RESUMO
BACKGROUND: Corynebacterium mucifaciens has been mainly isolated from skin, blood and from other normally-sterile body fluids. It has rarely been described as a human pathogen since its description. CASE PRESENTATION: We herein report the first case of cavitary pneumonia due to C. mucifaciens in an immunocompetent man returning from Maghreb. CONCLUSION: C. mucifaciens should be considered as important human pathogen in patients with severe illness and compatible history of exposure even in individuals with no clearly identified immunosuppression.
Assuntos
Infecções por Corynebacterium/diagnóstico , Corynebacterium/isolamento & purificação , Pneumonia Bacteriana/diagnóstico , África do Norte , Humanos , Masculino , Pessoa de Meia-Idade , Pneumonia Bacteriana/microbiologiaRESUMO
Engagement of surface receptors contributes to the antimicrobial activity of human immune cells. We show here that infection of human monocyte-derived macrophages (MDM) with live Mycobacterium avium induced the expression of CD23 on their membrane. Subsequent cross-linking of surface CD23 by appropriate ligands induced a dose-dependent antibacterial activity of MDM and the elimination of most infected cells. The stimulation of inducible nitric oxide synthase-dependent generation of NO from MDM after CD23 activation played a major role during their anti-M. avium activity. CD23 activation also induced tumor necrosis factor alpha (TNF-alpha) production from MDM. Mycobacteria reduction was partially inhibited by the addition of neutralizing anti-TNF-alpha antibody to cell cultures without affecting NO levels, which suggested the role of this cytokine for optimal antimicrobial activity. Finally, interleukin-10, a Th2 cytokine known to downregulate CD23 pathway, is shown to decrease NO generation and mycobacteria elimination by macrophages. Therefore, (i) infection with M. avium promotes functional surface CD23 expression on human macrophages and (ii) subsequent signaling of this molecule contributes to the antimicrobial activity of these cells through an NO- and TNF-alpha-dependent pathway. This study reveals a new human immune response mechanism to counter mycobacterial infection involving CD23 and its related ligands.
Assuntos
Macrófagos/imunologia , Macrófagos/microbiologia , Mycobacterium avium/imunologia , Receptores de IgE/biossíntese , Células Cultivadas , Humanos , Interleucina-10/imunologia , Viabilidade Microbiana , Óxido Nítrico/metabolismo , Fator de Necrose Tumoral alfa/metabolismoRESUMO
BACKGROUND: Chikungunya fever is an emerging arboviral disease characterized by an algo-eruptive syndrome, inflammatory polyarthralgias, or tenosynovitis that can last for months to years. Up to now, the pathophysiology of the chronic stage is poorly understood. CASE PRESENTATION: We report the first case of CHIKV infection with chronic associated rheumatism in a patient who developed progressive erosive arthritis with expression of inflammatory mediators and persistence of specific IgM antibodies over 24 months following infection. CONCLUSIONS: Understanding the specific features of chikungunya virus as well as how the virus interacts with its host are essential for the prevention, treatment or cure of chikungunya disease.
Assuntos
Infecções por Alphavirus/complicações , Anticorpos Antivirais/sangue , Artrite Infecciosa/etiologia , Vírus Chikungunya/imunologia , Imunoglobulina M/sangue , Infecções por Alphavirus/sangue , Infecções por Alphavirus/imunologia , Artrite Infecciosa/sangue , Artrite Infecciosa/imunologia , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
In contrast to young rats, adult rats given i.p. Plasmodium berghei Anka (PbA) control the parasitaemia and repair their anaemia. Here, we investigated whether IgE and CD23/NO immune pathway could be implicated in this age-related resistance of adult rats to PbA. Eight-week-old rats displayed significantly higher levels of plasma total IgE (p=0.01) and soluble CD23 (p=0.003) during the peak of parasitaemia, compared to 4-week-old rats. IgE Fc-binding antibody or aminoguanidine administration to parasitized 8-week-old rats slightly delayed blood parasite clearance or exacerbated anaemia. These data suggest that IgE and CD23/NO could play an important role in the resistance of adult rats experiencing PbA primary intraerythrocytic development.
