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A biologically active neutral peptide mediator is cleaved from a plasma protein substrate by an alpha-1-antitrypsin-inhibitable serine protease apparently residing on the membrane of the human neutrophil. The peptide mediator has an approximate mol wt of 1,000, and is distinguished from the kinin peptides by a neutral isoelectric point, susceptibility to inactivation by trypsin as well as chymotrypsin and activity on the isolated, atropinized, and antihistamine-treated guinea pig ileum with relatively little action on the estrous rat uterus. The neutrophil protease is fully inhibitable by DFP, trypsin inhibitors from lima or soy bean, and alpha-1-antitrypsin and is associated with the high mol wt fragments of the neutrophil and not the nuclear, lysosomal, or cytoplasmic subcellular fraction. The substrate has an approximate mol wt of 90,000 and is chromatographically separable from kininogen. The exquisite sensitivity of the neutrophil protease to alpha-1-antitrypsin was established both by inhibition with highly purified alpha-1-antitrypsin and by the inability of the protease to generate detectable neutral peptide in a homozygous (ZZ) alpha-1-antitrypsin-deficient patient without heat inactivation of the residual inhibitor. On the other hand, plasma from a (null) alpha-1-antitrypsin-deficient patient supported neutral peptide generation and revealed an additional factor which inactivated neutral peptide.

A factor has been derived from human leukocytes which irreversibly inhibits the response of human neutrophils to diverse chemotactic stimuli without impairing their viability. It is released by both polymorphonuclear and mononuclear leukocytes during incubation in acidic medium, after endotoxin exposure and subsequent incubation in low potassium medium, and during phagocytosis of particles. It is extractable from both leukocyte types and therefore must be preformed. This chemotactic inhibitor is completely separable from contaminating chemotactic activity in the crude supernatants, has a mol wt of 5000, and is inactivated by digestion with trypsin or chymotrypsin. It has been termed a neutrophil-immobilizing factor because it inhibits neutrophils directly and independently of the chemotactic stimulus, and has relatively little effect on human monocyte chemotaxis.

A factor has been derived from human leukocytes which irreversibly inhibits the response of human neutrophils to diverse chemotactic stimuli without impairing their viability. It is released by both polymorphonuclear and mononuclear leukocytes during incubation in acidic medium, after endotoxin exposure and subsequent incubation in low potassium medium, and during phagocytosis of particles. It is extractable from both leukocyte types and therefore must be preformed. This chemotactic inhibitor is completely separable from contaminating chemotactic activity in the crude supernatants, has a mol wt of 5000, and is inactivated by digestion with trypsin or chymotrypsin. It has been termed a neutrophil-immobilizing factor because it inhibits neutrophils directly and independently of the chemotactic stimulus, and has relatively little effect on human monocyte chemotaxis.

A system has been developed for the evaluation of hematological data in the newborn. Normal values were established for segmented (neutrophil) and nonsegmented (band) polymorphonuclear leukocytes in the peripheral blood of 169 healthy full-term infants during the first five days of life. This normal data is displayed graphically and the values for infants under study are compared directly. Newborn infants with proven bacterial infections had normal neutrophil counts; however, the band counts increased significantly beyond the normal range. Therefore, the simple laboratory procedure for ennumerating band neutrophils with comparison of absolute values and changes to a normal range represents a hematologic technique of major importance in the diagnosis of sepsis of the newborn infant. [ABSTRACT FROM AUTHOR]

The role of complement in the emigration of blood neutrophils and monocytes into gingival crevices has been studied in dogs by experimental decomplementation. Blood neutrophils and monocytes were labeled by intravenously injected colloidal carbon. At various time intervals after labeling and decomplementation by carragheenan cellular samples were taken from the gingival crevices. Labeled neutrophils and monocytes were found in both healthy and inflamed gingival crevices indicating a continuous migration of these cells in spite of decomplementation. For comparison the migration of leukocytes into skin window lesions was studied. Early leukocyte emigration was found to be inhibited in the decomplemented animals, but could be initiated by the application of dental plaque to the lesions. The results suggest that complement activation by immune complexes is not necessary for the continuous migration of leukocytes into gingival crevices. Other factors such as chemotactic substances present in dental plaque may be responsible for the continuous migration of leukocytes into gingival crevices. [ABSTRACT FROM AUTHOR]

Human dental plaque extracts in Gey's medium and whole human saliva were tested in the Boyden Chamber system for chemotactic activity against rabbit peritoneal polymorphonuclear leukocytes. Positive controls were a semipurified bacterial chemotactic factor and a complement derived chemotactic factor. Gey's medium alone served as a negative control. The method appeared to be very sensitive as concentrations of less than one mg plaque per ml medium gave a clear chemotacitc response. Whole human saliva was less chemotactic than dental plaque. Incubation of equal concentrations of plaque extract in both the upper and lower compartments of the Boyden chamber did not result in a large increase in cell migration, indicating that most cell migration in reaction to dental plaque extract in this system depends on a concentration gradient. The chemotactic factor or factors were present in plaque at the time it was collected, they were readily dissolvable in water, and they were heat stable. The dose range in which dental plaque extract would evoke a chemotactic response was variable and usually very narrow. Replicate tests yielded highly variable results indicating that this method is not suitable for quantitative comparisons. Extracts of the same dilution were prepared from dental plaque samples from a small population with varying degrees of gingivitis. Differences in chemotactic response to those samples could not be related to the intensity of gingivitis. [ABSTRACT FROM AUTHOR]

