IMMUNOLOGY (2)

Author: SÁNCHEZ TILLÓ ESTER.
Year: 2005.
University: BARCELONA [www.ub.es].
Place of defense: FACULTAD DE BIOLOGÍA.
Place of preparation: FACULTAD DE BIOLOGÍA (UB) Y PARQUE CIENTÍFICO DE BARCELONA.

Summary: The macrophages are cells that have a critical role in the immune response. In the presence of the growth factor M-CSF proliferate and if one adds the bacterial lipopolysaccharide (LPS) induces its activation is blocked proliferation. The ERK MAPK is necessary for proliferation and for the activation of macrophages. However, their kinetic phosphorylation is different. In response to factors proliferation, the phosphorylation of ERK starts early and lasts for a very limited time. In response to LPS, the phosphorylation of ERK starts later and lasts longer. Thus, the response of macrófago depend on the initiation and spread of the activation of ERK. The phosphatase MKP-1 is that defosforila to MAPKs and kinetics of induction is parallel to the deactivation of MAPKs. The induction by M-CSF or LPS are mediated by PKC. Our studies in primary cultures of murine macrophages derived from bone marrow, show that the expression of MKP-1 by the two stimuli is dependent on the activation of the kinase Raf-1 and the interaction of PKC with it. The kinetics of activation of ERK is parallel to the activation of Raf-1 and MEK-1 / 2. Using inhbidores specific and RNA interference, has shown that the activation of ERK when induces proliferation is totally dependent on Raf-1 while stimuli activators can use alternate routes. The inhibition of the activity Raf-1 generates a cell cycle arrest in G1 caused by an increase in the expression of the kinase inhibitors dependent ciclina (Cdks) p21WALF1 and p24Kip1. With respect to the activation, there are no defects in the expression of proinflammatory cytokines in the induction of sinatasa of nitric oxide. Induction of MKP-1 by M-CSF by LPS, is not mediated by ERK and p38. However, inhibition of JNK blocks the expression of MKP-1. As a result of the inactivation of this phosphatase, there is a lengthening of the activity of the other MAPKs, ERK and p38. In macrophages are expressed constitutively, isoforms JNK1 and JNK2, while JNK3 not found. Most dela JNK activity was detected due to the isoform JNK1. Using mice in which it has been inactivated genes jnk1 and jnk2, we have shown that the induction of MKP-1 is mediated by the isoform JNK1. Moreover, this isoform is necessary for the biosynthesis of proinflammatory cytokines (TNF-alpha, IL-1beta and IL-6) and for the induction of NOS2.Esta activity is separate from the role of JNK1 as regulator MKP-1, as with mice have demonstrated that this gene inactivated. Activating the ciclofilina To produce inhibition of proliferation by blocking cell cycle in phase G1 without affecting cell viability. This blockade is drunk a decrease in expression of c-myc and the inactivation of the activity Cdk2 through the transcriptional induction of p27Kip1. The ciclofilina A intervenes in the early stages of signaling by M-CSF, inhibiting the phosphorylation of Raf-1 and ERK and the expression of MKP-1. Activating the ciclofilina A does not affect the activation of macrophages mediated by LPS. However, the ciclofilina alter the induction of the molecules increased histocompatibility complex (MHC) class II mediated IFN-gamma. This inhibition occurs as a consequence of blocking the expression of transactivador CIITA. Surprisingly, the ciclofilina not involved in the phosphorylation of STAT - 1 at tyrosine its nuclear translocation nor its binding capacity to Dna. However, we found that the phosphorylation at serine STAT - 1 mediated by PKC and ERK is inhibited, these being probably responsible for the blocking of the transcript of CIITA.

Author: YERAMIAN ANDREE.
Year: 2005.
University: BARCELONA [www.ub.es].
Place of defense: UNIVERSIDAD DE BARCELONA.
Place of preparation: UNIVERSIDAD DE BARCELONA.

