífero. Para expressão em E. coli, utilizou-se a construção pRSET-calL- 2, anteriormente gerada por nosso grupo. Para expressão em células de mamíferos, foi realizada a clonagem do cDNA de IL-2, sintetizado por reação de transcrição reversa seguida de reação da polimerase em cadeia (RT-PCR) a partir do RNA total de células mononucleares do sangue periférico (CMSP) de cão estimuladas com concanavalina A (Con-A), no vetor pcDNAS.l, gerando a construção pcDNA3.1-caIL-2. O sucesso da clonagem em ambos os vetores de expressão foi confirmado a partir do sequenciamento de DNA e comparação dos resíduos de nucleotídeos com a seqüência de IL-2 canina previamente descrita por outro grupo de investigadores. A IL-2 canina recombinante (rcaIL-2) foi obtida como proteína de fiisão contendo cauda de histidina a partir da transformação de E. coli BL21(DE3)pLysS com pRSET- caIL-2, purificada por cromatografia de afinidade e renaturada por diálise. Além disso, a forma nativa de rcaIL-2 foi secretada no sobrenadante de cultura de células COS-7 transfectadas com a construção pcDNA3.1-caIL-2. A atividade proliferativa de rcaIL-2 sobre células CTLL-2 foi demonstrada em concentrações de até 220 pg/mL da citocina purificada a partir da expressão em E. coli e até a diluição de 1:256 do sobrenadante de COS-7 contendo rcaIL-2. A proteína biologicamente ativa foi capaz de manter a proliferação de CMSP de cães sadios por até 12 dias de cultivo quando as células foram tratadas com 50 ng/mL de IL-2 canina obtida de E. coli e por 10 dias com diluições de até 1:200 do sobrenadante de COS-7 contendo a citocina, na ausência de estímulo prévio ou concomitante. A proliferação foi dose-dependente, com ponto máximo ocorrendo no 8° dia de cultivo. A produção de interferon gama (IFN-y) por CMSP de cães sadios estimuladas com sobrenadante de COS-7 contendo IL-2 ou contendo IL-12 não foi significantemente maior que a produção basal. No entanto, um efeito sinérgico sobre a produção in vitro de IFN-y ocorreu quando concentrações subótimas de ambas as citocinas foram associadas. Diante dos resultados obtidos, a construção pcDNA3.1-caIL-2 e ambas as formas de IL-2 canina recombinante obtidas, assim como as condições experimentais aqui descritas, poderão ser utilizadas no futuro para o estudo do potencial de IL-2, associada ou não a IL-12, como modulador da resposta imune in vitro e in vivo de cães, durante o desenvolvimento de uma vacina ou imunoterapia para leishmaniose visceral canina. / Aiming to study in the fiiture the role of cytokines in inducing specific ThI cellular immune response when associated to Leishmania chagasi/infantum recombinant antigen(s) in dogs, the combination of recombinant canine IL-2 and IL-12 is analysed in the present study. Herein, we describe complementary DNA (cDNA) cloning and, for the first time, the expression of biologically active recombinant canine IL-2 in Escherichia coli and mammal cells. The construction pRSET-caIL-2, previously generated by our group, was used for E. coli expression. For mammalian expression, canine IL-2 cDNA was synthesized by reverse transcription followed by polymerase chain reaction (RT-PCR), using cDNA from a healthy dog’s peripheral blood mononuclear cells (PBMC) stimulated with concanavalin A (Con-A) and cloned into pcDNA3.1, generating the construction pcDNA3.1-caIL-2. Success in canine IL-2 cDNA cloning was accessed in both expression vectors by DNA sequencing and was confirmed by comparing nucleotides residues with canine IL-2 sequence previously described by other investigators. Recombinant canine IL-2 (rcaIL-2) was expressed as His-tag fiision protein after E. coli BL21(DE3)pLysS transformation with pRSET-caIL-2, purified by affinity chromatography and renatured through dialysis. In addition, the native form was secreted in culture supematants of pcDNA3.1-caIL-2 transfected COS-7 cells. A proliferative activity was demonstrated in CTLL-2 cells when the recombinant protein was diluted at 220 pg/mL of purified cytokine from E. coli expression or when COS-7 supernatant was diluted at 1:256. The biologically active protein was able to induce proliferation of PBMC of six healthy dogs until 12 days of culture when cells were treated with 50 ng/mL of E. coli expressed-IL-2 or until 10 days when treated with 1:200 COS-7 supernatant dilution containing the cytokine, without previous or concomitant stimulus. The proliferative effect was dose-dependent and maximum at 8th day of culture. Interferon-gamma (IFN-y) production by PBMC of eight healthy dogs induced by COS-7 IL-2 or IL-12-containing supematants was not significantly higher than the baseline production. However, the association of suboptimal concentrations of both cytokines induced synergistic effect upon in vitro IFN-y production by PBMC. Based on the presented results, the construction pcDNA3.1-caIL-2 or both recombinant protein forms obtained, as well as the experimental conditions described here, can be used in the future to evaluate the potential role of IL-2, associated or not to canine IL-12, as a modulator of in vitro and in vivo immune response of dogs during the development of a vaccine or immunotherapy for canine visceral leishmaniasis.