Assuntos
Envelhecimento/imunologia , Imunoglobulina E/sangue , Malária/imunologia , Óxido Nítrico/metabolismo , Parasitemia/imunologia , Plasmodium berghei/patogenicidade , Receptores de IgE/sangue , Animais , Eritrócitos/parasitologia , Feminino , Malária/parasitologia , Parasitemia/parasitologia , Ratos , Ratos Endogâmicos Lew , Organismos Livres de Patógenos EspecíficosRESUMO
The efficiency of a monomolecular film of (3-glycidoxypropyl) trimethoxysilane (GPTS) on a shear horizontal guided (Love) acoustic wave immunosensor to detect whole Escherichia coli (E. coli) bacteria is demonstrated. Direct anti-E. coli antibodies grafting onto the sensor surface did not lead to a significant bacteria immobilisation, partially attributed to the SiO2 sensor surface roughness. An innovative method has been set up to get around this difficulty and to detect whole bacteria. It consists in grafting goat anti-mouse antibodies (GAM) onto the sensor surface in a first step and introducing E. coli bacteria mixed with anti-E. coli antibodies onto the sensor in a second step. We describe the characteristics of such a technique like sample preparation time (lower than 30 min) and temperature improvements. A 37 degrees C experimental temperature led to the fastest bacteria binding kinetic, reducing the total analysis time. This method enables to keep the specificity of the antibody/antigen interaction and provides significant results in less than 1h. This leads to a detection threshold of 10(6) bacteria/ml in a 500 microl chamber.
Assuntos
Técnicas Biossensoriais/instrumentação , Escherichia coli/isolamento & purificação , Animais , Escherichia coli/imunologia , Imunoensaio/instrumentação , CamundongosRESUMO
The engineering of nanomaterials, because of their specific properties, is increasingly being developed for commercial purposes over the past decades, to enhance diagnosis, cosmetics properties as well as sensing efficiency. However, the understanding of their fate and thus their interactions at the cellular level with bio-organisms remains elusive. Here, we investigate the size- and charge-dependence of the damages induced by silica nanoparticles (SiO2-NPs) on Gram-negative Escherichia coli bacteria. We show and quantify the existence of a NPs size threshold discriminating toxic and inert SiO2-NPs with a critical particle diameter (Φc) in the range 50nm-80nm. This particular threshold is identified at both the micrometer scale via viability tests through Colony Forming Units (CFU) counting, and the nanometer scale via atomic force microscopy (AFM). At this nanometer scale, AFM emphasizes the interaction between the cell membrane and SiO2-NPs from both topographic and mechanical points of view. For SiO2-NPs with Φ>Φc no change in E. coli morphology nor its outer membrane (OM) organization is observed unless the NPs are positively charged in which case reorganization and disruption of the OM are detected. Conversely, when Φ<Φc, E. coli exhibit unusual spherical shapes, partial collapse, even lysis, and OM reorganization.
Assuntos
Antibacterianos/farmacologia , Escherichia coli/efeitos dos fármacos , Nanopartículas , Dióxido de Silício/química , Antibacterianos/química , Membrana Celular/efeitos dos fármacos , Difusão Dinâmica da Luz , Escherichia coli/ultraestrutura , Microscopia de Força Atômica , Nanopartículas/química , Tamanho da Partícula , Dióxido de Silício/farmacologiaRESUMO
Pentahydroxyflavone dihydrate, quercetin (QU) is one of common flavonols biosynthesized by plants and has been suggested to modulate inflammatory responses in various models. In the present study, we investigated in vivo effects of oral or intra-cutaneous QU in chronic rat adjuvant-induced arthritis (AA). Growth delay and arthritic scores were evaluated daily after AA induction in Lewis rats. Oral administration of QU (5 x 160 mg/kg) to arthritic rats resulted in a clear decrease of clinical signs compared to untreated controls. Intra-cutaneous injections of lower doses (5 x 60 mg/kg) of QU gave similar anti-arthritic effects, while 5 x 30 mg/kg concentrations were inefficient in this respect. Finally, injection of relatively low QU doses (5 x 30 mg/kg) prior to AA induction significantly reduced arthritis signs. As QU was suggested to inhibit macrophage-derived cytokines and nitric oxide (NO), we then analyzed macrophage response ex vivo. Anti-arthritic effects of QU correlated with significant decrease of inflammatory mediators produced by peritoneal macrophages, ex vivo and in vitro. These data indicate that QU is a potential anti-inflammatory therapeutic and preventive agent targeting the inflammatory response of macrophages.
Assuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Artrite Experimental , Mediadores da Inflamação/imunologia , Macrófagos Peritoneais/efeitos dos fármacos , Quercetina/uso terapêutico , Administração Oral , Animais , Anti-Inflamatórios não Esteroides/administração & dosagem , Artrite Experimental/tratamento farmacológico , Artrite Experimental/imunologia , Artrite Experimental/prevenção & controle , Feminino , Injeções Intradérmicas , Macrófagos Peritoneais/enzimologia , Macrófagos Peritoneais/imunologia , Óxido Nítrico Sintase Tipo II/metabolismo , Quercetina/administração & dosagem , Ratos , Ratos Endogâmicos LewRESUMO
The present study aims at evaluating intrinsic changes in Escherichia coli (E. coli) surface over time, by Atomic Force Microscopy (AFM). For that purpose, bacteria were immobilized on mica or on mica previously functionalized by the deposition of a polyelectrolyte multilayer cushion. AFM images reveal that E. coli population goes through different stages. Firstly, after a week, the number of healthy bacteria decreases resulting in a release of cellular components which likely become, in turn, a nutrition source for increasing the healthy population after around two weeks. Finally, after one month, most of the bacteria is dead. Our study shows a transition of a healthy rod-shaped bacterium to a dead collapsed one. Most importantly, along with the morphological evolution of bacteria, are the structure changes and the mechanical properties of their outer membrane, emphasized by AFM phase images with very high resolution. Indeed, the surface of healthy bacteria is characterized by a phase separation pattern, thereafter mentioned as "ripples". Bacterial ageing goes along with the loss of this organized structure, turning into circular areas with irregular boundaries. These changes are likely caused by a re-organization, due to external stress, of mainly lipopolysaccharides (LPS) present in the outer membrane of E. coli.
Assuntos
Silicatos de Alumínio/química , Membrana Celular/química , Escherichia coli/química , Microscopia de Força Atômica/métodos , Biofilmes , Escherichia coli/fisiologia , Escherichia coli/ultraestrutura , Lipopolissacarídeos/química , Viabilidade Microbiana , Nanoestruturas/química , Propriedades de Superfície , Fatores de TempoRESUMO
Human actinomycosis with involvement of the spine is a rare condition although it has been first described a long time ago. It is probably underrecognized since its clinical presentation is often misleading and accurate bacteriological diagnosis is challenging. We herein report a rare case of cervical actinomycosis with paravertebral abscess and spondylitis imputed to an infection by Actinomyces meyeri in a 52-year-old immunocompetent Caucasian man. A. meyeri should be considered as a potential cause for subacute or chronic spondylitis, even in immunocompetent subjects. Modern diagnostic tools such as Matrix-Assisted Laser Desorption-Ionization Time of Flight mass spectrometry and 16S rRNA sequencing are efficient for accurate microbiological identification.
Assuntos
Abscesso/microbiologia , Actinomyces/isolamento & purificação , Actinomicose/diagnóstico , Vértebras Cervicais/microbiologia , Doenças da Coluna Vertebral/microbiologia , Espondilite/microbiologia , Abscesso/diagnóstico , Actinomyces/genética , DNA Bacteriano/genética , Humanos , Imunocompetência , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , RNA Ribossômico 16S/genética , Doenças da Coluna Vertebral/diagnóstico , Espondilite/diagnósticoRESUMO
Polyphenolic anthocyanins are major colorful compounds in red fruits, known to prevent cardiovascular and other diseases. Grape polyphenols are a mixture of various molecules and their exact contribution to above bioactivities remains to be clarified. In the present study, we first analyzed the effect of purified grape-derived compounds on human peripheral blood mononuclear cell (PBMC) survival, proliferation, as well as for their ability to inhibit the activation of human normal macrophages. Data indicated that malvidin-3-O-ß glucoside (Malßg), the major grape anthocyanin, is bioactive with no toxicity on human PBMC. Malßg decreased the transcription of genes encoding inflammatory mediators, confirmed by the inhibition of TNFα, IL1, IL-6 and iNOS-derived nitric oxide (NO) secretion from activated macrophages. As Malßg also inhibited inflammatory response of rat macrophages, we investigated the anti-inflammatory potential of Malßg in chronic rat adjuvant-induced arthritis (AIA). Malßg significantly diminished inflammatory cachexia and arthritic paw scores in AIA rats at both therapeutic and preventive levels. In vivo effects of Malßg correlated with down-regulation of NO generation from AIA rats' peritoneal macrophages ex vivo. These data indicate that Malßg, major grape anthocyanin, is a potent anti-inflammatory agent in vitro and in vivo, without detectable toxic effect.