Summary: The bone marrow derived macrophages stimulated by cytokines type Th1 as interferon range (IFN-gamma), are triggered by the "classical route" through induction of NOS2 and use the L-arginine to produce NO. In contrast, macrophages stimulated with cytokines type Th2 cells, such as interleukin 4 (IL-4), are triggered by the "alternative route" through induction of arginase and use the L-arginine to produce polyamines. Our results suggest that activation of classic and alternative transportation arginine increases through the system and +, inducing the expression of Slc7A2. At baseline conditions transport system and + is solely CAT1, while in macrophages activated by the tracks for the classical route, the increase in transport is mediated by Slc7A2 or CAT2. Indeed, in the absence of SlcA2, activation of macrophages is limited. The transport of arginine is independent of the expression levels of NOS2 or arginase. The regulation of the expression of genes involved in the activation classical (MHC class II, IA-beta, NOS2 or TNF-alpha), in the alternative activation (Arginasa Io recipient of the manosa) or in the same carriage (Slc7A1 or Slc7A2 ) is independent of the presence of extracellular arginine in the middle. The macrophages are able to proliferate in tissues in response to M-CSF. The proliferation of macrophages depends on the presence of extracellular arginine in the middle. The proliferation and activation of macrophages induce different routes catabolios of arginine. Unlike macrophages treated with IFN-gamma + LPS or IL-4 + IL-10, for the proliferation macrophages use a part of arginine for protein synthesis, and excessive exchange with the extracellular environment. At the controls or macrophages stimulated with M-CSF, L-Leucina in the presence of sodium is able to inhibit the bulk transport of arginine, indicating that takes place mostly through the system y + L, while system activity y + is very modest. Slc7A1 is expressed at low levels in the macrophages controls as macrophages stimulated with M-CSF or cytokines type Th1 or Th2. Slc7A2 not detected in macrophages controls or in treaties with M-CSF but is induced in the stimulated with cytokines. Slc7 $ 3 is not detected in macrophages. The expression of these genes is independent of the presence of extracellular arginine in the middle. The macrophages of the stock control and the strain Slc7A2 mutda stimulated with M-CSF, metabolize arginine in a similar way, producing equal amounts of citrulina or orinitina, indicating that the proliferation of macrophages is independent of Slc7A2. The lesihmaniasis is a disease caused by an intracellular parasite whose progress depends on the susceptiblidad genetics. BALB / c. The phenotype of mice susceptible to Leishmania major as BALB / c is associated with levels of IL-4 and a response TH2, while the resistant phenotype to L.major as CBA is associated with a response dominated by cytokines IFN range, IL-12 that are responsible for the elimination of the parasite through induction of NOS2 in macrophages. The induction of NOS2 and subsequent NO production is critical in eliminating the parasite. In our model of macrophages derived from bone marrow, infected with Leishmania Better we see no differences in mRNA expression of the NOS2 or production of NO in macrophages activated by cytokines TH1 from strains CBA and BALB / c. Nor are there any difference in mRNA expression of arginase I and the activity of this enzyme in macrophages from both strains with stimulating cytokine type TH2. Different levels of transport and catabolism of arginine between macrophages from the sensitive and resistant strains. We note that treatment with IL-4 as infection with Leishmania increased catabolism of arginine in macrophages. This increase in the catabolism of arginine and 8 ¢ ma 658 crófagos stimulated with IL-4 or infected with Leishmania is significantly more important in the sensitive strain BALB / c strain resistant to the CBA. This increase is mediated transport system and +, which is due to the increase in the induction of gene expression SlcA2. Treatment with nor-NOHA of macrophages stimulated with IL-4 induced a significant reduction in the viability of the parasites you macrophages of the two strains, stressing the role of arginase I in the control of growth in Leishmania. When we remove the extracellular environment arginine and ornithine substitute for seeing an increase in the 50% of the viability of the parasite in the macrophages of the two strains. However, this increase is more important in the sensitive strain than in the resistant strain. These data demonstrate that susceptibility to Leishmania amongst the transport system arginine / ornithine.

Author: MOSCOSO DEL PRADO UCELAY JUAN.
Year: 2005.
University: COMPLUTENSE DE MADRID [www.ucm.es].
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: DPTO. INMUNOLOGÍA, FACULTAD DE MEDICINA, UNIVERSIDAD COMPLUTENSE DE MADRID.

