Epidemiologische Daten belegen eine Assoziation von Infektionen mit der Exazerbation von Autoimmunerkrankungen, wobei man aber die Wege, die dahin führen noch nicht voll versteht. Die gängigste Hypothese sieht die Ursache für die Entstehung von Autoimmunerkrankungen in der Kreuzreaktivität zwischen mikrobiellen und Selbst-Antigenen, die von der selben T-Zelle erkannt werden. Dieser Antigen-spezifische Mechanismus konnte allerdings bisher nicht durch die Forschung belegt werden. In dieser Arbeit wird gezeigt, dass im Modell der experimentellen autoimmunen Enzephalitis durch die Aktivierung mit Lipopolysacchariden gramnegativer Bakterien (LPS) Schübe in normalen Mäusen ausgelöst werden können. Diese Art der Behandlung führt in-vitro zur Proliferation und zur Zytokinproduktion bei einem Teil der T-Helfer (Th)-Effektor/Gedächtniszellen. Dabei ist zwar der physische Kontakt zwischen Th-Zellen und CD4--LPS-responsiven Zellen essentiell, jedoch geschieht die Aktivierung nicht über den T-Zellrezeptor. Als entscheidend hat sich die Bindung von kostimulatorischen Rezeptoren auf Th-Zellen durch kostimulatorische Moleküle auf CD4--Zellen erwiesen. Diese Form der Bystander-Aktivierung bietet eine Antigen-unabhängige Erklärung für die Zusammenhänge von Infektion und Autoimmunität, die mit den klinischen und epidemiologischen Daten besser vereinbar ist als die Antigen-spezifischen Modelle. / Infections sometimes associate with exacerbations of autoimmune diseases through pathways that are poorly understood. Antigen-specific mechanisms such as cross-reactivity between a microbial antigen and a self-antigen have received no direct support. Here it is shown that activation by lipopolysaccharide (LPS) induces relapses of experimental autoimmune encephalitis (EAE) in normal mice. This form of treatment induces proliferation and cytokine production in a fraction of effector/memory T helper (Th) lyphocytes in vitro via physical contact of Th cells with CD4--LPS-responsive cells. TCR mediated signals are not necessary; rather what is required is ligation of costimulatory receptors on Th cells by costimulatory molecules on the CD4- cells. This form of bystander activation provides an antigen-independent link between infection and autoimmunity that might fit the clinical and epidemiological data on the connection between infection and autoimmunity better than the antigen-specific models.

In der Arbeit wurde die Rolle von Bakterien für das Entstehen von Autoimmunität untersucht. Insbesondere wurde untersucht, inwieweit Bakterien entweder spezifisch (über "Kreuzreaktivität") oder antigenunabhängig (über "Bystander-Aktivierung") eine Aktivierung von autoreaktiven CD4+- T-Zellen induzieren können. Es konnte gezeigt werden, dass es bei dem untersuchten, MBP-spezifischen T-Zellrezeptor multiple, natürlich vorkommende, kreuzreaktive Peptide mikrobiellen Ursprungs gibt, die eine Aktivierung der T-Zellen hervorrufen und in vivo experimentelle autoimmune Enzephalomyelitis (EAE) induzieren können. Weiterhin wurde untersucht, inwieweit Lipopolysaccharid (LPS) als unspezifischer Aktivator des Immunsystems eine Aktivierung der autoreaktiven T-Zellen in vitro hervorrufen kann und inwieweit in vivo EAE durch LPS hervorgerufen werden kann. Es wurde gezeigt, dass LPS in vitro einen kleinen Anteil der CD4+ - T-Zellen aktiviert. Wurden den transgenen T+alpha- -Mäusen LPS appliziert, erkrankten diese an EAE. Somit gibt es sowohl in vitro als auch in vivo in den T+alpha- -Mäusen Hinweise für eine Relevanz von "Bystander-Aktivierung". Abschließend wurde diskutiert, inwieweit entweder "Kreuzreaktivität" oder "Bystander-Aktivierung" als Auslöser für Autoimmunität unter physiologischen Bedingungen in Frage kommt. Aufgrund der in dieser Arbeit gezeigten Ergebnisse wurde postuliert, dass keine der beiden Mechanismen alleiniger Auslöser sei, da es aufgrund der Häufigkeit von Infektionen, kreuzreaktiven Peptiden und des Vorkommens von autoreaktiven T-Zellen auch in gesunden Individuen ansonsten sehr viel häufiger zu Autoimmunität kommen müsste. Unter bestimmten Bedingungen könnte die Aktivierung von T-Zellen über Kreuzreaktivität oder über "Bystander-Aktivierung" Autoimmunität auslösen oder verstärken, wenn bereits andere Mechanismen des Immubnsystems, die Autoimmunität