bluuebunny/crossref_metadata_2025 huggingface dataset:
Cytokines in Clinical and Experimental Transplantation
Authors: Vossen Ann C. T. M. , Savelkoul Huub F. J.
Published: March 2007

Allograft rejection is a complex process, which requires
interactions between different cell types and a variety of soluble
factors, such as cytokines. In this review we discuss the role of
cytokines in the induction and effector phases of the rejection
process and in the induction and maintenance of allospecific graft
tolerance. Furthermore, we discuss the feasibility of clinical graft
function monitoring by measuring cytokines and the possibilities for
intervention in the cytokine network in order to inhibit graft
rejection and eventually obtain graft acceptance.
A comparison of the Antileukaemic Effects of Recombinant HumanTumour Necrosis Factor‐α and its Muteins on Leukaemia L1210 and Leukaemia P388 in Mice
Authors: Warzocha K. , Góra-Tybor J. , Kwinkowski M. , Szymańska B. , Robak T.
Published: March 2007

We investigated the influence of recombinant human tumour necrosis
factor alpha (TNF‐α) and its derivatives termed muteins III, V,
VI—in which the first 3 to 7 amino acids of native TNF‐α have been
replaced—on the survival time of mice inoculated with leukaemia
L1210 or leukaemia P338. TNF‐α prolonged the survival of mice with
leukaemia L1210 but did not have any therapeutic activity in
leukaemia P388‐bearing mice. Muteins‐treated mice with leukaemia
P388 lived longer than animals receiving TNF‐α, while those
inoculated with leukaemia L1210 did not show any significant
prolongation of life compared with the TNF‐α treated group. The
results presented in this report indicate that the antileukaemic
activity of TNF‐α is governed at least in part by the nature of the
N‐terminal amino acids.
Analysis of Signals Leading to Differentiation of Immature HL‐60 after Administration of IL‐4
Authors: Vassiliadis Simon
Published: March 2007

Treatment of acute myeloid leukaemia (AML) cells with
differentiation agents leads not only to the acquisition of normal
phenotypes but also contributes to the understanding of special
immuno‐haematology issues. For instance, induction of HLA‐DR
antigens on human promyelocytic leukaemia HL‐60 cells by
interleukin‐4 (IL‐4) is of pivotal importance in immunology not
only because class II expression is prerequisite to antigen
recognition and response but also because IL‐4 participates in a
plethora of inflammatory or non‐inflammatory reactions. At the same
time, the same observation coupled with an increase in Mac‐1, mature
monocyte marker, is revealing ways to haematologists for converting
malignant cases to normal situations. Based on previous reports that
HLA‐DR induction by IL‐4 in the HL‐60 system is mediated via the
G‐protein system (p21ras), this study was undertaken in order to
define the intermediate signalling steps followed by this agent from
the moment it is added to cultures to the differentiated cellular
form obtained. It is proposed that IL‐4 increases p21ras which in turn
suppresses the HL‐60 cells′ p34cdc2 constitutive expression. This
inhibition appears to be responsible for the subsequently.observed
cessation ofgrowth. Concomitant to decreased cellular proliferation,
HI‐DR antigen expression increases, a finding that matches the
initially mentioned induction of p21ras since its inhibition abolishes HI‐DR upregulation.
Role of Acetylsalicylic Acid in Cytokine Stimulation of Macrophagesin Antibody‐Dependent Cellular Cytotoxicity (ADCC)
Authors: Jäpel M. , Lötzerich H. , Rogalla K.
Published: March 2007

In addition to the spectrum of biological action already known to be
exhibited by acetylsalicylic acid (ASA) as an analgesic,
anti‐inflammatory and platelet aggregation inhibitor, there is
growing evidence of a stimulatory effect on the immune system. ASA
has been found to increase the production ofcytokines and to
increase the activity of various leukocytes. The action of ASA on
the activity of mouse peritoneal macrophages was therefore
investigated in the present study. Therapeutically effective
concentrations of ASA, which are known to decrease levels of
prostaglandins, had neither a stimulating nor an inhibiting
influence on antibody‐dependent cellular cytotoxicity (ADCC) or on
the binding capacity of macrophages with regard to SW 948 tumour
cells. Likewise ASA had little or no adverse effect on the capacity
of the macrophages for stimulation by interferon‐gamma (IFN‐gamma)
and interleukin‐4 (IL‐4). Taken together, the immunostimulant effect
of ASA shown in the literature as an increased production of
interleukin‐2 (IL‐2) and IFN, could not be confirmed on the basis of
the macrophage cytotoxiclty.
Acute Phase Protein Response and Polymorphonuclear LeukocyteCathepsin G Release After Slow Interleukin‐1 Stimulation in the Rat
Authors: Björk Peter , Gudmundsson Thorarinn , Ohlsson Kjell
Published: March 2007

In this work we have studied the acute phase protein response and
degranulation of polymorphonuclear leukocytes in vivo in the rat
after a slow interleukin‐1β stimulation. A total dose of 1 μg, 2 μg,
4 μg and 0 μg (controls with only vehicle) of interleukin‐1β was
released from osmotic minipumps over a period of 7 days. The pumps
were implanted subcutaneously. A cystic formation was formed around
the pumps that contained interleukin‐1β whereas no tissue reaction
was seen around pumps containing only vehicle. Besides flbroblasts
the cyst wall contained numerous polymorphonuclear leukocytes which
were positively stained for cathespin G. α2‐macroglobulin,
α1‐inhtbitor‐3, α1‐proteinase inhibitor, albumin and C3 were
measured by electroimmunoassay and all showed plasma concentration
patterns that were dose‐dependent to the amount of interleuktn‐1β
released. Fibrinogen in plasma was elevated in the control group but
showed decreased plasma values with higher doses of interleukin‐1β
released. All animals showed increased plasma levels of cathespin G
but the lowest levels for cathespin G were seen for the highest
interleukin‐1β dose released. It was clearly seen that a slow
continuous release of interleukin‐1β in vivo caused an inflammatory
reaction. Plasma levels for the proteins analysed all showed a
similar pattern, namely an initial increase or decrease of plasma
concentration followed by a tendency to normalization of plasma
values. It was concluded that a long‐term interleukin‐1β release
could not sustain the acute phase protein response elicited by the
initial interleukin‐1β release.
Endothelin‐1 Production by Normal Human Cultured Keratinocytes and its Regulation
Authors: Inoue Hajime , Wakisaka Nagaoki , Tane Nobuhiro , Ando Kazumasa , Isono Emiko , Yamanaka Miwa , Aihara Masaki , Ishida Hirotomo
Published: March 2007