Summary: This thesis belongs to the framework of the studies of the genetics of human populations. We have based on the genes System Main Histocompatibility (HLA) as a genetic marker. It is the most polymorphic described in humans, the genes are responsible for tolerance / rejection in organ transplants, and has been recently that many alleles in different loci have a strong geographic correlation. Thanks to these characteristics, we have studied the genetic profile of the population HLA Maya of Guatemala for the purpose of studying the following objectives. Studying the genetic profile of the population HALA Maya study HLA haplotypes extended to check whether genetically belongs to all Amerindian populations and, finally, and with the help of other studies conducted in populations of the Americas, known as the continent pobló America since there is a lot of uncertainty about it. We obtained 132 blood samples from healthy individuals, and not related to a Guatemalan Mayan population, specifically of Xela (Quetzaltenango), which have remained isolated. We extracted DNA with conventional techniques, we conducted of the tipaje HLA loci-A, - B, -DRB1 and -DQB1 using molecular biology techniques, and finally, along with other populations, we conducted dendrogramas of Neighbor-Joining, studies of genetic distances and correspondence analysis to conduct comparisons with other populations and explain the settlement of America. We note that the Mayan population, in view of the allelic frequencies, their extended HLA haplotypes, dendrogramas of emparentamiento of Neighbor-Joining, genetic distances from the rest of the Maya people and correspondence analysis, belongs to a group of people defined as Amerindians. With regard to how pobló the American continent, we concluded that took place in different migration at the time from different parts of the Asian continent, as well as the existence of several population movements within the same continent once entered America.

Author: RODRÍGUEZ PENA REBECA.
Year: 2005.
University: COMPLUTENSE DE MADRID [www.ucm.es].
Place of defense: MEDICINA.
Place of preparation: FACULTAD DE MEDICINA DE LA UNIV. COMPLUTENSE DE MADRID.

Summary: The dendritic cells (DCs) comprise a population essential to obtain specific immune responses, both of tolerance so as effector versus pathogens, as they are the only cells that stimulate T lymphocytes virgin wing after the modulated to obtain the answer more effective against each pathogen. Knowing its biology will establish therapies that reconduzcan immune responses in diseases such as cancer, autoimmunity, transplantation, allergy or infection. This thesis has been studied how the DCs molecules derived from monocytes exchanged among themselves the consequences of this exchange. By flow cytometry and microscopy states that this phenomenon affects molecules of the material already own cell exogenous endocitado. Requires the contac5to direct intercellular and a traffic system endocítico correct, as well as lipid rafts cholesterol functioning. The model is enhanced during the maturation of DCs, both by increasing intracellular calcium and through LPS-TLR4 or after endocytosis of exogenous molecules. The exchange is facilitated if DCs involved in various stadiums madurativos, generating at the same time the maturation of the speedy and effective immature DCs. The exchange occurs between DCs and linfoblastos so bidirectional, affecting the maturation of DCs. The exchange of molecules, therefore, would be a process involved in the amplification of immune responses and in the activation of the system, since immature DCs molecules and receive signals from other maturing DCs mature prior to having direct contact with the pathogens.

Author: ESCRIBESE ALONSO MARÍA MARTA.
Year: 2005.
University: COMPLUTENSE DE MADRID [www.ucm.es].
Place of defense: FACULTAD DE CIENCIAS BIOLÓGICAS.
Place of preparation: FACULTAD DE CIENCIAS BIOLÓGICAS.

