Neutrophil chemotactic factors secreted fromToxoplasma gondii

Neutrophil chemotactic activity was detected in the fluid ofToxoplasma gondiicultures by the agarose plate and the Boyden chamber methods.Toxoplasmaculture fluid was obtained by incubating the tachyzoites at 37 °C in a 5% CO2atmosphere for 6 h in Dulbecco's modified Eagle's minimum essential medium containing 10% heat-inactivated foetal calf serum. Soluble extracts from tachyzoites had negligible activity, indicating that the chemotactic factors were metabolites secreted from tachyzoites. The chemotactic activity inToxoplasmaculture fluid depended on the number of tachyzoites and the period of incubation. The substances responsible for neutrophil chemotaxis were two heat-labile peptides with estimatedMr4·5 and 14 kDa with N-terminal groups free.

Download Full-text

The Inhibitory Effect of Heparin and Related Glycosaminoglycans on Neutrophil Chemotaxis

Thrombosis and Haemostasis ◽

10.1055/s-0038-1661157 ◽

1984 ◽

Vol 52 (02) ◽

pp. 134-137 ◽

Cited By ~ 69

Author(s):

Yaacov Matzner ◽

Gerard Marx ◽

Ruth Drexler ◽

Amiram Eldor

Keyword(s):

Specific Binding ◽

Anticoagulant Activity ◽

Neutrophil Migration ◽

Inhibitory Effect ◽

Chemotactic Factor ◽

Human Neutrophils ◽

Boyden Chamber ◽

Neutrophil Chemotaxis ◽

Inhibitory Effects ◽

Chemotactic Factors

SummaryClinical observations have shown that heparin has antiinflammatory activities. The effect of heparin on neutrophil chemotaxis was evaluated in vitro in the Boyden Chamber. This method enabled differentiation between the direct effects of heparin on neutrophil migration and locomotion, and its effects on chemotactic factors. Heparin inhibited both the random migration and directed locomotion of human neutrophils toward zymosan-activated serum (ZAS) and F-met-leu-phe (FMLP). Inhibition was found to be dependent on the concentrations of the heparin and of the chemotactic factors. No specific binding of heparin to the neutrophils could be demonstrated, and heparin’s inhibitory effects were eliminated by simple washing of the cells. When added directly to the chamber containing chemotactic factor, heparin inhibited the chemotactic activity of ZAS but not that of FMLP, suggesting a direct inhibitory effect against C5a, the principal chemotactic factor in ZAS.Experiments performed with low-molecular-weight heparin, N-desulfated heparin, dextran sulfate, chondroitin sulfate and dextran indicated that the inhibitory effects of heparin on neutrophil chemotaxis are not related to its anticoagulant activity, but probably depend on the degree of sulfation of the heparin molecule.

Download Full-text

Neutrophil chemotaxis, random migration, and adherence in patients with hyperthyroidism

Acta Endocrinologica ◽

10.1530/acta.0.1210817 ◽

1989 ◽

Vol 121 (6) ◽

pp. 817-820 ◽

Cited By ~ 5

Author(s):

B. Wolach ◽

B. Lebanon ◽

A. Jedeikin ◽

M. S. Shapiro ◽

L. Shenkman

Keyword(s):

Thyroid Function ◽

Chemotactic Activity ◽

Neutrophil Chemotaxis ◽

Phagocytic Cells ◽

Random Migration ◽

Significant Difference ◽

Neutrophil Adherence ◽

Hyperthyroid Patients ◽

Normal Controls ◽

Neutrophil Chemotactic Activity

Abstract. We have examined neutrophil adherence, chemotactic activity, and random migration in 35 hyperthyroid patients with Graves' disease and 106 normal volunteers. No statistically significant differences were found between granulocyte adherence of 17 hyperthyroid subjects (67 ± 15.6%) and 81 healthy volunteers (63.1 ± 17%). In 3 thyrotoxic patients, impaired neutrophil adherence was found, which resolved when thyroid function returned to normal. The neutrophil chemotactic activity of 32 normal controls was 107.5 ± 21.4 cells, and the random migration 36 ± 15.5 cells. No statistically significant difference was demonstrated in 13 hyperthyroid patients who had a neutrophil chemotactic activity of 102 ± 14.6 cells and a random migration of 31.2 ± 13.2 cells. Defective chemotactic activity and random migration was found in 2 patients. Neutrophil functions returned to normal in one of the two subjects who were re-evaluated when thyroid function recovered. In summary, 14% of hyperthyroid patients had impaired leukocyte functions. However, severe pyogenic infections are quite rare in hyperthyroid patients, indicating that the observed alterations in function of phagocytic cells are not clinically important.