The possibility that cultured keratinocytes produce
endothelins were investigated. The results showed that
cultured keratinocytes derived from normal human
skin produce endothelin‐1. Moreover, keratinocyte
endothelin‐1 production was completely inhibited by the
presence of actinomycin D in the medium. As in the case
of endothelial cells, recombinant interleukin‐1β was
capable of promoting endothelin‐1 production in
keratinocytes, whereas herapin inhibited it. Thrombin
also inhibited endothelin‐1 production. These results indicate
that the mechanism of endothelin‐1 production in
keratinocytes is slightly different from the mechanism in
vascular endothelial cells.
Inhaled Nitric Oxide Reverses Vascular and Respiratory Effects of ET‐1 and PAF in Pigs
Authors: Albertini Mariangela , Clement Maria Giovanna , Dimori Milena
Published: March 2007

In anaesthetized paralysed, mechanically ventilated pigs, the
vascular and respiratory effects of 80 ppm nitric oxide (NO) inhaled
for 6 min were evaluated. To evoke different levels of smooth
muscle contraction ET‐1 or PAF, mediators involved in pulmonary
disorders, were used. In control conditions, inhaled NO caused
selective pulmonary vasodilatation without affecting respiratory
resistances. This pulmonary vascular activity influenced the
distensibility of the respiratory system and decreased inspiratory
work. ET‐1 administration significantly increased pulmonary arterial
pressure and modestly changed mechanical properties of the
respiratory system, while PAF caused potent vasoconstriction and
bronchoconstriction associated with a marked change in
volume‐pressure relationship. In both cases, the changes in vascular
and mechanical properties of the respiratory system increased
inspiratory work. The vascular and respiratory activities of inhaled
NO were correlated with preconstriction levels. The data show that
the combination of vascular and respiratory effects improves
pulmonary function, suggesting that inhalation of NO is a possible
therapeutic approach for obstructive and inflammatory pulmonary
diseases.
Differential Proliferative Characteristics of Alveolar Fibroblastsin Interstitial Lung Diseases: Regulative Role of IL‐1 and PGE2
Authors: Fireman Elizabeth , Ben Efraim Shlomo , Greif Joel , Peretz Hava , Kivity Shmuel , Topilsky Marcel , Rodrig Yosef , Yellin A. , Apte Ron N.
Published: March 2007

Fibroblasts (Fb) from patients with sarcoidosis (SA) and
hypersensitivity pneumonitis (HP) exhibited a lower proliferative
capacity compared with Fb obtained from control (CO) and diffuse
interstitial fibrosis patients (DIF). Proliferation of Fb from SA or
lip patients was suppressed by autologous LPS‐stimulated alveolar
macrophages (AM) supernatants but not by those from CO patients.
Similarly, alveolar macrophages (AM) derived supernatant, obtained
from CO, did not suppress the proliferation of SA and HP Fb. AM from
SA and HP patients secreted higher amounts of IL‐1α and β
compared with controls and compared with Fb from SA and HP patients.
Steady levels of IL‐1α and βmRNA were expressed in
unstimulated and stimulated cultures. Fb from SA and HP patients
could be stimulated by LPS to secrete significantly higher levels of
PGE2 than those detected in supernatants from LPS
stimulated Fb of DIF patients. Only the proliferation of Fb from SA
and HP patients was sensitive to amounts of IL‐1 equivalent to those
detected in the lung of these diseases. As SA and HP are two
diseases where irreversible deterioration occurs in only 20%
of the patients, we hypothesize that mediators in the lung may
modulate Fb proliferation. IL‐1 of AM origin and PGE2 of
Fb origin secreted at high levels, may be candidates for this
suppression because it was abrogated by anti IL‐1β and indomethacin.
Inhibition of Immune Complex‐Induced Inflammation by A small Molecular Weight Selectin Antagonist
Authors: Nair X. , Todderud G. , Davern L. , Lee D. , Aruffo A. , Tramposch K. M.
Published: March 2007

The anti‐inflammatory effect of a small molecular weight antagonist
of P‐ and E‐selectin‐dependent cell adhesion was examined. The
glycolipid sulphatide was shown to block the adherence of
thrombin‐activated rat platelets to HL‐60 cells. This interaction is
known to be dependent on P‐selectin. The rat dermal reverse passive
Arthus reaction was used to assess the effect of sulphatide on a
neutrophil dependent inflammatory response. Sulphatide
dosedependently blocked both the vascular permeability increase and
cell infiltration after intraperitoneal administration. These
results show that a small molecular weight compound which blocks P‐
and E‐selectin dependent adhesion in vitro can
effectively block the inflammation due to immune complex deposition.
A compound with this type of profile may have therapeutic potential
in the treatment of immune complex mediated diseases.
Effect of Nedocromil Sodium on Polymorphonuclear Leukocyte Plasma Membrane
Authors: Kantar A. , Oggiano N. , Giorgi P. L. , Coppa G. V. , Gabbianelli R. , Bruni S. , Cutrona F. M. , Fiorini R.
Published: March 2007