verhindern, versagt haben. / In this thesis the role of bacteria for the induction of autoimmunity was investigated. In detail, it was examined whether bacteria are able to activate autoreactive CD4+-T-cells antigen-specific ("cross-reactivity") or antigen-unspecific ("bystander-activation"). It was shown that the examined transgenic MBP-peptide specific T-cell-receptor recognized many natural occurring cross-reactive peptides of microbial origin, which induced an activation of the T-cells in vitro and which could induce autoimmune encephalomyelitis (EAE) in the T-cell-receptor transgenic mice in vivo. Furthermore, it was examined, whether lipopolysaccharide (LPS) as activator of the innate immune system could induce an unspecific activation of the autoreactive T-cells in vitro and whether administration of LPS in the transgenic mice could induce EAE in vivo. It was shown that LPS activates a small percentage of CD4+ - T-cells. Application of LPS to the transgenic T+alpha- mice induced EAE. Therefore, the role of bystander-activation was indicated in vitro and in vivo. Finally, it was discussed, whether either cross-reactivity or bystander-activation could be sufficient for inducing autoimmunity under physiologic conditions. Due to the results presented in this work, it is postulated that none of the both mechanisms could be inductor of autoimmunity alone. If one of these mechanisms was sufficient, autoimmunity in humans should be a frequent event, because infections and autoreactive T cells are both findings which occur in healthy humans very often. However, under certain conditions either cross-reactivity or bystander-activation could trigger or exacerbate autoimmunity, when other mechanisms which inhibit autoimmunity have failed.

Les Formes réactives de l’oxygène (FRO) ont un rôle bien établi dans l’oncogénèse en augmentant les capacités de prolifération et d’invasion des cellules tumorales. Mais les FRO exercent aussi un effet important sur le microenvironnement tumoral, en particulier en participant à l’échappement des tumeurs au système immunitaire. Une modulation pharmacologique de l’équilibre oxydo-réductif tumoral est donc susceptible d’influencer la progression tumorale. Il a été montré que l’induction pharmacologique d’un stress oxydant dans les cellules tumorales peut induire un effet cytotoxique mais ses effets sur le microenvironnement sont moins bien connus. L’objectif de nos travaux était d’étudier l’effet d’une modulation du stress oxydant sur les cellules immunitaires du microenvironnement tumoral et in fine de préciser les conséquences potentielles sur la progression tumorale. L’arsenic trioxyde (As2O3), inducteur de FRO, présentait à faible dose, dénuée d’effet cytotoxique direct sur les cellules malignes, un effet antitumoral dans un modèle de cancer colique murin. Cet effet était lié à une déplétion sélective en lymphocytes T régulateurs (Tregs) et était médié par la génération d’anions superoxyde (O2°-) et de monoxyde d’azote (NO), eux même responsables de la formation de peroxynitrite. Les Tregs présentaient un niveau basal de FRO plus élevé que les autres populations lymphocytaires, qui pourrait expliquer leur plus grande sensibilité à un surcroît de stress oxydant induit par l’As2O3. Un cytotoxique antitumoral, la vinorelbine, s’est également montré capable d’exercer un effet sur le microenvironnement tumoral. Par co-cultures, nous avons montré que la vinorelbine induisait un effet bystander toxique sur les cellules immunitaires effectrices voisines des cellules tumorales. In vivo, le prétraitement par vinorelbine de cellules malignes implantées à la souris était responsable d’une perte de la réactivité anti-tumorale des cellules mono-nuclées. Cet effet était dépendant de la production d’O2°- et de NO par les cellules malignes. Un modulateur du stress oxydant, le mangafodipir, inhibait cet effet, permettant ainsi de restaurer la réponse immunitaire antitumorale locale. Notre travail a donc permis de mettre en évidence que des modulateurs du stress oxydant peuvent agir sur le microenvironnement, et spécialement sur les cellules immunitaires. Ils pourraient être utilisés en clinique pour restaurer la réponse immunitaire antitumorale. Une meilleure compréhension du rôle du stress oxydant dans la