Summary: Treatment with mercury induced in the Brown Norway rats induced a polyclonal response, which leads to the production of auto anti-membrana baseline glomerular (MBG) mostly as both circulating deposited production of proteinuria and a prominent interstitial nephritis. In this paper we have raised two key objectives: 1-Studying the role of integrina VLA-4 in the development of autoimmune nephritis. 2-To study the effect of AR-tt in nephritis model of autoimmune-induced HgCl2. Q Prior has been described that treatment of this disease with antibody against alpha chain of integrina VLA-4 showed the relevance of this integrina in the development of the disease. This work also demonstrated that the protective effect is mediated by the ability VLA-4 to divert the answer inmunehacia Th1 a model of autoimmune disease Th2. Another aim has been to study the effect of tt-AR in nephritis model of autoimmune-induced HgC2. Our results indicate that this metabolite of vitamin A acts as anti-inflammatory in this model because it is capable of inhibiting the production of inflammatory mediators such as cytokines, quimioquinas, endothelial adhesion molecules and release of free radicals. Well Contributing to the maintenance of renal function. Within this objective raised explore the effect of AR-tt on the expression and binding of integrina VLA-4, and the results showed that AR-tt is able to reduce the expression of this integrina in circulating leukocytes. and thus to migrate to the renal interstitium, thus impeding the development of the inflammatory response. In conclusion if we could say that this is working is demonstrated the importance of lintegrina VLA-4 in the development of autoimmune nephritis induced HgCl2 and that this integrina is a target for therapeutic treatment AR-tt.

Author: ÁLVAREZ ERRICO DAMIANA.
Year: 2005.
University: POMPEU FABRA [www.upf.edu].
Place of defense: DEPARTAMENTO DE CIENCIAS EXPERIMENTALES Y DE LA SALUD.
Place of preparation: DEPARTAMENTO DE CIENCIAS EXPERIMENTALES Y DE LA SALUD.

Summary: This paper describes the cloning and characterization of a new receptor inhibitor of the Ig superfamily, which we have called IREM-1 (Immune Receptor Expressed by Myeloid Cells). The analysis of its sequence shows that this molecule is part of the family multigénica known as CMRF35 or CD300. IREM-1 was cloned from cDNA derived from peripheral blood mononuclear cells (PBMCs), and is a glucoproteina type I membrane with a single Ig Extracellular Domain, a fragment transmembrane and cytoplasmic tail with five tyrosine residues that are different reasons for intracellular signaling. Two of these residues, tirosinas 205 and 249 relate to ITMs, while the tirosinas 236 and 263 are part of the grounds YxxM associated with the recruitment of Pl3K. The tyrosine distal (Y284) could be considered as part of a plea ITIM-like. IREM-1 is able to associate itself with the fosfatas SHP-1, and we have identified the residue Y205 as primarily responsible for such interaction. IREM-1 is capable of producing signals on inhibitory receptors activators as FceRI, in a rat model with heterologous line basofílica RBL. This inhibition is mediated by both ITIMs. IREM-1 is also capable of uniting subunit p85 of the Pl3K through its two reasons YxxM. Both pleas have the ability to unite this molecule. The triple mutant IREM-1 of the 2 sites ITIMs and ITIM-like, which unites SHP-1, is capable of producing signals activating the induction of degranulation in RBLs. This response is dependent on PI3K and is canceled inhibitor of this enzyme. The expression of IREM-1 is restricted to myeloid cells hematopoiéticas lineage precursors including bone marrow CD34 +. In conclusion, we presented the identification and molecular characterization and functional receptor IREM-1.

Author: GUMÁ URIEL MÓNICA.
Year: 2005.
University: POMPEU FABRA [www.upf.edu].
Place of defense: DEPARTAMENTO DE CIENCIAS EXPERIMENTALES Y DE LA SALUD.
Place of preparation: DEPARTAMENTO DE CIENCIAS EXPERIMENTALES Y DE LA SALUD.

Summary: The objectives of this study were to study the expression of receptors of NK cells (NKR), in particular CD94/NKG2C, in connection with human cytomegalovirus (HCMV). The results described are the first evidence that infection with HCMV amending the code of NKR. The increase in the proportion of cells CD94/NKG2C + in seropositive donors for HCMV suggests involved in the response to the pathogen. The receiver CD94/NKG2C not only stimulates effector functions and the proliferation of NK cells, but also to an active minority subpopulation of T cells CD8 +. The study of in vitro expansion of the NK CD94/NKG2C + subpopulation after interaction with fibroblasts infected with HCMV, suggests that the receiver itself is involved in proliferation.

Author: SABORIT VILLARROYA IFIGÉNIA.
Year: 2005.
University: BARCELONA [www.ub.es].
Place of defense: FACULTAT DE MEDICINA.
Place of preparation: FACULTAT DE MEDICINA.

