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PubMed1976

Antimicrobial factors in the exudates of skin windows in human subjects.

The "skin window" technique in which the horny layer of the skin is abraded with a high speed grinder has been used to study the appearance of proteins with antimicrobial activity in the fluid accumulating in damaged human skin. The fluid was absorbed into paper discs and protein levels measured by radial diffusion. The skin exudates contained about 45% as much IgG and IgM as the subjects' serum, but the amount of IgA (68% of the serum level) in the exudate was significantly greater, suggesting selective transport into the lesion. The fluid also contains complement proteins, lysozyme and lactoferrin. The methods used in this study may provide useful information about clinical situations in which susceptibility to cutaneous infection is increased.

Complement System Proteins↗
PubMed1982

Severe Fletcher factor (plasma prekallikrein) deficiency with partial deficiency of Hageman factor (factor XII): report of a case with observation on in vivo and in vitro leukocyte chemotaxis.

A case of cross-reacting material-negative Fletcher trait with additional partial deficiency of Hageman factor (HF, Factor XII) is described. Although the patient presented with a recent history of frequent epistaxis, he had no other personal or family history of a tendency toward bleeding or infection. Similar to other cases of Fletcher trait, his plasma showed a markedly prolonged partial thromboplastin time which could be corrected by prolonged incubation with the surface-activator kaolin. Surface-induced fibrinolysis, amidolysis of alpha-N-benzoyl-proline-L-phenylalanine-L-arginine-p-nitroanilide, and cold-promoted enhancement of factor VII activity, reactions requiring the presence in the plasma of fletcher factor (prekallikrein), in addition to Hageman factor and Fitzgerald factor (high-molecular weight kininogen), were also defective. In vivo chemotaxis of polymorphonuclear leukocytes and monocytes (Rebuck's skin window technique) in response to skin abrasions was defective, but was normal when diphtheria-tetanus toxoid was also applied. In vitro leukocyte chemotaxis (Boyden chamber technique) in response to normal or patient's own serum activated with zymosan was normal. Together with previous observations that kallikrein generated chemotactic activity, possibly via activation of C5, the present observations suggest that prekallikrein activation may be important for in vivo leukocyte chemotactic response to skin abrasion. The inheritance of Fletcher trait in this patient is unclear.l Although the father was an apparent heterozygote, the mother was completely normal for Fletcher factor procoagulant activity and antigen. The mild Hageman factor deficiency in the patient did not contribute significantly to the plasma defects described and was likely inherited from the father who had a low HF procoagulant activity.

Blood Coagulation↗
PubMed1979

Skin-window study on the migration of leukocytes of newborns and infants.

Migration of leukocytes of newborns and of infants up to the age of 6 months was studied using the in vivo skin-window technique according to Rebuck. Using the non-specific stimulation (abrasion of the skin only) a slight age-dependent physiological increase of migration of cells was observed within the observation period; after a strong local irritation with diphtheria-tetanus-pertussis vaccine (Alditepera) there was a vigorous migratory response of cells in the skin lesion. Both quantitatively and qualitatively, the migration pattern (i.e. shift from PMN leukocyte to mononuclear cells in the exudate within a 1 d period after abrasion) was not influenced by immunization of infants with Alditepera, suggesting thus the nonspecific nature of this cellular response. The "normal" values of the chemotactic response of leukocytes of newborns and infants are given as a basis for evaluation of defects of this functional activity of leukocytes.

Age Factors↗
PubMed1981

Serum zinc levels in lung cancer patients.

Serum zinc concentrations were determined in 26 extensive squamous cell lung cancer patients and were tested for correlations with survival, response to therapy, nutritional status indices, and various host defense characteristics. Subnormal serum zinc levels were found in 24 of the 26 patients. The mean serum zinc concentration was 43.2/micrograms 100 ml +/- 3.6 S.E.M. (normal = 80-100 micrograms/100 ml). A significant (P = 0.007) survival advantage was demonstrated for those patients with pretreatment zinc concentrations greater than 45 micrograms%, but serum zinc levels did not correlate with response to chemotherapy (also significantly affecting survival). Decreased serum zinc concentrations were significantly associated with decreased neutrophil migration measured by the skin window technique and with decreased triceps skin fold thickness but not with any of the other host defense and nutritional induces measured. These data suggest that further studies are indicated to examine the role of serum zinc concentration as a possible sensitive prognostic characteristic and to determine if zinc administration may be of therapeutic benefit in cancer patients.