Download Full-text

Circulating Histamine and Neutrophil Chemotactic Activity During Allergen-Induced Asthma: The Effect of Inhaled Antihistamines and Anti-Allergic Compounds

Clinical Science ◽

10.1042/cs0690063 ◽

1985 ◽

Vol 69 (1) ◽

pp. 63-69 ◽

Cited By ~ 7

Author(s):

D. J. R. Morgan ◽

I. Moodley ◽

D. R. Cundell ◽

B. D. Sheinman ◽

W. Smart ◽

...

Keyword(s):

Airflow Obstruction ◽

Isotonic Saline ◽

Forced Expiratory Volume ◽

Chemotactic Activity ◽

Boyden Chamber ◽

Plasma Histamine ◽

Double Blind ◽

Provocation Testing ◽

Bronchial Provocation ◽

Neutrophil Chemotactic Activity

1. Plasma histamine and serum neutrophil chemotactic activity (S-NCA) were measured in ten atopic asthmatic patients on four separate occasions after allergen bronchial provocation testing (BPT). 2. Single doses of inhaled sodium cromoglycate (SCG; 20 mg), clemastine (0.5 mg), ketotifen (0.5 mg) and isotonic saline (0.9% NaCl) placebo were administered 30 min before bronchial provocation testing in random order and double-blind. 3. The airflow obstruction after BPT was monitored by measurement of forced expiratory volume in 1s (FEV1). Plasma histamine was measured by the double-isotope radioenzymatic assay and S-NCA by a modified Boyden chamber technique. 4. A highly significant decrease in FEV1 after BPT occurred on the placebo pre-treatment visit (P < 0.001). Prior administration of inhaled SCG, clemastine and ketotifen significantly reduced the decrease in airflow obstruction seen after BPT when compared with placebo treatment (P < 0.01, P < 0.02, P < 0.05 respectively). 5. No significant alteration in plasma histamine was detected during allergen-induced airflow obstruction. 6. Levels of S-NCA were significantly higher 5, 10 and 15 min after BPT when compared with the pre-challenge level (P < 0.01, P < 0.01, P < 0.001 respectively). These levels were not significantly decreased when airflow obstruction was inhibited by the prior inhalation of SCG, clemastine or ketotifen.

Download Full-text

Bronchial epithelial cells release neutrophil chemotactic activity in response to tachykinins

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1992.263.2.l226 ◽

1992 ◽

Vol 263 (2) ◽

pp. L226-L231 ◽

Cited By ~ 2

Author(s):

S. G. Von Essen ◽

S. I. Rennard ◽

D. O'Neill ◽

R. F. Ertl ◽

R. A. Robbins ◽

...

Keyword(s):

Epithelial Cells ◽

Proteinase Inhibitors ◽

Bronchial Epithelial Cells ◽

Chemotactic Activity ◽

Induced Response ◽

Neurokinin A ◽

Maximal Effect ◽

Neutrophil Chemotaxis ◽

Bronchial Epithelial ◽

Neutrophil Chemotactic Activity

The purpose of this study was to determine whether substance P (SP), neurokinin A (NKA), and neurokinin B (NKB) induce the release of neutrophil chemotactic activity (NCA) from bovine bronchial epithelial cells (BBEC) and whether neutral endopeptidase (NEP), a membrane-bound metalloenzyme that hydrolyzes tachykinins, modulates these effects. BBEC monolayers were exposed to SP, NKA, and NKB in the absence or presence of phosphoramidon (10(-6) M), a selective NEP inhibitor, for 72 h. Using a modified blind-well in vitro neutrophil chemotaxis assay, we found that tachykinin-exposed BBEC culture supernatant fluids induced significant neutrophil chemotaxis compared with supernatants obtained from unstimulated BBEC. Maximal effect was observed after 48 h of incubation and at SP concentration of 10(-13) M [92 +/- 3 (SP) vs. 64 +/- 2 (media) cells/high-power field (HPF), mean +/- SE, n = 7, P less than 0.05]. Release of NCA was mediated by the COOH-terminal of the SP molecule. The rank order of potency of tachykinins in inducing release of NCA was SP greater than NKA = NKB. SP-induced response was significantly potentiated by phosphoramidon (109 +/- 3 vs. 92 +/- 3 cells/HPF, n = 7, P less than 0.05), whereas other proteinase inhibitors had no effect. The released NCA was composed of protein and lipid-soluble components. These data indicate that mammalian tachykinins induce the release of NCA from BBEC and that NEP modulates these effects. We suggest that tachykinins regulate neutrophil recruitment into the lower respiratory tract, in part, by inducing the release of NCA from airway epithelial cells.