défaillance de l’immunité antitumorale est nécessaire. / Several reports have demonstrated the involvement of reactive oxygen species (ROS) in carcinogenesis, through promotion of cancer cell proliferation and invasion. But ROS could also have consequences on non cancerous cells which are part of the tumor microenvironment, such as immune cells. Therefore, a pharmacological modulation of oxidative stress can induce a cytotoxic effect on tumor cells but its consequences on microenvironment are unknown. The aim of our studies was to evaluate the effects of a pharmacological modulation of oxidative stress on immune cells from the tumor microenvironment. At low dose, Arsenic trioxide (As203), an oxidative stress modulator, was shown to exert antitumor effects in colon tumor-bearing mice. We observed that this effect was related to As203-induced regulatory T cells (Tregs) -selective depletion in vitro and in vivo and was mediated by oxidative and nitrosative bursts. The differential effect of As203 on Tregs versus other CD4 cells was related to difference in the cells’redox status. We also observed that vinorelbine, an anticancer agent, could interfere with the antitumor immune response. We showed that vinorelbine could alter the function of immune cells surrounding tumor cells by a bystander toxic effect against tumor effector cells. In vivo experiment in A549 tumor bearing nude mice showed that adoptive transfer of A549 immune splenocytes was not able to delay tumor growth when vinorelbine-pretreated A549 cells were used for immunization. This effect was mediated by ROS and was inhibited by an oxidative stress modulator, mangafodipir, which restored antitumor immune function. Therefore, our work showed that oxidative stress modulators can influence tumor microenvironment and more specifically, immune cells. They could be used to restore antitumor immune response.

No description available.

The aim of the present thesis was to listen to, examine and conceptualise students’ perspectives on bullying. Students’ perspectives have not been commonly heard in research and less qualitative research has been conducted. This study contributes with students’ perspectives on bullying using semi-structured interviews with students from fourth-to eighth grade. This thesis includes four studies. The aim with paper I was to investigate how bystander actions in bullying situations and reasons behind these actions were articulated. Paper II was a comparison study between Sweden and US, focused on how students articulate and discuss what factors influence students’ decisions to defend or not defend victims when witnessing bullying. The aim in Paper III was to study how students themselves discuss, reason and make sense of how and why bullying processes emerges in their social worlds. In paper IV the aim was to study how junior high school girls discuss and understand bullying. Findings reveal that students’ reactions as bystanders to bullying depend on how they define the situation. Explanations to the emergence of bullying were understood through a complex social ordering of belonging process. Students position themselves and others in striving to belong, and when defining victims as responsible for bullying. Social norms and negotiation of identities were also discussed among the students. Students discussed how gender and a normative peer structure, where a pressure to fit in, interlinked with how they understood bullying. / Syftet med denna avhandling är att lyssna på, utforska och konceptualisera elevers perspektiv på mobbning då elevers perspektiv utgör ett viktigt bidrag till vår fortsatta förståelse av mobbning. Det finns fortfarande få kvalitativa studier som fokuserat elevers perspektiv på mobbning. Denna studie utgör därför ett viktig bidrag och baseras på semi-strukturerade intervjuer med elever ifrån fjärde till åttondeklass. Avhandlingen består utav fyra studier. Syftet med studie I var att undersöka hur åskådarageranden i mobbningssituationer artiklueras av eleverna och skälen bakom dessa. Syftet med studie II var att jämföra hur elever i Sverige och USA artikulerar och diskuterar vilka faktorer som influerar deras beslut att ingripa eller inte ingripa när de blir åskådare till mobbning. Syftet med studie III var att undersöka