Summary: The family of CD 150, is formed by membrane receptors, expression in cells of the immune system that are a part of the superfamily of inmunolglubinas. They are characterized by a high share and locate structural homology in the same position 23 on the long arm of chromosome 1. Members CD84, CD150, CD229, CD244, CD319 and NTB-A have an intracellular region with at least one occasion tyrosine types of TV / IY-XX-V / R. This is why consensus union of a family of molecules called SAP adapter and EAT-2. In humans, deficiencies in SAP are responsible for an immune linked to chromosome X syndrome XLP, characterized by an extra susceptibility to the Epstein-Barr virus with infectious mononucleosis fulminant. The aim of this thesis has been to identify new ligands to intracellular receptor CD244. Activation of CD244 (or 2B4) involves activation of the citotoxicita increase in production of interferon g (IFNg) of NK cells and lymphocytes CD8 + cytotoxic. Through the system three times in yeast hybrid, we found that the molecule 3BP2 associates directly to the intracellular region of CD244 in a position to phosphorylation. The 3BP2 is an adapter molecule expression in hematopoietic cells, which, although possessing no catalytic activity has the capacity to be associated with different proteins, forming a signolosoma and positively participate in the activation of cells of hematopoietic lineage. Eh human point mutations in the 3BP2 are responsible for an autosomal dominant disease, called querubismo, characterized by a rapid degradation of the bones of maxilas, associated with a dysfunction of osteoclasts. The first objective was to verify and characterize the interaction between the receptor CD244 i adapter 3BP2 in human NK cells. In terms of activation was detected interaction between the two proteins in a line called tumor YT in what sobreexpresamos the 3BP "pattern with the green fluorescent protein (EGFP). Also show this interaction in human cells NL, obtained from lymphocyte peripheral blood following a protocol enrichment, selection and expansion with Interleukin 2. was necessary in this case to produce a monoclonal antibody against 3BP2 human following a protocol immunization in mice with a peptide of 3BP2. The second goal pass characterize the signaling pathways and functional responses to the activation of CD244 in NK cells in a background of involvement 3BP2.Encatamos that 3BP2 is affecting and enhancing ERK pathways and the ability of lithic NK cell. however, neither the activity of the kinase p38 or production of IFNg are modified by the presence or absence of the adapter. propose the 3BP2 as the molecule that gives the dichotomous role of the role of NK cells activated via CD244. also analyzed the interaction of 3BP2 with the other members of the family of CD150, by the technique of triple hybrid. found that the region intracellular receptor CD229 (or Ly9) interacts directly with the 3BP2 in a position to phosphorylation. has been reported that the recipient CD229 plays a role in attenuator TCR activation via the ERK pathways i the transcription factor NFAT in T lymphocyte Studies in this thesis, indicate that 3BP2 power of CD229-mediated inhibition on the TCR. This thesis has characterized the importance biochemical and functional the molecule adapter 3BP2 in two signaling receptor family of CD150. On the one hand, by partnering with the Cd229, the adapter 3BP2 increases attenuation of the activation of lymphocyte T. Therefore, in terms of domestic partners union sets, 3BP2 can play a positive or negative role in the activation of these two cell types.

Author: LORENZO ABALDE SILVIA.
Year: 2005.
University: VIGO [www.uvigo.es].
Place of defense: FACULTAD DE BIOLOGÍA.
Place of preparation: FACULTAD DE BIOLOGÍA, UNIVERSIDAD DE VIGO.