The effect of nedocromil sodium on the plasma membrane fluidity of
polymorphonuclear leukocytes (PMNs) was investigated by measuring
steady‐state fluorescence anisotropy of
1‐[4‐trimethylammonium‐phenyl]‐6‐phenyl‐ 1,3,5‐hexatriene (TMA‐DPH)
incorporated in the membrane. Our results show that nedocromil
sodium 300 μM significantly decreased membrane fluidity of PMNs.
The decrease in membrane fluidity of PMNs induced by fMLP was
abolished in the presence of nedocromil sodium. These data suggest
that nedocromil sodium interferes with the plasma membranes of PMNs
and modulates their activities.
Corticosteroid‐Sparing Effect of Chromoglycate Sodium and Nedocromil
Authors: Capristo A. F. , del Giudice Jr M. Miraglia , Alfaro C. , Maiello N.
Published: March 2007

The most appropiate management for bronchial asthma is the control
of airway inflammation. Corticosteroids are the most effective
anti‐inflammatory drugs available, but they have a number of side
effects; most of these are dose‐dependent. In children, asthma
control should be accomplished with low steroid doses possibly given
by inhalation. In a double‐bind placebo‐controlled crossover study a
group of children with mild to moderate asthma received NED 16
mg/day or BDP 400 μg/day. Values for FEV1, PEF, symptoms use
ofbronchodilators overlapped, whereas bronchial hyper‐responsiveness
assessed by histamine bronchoprovocation challenge was better with
BDP than NED. In another case, one boy with high bronchial
hyper‐reactivity assessed by provocation test with hypertonic
solution, experienced a significant improvement only after 2 weeks
of therapy with Deflazacort (2 mg/Kg/day) followed by 4 months on
combined treatment with NED (16 mg/day) and BDP (300 μ/day). Authors
conclude that NED could have a steroidsparing effect over long‐term use.
The Effect of Cromolyn Sodium and Nedocromil Sodium Administered by A pressurized Aerosol with A spacerDevice on Exercise‐Induced Asthma in Children
Authors: de Benedictis F. M. , Tuteri G. , Niccoli A. , Mezzetti D. , Rossi L. , Bruni L.
Published: March 2007

To compare the effectiveness of cromolyn sodium (CS)
(10 mg) and nedocromil sodium (NS) (4 mg) administered
by a metered dose inhaler (MDI) with a spacer
device in preventing exercise‐induced asthma (EIA), eight
asthmatic children with EIA were studied in a
randomized double‐blind, cross‐over, placebo‐controlled
study, CS and NS provided significant, comparable protection
from EIA and both were better than placebo. We
conclude that CS and NS administered by a pressurized
aerosol with a spacer device provide equal protection
against EIA in children.
Platelet‐activating factor mediates trinitrobenzene induced dysmotility in the left colon
Authors: Longo W. E. , Standeven J. , Chandel B. , Deshpande Y. , Vernava A. M. , Kaminski D.
Published: March 2007

We investigated whether TNB alters colonic tissue levels of PAF at the expense of changes in colonic motility in the isolated perfused rabbit left colon. Colonic inflammation was induced by the intracolonic administration of 0.25 ml of 50% ethanol containing 30 mg of trinitrobenzene. Strain gauge transducers were sewn onto the serosal surface of the colon to evaluate colonic motility. TNB administration increased tissue levels of PAF and increased the average force of each colonic contraction. Pretreatment with the PAF receptor antagonist WEB‐2170 prior to TNB infusion decreased tissue concentrations of PAF and ameliorated the altered motility parameters seen with TNB alone. These results suggest that PAF is stimulated by TNB and may participate in colonic dysmotility during inflammatory states.
Inducible nitric oxide synthase and guinea‐pig ileitis induced by adjuvant
Authors: Seago N. D. , Thompson J. H. , Zhang X.-J. , Eloby-Childress S. , Sadowska-Krowicka H. , Rossi J. L. , Currie M. G. , Manning P. T. , Clark D. A. , Miller M. J. S.
Published: March 2007

We sought to establish a model of inflammatory bowel disease by augmenting the activity of the local immune system with Freund′s complete adjuvant, and to determine if inducible nitric oxide synthase (iNOS) expression and peroxynitrite formation accompanied the inflammatory condition. In anaesthetized guinea‐pigs, a loop of distal ileum received intraluminal 50% ethanol followed by Freund′s complete adjuvant. Control animals were sham operated. When the animals were killed 7 or 14 days later, loop lavage fluid was examined for nitrite and PGE2 levels; mucosal levels of granulocyte and macrophages were estimated by myeloperoxidase (MPO) and N‐acetyl‐D‐glucosaminidase (NAG) activity, respectively. Cellular localization if iNOS and peroxynitrite formation were determined by immunohistochemistry with polyclonal antibodies directed against peptide epitopes of mouse iNOS and nitrotyrosine, respectfully. Adjuvant administration resulted in a persistent ileitis, featuring gut thickening, crypt hyperplasia, villus tip swelling and disruption, and cellular infiltration. Lavage levels of PGE2 and nitrite were markedly elevated by adjuvant treatment. Immunoreactive iNOS and nitrotyrosine bordered on detectability in normal animals but were markedly evident with adjuvant treatment at day 7 and particularly day 14. Immunohistochemistry suggested that enteric neurons and epithelia were major sites of iNOS activity and peroxynitrite formation. We conclude that local administration of adjuvant establishes a chronic ileitis. Inducible nitric oxide synthase may contribute to the inflammatory process.
Role of lysosomal enzymes released by alveolar macrophages in the pathogenesis of the acute phase of hypersensitivity pneumonitis
Authors: Pérez-Arellano J. L. , Barrios M. N. , Martín T. , Sánchez M. L. , Buitrago J. M. González , Jiménez A.
Published: March 2007