hur elever diskuterar, resonerar och förstår vad som producerar mobbning. Syftet med studie IV var att undersöka hur högstadietjejer diskuterar och förstår mobbning. Resultaten visar att elevers reaktioner som åskådare till mobbning är situationsbundna utifån hur de definierar situationen. Förklararingar till vad som producerar mobbning sammanlänkas med en komplex socialt ordnande process där eleverna positionerar sig själv och andra i strävan att tillhöra och den utsatta ofta görs ansvarig för mobbningen. Även sociala normer och förhandlade av identiteter kommer till uttryck när eleverna diskuterar hur kön och normativa kamratnormer, där strävan att passa in, sammanlänkas med deras förståelse av mobbning.

O melanoma representa menos de 5% de todos os cânceres de pele, porém, quando em estádio metastático possui prognóstico ruim. Entretanto, o genótipo dos melanomas pode prover uma oportunidade para intervenção terapêutica pelo fato de 90% dos casos de melanoma possuem p53 selvagem e grande parte destes possuem deleção na região cromossômica codificadora de interferon beta. Em prévios estudos, desenvolvemos o vetor adenoviral AdRGD-PG que fornece expressão do transgene em resposta à p53 através do promotor PG e ainda o tripeptídeo RGD, que possibilita que o adenovírus transduza uma maior gama de células pela alteração de seu mecanismo de entrada. Temos utilizado este vetor para entrega da versão murina de interferon beta em modelos de terapia gênica e imunoterapia de melanoma murino, revelando uma significativa habilidade do interferon beta em inibir a proliferação celular in vitro e in vivo e promover resposta imune antitumoral. No presente trabalho, os esforços se aplicam em adaptar essa estratégia em modelo de melanoma humano para observar se a mesma interação é encontrada. O vetor AdRGD-PGhIbeta, portador do cDNA de interferon beta humano (hIbeta) foi construído e expressão do transgene observada após transdução das linhagens estabelecidas de melanoma humano SK-MEL-05 e SK-MEL-147 (ambas p53 selvagem). Foi observado um robusto efeito antitumoral in vitro onde transferência de hIbeta promoveu acumulo de células hipodiploides (mais que 80% da população celular 96 horas após transdução) e evidências de morte por apoptose (exposição de fosfatidilserina e atividade de caspases 3/7) em ambas as linhagens. Nas duas linhagens, o efeito bystander foi demonstrado quando a presença de poucas células transduzidas (ex., 10%) foi suficiente para promover o acumulo significativo de células hipodiploides (mais que 40% neste exemplo). Em modelo de terapia gênica in situ utilizando células SK-MEL-147, também foi observado forte efeito antitumoral da hIbeta com total remissão do tumor de todos os animais tratados sem recidiva durante noventa dias. A presença de hIbeta na circulação dos animais foi confirmada 48h após o tratamento com AdRGD-PG hbeta mas presente em somente dois de sete animais 90 dias após o tratamento, sugerindo que o tratamento inicial e não um efeito off target foi responsável pela resposta. Com a finalidade de investigar efeitos colaterais do sequestro do vetor adenoviral pelo fígado, observamos a concentração circulante das enzimas aminotransferase de aspartate e aminotransferase de alanine (AST e ALT, respectivamente), que se mostrou não alterada quando comparadas entre animais que receberam injeção do vetor tratamento, vetor controle e solução salina. Com nossos resultados concluímos que vetores adenovirais carreando interferon-beta humano são capazes de transduzir a linhagem de melanoma SK-MEL-147 in vitro e in vivo, promovendo efeito bystander e remissão tumoral sem indução de efeitos adversos / Melanoma represents less than 5% of all cases of skin cancer, although, when metastatic, prognosis is dire. However, the genotype of melanomas might provide an opportunity for therapeutic intervention since 90% of melanoma cases possess wild-type p53 and a great portion of these possess deletion of the chromosomal region encoding interferon beta. In previous studies, we developed the adenoviral vector AdRGD-PG that supplies expression of the transgene in response to p53 through the PG promoter and that utilizes the RGD tripeptide, allowing the adenovirus to transduce a wider range of cells due to the alterated