Assuntos
Antocianinas/farmacologia , Anti-Inflamatórios/farmacologia , Artrite Experimental/tratamento farmacológico , Citocinas/antagonistas & inibidores , Glucosídeos/farmacologia , Macrófagos Peritoneais/efeitos dos fármacos , Óxido Nítrico/antagonistas & inibidores , Vitis , Animais , Antocianinas/uso terapêutico , Anti-Inflamatórios/uso terapêutico , Artrite Experimental/metabolismo , Artrite Experimental/fisiopatologia , Doença Crônica , Citocinas/metabolismo , Feminino , Glucosídeos/uso terapêutico , Humanos , Interleucina-1/antagonistas & inibidores , Interleucina-1/metabolismo , Interleucina-6/antagonistas & inibidores , Interleucina-6/metabolismo , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/metabolismo , Ativação de Macrófagos , Macrófagos Peritoneais/metabolismo , Óxido Nítrico/biossíntese , Óxido Nítrico Sintase Tipo II/antagonistas & inibidores , Óxido Nítrico Sintase Tipo II/metabolismo , Polifenóis/farmacologia , Ratos , Ratos Endogâmicos Lew , Fator de Necrose Tumoral alfa/antagonistas & inibidores , Fator de Necrose Tumoral alfa/metabolismoRESUMO
BACKGROUND: In addition to helminthic infections, elevated serum IgE levels were observed in many protozoal infections, while their contribution during immune response to these pathogens remained unclear. As IgE/antigen immune complexes (IgE-IC) bind to human cells through FcεRI or FcεRII/CD23 surface molecules, the present study aimed to identify which functional receptor may be involved in IgE-IC interaction with human macrophages, the major effector cell during parasite infection. METHODOLOGY/PRINCIPAL FINDINGS: Human monocyte-derived macrophages were infected with Toxoplasma gondii before being incubated with IgE-IC. IgE receptors were then identified using appropriate blocking antibodies. The activation of cells and parasiticidal activity were evaluated by mediator quantification and direct counting of infected macrophages. RNAs were extracted and cell supernatants were also collected for their content in tumor necrosis factor (TNF)-α, interleukin-10 (IL-10) and nitrites. Sera from symptomatic infected patients were also tested for their content of IgE, IL-10 and nitrites, and compared to values found in healthy donors. Results showed that IgE-IC induced intracellular elimination of parasites by human macrophages. IgE-mediated effect was FcεRI-independent, but required cross-linking of surface FcεRII/CD23, cell activation and the generation of nitric oxide (NO). Although TNF-α was shown to be produced during cell activation, this cytokine had minor contribution in this phenomenon while endogenous and exogenous IL-10 down-regulated parasite killing. Inverse relationship was found between IL-10 and NO expression by infected human macrophages at both mRNA and mediator levels. The relationship between these in vitro data and in vivo levels of various factors in T. gondii infected patients supports the involvement of CD23 antigen and IL-10 expression in disease control. CONCLUSION: Thus, IgE may be considered as immune mediator during antiprotozoal activity of human macrophages through its ability to trigger CD23 signaling. Increased cell activation by IgE-IC may also account for chronic inflammatory diseases observed in some patients.
Assuntos
Imunoglobulina E/imunologia , Interleucina-10/farmacologia , Espaço Intracelular/parasitologia , Macrófagos/imunologia , Receptores de IgE/metabolismo , Transdução de Sinais/efeitos dos fármacos , Toxoplasma/citologia , Animais , Morte Celular/efeitos dos fármacos , Humanos , Imunoglobulina E/sangue , Interleucina-10/sangue , Espaço Intracelular/efeitos dos fármacos , Ativação de Macrófagos/efeitos dos fármacos , Macrófagos/enzimologia , Macrófagos/parasitologia , Camundongos , Camundongos Endogâmicos BALB C , Óxido Nítrico/metabolismo , Óxido Nítrico Sintase Tipo II/metabolismo , Nitritos/sangue , Parasitos/citologia , Parasitos/efeitos dos fármacos , Toxoplasma/efeitos dos fármacos , Fator de Necrose Tumoral alfa/biossínteseAssuntos
Arginase/metabolismo , Parasitos/enzimologia , Doenças Parasitárias/enzimologia , Animais , Arginase/antagonistas & inibidores , Arginase/classificação , Arginina/metabolismo , Humanos , Macrófagos Peritoneais/enzimologia , Macrófagos Peritoneais/imunologia , Camundongos , Modelos Biológicos , Óxido Nítrico/biossíntese , Óxido Nítrico/toxicidade , Óxido Nítrico Sintase/metabolismo , Doenças Parasitárias/imunologia , Doenças Parasitárias/parasitologia , Trypanosoma brucei brucei/crescimento & desenvolvimento , Trypanosoma brucei brucei/imunologia , Tripanossomíase/imunologiaRESUMO
Human actinomycosis with involvement of the spine is a rare condition although it has been first described a long time ago. It is probably underrecognized since its clinical presentation is often misleading and accurate bacteriological diagnosis is challenging. We herein report a rare case of cervical actinomycosis with paravertebral abscess and spondylitis imputed to an infection by Actinomyces meyeri in a 52-year-old immunocompetent Caucasian man. A. meyeri should be considered as a potential cause for subacute or chronic spondylitis, even in immunocompetent subjects. Modern diagnostic tools such as Matrix-Assisted Laser Desorption–Ionization Time of Flight mass spectrometry and 16S rRNA sequencing are efficient for accurate microbiological identification.