Extracts of 4-day-old dental plaque were fractionated on Sephadex® columns (G- 10; G-25; G-200). Five fractions were selected from the protein and conductivity peaks obtained in chromatograms. The chemotactic activity elaborated by the fractions was tested in the Boyden's in vitro model system. The permeability altering capacity of the plaque fractions was tested in vivo in a wound chamber. The results from the in vitro experiments demonstrated that all fractions were chemotactic for rabbit neutrophil leukocytes. Substances of a low molecular size (mol. wt. ≈ 5.000) elaborated, however, the peak chemotactic activity. The results from the in vivo experiments showed that the plaque fractions induced an increased (1) vascular permeability, and (2) emigration of leukocytes from a granulation tissue vasculature. The most pronounced emigration and exudation was induced by the low molecular size fraction (mol.wt. ≈ 5.000). [ABSTRACT FROM AUTHOR]

The present investigation was performed to study whether a filtrate from human dental plaque when applied topically to the gingival margin enhances gingivitis. Four-day- old plaque was sampled from 20 dental students. The plaque samples were pooled, suspended in NaCl, homogenized, centrifuged and filtered. The capacity of the plaque filtrate to attract neutrophil leukocytes was determined by measuring the number of leukocytes (a) which migrated into wound chambers containing the filtrate and (b) which could be sampled from the gingival crevices of dogs and monkeys after repeated topical application of the filtrate to the gingival margin. Gingivitis was assessed by gingival exudate measurements. The results showed that substances present in human dental plaque are highly chemotactic for neutrophil leukocytes. The filtrate stimulated enhanced emigration of leukocytes into the gingival crevices and enhanced gingival exudation. The observations indicate that a plaque filtrate may enhance vascular and cellular changes similar to those seen in incipient gingivitis. [ABSTRACT FROM AUTHOR]

The morphology of oral leukocytes was studied in healthy subjects. The different stages of degeneration of neutrophils were classified and a differential count was made in stimulated and unstimulated saliva and in the mobile mucus phase, collected by serial mouth rinses. The majority of cells in mouth rinses were neutrophils with multi-lobular nuclei, whilst in saliva the mononuclear degenerated cell form was predominant. However, all the media contained equal numbers of identifiable but degenerated neutrophils (group III, about 35 %). The pH values of stimulated and unstimulated saliva were measured in the mouth with a microelectrode system. It was observed that the morphologic appearance of the cells was similar in the two types of saliva in spite of the difference in pH. The pH of stimulated saliva was 7.20 and of whole saliva 6.76. [ABSTRACT FROM AUTHOR]

The differential count of oral leukocytes was estimated from a group of 43 healthy dental students. A modified Millipore® technique was used. The differential count was made from samples of stimulated saliva, whole saliva, and sequential mouth rinses with hypertonic saline. The media from which the leukocytes were obtained did not influence the results and the differential counts in all the samples were consistent. The predominating cell type was a polymorphonuclear (PMN) neutrophil (98%-99%). Lymphocytes were found in all samples (average proportion 1%). Some monocytes and eosinophils were also observed, but they were so few in number that their precise proportion in the oral leukocyte count could not be determined. Basophilic leukocytes were not observed in the samples.

Culture of human mononuclear leucocytes in the presence of hydrocortisone results in the appearance in the culture medium of a factor which markedly stimulates the migration of human polymorphonuclear leucocytes in vitro. This effect is not due directly to hydrocortisone, which has an inhibitory effect on polymorph migration. The steroid-mononuclear cell interaction is dependent on the duration and temperature of the culture, and on the concentration of hydrocortisone. It is there- fore suggested that an active biological process is involved which may be relevant to the immunosuppressive action of corticosteroids. [ABSTRACT FROM AUTHOR]

Neutrophils from three patients with recurrent infections were studied for defects in the phagocytic process. In one, random and directed migration, adherence and ability to kill Gram-negative bacteria were defective. In a second patient random and directed migration was abnormal, but adherence was unimpaired; this patient also had an impaired ability to kill Gram-negative bacteria. The third patient had defective leucocyte motility, inability to reduce nitro-blue tetrazolium dye, and deficient killing of both Staphylococcus aureus and Gram-negative organisms. This patient's mother showed an intermediate bacterial killing defect consistent with the heterozygous state. The previously unrecognized patterns of defects in these patients emphasize the importance of early cell-membrane associated events to the ultimate success of the bactericidal process, and illustrate the heterogeneity of defects responsible for impaired neutrophil function. [ABSTRACT FROM AUTHOR]

A patient is presented who has recurrent infections associated with a cellular defect of PMN chemotaxis and phagocytosis, as well as a sex-linked form of congenital agammaglobulinaemia. The impairments of PMN function were demonstrated in vitro by an inability of the patient's PMNs to respond to chemotactic factors, by an inability of the PMNs to phagocytize S. aweus, and by abnormal NBT tests. These abnormalities were not corrected by the addition of normal serum or plasma to the patient's PMNs. No evidence was obtained for a plasma inhibitor of PMN function. [ABSTRACT FROM AUTHOR]