mechanism of entrance. We have used this vector to deliver the murine version of interferon beta in murine models of melanoma gene therapy and immunotherapy, revealing a significant ability of interferon beta to inhibit cellular proliferation in vitro and in vivo and promote an anti-tumor immune response. In the present project, we aimed to adapt this strategy for a human melanoma model in order to reveal if the same impact will be observed. The AdRGD-PGhIbeta vector encoding the human interferon beta (hIbeta) cDNA was constructed and expression of the transgene confirmed after transduction of the established human melanoma cell lines SK-MEL-05 and SK-MEL-147 (both wild-type p53). A striking anti-tumor effect was observed in vitro where the transfer of hIbeta promoted an accumulation of hypodiploid cells (over 80% of the cellular population 96 hours after transduction) and evidence of death by apoptosis (exposure of phosphatidylserine and activity of caspases 3/7) in both cell lines. In these cell lines, a bystander effect was demonstrated when the presence of few transduced cells (ex., 10%) was enough to promote significant accumulation of hypodiploid cells (over 40% in this example). In a model of in situ gene therapy using SK-MEL-147 cells, hIbeta induced a strong anti-tumor effect including total tumor remission in all treated animals without relapse during ninety days. The presence of hIbeta in the circulation of the animals was confirmed 48h after treatment with AdRGD-PGhIbeta, but was present in only two of the seven animals 90 days post-treatment, suggesting that the initial treatment, not off target effects, was responsible for the response. With the goal of investigating collateral effects of adenoviral sequestration by the liver, we assayed the circulating concentration of aspartate aminotransferase and alanine aminotransferase (AST and ALT, respectively), which showed no alteration when compared with animals that received the treatment with a control vector or saline solution. We conclude that our adenoviral vector carrying human interferon-beta is capable of transducing the human melanoma cell line SK-MEL-147 in vitro and in vivo, promoting a bystander effect and tumor remission without inducing adverse effects

L'effet de voisinage radio-induit s'observe dans les cellules voisines de cellules irradiées mais non directement touchées par l'irradiation. A ce jour, les espèces réactives de l'oxygène (EROs) sont considérées comme ayant un rôle actif dans la survenue de réponse au voisinage, mais leur implication n'est pas encore totalement définie. Afin de déterminer leur impact dans la réponse au voisinage, à la fois temporellement et spatialement, des irradiations par microfaisceau de particules sont mises au point afin de cibler une fraction définie de cellules au niveau du noyau dans une culture cellulaire. Les irradiations sont pratiquées sur des cellules normales ostéoblastiques, à sous-confluence, nommées MC3T3-E1. L'observation directe de la génération d'EROs cellulaires et mitochondriales révèle que la réponse bystander est caractérisée par un stress cellulaire d'origine double et temporellement distinct. A court terme, la signalisation issue de la membrane induit une importante production d'EROs impliquée dans l'apparition de cassures double brin de l'ADN retardées. Les mitochondries produisant des EROs jusque 6 heures après l'irradiation semblent être impliquées dans un processus à long terme différent. L'étude indépendante de la réponse des cellules ciblées et voisines met en évidence un phénomène nouveau. L'effet de voisinage induit une réponse des cellules voisines mais est aussi impliqué dans une amplification de la réponse des cellules ciblées par l'irradiation. Ainsi toute cellule, irradiée ou non, peut recevoir ces signaux bystander et répondre. L'ensemble de cette étude mène à considérer les cultures cellulaires comme des réseaux complexes de communication; leurs réponses à l'irradiation indiquent une relation mêlée entre signaux relatifs à l'irradiation elle-même et réponse au voisinage. Cette complexité du phénomène d'effet de voisinage peut justifier que son incidence sur l'établissement de règles de radioprotection et sur la courbe linéaire sans seuil n'est pas encore clairement déterminée.

