Olle Stendahl - Academia.edu (original) (raw)

Papers by Olle Stendahl

The Journal of Immunology, 2007

Journal of Leukocyte Biology

The interaction of platelets with neutrophil granulocytes is considered to play an important role... more The interaction of platelets with neutrophil granulocytes is considered to play an important role in the inflammatory process, and the present study was focused on platelet-induced modulation of Fcgamma receptor-mediated functions in neutrophils. We found that phagocytosis and the respiratory burst (measured as luminol-enhanced chemiluminescence), triggered in neutrophils by immunoglobulin G (IgG)-opsonized yeast particles, were potentiated by platelets and that maximal enhancement was achieved at a physiological neutrophil/platelet ratio of about 1:50 to 1:100. Platelets both increased the intra- and extracellular generation of oxygen radicals as well as the release of myeloperoxidase from stimulated neutrophils. The presence of platelets also induced a cortical actin polymerization in neutrophils, which might explain the increased phagocytic capacity. Platelets appear to affect neutrophil function in a contact-independent manner that most likely involves ATP, indicated by the foll...

Blood

The effect of human platelets on chemoattractant-induced generation of oxygen metabolites in neut... more The effect of human platelets on chemoattractant-induced generation of oxygen metabolites in neutrophils was investigated, using luminol-enhanced chemiluminescence (CL). Resting platelets inhibited the extracellular, but not the intracellular, production of oxygen radicals in formyl-methionyl-leucyl-phenylalanine (fMet-Leu-Phe)-stimulated neutrophils. Maximal effect was obtained at the physiological neutrophil/platelet ratio of 1/50. Similar results were acquired by adding supernatants of platelets, indicating a role for a soluble factor. Removal of extracellular adenosine by adenosine deaminase (ADA), or blocking of adenosine-receptors by theophylline, antagonized the inhibitory effects of platelets (or the equivalent supernatant) on the neutrophil respiratory burst. In contrast, accumulation of adenosine by apyrase enhanced the inhibition. Exogenous adenosine mimicked the effects of platelets on the fMet-Leu-Phe-induced respiratory burst. To further assess the role of platelet-der...

The Journal of Immunology

Ingestion of aged or irradiated apoptotic neutrophils actively suppresses stimulation of macropha... more Ingestion of aged or irradiated apoptotic neutrophils actively suppresses stimulation of macrophages (Mphi). Many bacterial pathogens can also provoke apoptosis in neutrophils, but little is known about how such apoptotic cells influence Mphi activation. We found that neutrophils undergoing apoptosis induced by UV irradiation, Escherichia coli, or Staphylococcus aureus could either stimulate or inhibit Mphi activation. In contrast to Mphi that had ingested irradiated apoptotic neutrophils, Mphi that had phagocytosed bacteria-induced apoptotic neutrophils exhibited markedly increased production of the proinflammatory cytokine TNF-alpha, but not the anti-inflammatory cytokine TGF-beta. Moreover, ingestion of bacteria, but not UV-induced apoptotic neutrophils, caused increased expression of FcgammaRI on Mphi, and this effect was not provoked directly by bacteria associated with the apoptotic neutrophils. Instead, we found that a link between pathogen-induced apoptotic neutrophils and u...