Chemotactic activity for eosinophils and neutrophils has been studied using guinea-pig serum activated by preformed antigen-antibody complexes. Rabbit complexes or complexes prepared either with guinea-pig IgG2 or IgG1 were equally capable of generating a heat stable activity from guinea-pig serum that was chemotactic for guinea-pig eosinophils and for neutrophils of both the guinea-pig and the rabbit. This was distinguished from a relatively heat-labile chemotactic activity present in untreated guinea-pig serum. The heat-stable complex-mediated chemotactic activity was thought to be complement dependent since chemotaxis for eosinophils or neutrophils could not be generated from heated serum, ammonia treated serum or from serum treated with complexes in the presence of 0.01 M EDTA. Guinea-pig sera, activated either with rabbit, guinea-pig IgG1 or IgG2 complexes, was fractionated by sucrose-density grad lent ultracentrifugation and by Sephadex chromatography. In all experiments two peaks of cell specific chemotactic activity could be separated. The peak of activity for guinea-pig neutrophils was approximately 4.5S and in the fractionation range of proteins having a molecular weight of between 65,000 and 85,000. The peak of guinea-pig eosinophil chemotactic activity was 1.5S-2S and in the molecular weight range of between 15,000 and 20,000. Those Fractions which were chemotactic for guinea-pig neutrophils did not attract rabbit neutrophils. Rabbit neutrophils migrated towards those fractions of guinea-pig serum chemotactic for guinea-pig eosinophils; therefore, the properties associated with guinea-pig eosinophil chemotactic activity were similar to previously published data for a fragment cleaved from the 5th component of complement. [ABSTRACT FROM AUTHOR]

The basic proteins of rat neutrophils possessing the capacity to induce vascular Permeability were analysed. Evidence indicating the presence of four such permeability proteins was presented. One of these induced the release of' histamine from rat mast cells. [ABSTRACT FROM AUTHOR]

Eosinophil leucocytes migrated into the site of PCA reactions mediated by those fractions of 7S guinea-pig lgG containing lgG1; neutrophils were associated with IgG1 and IgG2. Maximal eosinophil infiltration is seen at 12 hr and was associated with degranulation. Intradermal histamine was not eosinophilotactic in guinea-pigs. Performed antigen-antibody complexes of IgG1 and IgG2 both promoted eosinophils and neutrophil migration in guinea-pig skin but slightly more eosinophils were seen following injections of complexes containing IgG1. Local eosinophilia and PCA activity were mediated by a relatively heat-stable element since these effects were demonstrable even after prolonged heating of fractions containing IgG1. Eosinophils were seen following injections of Compound 48/80 and this was accompanied by low mast cell counts; however, there was also some associated tissue destruction. [ABSTRACT FROM AUTHOR]

It was possible to demonstrate complement fixation by IgG1 and IgG3 sub-class myeloma proteins which had been heat aggregated under standard conditions. Phagocytosis of bacteria by neutrophils was inhibited by IgG 1, IgG2 and IgG3. Lysis of antibody-sensitized target cells by cytotoxic B lymphocytes was inhibited by all four sub-classes of IgG. IgG interacted with cytotoxic B lymphocytes, neutrophils and complement only after physical alteration. Cytotoxic B cell reacting activity was acquired during protein purification procedures and was only slightly increased by heating. Neutrophil inhibition activity appeared only after the IgG sub-classes were heated and aggregation had occurred. Very large aggregates, however, were inefficient inhibitory units. Complement fixation by IgG1 and IgG3 was markedly increased by heating and the largest aggregates showed greatest activity. [ABSTRACT FROM AUTHOR]

ECF-A and C5a were chemotactic both for eosinophils and neutrophils. However, when eosinophils comprised approximately 10 per cent or more of a mixed leucocyte population, they were preferentially attracted by both of these agents. Marked synergism was observed between ECF-A and C5a in their ability to attract eosinophil leucocytes. [ABSTRACT FROM AUTHOR]

More eosinophils accumulate in sites of cutaneous anaphylaxis mediated by homologous IgGl or reagin, than in skin treated with IgG2, 18 hours after intravenous challenge. The increased number of eosinophils becomes apparent at 4 hours and reaches a maximum at 12-24 hours, whereas neutrophils infiltrate skin sensitized with IgG1 within 15 minutes of challenge and are most numerous at 4-8 hours. There is a much higher ratio of eosinophils to neutrophils in skin passively sensitized with reagin and challenged after 14 or 28 days. In anaphylactic skin, eosinophils accumulate round changed mast cells. The numbers of eosinophils in anaphylactic skin reflect the numbers in the blood when challenged, and no increase in the number of haematogenous eosinophils occurs between the time of challenge and sampling. The behaviour of eosinophils in vitro appears to differ from that in vivo, in that they, like neutrophils, are attracted more strongly to complexes containing IgG2 than igGI antibody. They are also attracted to damaged neutrophils. It is suggested that eosinophils are not selectively attracted to sites of cutaneous anaphylaxis, but enter them with the neutrophils in the relative proportions in which they are present in the blood. They are however selectively retained in anaphylactic, or anaphylactoid tissue, while the neutrophils continue to emigrate. [ABSTRACT FROM AUTHOR]

Rabbits rendered leucopenic by means of nitrogen mustard reject kidney xenografts more slowly than normal controls It is therefore probable that polymorphonuclear leucocytes play an important role in the acute rejection of xenografts. The previously reported complement requirement for xenograft rejection was also confirmed in this work, by depleting the graft recipients of C3 with cobra venom factor. [ABSTRACT FROM AUTHOR]