No description available.

abstract: Sexual harassment has emerged as a widespread problem facing women in public space in Egypt. Activism to combat sexual harassment began in 2005. However, just prior to and in the years following the January 25, 2011 Egyptian Revolution, which witnessed an increase in the collective sexual harassment, assault and rape of women, this activism has increased. Subsequently, scholarly attention to sexual harassment and public sexual violence has also expanded. Much of the attention in scholarly analyses has been directed toward politically motivated sexual violence, focused on understanding the state commissioning of sexual violence against female protestors to drive them from protest participation. There is an emerging critique of activist approaches that seems to ignore the politicalized nature of sexual harassment to focus instead on “cultural” targets. The early work of the Egyptian Center for Women’s Rights (ECWR) and current work of HarassMap have been criticized for depoliticizing sexual harassment by failing to include an analysis of state-commissioned sexual violence in their work. Similarly, both have been accused of expanding the scope of the security state by calling for increased policing of public space to protect women from “culturally-bad” men. With data collected through one year of participant observation with HarassMap, interviews with activists from eleven anti-sexual harassment initiatives and advocacy NGOs, and community-level surveys with non-activist individuals, this dissertation argues that “cultural” work undertaken through the community-based approaches by entities like ECWR and HarassMap is, in fact, an inherently political process, in which political engagement represents both an attempt to change political culture and state practice and a negotiative process involving changing patriarchal gender norms that underpin sexual harassment at a society-wide level. New conceptualizations of sexual harassment promoted by anti-sexual harassment initiatives and NGOs in Egypt frame it as a form of violence against women, and attempt to make sexual harassment an offense that may be criminalized. Yet, this dissertation contends there is a tension between activist and widespread public understandings of sexual harassment, predicated on the incomplete framing of sexual harassment as a form of violence. / Dissertation/Thesis / Doctoral Dissertation Anthropology 2016

O melanoma representa menos de 5% de todos os cânceres de pele, porém, quando em estádio metastático possui prognóstico ruim. Entretanto, o genótipo dos melanomas pode prover uma oportunidade para intervenção terapêutica pelo fato de 90% dos casos de melanoma possuem p53 selvagem e grande parte destes possuem deleção na região cromossômica codificadora de interferon beta. Em prévios estudos, desenvolvemos o vetor adenoviral AdRGD-PG que fornece expressão do transgene em resposta à p53 através do promotor PG e ainda o tripeptídeo RGD, que possibilita que o adenovírus transduza uma maior gama de células pela alteração de seu mecanismo de entrada. Temos utilizado este vetor para entrega da versão murina de interferon beta em modelos de terapia gênica e imunoterapia de melanoma murino, revelando uma significativa habilidade do interferon beta em inibir a proliferação celular in vitro e in vivo e promover resposta imune antitumoral. No presente trabalho, os esforços se aplicam em adaptar essa estratégia em modelo de melanoma humano para observar se a mesma interação é encontrada. O vetor AdRGD-PGhIbeta, portador do cDNA de interferon beta humano (hIbeta) foi construído e expressão do transgene observada após transdução das linhagens estabelecidas de melanoma humano SK-MEL-05 e SK-MEL-147 (ambas p53 selvagem). Foi observado um robusto efeito antitumoral in vitro onde transferência de hIbeta promoveu acumulo de células hipodiploides (mais que 80% da população celular 96 horas após transdução) e evidências de morte por apoptose (exposição de fosfatidilserina e atividade de caspases 3/7) em ambas as linhagens. Nas duas linhagens, o efeito bystander foi demonstrado quando