Hydrolytic enzymes are the major constituents of alveolar macrophages (AM) and have been shown to be involved in many aspects of the inflammatory pulmonary response. The aim of this study was to evaluate the role of lysosomal enzymes in the acute phase of hypersensitivity pneumonitis (HPs). An experimental study on AM lysosomal enzymes of an HP‐guinea‐pig model was performed. The results obtained both in vivo and in vitro suggest that intracellular enzymatic activity decrease is, at least partly, due to release of lysosomal enzymes into the medium. A positive but slight correlation was found between extracellular lysosomal activity and four parameters of lung lesion (lung index, bronchoalveolar fluid total (BALF) protein concentration, BALF LDH and BALF alkaline phosphatase activities). All the above findings suggest that the AM release of lysosomal enzymes during HP is a factor involved, although possibly not the only one, in the pulmonary lesions appearing in this disease.
Defective antigen presentation by monocytes in ESRD patients not responding to hepatitis B vaccination: impaired HBsAg internalization and expression of ICAM‐1 and HLA‐DR/Ia molecules
Authors: Stachowski J. , Barth C. , Pollok M. , Michałkiewicz J. , Madaliński K. , Maciejewski J. , Baldamis C. A.
Published: March 2007

This study was undertaken to evaluate the monocyte function of uraemic non‐responders to hepatitis B vaccination. Therefore, some parameters concerning antigen processing by monocytes (Mo) as antigen presenting cells (APC) were analysed. It was found that in uraemic non‐responders, (1) the internalization of HBsAg by monocytes was significantly decreasjed—HBsAg complexed with specific IgG or as immune complex isolated from patients is better internalized compared with free HBsAg; (2) during antigen presentation the expression of adhesion (ICAM‐1) and accessory (HLA‐DR/Ia) molecules was significantly decreased in uraemic patients, especially in non‐responders; and (3) impaired internalization of HBsAg as well as a decrease in ICAM‐1 and HLA‐DR/Ia expression, correlated well with the blunted proliferation of CD4+ T cells stimulated by autologous monocytes induced by HBsAg.
Possible bi‐directional link between ETA receptors and protein kinase C in rat blood vessels
Authors: Northover A. M. , Northover B. J.
Published: March 2007

Possible links have been investigated between activation of protein kinase C (PKC) and endothelin (ET) production by small blood vessels. Perfusion pressures were recorded from rat isolated mesenteric artery, with or without the small intestine attached, before and after addition to the perfusate of either ET‐1, ET‐3 or the PKC activator 12‐deoxyphorbol 13‐phenylacetate (DOPPA). Rises in perfusion pressure in response to ET‐1 (10−8 M)or DOPPA (10−6 M) were reduced significantly by pre‐treatment with either the ETA receptor antagonist PD151242 (10−6 M) or the PKC inhibitor Ro 31‐8220 (10−6 M). ET‐3 (10−8 M) had a significant, albeit small, effect only when the gut was still attached to the mesentery. Inthis latter preparation ET‐1 and DOPPA increased the permeability of villi microvessels to colloidal carbon in the perfusate. This effect of DOPPA was reduced by pre‐treatment with either PD151242 or Ro 31‐8220, but the effects of ET‐1 were reduced significantly only by Ro 31‐8220. ET‐3 (10−8 M) was without effect. The results suggest a possible bi‐directional link between ETA receptors and PKC in the intestinal vasculature.
Modulation of nitric oxide synthase activity in macrophages
Authors: Jorens P. G. , Matthys K. E. , Bult H.
Published: March 2007

L‐Arginine is converted to the highly reactive and unstable nitric oxide (NO) and L‐citrulline by an enzyme named nitric oxide synthase (NOS). NO decomposes into other nitrogen oxides such as nitrite () and nitrate (), and in the presence of superoxide anion to the potent oxidizing agent peroxynitrite (ONOO−). Activated rodent macrophages are capable of expressing an inducible form of this enzyme (iNOS) in response to appropriate stimuli, i.e., lipopolysaccharide (LPS) and interferon‐γ (IFNγ). Other cytokines can modulate the induction of NO biosynthesis in macrophages. NO is a major effector molecule of the anti‐microbial and cytotoxic activity of rodent macrophages against certain micro‐organisms and tumour cells, respectively. The NO synthesizing pathway has been demonstrated in human monocytes and other cells, but its role in host defence seems to be accessory. A delicate functional balance between microbial stimuli, host‐derived cytokines and hormones in the microenvironment regulates iNOS expression. This review will focus mainly on the known and proposed mechanisms of the regulation of iNOS induction, and on agents that can modulate NO release once the active enzyme has been expressed in the macrophage.
The effect of lysolecithin on prostanoid and platelet‐activating factor formation by human gall‐bladder mucosal cells
Authors: Nag M. K. , Deshpande Y. G. , Beck D. , Li A. , Kaminski D. L.
Published: March 2007

It has been demonstrated that lysolecithin (lysophosphatidyl choline, LPC) produces experimental cholecystitis in cats mediated by arachidonic acid metabolites. LPC is a cytolytic agent that has been postulated as a contributing factor in the development of cholecystitis in humans. The purpose of this research was to evaluate the effect of LPC on human gall‐bladder mucosal cell phospholipase A2 and cyclooxygenase activity. Gall‐bladder mucosal cells were isolated from the gall‐bladders of patients undergoing routine cholecystectomy. Fresh, isolated cells were maintained in tissue culture and stimulated with varying doses of LPC. Platelet‐activating factor concentration was quantitated as an index of phospholipase A2 activity and prostanoids were measured as an index of cyclooxygenase activity. Also, the effect of LPC on cyclooxygenase 1 and 2 expression in microsomal protein was evaluated. LPC caused dose related increases in 6‐keto‐PGF1α and PAF produced by human gall‐bladder mucosal cells. Exposure of human gall‐bladder mucosal cells to LPC failed to elicit expression of constitutive cyclooxygenase‐1, while the expression of inducible cyclooxygenase‐2 was increased. The results of this study indicate that LPC induces the formation of prostanoids and PAF by human gall‐bladder mucosal cells, suggesting that this substance may promote the development of gall‐bladder inflammation.
Relevance of histamine and tryptase concentrations in nasal secretions after nasal challenges with phosphate buffered saline and allergen
Authors: Wang D. , Clement P. , Smitz J. , Derde M.-P.
Published: March 2007