Microbiology Discovery, 2013

The role of vitamin D supplementation as adjuvant treatment of tuberculosis (TB) has lately attra... more The role of vitamin D supplementation as adjuvant treatment of tuberculosis (TB) has lately attracted increasing interest. Our aim was to investigate the capacity of alveolar macrophages (AMs) from patients with or without exposure to TB to control intracellular growth of virulent Mycobacterium tuberculosis (Mtb). Methods: AMs were freshly harvested from the bronchoalveolar lavage fluid of 7 patients with a history of TB (4 patients with previous TB and 3 patients with current TB) and 4 non-TB subjects. The H37Rv strain, genetically modified to express Vibrio harveyi luciferase, was used to determine the growth of Mtb by luminometry in the AMs from study subjects. Cytokine levels in culture supernatants were determined using a flow cytometry-based bead array technique. Results: AMs from patients with a TB history were less efficient in restricting Mtb growth. Stimulation with 100 nM1, 25-dihydroxyvitamin D (1,25D 3 ) did not significantly influence the capacity of AMs from any study subjects to control the infection. Out of the cytokines evaluated (TNF-α, IL-1β, IL-10 and IL-12p40) only TNF-α demonstrated detectable levels in culture supernatants, but did not respond to stimulation with 1,25D 3 .

Cell biophysics, 1980

The capacity of synthetic amphiphiles poly(ethyleneglycol) 6000 (PEG) esterified with saturated f... more The capacity of synthetic amphiphiles poly(ethyleneglycol) 6000 (PEG) esterified with saturated fatty acids (C2-C18), to modify polymorphonuclear leukocyte (PMNL) locomotion has been investigated. It was noticed that PEG-myristate (M-PEG; C14) stimulated the random locomotion of PMNL populations in concentrations up to about 1 g/L. The esters with shorter aliphatic chains had negligible effects, whereas those with longer chains, PEG-palmitate (P-PEG; C16) and PEG-stearate (S-PEG; C18) reduced the locomotion, irrespectively of concentration. The ability of the PMNL to be stimulated by an attractant liberated from normal human serum was slightly impaired by M-PEG, but not by P-PEG. The response to M-PEG of individual PMNL was heterogeneous in that some cells were stimulated and others were inhibited. However, the average result was a reduction of the motility. This indicates that methods used for the study of the locomotion of cell populations may not always reflect the average behavi...

Journal of free radicals in biology & medicine, 1986

Superoxide production and chemiluminescence induced in differentiated HL-60 cells by the chemoatt... more Superoxide production and chemiluminescence induced in differentiated HL-60 cells by the chemoattractant formylmethionyl-leucyl-phenylalanine: In order to study the generation of oxidative metabolites in relation to cell differentiation, dimethyl sulfoxide (DMSO) and retinoic acid (RA) differentiated HL-60 cells were stimulated with the chemotactic peptide formylmethionyl-leucyl-phenylalanine (FMLP). The oxidative response was measured as luminol-dependent chemiluminescence, lucigenin-dependent chemiluminescence, and cytochrome c reduction. Cells grown in the presence of DMSO or RA progressively expressed morphological changes, and when the mature cells were exposed to FMLP the cells produced oxidative metabolites. Quantitatively the HL-60 cells grown in the presence of DMSO gave rise to the most pronounced response. No correlation was obtained between superoxide production, luminol-chemiluminescence and lucigenin-dependent chemiluminescence, indicating that different aspects of the...

Acta pathologica, microbiologica, et immunologica Scandinavica. Section B, Microbiology, 1985

The influence of culture period on mannose-specific and hydrophobic properties of the bacterial s... more The influence of culture period on mannose-specific and hydrophobic properties of the bacterial surface and on bacteria/polymorphonuclear leukocyte (PMNL) interaction was studied. Four E. coli strains, PN7 (01:K1), ABU2 (ON:K14), CU9 (06:K14) and CU13 (08:KN) and two Salmonella typhimurium strains 395 MR10 and 395 MS, well characterized according to physicochemical surface properties, presence of type 1 fimbriae and interaction with PMNL, were used in the study. The results show that with prolonged culture period, the liability to hydrophobic interaction increases, the agglutination-strength of mannose-specific maltobionamide liposomes increases, while the agglutination-titer with guinea-pig erythrocytes remains constant. Furthermore, the mannose-specific association with and metabolic activation of PMNL is augmented, while the ingestion is unchanged. In addition, our results demonstrate differences in sensitivity between the methods used to detect exposure of mannose-specific struc...