Summary: The bivalve molluscs are morphologically very similar in their early stages of development. The current method of identification larval is based on the analysis of certain morphological features of the ultrastructural charnela, with a slow and impractical for the larval discrimination in complex samples of plankton. To develop techniques faster, more sensitive and specific for discrimination larvae Mytilus gallo provincialis, we decided to generate monoclonal antibodies specific (AcsMo) following the protocol described by Drs. Milsteiny KÃ ¶ hler. Obtaining larvae was conducted through in vitro fertilization. Larvae of two days of age were used for immunization of mice, and splenocytes were fused with myeloma cells. Several hybridomas were obtained secretores of AcsMo face mussel larvae, which have been studied more extensively two of them: 36.5 and 22.8. These two AcsMo are specific versus mussel. Not recognize larvae of other species of bivalves (cockle, oyster, volandeira, lameja), but recognize mussel larvae of other populations; M.galloprovincialis Mediterranean (genetically divergent from galician), and M.edulis. Both AcsMo recognized by indirect immunofluorescence, mussel larvae in various stages of development, including postlarvas. To confirm the utility of antibodies in the samples of plankton, AcsMo were tested in different locations (Galicia, Italy and the Netherlands), by comparing the results obtained with the AcsMo, with the traditional morphological studies. The results showed that AcsMo stained exclusively to mussel larvae, not detected any cases of false positive in samples made. In this paper we show that these antibodies used in immunological techniques provide a tool for identifying specific and reliable mussel larvae. Automation using image analysis techniques, separation inmunomagnética or flow cytometry, allow for the incorporation of routine monitoring larvae den plankton.

Author: LÓPEZ SANTALLA MERCEDES.
Year: 2005.
University: COMPLUTENSE DE MADRID [www.ucm.es].
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: ÁREA DE INMUNOLOGÍA (DEPARTAMENTO DE MICROBIOLOGÍA I).

Summary: Recent research conducted in cancer patients and in animal models try to elucidate the delicate network connection established between the immune system cells and tumor cells. The term "immunological surveillance" describes the ability of the immune system to detect and destroy tumor cells. The role of the immune system in controlling tumor growth has been the target of investigaicón of active recent decades. The T lymphocytes appear to be central in the response to tumor cells and, in fact, have found tumor-specific T cells in the circulation of patients with cancer or infiltrating the tumor tissue. These assets have been T lymphocytes stimulated by neoplastic antigens on the surface of tumor cells and thus could eliminate them. However, it is not clear why T lymphocytes fail to exercise this control, and the tumor progresses and spreads. This failure of the immune system has been explained, among other causes, for the secretion of a factor secreted by the tumor can affect the functionality of T lymphocytes leading to a state inmunodeficiente that it would not be able to control the spread of the tumor. It has been described several malfunctions on T cells in patients with cancer, including poor responses proliferativas, some alterations of cell subpopulations, decreased production of cytokines, and finally absence of cell surface molecules associated with the TCR, as CD3. The objective of this paper is twofold. On the one hand, there has been a caracterizaicón phenotypic and functional peripheral blood mononuclear cells (CMSP) of patients, classified according to their clinical situation, and second, to analyze the role of T lymphocyte, using a model of transformation lymphotropic virus Herpesvirus saimiri, thus obtaining stable lines, which are grown in the laboratory. The results reveal that the presence of CD45 and response to CD3 can be used as markers to identify patients with a clinical situation worse. Furthermore, analysis of the lines showed the existence of a defect in the expression of the chain CD3 on the lines of patients but not in healthy cultured in the absence of IL-2. Finding the flaw in the lines suggests that is inherent in the cells of patients and not due to factors absent in tumor growth medium.

Author: CAMPOS MARTÍN YOLANDA.
Year: 2005.
University: COMPLUTENSE DE MADRID [www.ucm.es].
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: FACULTAD DE MEDICINA.

Summary: The cytolytic activity of NK cells can be inhibited by the recognition of MHC class I molecules and non-classical classics. The system CD1 consists of a family of glycoproteins related to the MHC class I molecules classic. CD1a, -by c lipids or glycolipids presented to lifnocitos T, has recently been described its ability to inhibit the activity of lithic NK cells, as with DC1d1 mouse. This paper describes how CD1d human is capable of inhibiting the activity lithic cell NKs. Treatment with alfa-GalCer or beta-GalCer, ligands CD1d, reverses the inhibition observed initially. The dendritic cells (DC) expressed physiological levels of CD1d and have an important role in immune response, which led us to study their interaction with NK and NKT cells. In addition, parasites of the genus Leishmania can decrease the expression of molecules CD1 Group I. In our work are shown as iDCs infected L.infantum, despite not increase the expression of HLA class I molecules, are resistant to NK lysis of cells, due to increased expression of HLA-E, ligand receptor inhibitor CD94/NKG2A. The iDCs infected with a greater expression of CD1d, being effectively recognized and lisadas cell iNKT, besides producing high concentrations of Infama. These results suggest an important role of NKT cells in the innate immune response compared with Leishmania.