In this prospective study, a quantitative determination of histamine and tryptase in nasal secretions after nasal phosphate buffered saline (PBS) and allergen challenge was performed in 18 atopic patients who were compared with ten non‐allergic healthy volunteers. The aim of the study was to determine the normal and pathological concentrations of these important mediators in nasal secretions. The second objective was to test the relevance of these two mast cell secreted mediators after nasal challenge. Results showed that the concentrations of tryptase in almost all samples were under the minimal detection limit (< 0.5 μU/g) and only a sigrtificant increase of tryptase (median, 28 μU/g) occurred immediately after nasal allergen challenge in the patient group. Histamine concentration significantly increased after every nasal PBS challenge (median, 69 ng/g after first PBS challenge and 165 ng/g after second PBS challenge) in the control group, as well as in the patient group after both PBS (median, 69 ng/g) and allergen (median, 214 ng/g) challenge. On the other hand, a rapid onset of sneezing and increase in nasal airway resistance was experienced only in the patient group after nasal allergen challenge, but did not occur after PBS challenge even though the histamine concentrations significantly increased in both groups. This study suggests that tryptase is a more preferable marker than histamine in quantitative monitoring of mast cell activation especially during the early phase nasal allergic reaction.
Nitric oxide mediates interleukin‐1 induced inhibition of glycosaminoglycan synthesis in rat articular cartilage
Authors: Järvinen T. A. H. , Moilanen T. , Järvinen T. L. N. , Moilanen E.
Published: March 2007

Interleek‐1β (IL‐1) is a key mediator of cartilage matrix degradation in osteoarthritis and rheumatoid arthritis. It was found that the IL‐1‐induced suppression of glycosaminoglycan (GAG) synthesis in rat articular cartilage occurred simultaneously with the accumulation of nitrite (a metabolite of nitric oxide (NO) in aqueous milieu) in the culture medium. NO‐synthase inhibitors, L‐NMMA and L‐NIO, inhibited both these IL‐1 effects. Dexamethasone suppressed GAG synthesis additively to IL‐1, but did not alter nitrite accumulation. Three NO‐donors (GEA 3175, SNAP and SIN‐1) also had an inhibitory effect on cartilage GAG synthesis. Therefore, it is concluded that IL‐1 induced suppression of GAG synthesis in rat articular cartilage is mediated by the production of NO.
Beneficial haemodynamic effect of indomethacin during endotoxin shock in anaesthetized pigsputative involvement of nitric oxide?
Authors: Mózes T. , Gelderen E. M. van , Mylecharane E. J. , Saxena P. R.
Published: March 2007

Endotoxin shock was induced in 31 anaesthetized pigs by infusion of 5 μg/kg of Escbeicbia coli endotoxin (LPS) over 60 min into the superior mesenteric artery. Fifteen of these pigs died within 30 min of the start of LPS infusion whereas the remaining 16 survived the experimental period of 2 h. In a group of nine pigs indomethacin (2 mg/kg, i.v.)was inected 20–25 rain after the start of LPS infusion at which time mean arterial blood pressure (MABP) had decreased below 40 mmHg indicating imminent death. Indomethacin immediately reversed the hypotension. In another group of five pigs, NG‐nitro L‐arginine‐methyl ester (L‐NAME, 1 and 3 mg/kg)was iniected 10 and 5 min, respectively, before the expected death without any beneficial effect on the hypotension. Three rain after the last dose of L‐NAME, indomethacin (2 mg/kg, i.v.) was iniected. In three animals the hypotension was reserved by indomethacin, although this beneficial effect was delayed in comparison with the LP‐Streated group not receiving L‐NAME. Four pigs were pretreated with L‐NAME, 3 mg/kg, i.v., 10 min prior to LPS infusion. All pretreated animals tended to die within 30 min of the start of the LPS infusion. Five rain before the expected death (20–25 rain after the start of LPS infusion) indomethacin (2 mg/kg) was inected. In three of these animals indomethacin reversed hypotenston and prevented death. Interestingly, this rise in the MABP developed very slowly. These results suggest that the beneficial effect of indomethacin in endotoxin shock might be related partially to interference with nitric oxide, which is not the only factor determining blood pressure levels during endotoxic shock.
Inhibition of neutrophil migration by a selective inhibitor of matrix metalloproteinase: analysis by intravital microscopy
Authors: Oda T. , Katori M. , Hatanaka K. , Nagai Y.
Published: March 2007

Observation of the microcirculation of the hamster cheek pouch by intravital microscopy revealed five steps of neutrophil migration from the venules after topical application of leukotriene B4 to the microvasculature: rolling along the venular wall (Step 1), adhesion to it (Step 2), disappearance from the vascular lumen (Step 3), presence between the endothelial cells and the subendothelial basement membrane (Step 4) and passage through the basement membrane (Step 5). The present study was performed to examine whether a metalloproteinase inhibitor inhibits neutrophil migration at any of the above five steps. Chymostatin and leupeptin did not inhibit any of these five steps. In contrast, FN‐439, a selective inhibitor of matrix metalloproteinase, reduced the number of neutrophils in the perivascular space without affecting Steps 1 to 3. It was concluded that neutrophils may use metalloproteinase (collagenase/gelatinase) to penetrate the subendothelial basement membrane.
Androgen metabolism and inhibition of interleukin‐1 synthesis in primary cultured human synovial macrophages
Authors: Cutolo M. , Accardo S. , Villaggio B. , Barone A. , Sulli A. , Balleari E. , Bason C. , Felli L. , Granata O. M. , Amodio R. , Castagnetta L.
Published: March 2007