Passive cutaneous anaphylaxis (PCA) was studied in guinea-pigs sensitized with rabbit antisera to bovine serum albumin or horse spleen ferritin and challenged 4-5 hours later with antigen. Ultrastructural studies of lesions obtained after sensitization or challenge indicated that substantial numbers of neutrophils had infiltrated into the skin. Following challenge these elements exhibited phagocytic activity, presumably representing the uptake of immune precipitates formed in the interstitial spaces. This was associated with degranulative and degenerative phenomena, suggesting that lysosomes were discharged extracellularly. A significant suppression of the intensity of the hyperpermeability response in PCA was noted when animals were pretreated with an antiserum to guinea-pig neutrophils. Antihistamines also suppressed PCA, and the combined effect of neutrophil depletion and antihistamine therapy was additive. It was concluded that a combination of anaphylactic (histamine release) and Arthus-like (neutrophil lysosome release) mechanisms mediated the vascular response in this type of PCA. [ABSTRACT FROM AUTHOR]

A chemotactic factor (leucoegresin) for polymorphonuclear leucocytes (PMN) of rabbit, guinea-pig, rat and mice was extracted in the pseudoglobulin fraction of Arthus skin lesions and then highly purified by chromatography using Sephadex G-50, DEAE-Sephadex A-50 and CM-Sephadex C-50 in this order. Chemotactic activity was estimated by Boyden's method during the process of purification. Leucoegresin behaved as a homogeneous substance on ultracentrifugation and on moving boundary electrophoresis. This factor was a protein free of nucleic acid and its molecular weight was approximately 140,000 when measured by gel filtration. The sedimentation coefficient of this substance was 6.58S and its isoelectric point was pH 5·0. It was relatively heat-stable. Leucoegresin induced pronounced PMN migration, but did not increase vascular permeability; PMN emigration occurred only at the site of venules. Leucoegresin satisfied such criteria for the natural mediator of Arthus leucotaxis as local availability, parallel between activity (amount) and time-course of the reaction, and histological resemblance to the Arthus reaction. [ABSTRACT FROM AUTHOR]

A rabbit anti-rat peritoneal cell (APCS) serum significantly suppresses (P<0.001) the immune response to Nippostrongylus brasiliensis infection in the rat and the infection can be prolonged by such an antiserum. The antiserum appeared to contain antibodies directed against the surface antigens of pharmacologically-active cells such as the mast cell. Prolongation of infection following treatment with APCS was accompanied by a reduction in the number of eosinophils, basophils and neutrophils in the peripheral circulation. The number of circulating peripheral lymphocytes was not significantly affected by the antiserum, nor did the antiserum affect the homografi response. This antiserum, however, suppressed the homologous PCA reaction and the response to an intradermal injection of histamine, and also provoked the complement-dependent release of histamine from mast cells in vitro up to a dilution of 1:512. It was estimated that the average rat mast cell contains 19.8 pg of histamine. These results provide evidence for the importance of eosinophils, basophils and mast cells, i.e. pharmacologically-active cells, in allowing the host to mount a homocytotropic antibody-allergen mediated reaction as part of the immune response. [ABSTRACT FROM AUTHOR]

In earlier work, a chemotactic factor (leucoegvesin) specific for polymorphonuclear (PMN) leucocytes had been isolated from the sites of Arthus reactions or cutaneous burns. The substance shared antigenic sites with IgG. The possible existence precursor of the chemotactic factor in the γ2-globulin fraction of normal rabbit sera is suggested since the protein fraction on incubation with a purified neutral SH-dependent protease from inflammatory tissue became strongly chemotactic. [ABSTRACT FROM AUTHOR]

C&567macr;, prepared by the interaction of purified C&56macr; with purified C7 in solution, has been shown to be chemotactic for neutrophils and eosinophils. This activity was not significantly affected by the presence of C8 and C9, even in substantial excess. However, if the interaction of C&56macr; with C7 took place in the presence of an excess of erythrocytes, these cells bound the C&567macr; and were converted to EC567; and no chemotactic factor was found. [ABSTRACT FROM AUTHOR]

Antibacterial activity against Escherichia coli was found in the cytoplasmic extracts of rabbit polymorphonuclear leucocytes. The lack of antibody activity was determined by agglutination, passive haemagglutination, bactericidal reaction in the presence of complement, bactericidal plaque technique and immunoelectrophoresis. Thermostability of the extract, higher activity in a sodium citrate buffer than in veronal buffer containing Ca++ and Mg++ and negative haemolysis give evidence against the participation of complement in this type of antibacterial activity. Inhibition of the activity following absorption with different bacterial suspensions is interpreted as involving electrostatic bonds between active cationic substances and the negative charge of the bacterial surfaces. In addition, inhibition of the antibacterial activity was proved in the presence of both native and inactivated piglet and rabbit whole sera and albumin, which suggests that there is no independent action of these cytoplasmic substances in serum in vivo. [ABSTRACT FROM AUTHOR]