Author: ESTHER TAMAYO REVUELTA.
Year: 2006.
University: CANTABRIA [www.unican.es].
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: FACULTAD DE MEDICINA.

Summary: For several years now investigating the potential use of enterotoxin termolábil E. Coli (LT) as a vaccine adjuvant manageable in mucosal surfaces. For this purpose develops an intense research activity that seeks to clarify the mechanisms involved in the adjuvant effect of LT, which have committed: the activation of antigen presenting cells, inducing molecules coestimuladoras in lymphocytes TyBo apoptosis of the immunocompetent cells. Another issue that commits the clinical use of LT is its high toxicity associated with the action ADP-ribosilasa its subunit A. That is why we have developed LT mutants with low or no toxicity and is evaluating its power mucosal adjuvant in vaccination. Methodology, assumptions and work plan: Our work focuses on the ability of LT to induce cell death aopotótica vivo, using an approximation several estripes murine experimental. We intend to explore what immunocompetent people involved in this effect aopotótico in different lymphoid tissues (thymus, bone marrow, spleen, lymph nodes, blood) depending on the route of administration of LT. Judge involvement of the different molecular pathways for the production of cell death by apoptosis, as well as the physiological mechanisms inducers (role of endogenous steroids). In addition examine whether apoptosis is linked to the activity enzimática-tóxica LT. Finally sie assess the effect of apoptosis of immune cells is a necessary process in the capacity of mucosal adjuvant in the LT. To develop these pilot studies will be used manipulation techniques of experimental animals (immunization ratone s by different routes: intraperitoneally, nose, intragástrica, ... taking samples of blood and tissue), techniques for detecting cell populations (cytometry flow) quantification antibody (ELISA) and the production of monoclonal antibodies through hybridomas.

Author: CASALS CASAS CRISTINA.
Year: 2006.
University: BARCELONA [www.ub.es].
Place of defense: FACULTAD DE BIOLOGÍA.
Place of preparation: FACULTAD DE BIOLOGIA UNIVERSIDAD DE BARCELONA PARQUE CIENTÍFICO DE BARCELONA.

Summary: The human genome is made up of more than 30000 genes. Not all cells will express all these genes, and will be expressed in a manner constituting or untold, depending on the cell type. These mechanisms need to be regulated thin and complex, which requires a great plasticity of both routes transucción signal and the modularity of the transcription factors. This thesis explores the plasticity of these two mechanisms through two concrete examples. LPS RAISES THE EXPRESSION OF CLASS II MHC IN CELLS DENDRÍTICAS AND CELL B ROUTE AP-1 NO AFFECT THE LEVELS OF CIITA The expression of genes in the MHC class II is strictly tissue specific. His expression is unspeakable, and only in certain tissues constitutively expressed in dendritic cells and B lymphocytes Although LPS block the induction of cytokine-dependent genes of MHC class II, in dendritic cells and B lymphocytes in the increases, both at the level of protein on the surface and at the level of mRNA. But this increase is not preceded by an increase in the expression of transactivador Class II, CIITA. In transient transfections, the LPS induces the expression is regulated by a fragment of 2035 bp containing a box AP-1 to -1722bp the start of transcription. If this box is lost mutates the effect produced by LPS, on the other hand it continues to respond to LPS if moves to -611 bp or invests in acting as a done. The LPS induces a complex union of the box AP-1, it will consist of a heterodímero c-Jun/c-Fos in dendritic cells, while in B lymphocytes, the complex will be a homodímero of c-Jun. These results are confirmed by inmunoprecipitación chromatin and the various inductions mRNA of c-Jun and c-Fos. From being described as a new mechanism for controlling gene expression of MHC class II by the LPS. THE M-CSF AND LPS REGULATING THE EXPRESSION OF MKP-1 IN MACRÓFAGOS ROUTE ONE BOX CRE/AP-1 The macrophages proliferate in response to growth factors such as MCSF, GM-CSF or IL-3. But when you add factors activators such as LPS or IFN.gamma, macrophages stop proliferate. Despite these two different answers, M-CSF and LPS induces the phosphorylation and activation of ERK-1 / 2 (External regulated kinase), but with different kinetics. So the activation of ERK-1 / 2 by M-CSF occurs at the 5 minute and induces proliferation, instead LPS active EERK-1 / 2 the 15 minutes activating macrophages. Both stimuli induce the expression of the phosphatase MKP-1, regardless of the activation of ERK, which desfoforila and thus inactivates the kinase ERK-1 / 2. In this paper we study the functional areas of the promoter of MKP-1 by transfection experiments with a reporter gene. Performing Gross deletions promoter's determined that the only area of the proximal promoter located between -380 and -180 bp was inducible by M-CSF and LPS. In addition, mutagenesis experiments delas boxes proximal promoter determined that the box CRE/AP-1 is necessary for induction of MKP-1, both M-CSF indicate that a heterodímero formed by c-Jun and CREB would join the box CRE/AP-1. These results indicate that the M-CSF and regulate LPS induction MKP-1 through a box AP-1.