Download Full-text

Infectivity of Theileria parva sporozoites following cryopreservation in four suspension media and multiple refreezing : evaluation by in vitro titration

Onderstepoort Journal of Veterinary Research ◽

10.4102/ojvr.v73i3.147 ◽

2006 ◽

Vol 73 (3) ◽

Cited By ~ 1

Author(s):

V. Mbao ◽

D. Berkvens ◽

T. Dolan ◽

N. Speybroeck ◽

J. Brandt ◽

...

Keyword(s):

East Coast ◽

Calf Serum ◽

Roswell Park Memorial Institute ◽

Foetal Calf Serum ◽

Theileria Parva ◽

Minimum Essential Medium ◽

Freezing Medium ◽

In Vitro Technique ◽

Phosphate Buffered Saline

Theileria parva sporozoite stabilates are used for immunizing cattle against East Coast fever and in in vitro sporozoite neutralization assays. In this study, we attempted to identify a cheaper freezing medium and quantified the infectivity loss of sporozoites due to refreezing of stabilates, using an in vitro technique. Pools of stabilates prepared using Minimum Essential Medium (MEM), Roswell Park Memorial Institute (RPMI 1640), foetal calf serum (FCS) and phosphate-buffered saline (PBS) were compared. All were supplemented with bovine serum albumin except the FCS. RPMI 1640 was as effective as MEM in maintaining sporozoite infectivity while the infectivity in PBS and FCS reached only 59 % and 67 %, respectively. In a second experiment, a stabilate based on MEM was subjected to several freeze-thaw cycles including various holding times on ice between thawing and refreezing. Refrozen stabilate gave an average sporozoite infectivity loss of 35 % per cycle. The results indicate that RPMI can be used as a cheaper freezing medium for T. parva stabilates and that refrozen stabilate doses need to be adjusted for the 35 % loss of infectivity.

Download Full-text

Cigarette Smoke Stimulates Release of Neutrophil Chemotactic Activity from Cultured Bovine Bronchial Epithelial Cells

Clinical Science ◽

10.1042/cs0880337 ◽

1995 ◽

Vol 88 (3) ◽

pp. 337-344 ◽

Cited By ~ 22

Author(s):

Shunsuke Shoji ◽

Ronald F. Ertl ◽

Sekiya Koyama ◽

Richard Robbins ◽

George Leikauf ◽

...

Keyword(s):

Epithelial Cells ◽

Cyclic Amp ◽

Cigarette Smoke ◽

Fetal Calf Serum ◽

Calf Serum ◽

Bronchial Epithelial Cells ◽

Chemotactic Activity ◽

Bronchial Epithelial ◽

Lipoxygenase Inhibitors ◽

Neutrophil Chemotactic Activity

1. Recruitment of neutrophils into the airway is a prominent feature of chronic bronchitis, a syndrome often associated with exposure to cigarette smoke. Since bronchial epithelial cells are the ‘first’ lung cells to come into contact with smoke, these cells may be responsible for neutrophil recruitment into the airway by release of neutrophil chemotactic activity in response to cigarette smoke. 2. To evaluate this hypothesis, we prepared bovine bronchial epithelial cells and measured their ability to release neutrophil chemotactic activity following exposure to cigarette smoke. Bronchial epithelial cells were prepared by overnight digestion with protease, filtered through 100-μm Nitex mesh and resuspended in Dulbecco's modified Eagle medium supplemented with 10% fetal calf serum and antibiotics and cultured at 2×106 cells in 2 ml of medium in 35-mm culture dishes. After 4 days, dishes were rinsed and refed with medium without fetal calf serum and incubated with and without 1:10 diluted smoke extract for 6 h at 37°C. The neutrophil chemotactic activity of the supernatant fluids was measured by the blindwell chamber technique. 3. Cigarette smoke itself added to medium did not stimulate chemotaxis. In contrast, cigarette smoke did stimulate the release of neutrophil chemotactic activity from bovine bronchial epithelial cells [15 ± 3 (control) versus 74 ± 5 (smoke), P < 0.01]. 4. This neutrophil chemotactic activity was dialysable, pepsin and acid stable, heat sensitive and lipid extractable. Sephadex G-75 column chromatography demonstrated two peaks of neutrophil chemotactic activity. 5. The lipoxygenase inhibitors diethylcarbamazine and nordihydroguaratic acid both diminished the release of chemotactic activity, suggesting that the activity may be a lipoxygenase product(s). 6. Dibutyryl cyclic AMP also blocked the smoke-stimulated release of neutrophil chemotactic activity, suggesting that its release may be regulated by intracellular cyclic AMP. 7. Thus, bovine bronchial epithelial cells release neutrophil chemotactic activity in response to cigarette smoke, suggesting that bronchial epithelial cells may be modulators of the airway inflammatory response caused by cigarette smoke.