An inflammatory reaction in the thyroid of guinea-pigs, induced by intravenous injection of rabbit anti-guinea-pig thyroglobulin serum, has been studied at time intervals ranging from ¼ hour to 20 days after injection. Specific staining methods for eosinophils have been used to demonstrate that the inflammatory infiltrate mainly consisted of eosinophil granulocytes, but that neutrophil granulocytes were also present at the earliest time intervals. PAS-staining revealed that the granulocytes contained rounded droplets of material with a PAS-reactivity comparable to that of the colloid. This was most clearly seen 12 and 24 hours after injection. The possibility that this material represents phagocytosed thyroglobulin perhaps in an antigen-antibody complex has been suggested. The number of mast cells was counted and degranulation, which was apparently most extensive 12 hours before maximum granulocyte infiltration, was observed. Possible mechanisms involved in eosinotaxis and uptake of antigen-antibody complexes in the granulocytes have been discussed. [ABSTRACT FROM AUTHOR]

The significance of the C′ system for chemotaxis of rabbit PMNs was studied in vitro. C′6-deficient rabbit sera and C′5-deficient mouse sera were compared with normal rabbit and mouse sera for their ability to generate chemotactic factors in the presence of antigen-antibody complexes and endotoxins. No difference could be detected between these C′-deficient and normal sera. Full chemotactic activity was observed even when a rabbit anti-C′6 serum was incubated with an antigen-antibody complex. These results are incompatible with C′ (5, 6, 7)a being the chemotactic factor in serum for polymorphonuclear leucocytes. [ABSTRACT FROM AUTHOR]

The chemotactic factor, generated in whole rabbit serum following treatment with immune precipitates, was found to be principally associated with fractions containing the fifth and sixth components of complement (C'5 and C'6) after electrophoretic separation of serum. The chemotactic factor could also be generated in the intact animal, adding credence to its importance in tissue reactions induced by immunological agents. The isolated and activated C'5 and C'6 complex was found to be at least 10-20 times more active in the chemotaxis of rabbit polymorphonuclear leucocytes (PMNs) in vitro than bradykinin, kallidin, histamine, serotonin and extracts of PMN granules. The amino acid derivatives N-CBZ-glycyl-L-phenylalanine and N-CBZ-α glutamyl-L-tyrosine inhibited formation of the chemotactic factor in serum. The latter derivative also caused loss of activity of the preformed chemotactic factor in rabbit serum and in density gradient ultracentrifugation it was found that the C'5 and C'6 complex dissociated in the presence of this inhibitor. Chemotactically active C'5 to C'6 fractions, when added to the suspension of PMNs, prevented the ability of these cells to migrate toward a chemotactic source. It was also found that the ratio of combination of C'5 and C'6 was critical for the full expression of chemotactic activity. Utilizing purified components of human C', very recently obtained data indicate the requirement of the seventh component of C' (C'7) for generation of chemotactic activity. No later reacting components of C' are required. Whether C'7 is incorporated into the chemotactically active complex which sediments rapidly in the ultracentrifuge is not yet established. Similar data with guinea-pig C' complexes were also obtained. [ABSTRACT FROM AUTHOR]

A modification of Boyden's method for evaluation of chemotaxis is described. It permits the measurement of chemotactic agents in serum-free medium. Its possible usefulness for the determination of isolated chemotactic mediator(s) is discussed. [ABSTRACT FROM AUTHOR]

Human polymorphs have been found to ingest a wide variety of particles without the aid of serum, indicating that the cells possess a serum-independent mechanism of phagocytosis. Polymorphs have also been shown to possess a different and complementary mechanism of phagocytosis which depends on the presence of serum and serum components in the medium. Ingestion of starch particles by human polymorphs in the absence of serum was unaffected by media at the extremes of physiological pH and at tonicities between 205 and 348 m-osmoles/1 and by the presence in media of neutral and acid mucopolysaccharides. Phagocytosis of starch particles was reversibly inhibited by media of high tonicity and irreversibly inhibited by low concentrations of bacterial lipopoly-saccharide, which had a lethal effect on the cells. Serum was unable to promote phagocytosis by polymorphs in media of high tonicity but both untreated serum and inactivated serum promoted phagocytosis in media containing endotoxin, probably by neutralizing the action of the lipopolysaccharide. Phagocytosis of starch particles by human polymorphs in the absence of serum was inhibited by prior treatment of the cells with iodoacetate, suggesting that serum-independent phagocytosis relies on glycolytic energy. Ingestion of starch particles by polymorphs, treated with iodoacetate, was largely contingent on the presence of serum in the suspending medium, but inactivated serum and individual plasma proteins also had a limited ability to promote phagocytosis. These results suggested that cell glycolysis is not the principal source of energy in the serum-dependent mechanism of phagocytosis. Using hydrocarbon test particles, it was shown that combinations of either of the heat-labile components of complement (C′1 or C′2) with the C′4 component were active in promoting phagocytosis. Evidence was also presented that the C′3 component had some activity and another serum factor, which was only present in the heat-inactivated sera of some individuals, also had a limited ability to promote phagocytosis. [ABSTRACT FROM AUTHOR]