The presence of androgen receptors on synovial macrophages in human normal and rheumatoid synovial tissues has been described previously. It is now reported that primary cultured human macrophages obtained from normal and rheumatoid synovia express functional androgen receptors. We have investigated the capacity of cultured macrophages to metabolize androgens and have found that these cells were capable of metabolizing testosterone to the bioactive metabolite dihydrotestosterone. Therefore, macrophages contain the key enzymes of steroidogenesis, in particular the 5α‐treductase. Furthermore, interleukin‐1β production by primary cultured rheumatoid macrophages was analysed, following exposure to physiological concentrations of testosterone (10−8 M). A significant decrease of IL‐1β levels in conditioned media after 24 h (p < 0.05) was observed. It is concluded that androgens may act directly on human macrophages and may interfere with some of their functions via receptor‐dependent mechanisms.
Interleukin‐2 receptor β chain as a possible target for low doses of mafosfamide
Authors: Pukhalsky A. , Toptygina A. , Khaidukov S.
Published: March 2007

The 7‐day cytotoxic lymphocytes (CTL) induced in mixed lymphocyte culture express only the chain of the interleukin‐2 receptor (IL‐2R). In the present study this fact has been confirmed in a murine semi‐allogeneic system. The ability of low doses of mafosfamide (Mf) to affect IL‐2‐induced CTL proliferation has been demonstrated. It was also shown that IL‐2 activated resting suppressor cells. The pretreatment of the suppressor cells with either monoclonal antibodies (mAbs) against the p75 chain of IL‐2R, or with Mf abolished the suppressive effect of these cells. No restoration of the proliferative response occurred when the anti‐IL‐2Rα mAb had been used. Flow cytometry analysis of 7‐day CTL was carried out with mAbs against the α and β chains of IL‐2R. CTL treatment with Mf inhibited anti‐IL‐2Rβ mAb binding. It may be assumed that the anti‐proliferative effects of Mf which have been demonstrated in this paper, were a result of blocking the IL‐2R β chain.
Production of lipopolysaccharide‐induced tumour necrosis factor during influenza virus infection in mice coincides with viral replication and respiratory oxidative burst
Authors: Masihi K. N. , Hintelmann H. , Madaj K. , Gast G.
Published: March 2007

Increased morbidity and mortality occur regularly during influenza epidemics. The exact mechanisms involved are not well defined but bacterial superinfection of influenza virus infected patients is considered to play an important role. In the present study, the effect of influenza virus infection on in vivo production of turnout necrosis factor (TNF) in response to bacterial stimuli was investigated. Release of TNF in mice infected by an aerosol of influenza virus was significant after administration of bacterial lipopolysaccharide (LPS) at 72 h, whereas administration of homologous influenza virus produced only modest amounts of TNF at 96 h. Significant production of TNF was observed 48 h after intravenous administration of infectious influenza in response to LPS but not with the homologous virus. TNF induced after influenza virus infection could be blocked by a specific murine anti‐TNF monoclonal antibody. Higher TNF production following aerosol influenza infection correlated with peak titres of influenza virus in the lungs of infected mice and with enhanced generation of luminoldependent chemiluminscence.
The influence of 2‐ chlorodeoxyadenosine in combination with tumour necrosis factor‐α or its mutein on murine leukaemias L1210 and P388
Authors: Góra-Tybor J. , Warzocha K. , Robak T.
Published: March 2007

We investigated the influence of 2‐chlorodeoxyadenosine (2‐CdA) in combination with tumour necrosis factor‐α (TNFα) or its mutein VI, which differs from the native molecule by N‐terminal amino acid composition, on the survival time of mice inoculated with leukaemia L1210 or P388. Groups of mice with leukaemia L1210 and P388 receiving 2‐CdA combined with TNFα had shorter survival times than animals treated with these agents separately. In contrast, the administration of 2‐CdA in conjunction with mutein VI, prolonged the survival of mice inoculated with these leukaemias as compared with animals receiving these agents separately. The results of the present study emphasize the importance of the biological activity of the TNFα molecule N‐terminus.
Analysis of the IFN‐γ‐induced signal transduction pathway in fetal rejection
Authors: Athanassakis Irene
Published: March 2007

The placenta, one of the most important fetal tissues during gestation, ensures nutrition, development and protection of the fetus. Although placenta lacks expression of class II MHC antigens, they can be induced either by interferon‐gamma (IFN‐γ) on the spongiotrophoblast zone, or by 5‐azacytidine (5‐azaC) on the labyrinthine trophoblast zone, two agents actively participating in a plethora of immunological and inflammatory reactions. This induction is correlated with fetal abortion and fetal developmental abnormalities. In this work the in vitro and in vivo signal transduction pathways followed by IFN‐γ or 5‐azaC to induce class H antigen expression on placental cells by using specific pathway inhibitors has been studied. It is shown that at least three intracellular pathways are implicated in the Ia induction, p21ras is the first protein activated by the two agents while further signalling requires Ca2+ mobilization and PKC activations. When the in vitro results are transferred to live animals using the same inducing agents and pathway inhibitors, it is found that theophylline (Ca2+/CaM inhibitor) and anti‐p21ras are the most potent suppressors of the IFN‐γ‐ and 5‐azaC‐induced side effects during pregnancy. The data presented here point to novel directions not only as to the intracellular signalling, but also to the use of pathway inhibitors in vivo to treat aberrant antigen expression associated with fetal loss.
The evidence for histamine H3 receptor‐mediated endothelium‐dependent relaxation in isolated rat aorta
Authors: Djuric D. M. , Andjelkovic I. Z.
Published: March 2007