Download Full-text

Cultivation of infective forms ofTrypanosoma congolensefrom trypanosomes in the proboscis ofGlossina morsitans

Parasitology ◽

10.1017/s0031182000041883 ◽

1981 ◽

Vol 82 (1) ◽

pp. 81-95 ◽

Cited By ~ 41

Author(s):

M. A. Gray ◽

I. Cunningham ◽

P. R. Gardiner ◽

A. M. Taylor ◽

A. G. Luckins

Keyword(s):

Culture Medium ◽

Primary Cultures ◽

Calf Serum ◽

Tsetse Flies ◽

Foetal Calf Serum ◽

Culture Vessel ◽

Minimum Essential Medium ◽

Glossina Morsitans ◽

Plastic Surface ◽

Dermal Collagen

SUMMARYTwo stocks ofTrypanosoma congolensewere established in culture at 28 °C using trypanosomes from the proboscides of infectiveGlossina morsitans. Successful primary cultures were initiated by placing an infected tsetse proboscis beside a bovine dermal collagen explant in Eagle's minimum essential medium supplemented with foetal calf serum. The trypanosomes multiplied rapidly in the medium and also gradually formed an adherent layer on the plastic surface of the culture vessel. Three primary cultures produced organisms infective for mice from 14, 20 and 35 days after initiation and thereafter continuously until days 76, 76 and 52 when they were discarded. Four attempts to initiate infective cultures using infected tsetse proboscides in medium without dermal explants were unsuccessful. When trypanosomes from primary cultures were placed in culture medium with proboscides from uninfected tsetse flies, the parasites multiplied, formed an adherent layer in the culture flasks and were seen in the proboscides within 24 h. A line of 1 stock was serially sub-passaged in this way 4 times during a period of 215 days. Infectivity titrations in mice indicated that primary and sub-passaged cultures each contained similar numbers of infective organisms. Another line of the same stock was also sub-passaged 4 times in medium alone over a period of 186 days. These sub-cultures again retained infectivity for mice, but titrations showed a decrease in infective organism production in the 4th sub-culture. Primary and sub-passaged cultures all included a variety of morphologically different developmental forms ofT. congolense, closely resembling those described in the labrum and hypopharynx ofGlossinaby previous workers. Short metacyclic-like trypanosomes and organisms with proteinaceous surface coats were present in infective cultures. Cultures were successfully re-established after cryopreservation at −196 °C and retained the ability to produce infective organisms.

Download Full-text

Exogeneous factors are not necessary in attachment, spreading and polarization of hela cells

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100082418 ◽

1978 ◽

Vol 36 (2) ◽

pp. 310-311

Author(s):

W. Liebrich

Keyword(s):

Hela Cells ◽

Calf Serum ◽

Glass Tube ◽

Serum Free Medium ◽

Nacl Solution ◽

Free Medium ◽

Minimum Essential Medium ◽

Serum Free ◽

All Solutions ◽

Glass Support

HeLa cells were grown for 2-3 days in EAGLE'S minimum essential medium with 10% calf serum (S-MEM; Seromed, München) and then incubated for 24 hours in serum free medium (MEM). After detaching the cells with a solution of 0. 14 % EDTA and 0. 07 % trypsin (Difco, 1 : 250) they were suspended in various solutions (S-MEM = control, MEM, buffered salt solutions with or without Me++ions, 0. 9 % NaCl solution) and allowed to settle on glass tube slips (Leighton-tubes). After 5, 10, 15, 20, 25, 30, 1 45, 60 minutes 2, 3, 4, 5 hours cells were prepared for scanning electron microscopy as described by Paweletz and Schroeter. The preparations were examined in a Jeol SEM (JSM-U3) at 25 KV without tilting.The suspended spherical HeLa cells are able to adhere to the glass support in all solutions. The rate of attachment, however, is faster in solutions without serum than in the control. The latter is in agreement with the findings of other authors.

Download Full-text