The phagocytic activity of human polymorphonuclear leucocyte preparations, which were free from plasma, has been estimated by direct determination under phase contrast of the number of living cells containing test particles. Spores of Aspergillusfumigatus were phagocytosed in the absence of added serum but phagocytosis of paraffin wax particles occurred only in the presence of serum containing the heat-labile and C′4 components of complement. In view of the unreactive nature of the paraffin hydrocarbons, it was considered unlikely that natural antibody played any part in the phenomenon. Although no phagocytosis of wax particles occurred in the absence of serum, almost 100 per cent of cells were phagocytic in preparations containing adequate concentrations of serum. It was therefore possible to determine the serum concentration necessary, for 50 per cent of the polymorphs to phagocytose wax particles. By this means it was demonstrated that the addition of the carbohydrate components of sputum had a small but significant inhibitory effect on phagocytosis and that dextran had no such effect. The sputum mucoprotein depressed the complement titre of serum and this might have accounted for the reduction in the ability of a serum to promote phagocytosis when this complex was added. The sputum mucopolysaccharide had no such effect on the complement titre of serum and must have exerted its inhibitor, action in some other way. [ABSTRACT FROM AUTHOR]

Serum from most normal persons contains specific antibodies which react with common bacterial species preparing their surfaces so that phagocytosis by leukocytes can take place. The Fab part of these antibodies reacts with immunologic specificity, with antigens on the surface of bacteria. Another part of the immunoglobulin molecule termed the Fc portion is activated during the attachment of the Fab portion to bacteria and becomes a site for attachment of bacteria to receptors on the surface of phagocytic cells. This activity is greatly amplified by heat-labile serum factors. Normally bacteria are rapidly killed by human polymorphonuclear leukocytes after engulfment occurs. However staphylococci and gram-negative species of bacteria survive in the leukocytes of patients with the syndrome "Chronic Granulomatous Disease of Childhood." These patients have suffered recurrent severe veto infections with bacterial species that are part of the body's resident bacterial flora. By contrast these patients are not at increased risk to infection from such pyogenic bacterial species a group A streptococci or pneumococci. The leukocytes from patients with chronic granulomatous disease produce little hydrogen peroxide during phagocytosis. Catalase-producing staphylococci and gram-negative bacteria are not killed, but hydrogen peroxide-producing streptococci and pneumococci are killed. A normal metabolic response to phagocytosis as well as release of lysosomal factor are essential for the bactericidal activity of human polymorphonuclear leukocytes. [ABSTRACT FROM AUTHOR]

Provides information on reversible neutrophil defect in ataxia telangiectasia. Results of the nitroblue tetrazolium dye test; Pathophysiology of the disease; Combinations of immune defects in ataxia telangiectasia.

Nitroblue tetrazolium (NBT) reduction by phagocytic leukocytes was investigated histochemically in 296 subjects with and without infections. Among 130 patients with noninfectious diseases, osteogenesis imperfecta patients and their relatives, and hemophiliacs frequently displayed high NBT-reduction values in their neutrophils. Four percent of the other noninfected patients yielded NBT-reduction values higher than normal in their neutrophils and were considered to have "false-positive" NBT-reduction tests. Most patients with acute bacterial infections (83%) showed an increased percentage of NBT-positive neutrophils. In patients who did not respond to a bacterial infection by increasing their percentage of NBT-positive neutrophils, there was suggestive evidence of a state of neutrophil dysfunction. Eighty-four percent of patients with viral or tuberculous infections demonstrated normal percentages of NBT-positive neutrophils. Acute bacterial infections were most often associated with a high percent of NBT-positive neutrophils, while low values predominated in viral infections regardless of the patient's total leukocyte counts. The NBT histochemical test would appear to be a useful adjunct to microbiologic techniques in establishing the diagnosis in patients with suspected infection. Pediatrics, 48:259, 1971, PHAGOCYTIC LEUKOCYTES, NITROBLUE TETRAZOLIUM DYE, BACTERIAL INFECTIONS, VIRAL INFECTIONS. [ABSTRACT FROM AUTHOR]

Polymorphonuclear leukocytes (PMNs) from children with chronic granulomatous disease of childhood have been shown to readily phagocytize staphylococci and certain gram-negative bacteria, but they demonstrate an impaired intracellular bactericidal capacity for these organisms. The in vitro phagocytic and bactericidal capacities of PMNs from three patients with this disease for four species is streptococci (Streptococcus faecalis, Streptococcus viridans, microaerophilic streptococci, and Streptococcus pyogenes) were tested by the modified method of Maaløe. The PMNs obtained from the patients phagocytized and killed the four species of streptococci in a normal manner while still showing the defect for Staph, aureus and S. marcescens. Morphologic examination of coverslip preparations of PMNs revealed minimal post-phagocytic degranulation and vacuole formation when either staphylococci and serratia or the streptococcal species were tested. This suggests that different intracellular mechanisms may be responsible for the streptococcal killing. These observations are in accord with the clinical courses of these patients, who rarely have serious streptococcal infections in contrast to the frequent and life-threatening infections caused by staphylococci and some gram-negative bacteria. [ABSTRACT FROM AUTHOR]

This article focuses on a research in which lipid composition and synthesis was studied in isolated polymorphonuclear (PMN) leucocytes in Down's syndrome (DS) and control patients. The subjects studied were 20 males aged 15 to 25 years all of whom bad physical signs typical of Down's syndrome. All 20 Down's syndrome subjects bad trisomy for chromosome 21. The results show that a decreased amount of total lipid is present in DS PMN leucocytes, primarily in cholesterol and in several phospholipid fractions.

Presents a case study of a child who developed mental deficiency, dwarfism and decreased segmentation of the neutrophilic leucocytes. Medical history of the patient; Findings of the laboratory and physical examinations conducted on the patient; Clinical presentation of the patient.