The presence of histamine H3 receptors was evaluated on the rat aorta endothelium. In the presence of pyrilamine (1 nM, 7 nM, 10 nM) or thioperamide (1 nM, 10 nM, 30 nM) the concentration–response curve for histamine‐induced (0.1 nM − 0.01 mM) endothelium‐dependent rat aorta relaxation was shifted to the right without significant change of the Emax indicating competitive antagonism by pyrilamine (pA2 = 9.33 ± 0.34, slope = 1.09 ± 0.36) or thioperamide (pA2 =9.31 ± 0.16, slope=0.94 ± 0.10). Cimetidine (1 μM) did not influence histamine‐induced endothelium‐dependent rat aorta relaxation. In the presence of thioperamide (1 nM, 10 nM, 30 nM) the concentration–response curve for (R)α‐MeHA‐induced (0.1 nM − 0.01 mM) endothelium‐dependent relaxation was shifted to the right without significant change of Emax indicated competitive antagonism by thioperamide (pA2 = 9.21 ± 0.4, slope = 1.03 ± 0.35). Pyrilamine (100 nM) or cimetidine (1 μM) did not influence (R)α‐MeHA‐induced endothelium‐dependent rat aorta relaxation. These results suggest the presence of a heterogenous population of histamine receptors, H1 and H3, on rat aorta endothelium.
Isolation and partial chemical characterization of macrophage‐derived neutrophil chemotactic factor
Authors: Dias-Baruffi M. , Roque-Barreira M. C. , Cunha F. Q. , Ferreira S. H.
Published: March 2007

Macrophages stimulated with lipopolysaccharide (LPS) release a factor (MNCF; macrophage‐derived neutrophil chemotactic factor) which induces neutrophil migration in vivo and in vitro. The in vivo chemotactic activity of crude MNCF is not affected by pretreating the animals with dexamethasone, an uncommon characteristic which discriminates MNCF from known chemotactic cytokines. We purified MNCF by affinity chromatography of the supernatant from LPS‐stimulated macrophages on immobilized D‐galactose, followed by gel filtration of the sugar‐binding material on Superdex 75. The activity was eluted in the volume corresponding to a MW of 54 kDa. SDS–PAGE of this preparation revealed a single band, also corresponding to a 54 kDa protein. MNCF is an acidic protein (pI < 4) as shown by chromatofocussing. Like the crude MNCF, the homogeneous protein induced neutrophil migration in vitro as well as in vivo. This was not modified by dexamethasone pretreatment.
Biological characterization of purified macrophage‐derived neutrophil chemotactic factor
Authors: Dias-Baruffi M. , Roque-Barreira M. C. , Cunha F. Q. , Ferreira S. H.
Published: March 2007

We have recently described the purification of a 54 kDa acidic protein, identified as macrophage‐derived neutrophil chemotactic factor (MNCF). This protein causes in vitro chemotaxis as well as in vivo neutrophil migration even in animals treated with dexamethasone. This in vivo chemotactic activity of MNCF in animals pretreated with dexamethasone is an uncommon characteristic which discriminates MNCF from known chemotactic cytokines. MNCF is released in the supernatant by macrophage monolayers stimulated with lipopolysaccharide (LPS). In the present study, we describe some biological characteristics of homogenous purified MNCF. When assayed in vitro, MNCF gave a bell‐shaped dose–response curve. This in vitro activity was shown to be caused by haptotaxis. Unlike N‐formyl‐methionylleucyl‐ phenylalanine (FMLP) or interleukin 8 (IL‐8), the chemotactic activity of MNCF in vivo and in vitro, was inhibited by preincubation with D‐galactose but not with D‐mannose. In contrast with IL‐8, MNCF did not bind to heparin and antiserum against IL‐8 was ineffective in inhibiting its chemotactic activity. These data indicate that MNCF induces neutrophil migration through a carbohydrate recognition property, but by a mechanism different from that of the known chemokines. It is suggested that MNCF may be an important mediator in the recruitment of neutrophils via the formation of a substrate bound chemotactic gradient (haptotaxis) in the inflamed tissues.
Serum levels of soluble IL‐2R, CD4 and CD8 in bronChial asthma
Authors: Di Lorenzo G. , Mansueto P. , Melluso M. , Morici G. , Norrito F. , Cigna D. , Candore G. , Caruso C.
Published: March 2007

The aim of the present study was to compare serum levels of soluble forms of interleukin‐2 receptor, CD4 and CD8, released by lymphocytes during activation of the immune system, in patients with allergic bronchial asthma, with those in healthy subjects. Significantly higher levels of soluble IL‐2R and soluble CD4 were found in patients with asthma compared with the control group. In contrast, lower levels of soluble CD8 values were found in patients with asthma compared to the control group. Significant correlations were found for both sIL‐2R and sCD4 and these two molecules, with lung function measured as bronchial responsiveness to inhaled methacholine. These results strengthen previous suggestions that in allergic bronchial asthma, activation of T cells plays a significant role in the disease pathogenesis.
Cyclic AMP stimulates platelet‐derived growth factor B chain mRNA expression in murine macrophage cell lines
Authors: Kovacs Elizabeth J. , VanStedum Susan
Published: March 2007