Adult↗
PubMed1990

Quantification of radiolabelled granulocyte migration in vivo.

Quantification of the tissue localisation of granulocytes is difficult, particularly in man. With the aim of facilitating such quantification, a technique is described which combines the micropore filter skin window technique with the infusion of autologous 111In-labelled granulocytes. The radioactivity in the removable micropore filters, placed on small forearm skin window abrasions, was compared with peripheral blood radioactivity following injection of 111In-labelled 'pure' granulocytes in normal subjects. Radioactivity in filters from abrasions which were made 8 h or more before injection of labelled cells followed the same time course as the cell-associated radioactivity in whole blood, but radioactivity in filters from abrasions made nearer to the time of labelled granulocyte injection increased to reach a peak 3-8 h after cell injection, at a time when cell-associated blood radioactivity had fallen to about 50% of its initial value. Exercise appeared to induce a transient decrease in radiolabelled granulocyte migration into the filters. This technique offers a means of studying the kinetics of granulocyte migration in vivo.

Cell Movement↗
PubMed1988

Lactoferrin, myeloperoxidase, lysozyme and eosinophil cationic protein in exudate in delayed type hypersensitivity.

A skin window technique was used to study the morphology of leucocytes in upper dermis and exudate during nickel challenge in patients with contact allergy to nickel. Contact allergic patients and healthy volunteers tested with a skin widow without addition of nickel to the chamber medium served as controls. The morphology of the leucocytes in dermis was studied in biopsies taken 8, 24, or 48 h after skin window application, and in a parallel test the morphology of the exudate was examined by sequential collection of the chamber medium during a 48 h period. The infiltrate in dermis of contact allergic patients with nickel challenge in the chamber medium showed a time-dependent increase of mononuclear cells, eosinophils and basophils and a concomitant decrease of polymorphonuclear granulocytes, characteristic of a combined specific and unspecific inflammation. The morphology of the exudate in contact allergic patients exposed to nickel showed a dominance of polymorphonuclear granulocytes throughout the study period, while mononuclear cells, eosinophils and basophils were detected at a much lower quantity and with a considerable delay. Further, we studied the kinetics of the leucocyte granule proteins: lactoferrin, myeloperoxidase, lysozyme and eosinophil cationic protein in exudate fluid in a parallel test. A significant higher flux was found for all during the second day of allergen exposure compared to contact allergic patients without allergen challenge as well as normal volunteers. The increased protein fluxes were not accompanied by an increased flux of polymorphonuclear granulocytes in the exudate.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗
PubMed1997

Studies of skin-window exudate human neutrophils: increased resistance to pentoxifylline of the respiratory burst in primed cells.

Human neutrophils were isolated both from peripheral blood (PB) and from aseptic inflammatory exudates obtained by the Senn's skin window technique (SW). The respiratory burst (O2- production) induced by in response to n-formyl-methionyl-lencyl-phenylalanine (fMLP) and by serum-treated zymosan (STZ) was investigated using a microplate assay. SW neutrophils were primed to enhanced fMLP-dependent O2- production in response to fMLP but not to STZ. Pentoxifylline, a cAMP-elevating drug, dose-dependently inhibited the respiratory burst in any experimental condition, but the dose-effect curves were markedly different according the stimulant used and the source of the cells. With fMLP as stimulant, a significant inhibition of the O2- production by PB neutrophils was obtained using doses of 10 micrograms/ml, while SW neutrophils were inhibited only by doses equal or higher than 100 micrograms/ml. With STZ as stimulant, the inhibition of the respiratory burst of PB neutrophils and of SW neutrophils was obtained only with doses higher than 400 micrograms/ml and 1 mg/ml respectively. Pentoxifylline dose-dependently (10 micrograms/ml to 1 mg/ml) increased the intracellular adenosine 3'-5'-cyclic monophosphate (cAMP) to the same extent in SW and in PB neutrophils. These data indicate that the priming of neutrophil oxidative metabolism by in vivo inflammation is associated with an increase in the resistance to the regulating effect of cAMP on the fMLP-dependent activation pathway of NADPH oxidase. The fact that therapeutic doses of pentoxifylline do not inhibit the respiratory burst of primed neutrophils may have relevance in the interpretation of the clinical effects of this drug.