This article presents information on an instance of mosaicism that included XY and XXXY cell lines. Cytogenetic studies were made on a mentally defective male patient who had duplication of sex chromatin in a proportion of nuclei in a buccal smear, skin biopsy specimen and Leydig cells of a testis biopsy specimen. Neutrophils in peripheral blood lacked drumstick nuclear appendages and the testes were normal in size and histological structure. The chromosome complement of leucocytes in peripheral blood and cells in bone marrow was that of a normal male. Two cell populations were present in cells of skin and testis grown in vitro.

The article presents a discussion related to the polymorphonuclear lobe count in mongolism and its relation to the total leucocyte count. A considerable amount of evidence has accumulated that in patients with mongolism the nuclei of the polymorphonuclear neutrophil cells have fewer lobes than those in normal people. While the shift to the left in mongols can therefore be regarded as well established, it has not so far been related to the rest of the blood picture. In particular, it has not been decided whether the lower lobe count of mongols is associated with an increased number of leucocytes.

In the present investigation an in vivo method is described for studying the chemotactic activity elaborated by factors in human dental plaque. Plaque was sampled from the buccal and lingual surfaces of teeth from dental students, who had refrained from mechanical tooth cleansing during one 4 and one 8 day period. The plaque samples were suspended in 0.15 M NaCl, homogenized, and centrifuged at 12,100 × g for 30 minutes at 4°C. The plaque supernatant was then separated from the cellular pellet and sterile filtered through a 0.45 μ Millipore filter. The chemotactic activity elaborated by the filtrates was examined in (i) Boyden's in vitro chamber (ii) a wound chamber model described by Ludgren and Lindhe (1970). The results showed (i) that the wound chamber method is well suited for studying leukocyte chemotactic activity elaborated by dental plaque (ii) that factors in dental plaque stimulate the ernigration mainly of neutrophils (iii) that there is no increase in the amount of chemotactic factors/mg plque with increasing age of the plaque. Data from the wound chamber experiments further revealed that the plaque filtrates tested do not markedly influence the permeability to plasmaproteins and water of granulation tissue vessels. [ABSTRACT FROM AUTHOR]

Hyaluronidase activity has been studied in gingival crevicular material from clinically healthy and chronically inflamed gingivae, and in neutrophil polymorphonuclear leukocytes (neutrophils) from peritoneal exudates. Activity was demonstrated by determining the release of N-acetyl-glucosamine from purified hyaluronic acid. Hyaluronidase activity was found in gingival crevicular material within the pH range, 3-7, with pH optima at 3.7 and 6.0. Material from inflamed gingivae showed greater enzyme activity than that from healthy gingivae. Hyaluronidase activity was present in homogenates of neutrophils from peritoneal exudates with a pH optimum of 3.7. Little enzyme activity was found above pH 5.0. This suggests that the hyaluronidase activity at pH 3.7 in gingival crevicular material might originate from neutrophils in the gingival crevice. [ABSTRACT FROM AUTHOR]

The presence of leukocytes in clinically healthy and chronically inflamed gingival crevices was studied in humans and in dogs. Neutrophils, lymphocytes and monocytes were regularly found in samples of crevicular contents from healthy gingivae, even when histological sections of the adjacent tissue were free from infiltrates of inflammatory cells. Differential cellular counts showed crevicular samples to consist of 95- 97% neutrophils, 1-2% lymphocytes and 2-3% monocytes. More leukocytes were obtained from inflamed gingival crevices compared to healthy gingivae. The proportions of the various leukocytes were the same in the healthy and inflamed states (I). The presence and relative proportions of leukocytes in dog gingival crevices was studied during the development of gingivitis. In animals with clinically healthy gingivae, inflammation was induced by the institution of a soft diet, and repeated sampling of the crevicular contents was undertaken over a period of 35 days. The numbers of crevicular leukocytes increased during this period. No acute phase, similar to that which is found in other early inflammatory reactions, was observed with the methods used. The relative proportions of the various leukocytes remained unaltered throughout the period being 95-98% neutrophils, 1-2% lymphocytes and 1-3% monocytes. Experiments on cellular emigration, utilizing the skin window technique, indicated that a sustained predominance of neutrophils in a cellular exudate occurs when dental plaque is continuously present on the surface of the lesion. The consistently high proportion of neutrophils in the gingival crevice, therefore, might be explained by the presence of dental plaque in this area (II). The migration of blood neutrophils and monocytes in non-mechanically irritated, (resting), healthy and chronically inflamed gingivae was studied in dogs. Intravenously injected colloidal carbon was used for studying vascular permeability and for labelling the leukocytes. The resting state of the gingivae was confirmed by the absence of carbon labelling of the vessels, Leukocytes, labelled with carbon were observed, however, in the extravascular tissue shortly after labelling. Crevicular samples taken at increasing time intervals after the labelling procedure, showed that the cells continuous migration of leukocytes in both resting healthy, and inflamed gingivae, in spite of the absence of increased vascular permeability, indicates that the two phenomena can operate independently in the gingival area (III, IV). The effect of a short-term reduction in the numbers of neutrophils at the dento-gingival junction was studied in dogs having chronically inflamed gingivae. Nitrogen mustard and heterologous anti-neutrophil serum were used to induce the leukopenia. During leukopenia, the number of neutrophils at the dento-gingival junction was reduced. Concomitant with this reduction a decrease in gingivitis, as judged by gingival fluid measurements, was observed. The activity of acid phosphatase, protease and hyaluronidase in crevicular samples was also reduced, After leukopenia the values all returned. After leukopenia the values all returned to normal (V, VI). These results indicate that neutrophils at the dento-gingival junction play a role in the maintenance of the level of gingivitis and that their effect might be due to the release of lysosomal enzymes. [ABSTRACT FROM AUTHOR]