During acute bacterial infections such as sepsis and meningitis, activation of inflammatory media... more During acute bacterial infections such as sepsis and meningitis, activation of inflammatory mediators such as nitric oxide (NO) plays a crucial role in both pathogenesis and host defense. We have previously reported that CNI-1493, a macrophage deactivator, reduced mortality in infant rats infected with Haemophilus influenzae type b (Hib) with associated decrease in the number of granulocytes in the infected tissue. The aim of the present study was to investigate how CNI-1493 affects granulocytes and macrophages in vitro. Murine macrophages (RAW 264.7) pre-incubated with CNI-1493 prior to activation with lipopolysaccharide (LPS)/interferon gamma (IFNgamma) had decreased NO production measured as NO(2)(-)/NO(3)(-) levels and reduction in inducible NO-synthase (iNOS) expression. Reactive oxygen species (ROS) production was increased in formylmethionyl-leucyl-phenylalanine (FMLP)-stimulated granulocytes following CNI-1493 treatment, whereas F-actin content, motility and chemotaxis were decreased under the same conditions. The effects of CNI-1493 on both NO production in LPS/IFNgamma-activated macrophages and ROS production, F-actin content, motility and chemotaxis in granulocytes, may contribute to the reduced inflammatory response and increased survival in Hib-infected animals treated with CNI-1493.

Journal of Cell Biology, 1990

Cytosolic free calcium ((Ca2+)0 and fusion of secondary granules with the phagosomal membrane (ph... more Cytosolic free calcium ((Ca2+)0 and fusion of secondary granules with the phagosomal membrane (phagosome-lysosome fusion, P-L fusion) were as- sessed in single adherent human neutrophils during phagocytosis of C3bi-opsonized yeast particles. Neutrophils were loaded with the fluorescent dye fura2/AM and (Ca2+)~ was assessed by dual excitation microfluorimetry. Discharge of lactoferrin, a secondary granule marker into the phagosome was verified by

Cell Biophysics, 1983

The promyelocytic leukemia cell line HL-60 and the histiocytic cell line U-937 were grown in susp... more The promyelocytic leukemia cell line HL-60 and the histiocytic cell line U-937 were grown in suspension culture. They were induced to differentiate during 5-d cultivation in the presence of dimethylsulfoxide (DMSO; 1.3% w/v) or phorbol-12-myristate-acetate (PMA; 10(-7) M), which yields granulocyte- and macrophage-like cells, respectively. Differentiation was evidenced by increased capacity to recognize and phagocytize IgG- or complement-coated yeast particles. Aliquots taken from the cultures with and without DMSO (or PMA) were spun down directly on glass microscope slides, washed, labeled with fluoresceinated wheat germ agglutinin (WGA), and directly examined at room temperature for the rate of fluorescence recovery after photobleaching (FRAP). It was found that cultivation of the HL-60 and the U-937 cells in the presence of DMSO, which yields granulocyte-like cells, reduced the average value of lateral diffusion coefficient D (X 10(10] from 1.72 +/- 0.13 cm2s-1 to 0.97 +/- 0.13 cm2s-1 and from 1.77 +/- 0.11 cm2s-1 to 0.82 +/- 0.13 cm2s-1, respectively. U-937 cells grown with PMA also showed a reduction of D(X 10(10] to 0.88 +/- 0.10 cm2s-1. There was a larger immobile fraction of fluorescence in the HL-60 cells than in the U-937 cells, viz., 70-80% compared to 10-50%. The total number of binding sites for WGA was not altered, but the surface density changed, since the HL-60 and the U-937 cells became smaller and larger, respectively, when grown in the presence of DMSO. It is concluded that differentiation reduces the average lateral mobility of the WGA-binding membrane component by a factor around 2.