Cyclic AMP↗
PubMed1990

In vivo effects of cetirizine on cutaneous reactivity and eosinophil migration induced by platelet-activating factor (PAF-acether) in man.

The aim of the study was to determine the effect of cetirizine, a new potent H1 antihistamine, on acute cutaneous inflammatory response and eosinophil accumulation induced in vivo by platelet-activating-factor (PAF-acether) and allergen. In a double-blind, crossover study, seven subjects allergic to grass pollen and three nonallergic control subjects received orally either cetirizine, 20 mg/day, or placebo for 4 days. On day 4, the subjects were skin tested with grass pollen and PAF-acether (400 and 40 ng per site). After the challenge, an evaluation of time-course cutaneous eosinophil infiltrations by a skin window technique was performed. Cetirizine pretreatment reduced skin wheal and erythema elicited by allergen and PAF, 400 and 40 ng, by 74.6% (p less than 0.001), 53.9% (p less than 0.001), and 47% (p less than 0.01), respectively. Skin reactivity induced by PAF-acether was also significantly reduced by cetirizine in nonallergic subjects. Cetirizine reduced at hour 24 eosinophil infiltrations induced by allergen and PAF, 400 and 40 ng, by 63% (p less than 0.001), 58.5% (p less than 0.001), and 57.8% (p less than 0.01), respectively. This inhibitory effect of cetirizine on allergen and PAF-induced eosinophil infiltration was already effective 2 hours after the challenge. PAF induced a nonsignificant eosinophil influx in all nonallergic subjects. In conclusion, cetirizine inhibited both the immediate cutaneous response and the eosinophil influx induced by allergen and by a potent eosinophil chemotactic factor, such as PAF-acether. Therefore, cetirizine, besides its anti-H1 effect, has the potential to modulate the allergic inflammatory response.

Adult↗
PubMed1975

Fitzgerald factor: a hitherto unrecognised coagulation factor.

A newly recognised asymptomatic coagulation-factor deficiency is reported in a 71-year-old man. This factor seems to operate at an early stage in the intrinsic coagulation pathway and participates in other Hageman-factor-mediated biological reactions. The patient has an atypical cellular inflammatory response as observed by the skin-window technique.

Aged↗
PubMed1985

Investigations of host defence in patients with sickle cell disease.

Parameters of host defence were investigated in 30 patients with sickle cell disease (SCD). A newly devised perfusion system was used to study the kinetics in whole blood of leucocyte adherence, phagocytosis, killing and solubilization of a mixture of Staph. aureus and Str. pneumoniae, and secretion of lactoferrin. A skin window technique was used to examine the accumulation of leucocytes at inflammatory foci and their subsequent rate of movement through a filter. Serum concentrations of C3, C4, total haemolytic complement and immunoglobulins were also measured. The rate of neutrophil migration into filters was slightly reduced in patients with SCD. The proportion of monocytes that emigrated from the skin windows and their rate of migration were markedly diminished. The adhesion of neutrophils and their ability to kill staphylococci were also reduced, particularly in patients of the haemoglobin (Hb) SS and Hb S-beta-thalassaemia genotypes. Neutrophil function was mostly impaired in patients with the greatest frequency of bacterial infection. The rate of clearance of pneumococci was related to the concentration of type specific immunoglobulin G but not M. Serum concentrations of immunoglobulins and complement were normal. We were unable to define a defect of host defence of sufficient magnitude to explain the susceptibility of these patients to severe infection.

