Micropore filter assay of human granulocyte locomotion: Problems and solutions

Maderazo, Eufronio G.; Woronick, Charles L.

doi:10.1016/0090-1229(78)90044-2

Cited by 67 publications

(21 citation statements)

References 8 publications

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“…Supernatants (0.5%, vol/vol) from cultures without ethanol failed to stimulate PMN migration beyond that seen with albumin-containing buffer. In contrast, significantly en- Since the "leading front" method of assaying chemotaxis measures only the distance traveled by the fastest migrating cells, PMN responses to culture supernatants were also examined using the method of Maderazo and Woronick (17). In a representative experiment, leukotactic indices calculated for PMN responses to albumin-containing buffer, culture supernatants (0.5%, vol/vol) of untreated hepatocytes, and culture supernatants (0.5%, vol/vol) of ethanol (10 mM)-treated hepatocytes were 8.3, 9.1, and 15.8, respectively.…”

Section: Resultsmentioning

confidence: 99%

Production of chemotactic activity for polymorphonuclear leukocytes by cultured rat hepatocytes exposed to ethanol.

Perez¹,

Roll²,

Bissell³

et al. 1984

J. Clin. Invest.

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Abstract. Acute alcoholic hepatitis is characterized by infiltration of the liver parenchyma with polymorphonuclear leukocytes. As a possible explanation for this phenomenon, we have found that ethanol stimulates cultured rat hepatocytes to generate potent chemotactic activity. Hepatocytes (>99% pure), isolated from the livers of Sprague-Dawley rats, responded to incubation with ethanol (2.0-10 mM) by releasing chemotactic activity for human polymorphonuclear leukocytes into culture supernatants in a time-and concentration-dependent fashion. Chemotactic activity was maximal after incubation of hepatocytes with 10 mM ethanol for 6 h. It was undetectable in the absence of ethanol and was reduced in the presence of either the alcohol dehydrogenase inhibitor, 4-methylpyrazole, or the acetaldehyde dehydrogenase inhibitor, cyanamide. Ethanol failed to stimulate generation of chemotactic activity by either rat dermal fibroblasts, hepatic sinusoidal endothelial cells, or Kupffer cells. The chemotactic activity generated by ethanol-treated rat hepatocytes was recovered from culture supernatants in the lipid phase after extraction with chloroform/methanol. Thin-layer Portions of this work were presented at

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Section: Resultsmentioning

confidence: 99%

Production of chemotactic activity for polymorphonuclear leukocytes by cultured rat hepatocytes exposed to ethanol.

Perez¹,

Roll²,

Bissell³

et al. 1984

J. Clin. Invest.

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“…Micropore filter assays have a variety of technical disadvantages (Maderazo & Woronick, 1978) and the distinction of drug effects on directed locomotion (chemotaxis) and on stimulated random migration (chemokinesis) requires the use of complicated chequerboard assays (Zigmond & Hirsch, 1973).…”

Section: Introductionmentioning

confidence: 99%

THE EFFECTS OF SOME ANTIRHEUMATIC DRUGS ON AN in vitro MODEL OF HUMAN POLYMORPHONUCLEAR LEUCOCYTE CHEMOKINESIS

Smith

Walker

1980

British J Pharmacology

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1 A rapid, reproducible in vitro assay for studying the chemokinetic movement of human polymorphonuclear leucocytes (PMNs) is described. Two synthetic peptides, formyl methionyl-leucylphenylalanine (FMLP) and formyl methionyl-phenylalanine (FMP), were used as standard chemokinesins. 2 Maximal chemokinetic movement was observed with peptide concentrations of 2.5 nm (FMLP) and 100 gM (FMP). EC50 values of 650.0 + 60.0 pM and 27.0 + 3.5 ILM respectively are similar to those reported for chemotactic activity of the peptides in micropore filter assays. 3 The PMN chemokinetic response to FMLP was enhanced by histamine (100 nM) and vitamin C (2.5 gM). 4 Human serum albumin was shown to induce chemokinesis but to antagonize the response to FMLP in a dose-related fashion. Fibrinogen similarly antagonized the cell response to peptide.5 Levamisole (250 nm to 2.5 gM) significantly potentiated the chemokinetic responses to FMLP and FMP in a dose-related manner. The chemokinetic response to FMLP was unaffected by D-penicillamine (250 tLM to 10 mM) while alclofenac (500 gM to 1 mM), salicylic acid (250 gM to 10 mM) and indomethacin (100 ItM to 1 mM) caused dose-related inhibition.