Assuntos
Humanos , Masculino , Pessoa de Meia-Idade , Abscesso/microbiologia , Actinomyces/isolamento & purificação , Actinomicose/diagnóstico , Vértebras Cervicais/microbiologia , Doenças da Coluna Vertebral/microbiologia , Espondilite/microbiologia , Abscesso/diagnóstico , Actinomyces/genética , DNA Bacteriano/genética , Imunocompetência , Reação em Cadeia da Polimerase , /genética , Doenças da Coluna Vertebral/diagnóstico , Espondilite/diagnósticoRESUMO
Novel glycidyl-terminated organosilicon coupling agents possessing a trialkoxysilyl head group and a very long hydrocarbon chain (C22) were synthesized. Their ability to afford densely packed self-assembled monolayers (SAMs) grafted on silica-based surfaces was investigated. Transmission FT-IR spectra showed that the most regular films were obtained by using trichloracetic acid as the catalyst (10 M%). Atomic force microscopy (AFM) and optical ellipsometry were consistent with well ordered monolayers exhibiting a marked decrease of the surface roughness. Epifluorescence microscopy revealed that these SAMs possessed a better surface reactivity than monolayers obtained with the commercially available (3-glycidoxypropyl) trimethoxysilane (GPTS) upon grafting of a fluorescent probe (dansylcadaverin). Moreover, direct attachment of fluorescent antibodies (RAG-TRITC) through covalent binding led to higher mean fluorescence intensities, showing that these new SAMs possess high potential for the immobilization of biological molecules.
Assuntos
Reagentes de Ligações Cruzadas/química , Membranas Artificiais , Silanos/química , Dióxido de Silício/química , Cadaverina/análogos & derivados , Cadaverina/química , Catálise , Corantes Fluorescentes/química , Microscopia de Força Atômica , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Propriedades de SuperfícieRESUMO
BACKGROUND: CD23 is a differentiation/activation antigen expressed by a variety of hematopoietic and epithelial cells. It can also be detected in soluble forms in biological fluids. Initially known as the low-affinity receptor for immunoglobulin E (Fc epsilonRII), CD23 displays various other physiologic ligands such as CD21, CD11b/c, CD47-vitronectin, and mannose-containing proteins. CD23 mediates numerous immune responses by enhancing IgE-specific antigen presentation, regulating IgE synthesis, influencing cell differentiation and growth of both B- and T-cells. CD23-crosslinking promotes the secretion of pro-inflammatory mediators from human monocytes/macrophages, eosinophils and epithelial cells. Increased CD23 expression is found in patients during allergic reactions and rheumatoid arthritis while its physiopathologic role in these diseases remains to be clarified. METHODOLOGY/PRINCIPAL FINDINGS: We previously generated heptapeptidic countrestructures of human CD23. Based on in vitro studies on healthy and arthritic patients' cells, we showed that CD23-specific peptide addition to human macrophages greatly diminished the transcription of genes encoding inflammatory cytokines. This was also confirmed by significant reduction of mediator levels in cell supernatants. We also show that CD23 peptide decreased IgE-mediated activation of both human and rat CD23(+) macrophages. In vivo studies in rat model of arthritis showed that CD23-blocking peptide ameliorates clinical scores and prevent bone destruction in a dose dependent manner. Ex-vivo analysis of rat macrophages further confirmed the inhibitory effect of peptides on their activation. Taken together our results support the role of CD23 activation and subsequent inflammatory response in arthritis. CONCLUSION: CD23-blocking peptide (p30A) prevents the activation of monocytes/macrophages without cell toxicity. Thus, targeting CD23 by antagonistic peptide decreases inflammatory markers and may have clinical value in the treatment of human arthritis and allergic reactions involving CD23.