Anemia, Sickle Cell↗
PubMed1985

Skin window examination according to Rebuck and cutaneous pathergy tests in patients with Behcet's disease.

Comparative in vivo examinations of skin (cutaneous pathergy test, skin window technique over 24 h according to Rebuck) were carried out in 12 patients with Behçet's disease (BD) and in 13 healthy volunteers serving as controls. 10 out of 12 patients at active phases of BD showed a positive papular (-pustular) skin reactivity to needle prick or venepuncture. These 10 patients simultaneously revealed an increased chemotactic activity of polymorphonuclear leukocytes (PMNL) in vitro. In the controls all pathergy tests were negative and chemotaxis of PMNL was within normal range. The proportion of PMNL initially coming up to 100% in the skin window preparations, very rapidly decreased in patients with BD as compared to a distinctly slower drop in the controls. Conversely to the number of PMNL, the skin window cellular pattern was built up by monophagocytic cells replacing the PMNL. The phenomenon of 'rapid depletion of PMNL', from skin window can be considered an additional hint on the functional disturbance of the PMNL involved in the pathogenesis of BD.

Behcet Syndrome↗
PubMed1989

In vivo leucocyte migration in Behçet's syndrome.

Serial studies of leucocyte migration in vivo were carried out in 15 patients with Behçet's syndrome using a skin window technique. Where possible, patients with and without active disease were studied during and in the absence of treatment. In patients with active disease neutrophil migration was frequently greater than normal, particularly with respect to numbers of cells migrating. There was also an increased frequency of emigrating neutrophils with less or more nuclear lobes than normal. In three patients in whom function of skin window neutrophils was studied nitroblue tetrazolium reduction and phagocytosis and killing of Candida guilliermondiae were normal. The monocyte component of the skin window was more often reduced in patients than in normal controls. Corticosteroid treatment did not exert a major effect on leucocyte migration, though the doses involved were relatively small. Neutrophil abnormalities were common in patients and particularly those with active disease. These results suggest that neutrophil hyperactivity may have an important role in the pathogenesis of Behçet's syndrome.

Adult↗
PubMed1975

Skin window studies in polycythaemia rubra vera.

The inflammatory response was studied in patients with primary polycythaemia by means of a modified skin window technique. In untreated patients, the overall cellularity was a prominent feature and, as compared with the controls, the 48 h preparations showed a significantly greater percentage of granulocytes with a corresponding decreased percentage of macrophages. In the peripheral blood of these patients, both total white cells and granulocyte counts were significantly higher than in the control subjects. A highly significant relationship between the blood and skin window neutrophils was demonstrated. In polycythaemia patients in remission, the results were generally similar to the controls, except that there was still a highly significant increase in the percentage of skin window basophils. The results provided no evidence of a defect in cell migration or in the phagocytic activity of macrophages in polycythaemia vera. Possible mechanisms of the cutaneous basophilia and its relevance to the manifestation of this disorder are discussed.

Adult↗
PubMed1988

Skin eosinophilia in atopic patients.

The responses of six atopic and six control patients to the intracutaneous administration of platelet-activating factor (PAF) acether, of the leukocyte secretagogue and chemoattractant f-methionyl-leucyl-phenylalanine (FMLP), and of allergens were studied. Immediate wheal-and-flare skin reactions to allergens and to PAF acether in atopic patients were the same, and when reactions were compared to reactions of nonatopic patients, these reactions to PAF acether and to FMLP were of equivalent intensity. Late-phase reactions were not observed macroscopically with allergen, PAF acether, or FMLP. The cellular responses, assessed by the Rebuck skin window technique, demonstrated a late (24 hours) eosinophilic response to the allergen and to both inflammatory agents, with 33% to 44% of intensively degranulated eosinophils. In comparison, very few, if there were any, eosinophils were observed in the nonatopic patients. Most infiltrated cells were neutrophils. The accumulation of eosinophils in atopic human skin indicates a potent eosinophilotactic activity restricted to the allergic patients. A similar cellular reaction has been observed in lungs from guinea pigs injected with PAF acether or with allergen, and in patients with severe asthma. The chemoattractant activity of FMLP toward eosinophils cannot be accounted for by its histamine-releasing activity and remains to be explained.

