RESUMEN
Malaria is one of the major parasitic killer diseases worldwide. Severe cases of malaria are mostly in children under the age of 5 years due to their naïve immune system and in pregnant women with weakened immune responses. Inflammatory immune responses against the parasite involve complement activation as well as the antibody and effector cell-mediated immune system. However, after an infection with Plasmodium falciparum (P. falciparum), the most dangerous malaria species, the host-derived immunity is often insufficient to completely inhibit the infection cycles of the parasite in red blood cells for yet unknown reasons. In the present chapter we aim to elucidate the role of the host's and the parasite's heat shock proteins (HSPs) in the development of a novel anti-malaria therapeutic approach.
Asunto(s)
Malaria Falciparum , Malaria , Preescolar , Eritrocitos , Femenino , Proteínas de Choque Térmico/genética , Humanos , Inmunidad , Malaria Falciparum/tratamiento farmacológico , Plasmodium falciparum , Embarazo , Proteínas Protozoarias/genéticaRESUMEN
A balanced proinflammatory cytokine response to Plasmodium ssp. infection is crucial to control the disease outcome. To elucidate the effect of cytokines and Plasmodium falciparum-infected erythrocytes on the regulation of interleukin (IL)-6 receptor (IL-6R), ciliary neurotrophic factor alpha (CNTFR-α) and glycoprotein (gp)130 in natural killer (NK) cells in the context of malaria, we assessed their gene expression and surface expression in NK92 cells. P. falciparum alone did not alter gene expression of the investigated receptors in NK92 cells. Analysis revealed a low effect of IL-6 on IL-6R surface expression in NK92 cells. However, at transcriptional level, a downregulation of IL-6R was observed following IL-6 stimulation. Thus, IL-6 might act within a negative feedback loop to terminate signal transduction by downregulating IL-6R expression. Additionally, we observed that IL-6R and CNTFR-α surface expression were regulated by a combination of IL-2, 12, and 18, and gp130 was influenced by interferon-α. Our results show that the IL-6 family receptors in NK92 cells are not directly influenced by P. falciparum. However, cytokines usually derived from accessory cells during malaria episodes may regulate IL-6 receptor signaling pathways. This finding encourages future studies in a more physiological context and with primary cells isolated from humans with and without malaria.
Asunto(s)
Receptor gp130 de Citocinas/metabolismo , Citocinas/farmacología , Eritrocitos/parasitología , Células Asesinas Naturales/metabolismo , Plasmodium falciparum/fisiología , Receptor de Factor Neurotrófico Ciliar/metabolismo , Membrana Celular/efectos de los fármacos , Membrana Celular/metabolismo , Receptor gp130 de Citocinas/genética , Eritrocitos/efectos de los fármacos , Eritrocitos/metabolismo , Humanos , Interferón-alfa/farmacología , Células Asesinas Naturales/efectos de los fármacos , Malaria Falciparum/inmunología , Malaria Falciparum/parasitología , Plasmodium falciparum/efectos de los fármacos , Unión Proteica/efectos de los fármacos , Receptor de Factor Neurotrófico Ciliar/genética , Transcripción Genética/efectos de los fármacos , Regulación hacia Arriba/efectos de los fármacosRESUMEN
In the early immune response to Plasmodium falciparum-infected erythrocytes (iRBC), Natural Killer (NK) cells are activated, which suggests an important role in innate anti-parasitic immunity. However, it is not well understood whether NK cells directly recognize iRBC or whether stimulation of NK cells depends mainly on activating signals from accessory cells through cell-to-cell contact or soluble factors. In the present study, we investigated the influence of membrane-bound host Heat shock protein (Hsp) 70 in triggering cytotoxicity of NK cells from malaria-naïve donors or the cell line NK92 against iRBC. Hsp70 and HLA-E membrane expression on iRBC and potential activatory NK cell receptors (NKG2C, CD94) were assessed by flow cytometry and immunoblot. Upon contact with iRBC, Granzyme B (GzmB) production and release was initiated by unstimulated and Hsp70-peptide (TKD) pre-stimulated NK cells, as determined by Western blot, RT-PCR and ELISPOT analysis. Eryptosis of iRBC was determined by Annexin V-staining. Our results suggest that presence of Hsp70 and absence of HLA-E on the membrane of iRBC prompt the infected host cells to become targets for NK cell-mediated cytotoxicity, as evidenced by impaired parasite development. Contact of iRBC with NK cells induced release of GzmB. We propose that following GzmB uptake, iRBC undergo eryptosis via a perforin-independent, GzmB-mediated mechanism. Since NK activity toward iRBC could be specifically enhanced by TKD peptide and abrogated to baseline levels by blocking Hsp70 exposure, we propose TKD as an innovative immunostimulatory agent to be tested as an adjunct to anti-parasitic treatments in vivo.
