An image processing workstation for automatic evaluation of human granulocyte motility

Azzarà, Antonio; Chimenti, M.; Azzarelli, L.; Fantini, E.; Carulli, Giovanni; Ambrogi, F

doi:10.1016/0022-1759(92)90155-m

Cited by 11 publications

(11 citation statements)

References 22 publications

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“…PMN from each of three uninfected controls were divided into three aliquots and diluted in PBS Dulbecco solution to a final cell concentration of 10 6 /ml: two aliquots were cultured with AgB at a concentration of 1 or 2 g/ml, and the third aliquot was cultured without antigen (blank control), and all were tested in triplicate. Chemotaxis and random migration were assayed by the modified Boyden method, as described elsewhere (1). In brief, PMN random migration and chemotaxis were assessed in Perspex chemotactic chambers (B.M.…”

Section: Methodsmentioning

confidence: 99%

Modulation of Human Immune Response byEchinococcus granulosusAntigen B and Its Possible Role in Evading Host Defenses

Riganò

Profumo

Bruschi³

et al. 2001

Infect Immun

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143

117

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). AgB skewed the Th1/Th2 cytokine ratios towards a preferentially immunopathology-associated Th2 polarization, predominantly in patients with progressive disease. AgB-stimulated patients' PBMC also proliferated less than SHF-stimulated PBMC (P ‫؍‬ 9 ؋ 10 ؊3 ). In vitro Th2 cytokine production was reflected in vivo by elevated specific immunoglobulin E (IgE) and IgG4 antibodies binding to AgB. These findings confirm that AgB plays a role in the escape from early immunity by inhibiting PMN chemotaxis. They also add new information on the host-parasite relationship, suggesting that AgB exploits the activation of T helper cells by eliciting a nonprotective Th2 cell response.Cystic echinococcosis (CE) is a widely endemic helminthic disease caused by infection with metacestodes (larval stage) of the tapeworm Echinococcus granulosus. It affects humans and a wide range of livestock species (28). The disease is characterized by the growth in the host internal organs, mostly liver and lungs, of steadily growing fluid-filled, unilocular cysts surrounded by a two-layered hydatid cyst wall. The main feature of the host-parasite relationship is the coexistence of the chronic infection with detectable humoral and cellular responses against the parasite. Parasite survival in vivo depends on efficient evasion mechanisms starting to operate as the parasite develops toward a hydatid cyst. A fibrotic host capsule of variable thickness usually develops, thus forming together with the parasite-derived acellular laminated layer a formidable cystic structure. As a consequence, the actively dividing germinal layer within the cyst along with its associated brood capsule and enclosed protoscoleces are effectively sequestered from the host immune responses. In addition to this physical barrier, the hydatid cyst has probably evolved other strategies for immune evasion. Although older models suggested a more passive role for parasites in immune evasion-for example sequestration, antigenic masking by host proteins, and global immunosuppression-later studies suggest in human parasitoses the active deployment of strategies that manipulate and exploit the host response (12, 30).

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Section: Methodsmentioning

confidence: 99%

Modulation of Human Immune Response byEchinococcus granulosusAntigen B and Its Possible Role in Evading Host Defenses

Riganò

Profumo

Bruschi³

et al. 2001

Infect Immun

Self Cite

143

117

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“…Data referring to the mean FP of the three tests performed in triplicate are summarized in figure 2a. In both cases, the mean interpolating curves displayed a typical gaussian pattern [21], even when the protein concentration was inhibitory. In fact, the values of adjustment R 2 relative to the linear regression confirm an exponential trend (0.904 for 1 µg/ml; 0.927 for 2 µg/ml; blank = 0.953) (data not shown).…”

Section: Resultsmentioning

confidence: 99%

“…The method, which has been described elsewhere [21], provides a completely new approach for the evaluation of cell migration throughout microporous filters. An area equal to 12 conventional microscopic fields is imaged onto a TV camera mounted on top of the microscope.…”

Section: Methodsmentioning

confidence: 99%

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Inhibitory Effects of Human Neutrophil Functions by the 45-kD Glycoprotein Derived from the Parasitic Nematode <i>Trichinella spiralis</i>

Bruschi

Carulli

Azzarà

et al. 2000

Int Arch Allergy Immunol

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Aim: We evaluated the effect of the 45-kD protein of Trichinella spiralis (gp45), purified by affinity chromatography, on random migration and chemotaxis, the oxidative metabolism of human neutrophils and on the CD11b upregulation induced by formyl-methionyl-leucyl-phenylalanine (f-MLP). Methods: Donor neutrophils incubated with different amounts of gp45 (0.5, 1, 1.5, 2 μg/ml) or buffer and the random migration and chemotaxis, evaluated by means of a special technique of image analysis, and the chemiluminescence response to f-MLP or phorbol myristate acetate (PMA) were analyzed. The effect on CD11b upregulation was assessed incubating cells with the protein, when activating them with f-MLP. Results: The results showed that gp45 inhibited both random and stimulated migrations, and reduced the response to f-MLP and PMA. Furthermore, gp45 significantly reduced the upregulation of the CD11b induced by f-MLP. Conclusion: The results show that gp45 inhibits PMN in different functions, suggesting an anti-inflammatory action.

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“…Computer-assisted analysis has been used to various extents to study the migration and morphology of many cell types, including neutrophils (Azzara et al, 1992;Cheung et al, 1987;Donovan et al, 1986;Dow et al, 1987;Pedersen et al, 1988;Moghe et al, 1995;Sadhu et al, 2003;Vollmer et al, 1992) and neurons (Bilsland et al, 1999;Distasi et al, 2002;Hossain and Morest, 2000;Hynds and Snow, 2002;Szebenyi et al, 2001). Migration and axon guidance studies rely upon phase-contrast or fluorescence microscopy to document the growth process.…”

Section: Introductionmentioning

confidence: 98%

Automated identification of axonal growth cones in time-lapse image sequences

Keenan¹,

Hooker²,

Spilker³

et al. 2006

Journal of Neuroscience Methods

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The isolation and purification of axon guidance molecules has enabled in vitro studies of the effects of axon guidance molecule gradients on numerous neuronal cell types. In a typical experiment, cultured neurons are exposed to a chemotactic gradient and their growth is recorded by manual identification of the axon tip position from two or more micrographs. Detailed and statistically valid quantification of axon growth requires evaluation of a large number of neurons at closely spaced time points (e.g. using a time-lapse microscopy setup). However, manual tracing becomes increasingly impractical for recording axon growth as the number of time points and/or neurons increases. We present a software tool that automatically identifies and records the axon tip position in each phase-contrast image of a time-lapse series with minimal user involvement. The software outputs several quantitative measures of axon growth, and allows users to develop custom measurements. For, example analysis of growth velocity for a dissociated E13 mouse cortical neuron revealed frequent extension and retraction events with an average growth velocity of 0.05 +/- 0.14 microm/min. Comparison of software-identified axon tip positions with manually identified axon tip positions shows that the software's performance is indistinguishable from that of skilled human users.

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An image processing workstation for automatic evaluation of human granulocyte motility

Cited by 11 publications

References 22 publications

Modulation of Human Immune Response byEchinococcus granulosusAntigen B and Its Possible Role in Evading Host Defenses

Modulation of Human Immune Response byEchinococcus granulosusAntigen B and Its Possible Role in Evading Host Defenses

Inhibitory Effects of Human Neutrophil Functions by the 45-kD Glycoprotein Derived from the Parasitic Nematode <i>Trichinella spiralis</i>

Automated identification of axonal growth cones in time-lapse image sequences

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