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“…Most of the tested dermatophyte concentrations were stimulatory for either neutrophil or monocyte chemotaxis (LI> 1.0) [40][41][42]. These data stress on the important role of neutrophil and monocyte in host defense against dermatophyte infection.…”

Section: Resultsmentioning

confidence: 74%

Effect of Dermatophytes on Neutrophils and Monocytes Chemotaxis

Nasser¹

2016

MOJI

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Introduction: Dermatophytes are a specialized group of fungi which affect keratinous tissue of humans and other vertebrates, causing superficial infections. Objectives:The current study aimed at investigating the chemotactic activity of a group of dermatophyte fungi towards neutrophils and monocytes. Material and methods:Fifty-three patients with superficial fungal infections of glabrous skin i.e. Tinea cruris, T. capitis, T. corporis and T. pedis were investigated. They were 28 females and 25 males aged 3-66 years old (average 32.7±15.3). In the present work, 26.6% (14/53) of these patients were suffering from T. corporis followed by T.pedis in 22.6% (12/53), T.versicolor in 22.6% (12/53), T.cruris in 18.8% (10/53) and T.capitis in 9.4% (5/53) of them. Fungal examination of the scaly lesions showed the presence of the following dermatophytes: Microsporum canis, Trichophyton rubrum, T. verrocosum and T.mentagrophytes. Neutrophils and monocytes separated from apparently healthy donors were tested for their migrating abilities using the under agarose (Nelson) technique. Different concentrations of fungal mycelia with or without normal serum were used in parallel to negative (tissue culture medium) and positive (zymosan activated serum) chemo attractants. Cell migration was measured by the leukotactic index (LI); the ratio of migration towards test/migration towards control.Results: Most of the tested dermatophyte concentrations were stimulatory for either neutrophil or monocyte chemotaxis (LI>1.0). It seemed that T.verrocosum was the most stimulatory one. Neutrophils were more actively migrating than monocytes as the differences between LIs of these cells were more than LSD at 5% (least significant difference). A non proportional dose-effect relationship between fungal concentration and chemotactic activity was observed. Conclusion:These data stress on the important role of neutrophil and monocyte in host defense against dermatophyte infection. The data also indicate that neutrophil is a more active responder to fungal infection than monocyte. Keywords Material and Methods Study groupFifty-three patients with superficial fungal infections of glabrous skin attended the Dermatology and Venereology outpatient clinic of Al-Hussien University Hospital, were studied during a period from May 2000 to September 2001. They were 28 females their age ranged from 5-55 (25.3±14) years and 25 males their age ranged from 3-66 (30.2±16.6) years old. A full history was taken from all patients including age, job, site of infection, presence of predisposing factors and antifungal treatment. The duration of the disease in 51 patients varied from 112 weeks (recent infection). In the remaining two patients, the duration of infection was more than 12 months (chronic infection). The patients were suffering from superficial dermatophytosis; no other skin infections or systemic diseases were detected.Mycological examination was done for all patients. Antifungal sensitivity testing and chemotaxis assay were done for positive culture...

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Micropore filter assay of human granulocyte locomotion: Problems and solutions

Cited by 67 publications

References 8 publications

Production of chemotactic activity for polymorphonuclear leukocytes by cultured rat hepatocytes exposed to ethanol.

Production of chemotactic activity for polymorphonuclear leukocytes by cultured rat hepatocytes exposed to ethanol.

THE EFFECTS OF SOME ANTIRHEUMATIC DRUGS ON AN in vitro MODEL OF HUMAN POLYMORPHONUCLEAR LEUCOCYTE CHEMOKINESIS

Effect of Dermatophytes on Neutrophils and Monocytes Chemotaxis

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