Asunto(s)
Eritrocitos/inmunología , Eritrocitos/parasitología , Granzimas/biosíntesis , Proteínas HSP70 de Choque Térmico/sangre , Células Asesinas Naturales/inmunología , Células Asesinas Naturales/metabolismo , Plasmodium falciparum/inmunología , Plasmodium falciparum/patogenicidad , Apoptosis/inmunología , Comunicación Celular/inmunología , Línea Celular , Técnicas de Cocultivo , Envejecimiento Eritrocítico/inmunología , Eritrocitos/metabolismo , Granzimas/genética , Proteínas HSP70 de Choque Térmico/inmunología , Células HeLa , Interacciones Huésped-Parásitos/inmunología , Interacciones Huésped-Parásitos/fisiología , Humanos , Inmunidad Innata , Modelos Inmunológicos , Subfamília C de Receptores Similares a Lectina de Células NK/metabolismo , Subfamília D de Receptores Similares a Lectina de las Células NK/metabolismo , Plasmodium falciparum/crecimiento & desarrollo , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
BACKGROUND: Mechanisms by which anti-malarial immune responses occur are still not fully clear. Natural killer (NK) cells are thought to play a pivotal role in innate responses against Plasmodium falciparum. In this study, the suitability of NK92 cells as models for the NK mechanisms involved in the immune response against malaria was investigated. METHODS: NK92 cells were assessed for several signs of activation and cytotoxicity due to contact to parasites and were as well examined by oligonucleotide microarrays for an insight on the impact P. falciparum-infected erythrocytes have on their transcriptome. To address the parasite side of such interaction, growth inhibition assays were performed including non-NK cells as controls. RESULTS: By performing microarrays with NK92 cells, the impact of parasites on a transcriptional level was observed. The findings show that, although not evidently activated by iRBCs, NK92 cells show transcriptional signs of priming and proliferation. In addition, decreased parasitaemia was observed due to co-incubation with NK92 cells. However, such effect might not be NK-specific since irrelevant cells also affected parasite growth in vitro. CONCLUSIONS: Although NK92 cells are here shown to behave as poor models for the NK immune response against parasites, the results obtained in this study may be of use for future investigations regarding host-parasites interactions in malaria.
Asunto(s)
Eritrocitos/inmunología , Eritrocitos/parasitología , Interacciones Huésped-Parásitos , Células Asesinas Naturales/inmunología , Plasmodium falciparum/inmunología , Plasmodium falciparum/patogenicidad , Perfilación de la Expresión Génica , Humanos , Análisis por MicromatricesRESUMEN
In this group we would like to answer the question why people show a different response against certain pathogens. In many infections the course of the disease can range from asymptomatic carriage to the severest forms even death. In the past we have analysed candidate genes and their role in the course of malaria and could detect some polymorphisms influencing infectious diseases in the genes encoding NOS2, MBL2, IFNa, FCN2, and receptors for IFNg and IFNa. Having worked initially mainly on malaria we broadened our spectrum also to other infectious diseases like hepatitis B, Leprosy, schistosomiasis. Here we give a short overview about ongoing projects.