Assuntos
Artrite/imunologia , Receptores de IgE/antagonistas & inibidores , Animais , Sequência de Bases , Western Blotting , Estudos de Casos e Controles , Primers do DNA , Feminino , Humanos , Macrófagos Peritoneais/imunologia , Ratos , Ratos Endogâmicos Lew , Receptores de IgE/fisiologia , Transdução de SinaisRESUMO
BACKGROUND: Dietary flavonols may play an important role in the adjunct therapy of chronic inflammation. The availability of therapeutic formulations of pentahydroxyflavone glycoside, rutoside (RU), led us to investigate the ability of this molecule to modulate the release of various proinflammatory mediators from human activated macrophages in vitro and to ameliorate arthritic markers in a rat model. METHODS: RU was added simultaneously to human macrophages during their activation. Cells were then analyzed for inflammation-related gene expression using a specific array, and cell supernatants were collected to measure inflammatory mediators. RU was also injected into adjuvant-induced arthritic rats, and disease progression and body weight were evaluated until 50 days after injection. Sera and peritoneal macrophages were also collected to quantify the RU effect on various inflammatory markers. RESULTS: RU inhibited inflammation-related gene expression in activated human macrophages and the release of nitric oxide, tumor necrosis factor-alpha, interleukin (IL)-1, and IL-6 from these cells. In a rat model, RU inhibited clinical signs of chronic arthritis, correlating with decreased levels of inflammatory cytokines detected in rat sera and macrophage supernatants. CONCLUSION: Thus, RU may have clinical value in reducing inflammatory manifestations in human arthritis and other inflammatory diseases.
Assuntos
Artrite Experimental/metabolismo , Artrite Experimental/fisiopatologia , Mediadores da Inflamação/antagonistas & inibidores , Macrófagos/metabolismo , Rutina/farmacologia , Animais , Artrite Experimental/genética , Artrite Experimental/patologia , Peso Corporal/efeitos dos fármacos , Células Cultivadas , Doença Crônica , Citocinas/antagonistas & inibidores , Citocinas/genética , Relação Dose-Resposta a Droga , Feminino , Humanos , Interleucina-1/antagonistas & inibidores , Interleucina-6/antagonistas & inibidores , Macrófagos/efeitos dos fármacos , Macrófagos Peritoneais/efeitos dos fármacos , Macrófagos Peritoneais/metabolismo , Óxido Nítrico/antagonistas & inibidores , Ratos , Ratos Endogâmicos Lew , Rutina/administração & dosagem , Transcrição Gênica/efeitos dos fármacos , Fator de Necrose Tumoral alfa/antagonistas & inibidoresRESUMO
B-cell chronic lymphocytic leukaemia (B-CLL) is a neoplastic disorder characterized by defective apoptosis, cell accumulation in G0/G1, and high expression of BCL2 oncogene. Intracellular cyclic adenosine monophosphate (cAMP) accumulation increases the chemosensitivity of B-CLL cells in vitro and in vivo. In the present study, we investigated the effects of beta2-adrenergic compounds, well known cAMP-inducing drugs, on the in vitro survival of leukaemia cells. In contrast to the short-acting beta2-mimetic (beta2Mim) salbutamol, a consistent pro-apoptotic effect was observed with the long-acting beta2Mim salmeterol and formoterol. Normal B cells isolated from control donors were totally resistant to the above molecules. These compounds also increased chlorambucil- and fludarabine-induced death of B-CLL cells. Blockade of beta-adrenergic receptor signalling or cAMP did not alter B-CLL apoptosis with beta2 Mimagents. Leukaemia cell apoptosis by beta2Mim correlated with an increase in calcium influx, decreased bcl-2 protein and mRNA levels, increase in BAX gene expression and a marked rise in BCL2/BAX mRNA ratios. Interleukin-4, a cytokine that increases bcl-2 expression in B-CLL cells, rescued leukaemia cell from apoptosis with beta2Mim. These data show that long-acting beta2-adrenergic agents promote apoptotic leukaemia cell death through an adrenoreceptor- and cAMP-independent, Ca2+-dependent mechanism.