The ultrastructure of clinically normal beagle sulcular epithelium and underlying connective tissue has been studied in order to obtain base line information for comparison with similar studies of overtly inflamed gingiva. Normal sulcular epithelium in the beagle was observed to undergo a cytodifferentiation from typical keratinocytes of the stratum basale and stratum spinosum to flattened superficial. cells which contained a relatively - prominent endoplasmic reticulum and Golgi system. These cells also contained many dense granules of an unknown function. Narrow intercellular spaces filled with a material of electron density were observed between flattened superficial cells. Moderate numbers of neutrophils were found in the epithelium along with "clear cells" of an unknown type. The underlying connective tissue was characterized by a preponderance of well organized collagen fibers and numerous fibroblasts. No unusual findings were noted in either blood vessels or peripheral nerves. Macrophages were prominently located in the lamina in propria and in perivascular locations: Lymphocytes, neutrophils and plasma cells were seen in moderate numbers, usually located in the lamina propria portion of the sulcular wall. These cells when present in moderate numbers do not appreciably alter the integrity of the tissues and should be considered as components of a normal defense system. [ABSTRACT FROM AUTHOR]

Leukopenia was induced in six beagle dogs with chronic gingivitis by the injection of heterologous anti-neutrophil serum. During leukopenia, the number of neutrophils in the gingival crevice and within the gingival tissues was reduced. Concomitant with the reduction of neutrophil the amount of gingival fluid decreased. The activity of acid phosphatase, hyaluronidase and protease in crevicular samples was determined and was found to decrease in parallel with the reduction in neutrophil number. All parameters returned to normal during the post-leukopenic period. The results of the present study support previous suggestions that neutrophils at the dento-gingival function contribute to the enzyme milieu in the gingival crevice and that these cells, through their lysosomal enzymes, may induce vascular damage in periodontitis. [ABSTRACT FROM AUTHOR]

Topically applied horseradish peroxidase (HRPO) failed to penetrate through keratinized rat gingiva. It rapidly entered the intercellular spaces of the non-kertainized epithelial attachment and within 10 minutes it spread as far as the attachment apical limit. Widening of the intercellular spaces occurred. Spread into the underlying connective tissue was associated with dilation of venules and, after 30 minutes, emigration of neutrophils into the epithelial attachment Macrophage and to a lesser extent, fibroblasts showed uptake of HRPO into vesicles and lysosomes Some macrophages containing HRPO were located close to the crest of the alveolus A few cells of the epithelial attachment also showed the presence of HRPO in lysosomes . [ABSTRACT FROM AUTHOR]

The migration of neutrophil polymorphonuclear leukocytes and monocytes into crevices of resting clinically healthy, and chronically inflamed gingivae has been studied in dogs. Cellular samples were obtained from crevices by the sue of the "Styroflex" technique. Labelling of blood neutrophils and moncytes was performed with intravenously injected colloidal carbon. Samples were collected from the crevices at various time intervals after the labelling procedure. Labelled neutrophils and monocytes were found in the crevices of both healthy and chronically inflamed gingivae. The cells appeared to migrate at a higher rate into the inflamed crevices than into the healthy crevices. The monocytes seemed to migrate somewhat more slowly than the neutrophils. [ABSTRACT FROM AUTHOR]

The presence of leukocytes within the crevices of clinically healthy and chronically inflamed gingivae has been studied in human and dog. cellular samples were obtained from the gingival crevices by the "Styroflex" technique. The area on the styroflex film showing the presence of leukocytes was determined and a differential count of the cells performed. Biopsies were also taken from dogs with clinically healthy gingivae. Neutrophils, lymphocytes and monocytes were regularly founding samples from clinically healthy gingivae even when histological sections failed to show any inflammatory infiltrates in the gingival connective tissue. The differential counts showed 95-97 % neutrophils, 1-2% lymphocytes and 2-3% monocytes. Increased numbers of leukocytes were found in the crevices of chronically inflamed gingivae. However, the proportions of the various leukocytes were the same as those found in the crevices of clinically healthy gingivae. The results of th present investigation support the view that only quantitative differences exist between clinically healthy and chronically inflamed gingivae. [ABSTRACT FROM AUTHOR]

Chronically inflamed human gingiva was studied by electron microscopy, with particular emphasis on alterations in the tissues adjacent to the gingival sulcus. In spite of dilated intercellular spaces in the crevicular epithelium recognizable bacteria remained confirmed to the most superficial layers of the epithelium. Such spaces did, however, contain a variety of emigrating cell types, cellular debris, lysosomes and a granular precipitate. Lysosomes were also detected in the connective tissues and were released from disrupting neutrophils. Morphological variants of the plasma cell series formed the majority of the inflammatory cell population. The synthesis and extra-cellular release of proteins (immunoglobulins) was suggested in light of their morphological appearance. [ABSTRACT FROM AUTHOR]