International Journal of Mycobacteriology, 2013

The emergence of multidrug-resistant strains of Mycobacterium tuberculosis (MTB), the bacterium r... more The emergence of multidrug-resistant strains of Mycobacterium tuberculosis (MTB), the bacterium responsible for tuberculosis (TB), has rekindled the interest in the role of nutritional supplementation of micronutrients, such as vitamin D, as adjuvant treatment. Here, the growth of virulent MTB in macrophages obtained from the peripheral blood of patients with and without TB was studied. The H37Rv strain genetically modified to express Vibrio harveyi luciferase was used to determine the growth of MTB by luminometry in the human monocyte-derived macrophages (hMDMs) from study subjects. Determination of cytokine levels in culture supernatants was performed using a flow cytometry-based bead array technique. No differences in intracellular growth of MTB were observed between the different study groups. However, stimulation with 100nM 1,25-dihydroxyvitamin D significantly enhanced the capacity of hMDMs isolated from TB patients to control the infection. This effect was not observed in hMDMs from the other groups. The interleukin (IL)-1β and IL-10 release by hMDMs was clearly increased upon stimulation with 1,25-dihydroxyvitamin D. Furthermore, the 1,25-dihydroxyvitamin D stimulation also led to elevated levels of TNF-α (tumor necrosis factor-alpha) and IL-12p40. It was concluded that vitamin D triggers an inflammatory response in human macrophages with enhanced secretion of cytokines, as well as enhancing the capacity of hMDMs from patients with active TB to restrict mycobacterial growth.

PLoS ONE, 2014

Macrophages in the lung are the primary cells being infected by Mycobacterium tuberculosis (Mtb) ... more Macrophages in the lung are the primary cells being infected by Mycobacterium tuberculosis (Mtb) during the initial manifestation of tuberculosis. Since the adaptive immune response to Mtb is delayed, innate immune cells such as macrophages and neutrophils mount the early immune protection against this intracellular pathogen. Neutrophils are short-lived cells and removal of apoptotic cells by resident macrophages is a key event in the resolution of inflammation and tissue repair. Since anti-inflammatory activity is not compatible with effective immunity to intracellular pathogens, we therefore investigated how uptake of apoptotic neutrophils modulates the function of Mtb-activated human macrophages. We show that Mtb infection exerts a potent proinflammatory activation of human macrophages with enhanced gene activation and release of proinflammatory cytokines and that this response was augmented by apoptotic neutrophils. The enhanced macrophage response is linked to apoptotic neutrophil-driven activation of the NLRP3 inflammasome and subsequent IL-1b signalling. We also demonstrate that apoptotic neutrophils not only modulate the inflammatory response, but also enhance the capacity of infected macrophages to control intracellular growth of virulent Mtb. Taken together, these results suggest a novel role for apoptotic neutrophils in the modulation of the macrophage-dependent inflammatory response contributing to the early control of Mtb infection.

Journal of Clinical Investigation - J CLIN INVEST, 1989

Phagocytosis of C3bior IgG-opsonized yeast particles in human neutrophils was found to be associa... more Phagocytosis of C3bior IgG-opsonized yeast particles in human neutrophils was found to be associated with an increased formation of inositol phosphates and diacylglycerol. Pertussis toxin only marginally affected phagocytosis of IgGand C3bi-opsonized particles and the associated formation of second messengers. Forskolin, which induced a threefold rise of cellular cAMP, however, markedly inhibited both C3bi-and IgG-mediated phagocytosis as well as the particle-induced formation of inositol phosphates and diacylglycerol. These observations are in contrast to what was found to occur with chemotactic factors and indicate that chemotactic and phagocytic signaling can be regulated independently in human neutrophils.