Author: MORENO ATANASIO CAROLINA.
Year: 2006.
University: BARCELONA [www.ub.es].
Place of defense: FACULTAD DE MEDICINA.
Place of preparation: FACULTAD DE MEDICINA.

Summary: The chronic lymphocytic leukemia (CLL), leukemia more common in Western countries, is a lymphoproliferative syndrome which is characterized by the progressive accumulation of mature lymphocytes ilk By CD5 + in lymphoid tissues, bone marrow and peripheral blood. The course of the disease is highly variable. Mutations in the genes of the IgVH distinguish two different forms of the disease with respect to prognosis, so that patients with forms "not mutadas" of the disease have poorer prognosis than those with the variety "mutated". This is independent of the type of treatment. At present, treatment with the purine analogues in combination with other agents and the transplantation of hematopoietic progenitors, they achieve a 60-80% RC. Moreover, many of these RC are accompanied by the eradication of ERM. Therefore, the study of the ERM in the LLC, its clinical significance and therapeutic implications is gaining considerable interest. However, despite some advances in the treatment of CLL, the disease remains incurable. This fact, together with the increasingly frequent diagnosis of this form of leukemia in patients still young, and the difficulties involved in the treatment of patients who fall or are resistant to existing treatments, makes them increasingly sick in which raises a transplant of hematopoietic progenitors. Doctoral Thesis This is based on research work that focus on the role of haematopoietic progenitor cell transplantation in the treatment of patients with LLC. The working hypotheses were as follows: 1 - The allogeneic haematopoietic progenitor cell in patients with LLC could eliminate negative prognostic significance of certain parameters, particularly the absence of mutations in genes lso the IgVH in these patients. 2-An analysis of the ERM LLC in patients undergoing transplantation of hematopoietic progenitors could provide interesting information both from the side as predicted with regard to mechanisms of action of various types of transplantation. FINDINGS AND CONCLUSIONS * The risk of clinical relapse was significantly higher in patients receiving an autologous transplant than in those undergoing an allogeneic (61% and 12% at 5 years, respectively, p less 0.05). The event-free survival and overall survival were significantly higher in alotrasplante. In Group LLC "not mutated, the risk of disease progression was significantly lower in those undergoing allogeneic. In conclusion, a transplanted allogeneic hematopoietic progenitors can eliminate the adverse effect that the absence of mutations VH carries in patients with lso LLC, which is very likely to end graft versus leukemia. * Detection of ERM correlates with time to progression in patients undergoing autologous transplantation. Similarly, the detection of MRA to 3-6 months of the transplant can identify those patients who will progress at an early stage in less than 2 years after the procedure. ERM levels are correlated with time to progression and survival. Therefore, quantitative methods (quantitative PCR and flow cytometry) are more accurate than qualitative methods (PCR consensus) to predict the evolution of the patients. In alotrasplante, the presence of MRA does not necessarily follow clinical relapse.