Prostaglandin E2 plays a role in cytokine production presumably by altering intracellular levels of cAMP. In this paper, we report on the differential expression of cytokine genes in murine macrophages in response to stimulation with activators of cAMP. Macrophages were cultured with or without cAMP activators in the presence or absence of LPS. Prior to treatment, macrophages do not express interleukin‐1β, but do express low levels of tumour necrosis factor α and platelet‐derived growth factor B chain mRNAs. After culture with cAMP‐inducers, including PGE2, dibutyryl cAMP and forskolin, PDGF B chain mRNA is induced. Forskolin, for example, induced expression PDGF B chain mRNA to a level ranging from 25% to 200% of the level induced by LPS in 6 h. In contrast, cAMP‐inducers enhance the expression of IL‐1β and TNF‐α mRNAs, but only in the presence of LPS. The combination of forskolin and LPS does not appear to act synergistically on PDGF B chain mRNA levels, suggesting that LPS‐stimulated effects are not mediated through a cAMP‐dependent pathway. Furthermore, macrophages differentially express cytokine genes in response to treatment with inducers of intracellular cAMP.
IL‐4 induces cAMP and cGMP in human monocytic cells
Authors: Dugas B. , Paul-Eugene N. , Yamaoka K. , Amirand C. , Damais C. , Kolb J. P.
Published: March 2007

Human monocytes, preincubated with IFN‐γ respond to IL‐4 by a cGMP increase through activation of an inducible NO synthase. Here, IL‐4 was found to induce an accumulation of cGMP (1 – 3 min) and cAMP (20 – 25 min) in unstimulated monocytes. This was impaired with NOS inhibitors, but also with EGTA and calcium/calmodulin inhibitors. These results suggest that: (1) IL‐4 may stimulate different NOS isoforms in resting and IFN‐γ activated monocytes, and (2) cAMP accumulation may be partially dependent on the NO pathway. By RT‐PCR, a type III constitutive NOS mRNA was detected in U937 monocytic cells. IL‐4 also increased the [Ca2+]i in these cells. Different NOS may thus be expressed in monocytic cells depending on their differentiation and the signals they receive.
Endothelial cell regulation of leukocyte infiltration in inflammatory tissues
Authors: Duperray A. , Mantovani A. , Introna M. , Dejana E.
Published: March 2007

Endothelial cells play an important, active role in the onset and regulation of inflammatory and immune reactions. Through the production of chemokines they attract leukocytes and activate their adhesive receptors. This leads to the anchorage of leukocytes to the adhesive molecules expressed on the endothelial surface. Leukocyte adhesion to endothelial cells is frequently followed by their extravasation. The mechanisms which regulate the passage of leukocytes through endothelial clefts remain to be clarified. Many indirect data suggest that leukocytes might transfer signals to endothelial cells both through the release of active agents and adhesion to the endothelial cell surface. Adhesive molecules (such as PECAM) on the endothelial cell surface might also ‘direct’ leukocytes through the intercellular junction by haptotaxis. The information available on the molecular structure and functional properties of endothelial chemokines, adhesive molecules or junction organization is still fragmentary. Further work is needed to clarify how they interplay in regulating leukocyte infiltration into tissues.
Tissue eosinophilia induced by recombinant human interleukin‐5 in the hamster cheek pouch membrane
Authors: Minnicozzi M. , Durán W. N. , Kim D. , Gleich G. J. , Wagner J. , Egan R. W.
Published: March 2007

Interleukin‐5 (IL‐5) is a cytokine that preferentially effects the development and function of eosinophils, and is considered important in the pathophysiology of allergic inflammation. In this study, we evaluated the ability of recombinant human IL‐5 (rHu IL‐5) to promote tissue eosinophilia and the importance of this eosinophilia to pathological alterations in vascular function. Repetitive subcutaneous administration for 18 days of rHu IL‐5 resulted in a 7‐fold increase in the number of eosinophils found in the ipsilateral hamster cheek pouch membrane. The contralateral cheek pouch membrane and peritoneum of these animals showed lesser but significant elevations in the number of eosinophils. In contrast, denatured rHu IL‐5 did not elevate eosinophils in these tissues. Through the use of intravital microscopy and fluorometric analysis, rHu IL‐5 treated hamster cheek pouch membranes were evaluated for alterations in microvascular permeability, using plasma clearance of FITC‐dextran 150 as an index. Despite promoting a prominent tissue eosinophilia, the repetitive subcutaneous injections of rHu IL‐5 did not alter the clearance of FITC‐dextran 150. Topical application of rHu IL‐5 to the cheek pouch, also, had no effect on the clearance of FITC‐dextran 150. Immunofluorescence observations using an antibody to the granule protein, eosinophil peroxidase, indicated that the recruited cells had not degranulated. Our results support the importance of IL‐5 in the recruitment of tissue eosinophils, but further stimulation is probably required to cause degranulation of these cells and the ensuing tissue damage.
Rate of perfusion modulates colloidal carbon leakage from rat intestinal microvessels in vitro
Authors: Northover A. M. , Northover B. J.
Published: March 2007

In order to investigate the effects of varying the rate of flow on endothelial integrity the rat isolated small intestinal vasculature was perfused at 1, 5, 10 or 20 ml/min with a gelatin‐containing physiological salt solution (GPSS), followed by an injection of colloidal carbon suspension (CC). Significantly greater microvascular CC leakage occurred at 1 or 5 ml/min than at 10 or 20 ml/ mitt. CC leakage at the two slower rates of flow was reduced by adding red blood cells to the GPSS, suggesting that the microvascular endothelium became hypoxic when perfused with GPSS at 1 or 5 ml/min. After perfusion at 20 ml/min with GPSS containing resiniferatoxin (1 μM) or 5‐hydroxytryptamine (100 μM), CC leakage was significantly lower than after similar perfusion at 10 ml/min. Two nitric oxide (NO) synthesis blockers, N‐nitro‐L‐arginine methyl ester (L‐NAME, 100 μM) and methylene blue (20 μM), and an NO scavenger CPTIO (100 μM) each increased CC leakage. This suggests that NO was being produced at perfusion rates of 10 or 20 ml/min. Sodium nitroprusside (10 μM), 8‐bromo‐cGMP (100 μM) and BN52021 (10 μM) each significantly reduced CC leakage in the presence of L‐NAME.