a presença de poucas células transduzidas (ex., 10%) foi suficiente para promover o acumulo significativo de células hipodiploides (mais que 40% neste exemplo). Em modelo de terapia gênica in situ utilizando células SK-MEL-147, também foi observado forte efeito antitumoral da hIbeta com total remissão do tumor de todos os animais tratados sem recidiva durante noventa dias. A presença de hIbeta na circulação dos animais foi confirmada 48h após o tratamento com AdRGD-PG hbeta mas presente em somente dois de sete animais 90 dias após o tratamento, sugerindo que o tratamento inicial e não um efeito off target foi responsável pela resposta. Com a finalidade de investigar efeitos colaterais do sequestro do vetor adenoviral pelo fígado, observamos a concentração circulante das enzimas aminotransferase de aspartate e aminotransferase de alanine (AST e ALT, respectivamente), que se mostrou não alterada quando comparadas entre animais que receberam injeção do vetor tratamento, vetor controle e solução salina. Com nossos resultados concluímos que vetores adenovirais carreando interferon-beta humano são capazes de transduzir a linhagem de melanoma SK-MEL-147 in vitro e in vivo, promovendo efeito bystander e remissão tumoral sem indução de efeitos adversos / Melanoma represents less than 5% of all cases of skin cancer, although, when metastatic, prognosis is dire. However, the genotype of melanomas might provide an opportunity for therapeutic intervention since 90% of melanoma cases possess wild-type p53 and a great portion of these possess deletion of the chromosomal region encoding interferon beta. In previous studies, we developed the adenoviral vector AdRGD-PG that supplies expression of the transgene in response to p53 through the PG promoter and that utilizes the RGD tripeptide, allowing the adenovirus to transduce a wider range of cells due to the alterated mechanism of entrance. We have used this vector to deliver the murine version of interferon beta in murine models of melanoma gene therapy and immunotherapy, revealing a significant ability of interferon beta to inhibit cellular proliferation in vitro and in vivo and promote an anti-tumor immune response. In the present project, we aimed to adapt this strategy for a human melanoma model in order to reveal if the same impact will be observed. The AdRGD-PGhIbeta vector encoding the human interferon beta (hIbeta) cDNA was constructed and expression of the transgene confirmed after transduction of the established human melanoma cell lines SK-MEL-05 and SK-MEL-147 (both wild-type p53). A striking anti-tumor effect was observed in vitro where the transfer of hIbeta promoted an accumulation of hypodiploid cells (over 80% of the cellular population 96 hours after transduction) and evidence of death by apoptosis (exposure of phosphatidylserine and activity of caspases 3/7) in both cell lines. In these cell lines, a bystander effect was demonstrated when the presence of few transduced cells (ex., 10%) was enough to promote significant accumulation of hypodiploid cells (over 40% in this example). In a model of in situ gene therapy using SK-MEL-147 cells, hIbeta induced a strong anti-tumor effect including total tumor remission in all treated animals without relapse during ninety days. The presence of hIbeta in the circulation of the animals was confirmed 48h after treatment with AdRGD-PGhIbeta, but was present in only two of the seven animals 90 days post-treatment, suggesting that the initial treatment, not off target effects, was responsible for the response. With the goal of investigating collateral effects of adenoviral sequestration by the liver, we assayed the circulating concentration of aspartate aminotransferase and alanine aminotransferase (AST and ALT, respectively), which showed no alteration when compared with animals that received the treatment with a control vector or saline solution. We conclude that our adenoviral vector carrying human interferon-beta is capable of transducing the human melanoma cell line SK-MEL-147 in vitro and in vivo, promoting a bystander effect and tumor remission without inducing adverse effects