Adult↗
PubMed1985

Neutrophil delivery to wounds of the upper and lower extremities.

Clinical observations and experimental evidence indicate that wounds of the lower extremity are more susceptible to infection than are wounds located elsewhere on the body. The details of this regional relative immunoincompetence are not described. In this initial study of regional inflammatory response, neutrophil delivery to standard wounds of the upper and lower extremities was measured in 15 human volunteer subjects using a quantitative skin-window technique. Neutrophil delivery proved to be relatively deficient in the lower extremity. Neutrophil delivery (mean +/- SEM) was significantly lower to the lower-extremity wounds (5,890 +/- 590 cells per cubic millimeter) than to the upper-extremity wounds (16,600 +/- 1,680 cells per cubic millimeter). This lower neutrophil response may be a part of the lower extremity's increased susceptibility to infection. Further functional study of regionally collected neutrophils may provide more details of differences in regional inflammatory response. The mechanisms underlying these differences remain undescribed.

Adult↗
PubMed1983

Quantitation by myeloperoxidase assay of neutrophil accumulation at the site of in vivo allergic reactions.

Skin window techniques to investigate neutrophil inflammatory reactions in human skin have been limited by cellular distortion and difficulties in quantitation. We have developed a quantitative approach based on the assessment of the myeloperoxidase (MPO) released from sonicated membrane filters to which exuding inflammative cells had adhered. Our studies have shown that (1) a standard curve can be derived for each subject from the linear relationship between amounts of MPO released and designated numbers of blood neutrophils which have been aspirated onto such filters; (2) the number of neutrophils adhering to filters appended to skin window sites can then be reliably estimated by comparing the amount of MPO released by sonication of such filters with the standard curve; and (3) neutrophil accumulation is greater at pollen-challenged than control sites in sensitive subjects.

Cell Count↗
PubMed1990

Does reversed-type anaphylaxis in healthy subjects mimic a real allergic reaction?

Immediate and late cutaneous reactions were induced in healthy and atopic subjects by anti-IgE challenge and a skin-window technique was used in order to verify if the pattern of cells observed at 24 hr was similar in both groups. The study was first performed under basal conditions, and in a second double-blind cross-over step, it was performed again during treatment with cetirizine 10 mg b.i.d. and terfenadine 60 mg b.i.d. Anti-IgE challenge was followed by a significant eosinophil accumulation in atopic subjects only. Cetirizine significantly inhibited this phenomenon while terfenadine showed a mild non-significant inhibitory effect.

Adolescent↗
PubMed1989

Granulocyte-macrophage colony-stimulating factor may inhibit neutrophil migration in vivo.

Recombinant human granulocyte-macrophage colony-stimulating factor (rh GM-CSF) was administered by constant intravenous infusion to eight patients with malignant disease prior to chemotherapy. rh GM-CSF was given at a dose of 15 to 25 micrograms m-2 h-1 for 1 or 2 h. Neutrophil migration into an inflammatory zone was monitored throughout this period using a micropore skin window technique. Neutrophil migration into the micropore membrane prior to the commencement of the rh GM-CSF infusion was almost identical to that in eight normal control individuals (leading front distance of migrating neutrophils in 20-min period 81.3 +/- 7 microns in the patients compared to 79.4 +/- 4 microns in the control individuals). During the rh GM-CSF infusions there was a significant fall compared to controls in the number of neutrophils entering the membrane and the leading front distance (P less than 0.05). In four out of nine patient studies (eight patients) there was a greater than 30% fall from the pre-infusion level. In the control individuals the largest fall recorded was less than 10% and overall there was a progressive rise in the number of neutrophils entering the membrane throughout the period of study. This study suggests that intravenous infusion of rh GM-CSF may impair the ability of neutrophils to infiltrate an inflammatory focus.

Colony-Stimulating Factors↗