Tuberculosis, 2011

In tuberculosis (TB), the production of nitric oxide (NO) is confirmed but its importance in host... more In tuberculosis (TB), the production of nitric oxide (NO) is confirmed but its importance in host defense is debated. Our aim was to investigate whether a food supplement rich in arginine could enhance clinical improvement in TB patients by increased NO production. Smear positive TB patients from Gondar, Ethiopia (n ¼ 180) were randomized to a food supplementation rich in arginine (peanuts, equivalent to 1 g of arginine/day) or with a low arginine content (wheat crackers, locally called daboqolo) during four weeks. The primary outcome was cure rate according to the WHO classification and secondary outcomes were sputum smear conversion, weight gain, sedimentation rate, reduction of cough and chest X-ray improvement as well as levels of NO in urine (uNO) or exhaled air (eNO) at two months. There was no effect of the intervention on the primary outcome (OR 1.44, 95% CI: 0.69e3.0, p ¼ 0.39) or secondary outcomes. In the subgroup analysis according to HIV status, peanut supplemented HIVþ/TB patients showed increased cure rate (83.8% (31/37) vs 53.1% (17/32), p < 0.01). A low baseline eNO (<10 ppb) in HIVþ/TB patients was associated with a decreased cure rate. We conclude that nutritional supplementation with a food supplement rich in arginine did not have any overall clinical effect. In the subgroup of HIV positive TB patients, it significantly increased the cure rate and as an additional finding in this subgroup, low initial levels of NO in exhaled air were associated with a poor clinical outcome but this needs to be confirmed in further studies.

Transactions of the Royal Society of Tropical Medicine and Hygiene, 2006

The kinetics of potential surrogate markers in HIV-positive (HIV+) and HIV-negative (HIV-), smear... more The kinetics of potential surrogate markers in HIV-positive (HIV+) and HIV-negative (HIV-), smear-positive tuberculosis (Tb+) patients in Gondar, Ethiopia (n = 60) was investigated. Clinical symptoms, sputum conversion, sedimentation rate (SR), HIV viral load and serum levels of TNF-alpha were determined before and 8 weeks after treatment initiation. The co-infection rate of HIV was 45%. There were significantly higher initial levels of SR and TNF-alpha in HIV+/Tb+ patients (79 +/- 29 mm/h and 13.5 +/- 7.6 pg/ml), than in HIV-/Tb+ patients (60 +/- 23 mm/h and 6.8 +/- 5.9 pg/ml, P&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt;0.001). In HIV-/Tb+ patients, there was a marked decrease in SR compared with co-infected patients (46% [33 +/- 24 mm/h at week 8] vs. 24% [61 +/- 27 mm/h at week 8]). The HIV viral load (4.99 [range 3.70-5.92] to 4.90 [range 3.96-5.78] log10 copies/ml from week 0 to 8) and TNF-alpha (13.5 +/- 7.6 to 12.0 +/- 6.0 pg/ml) remained high in HIV+/Tb+ patients. In Tb patients, SR was significantly increased in HIV+ compared with HIV- patients. Additionally, TNF-alpha and HIV viral load remained elevated in HIV+/Tb+ patients following treatment despite clinical improvement comparable to HIV-/Tb+ patients.

Science, 1994

Subcellular gradients of cytosolic free Ca2+ concentration, [Ca2+]i, are thought to be critical f... more Subcellular gradients of cytosolic free Ca2+ concentration, [Ca2+]i, are thought to be critical for the localization of functional responses within a cell. A potential but previously unexplored mechanism for the generation of gradients of [Ca2+]i is the accumulation of Ca2+ stores at the site of Ca2+ action. The distribution of the Ca2+ store markers Ca(2+)-dependent adenosine triphosphatase and calreticulin was investigated in resting and phagocytosing human neutrophils. Both proteins showed an evenly distributed fine granular pattern in nonphagocytosing cells, but became markedly concentrated in the filamentous actin-rich cytoplasmic area around the ingested particle during phagocytosis. This redistribution began at early stages of phagocytosis and did not depend on an increase in [Ca2+]i. Thus, accumulation of Ca2+ stores in a restricted area of the cell may contribute to the generation of localized increases in [Ca2+]i.