The effects of consuming eggs for lunch on satiety and subsequent food intake.

The aim of the present work was to investigate the effects of eggs consumed for lunch on satiety, satiation and subsequent energy intake at the next meal. Thirty-one healthy male and female subjects participated in a randomized, three-way, crossover study. Following consumption of a standard breakfast, participants were asked to consume three isocaloric test lunches: omelette, jacket potato and chicken sandwich. Subjective measures of satiety were recorded using visual analog scales at regular intervals throughout the day. Energy intake at the next meal was assessed 4 h after lunch with an ad libitum meal. The egg lunch showed a significantly stronger satiating effect compared with the jacket potato meal. No effect on energy intake was seen. These data indicate that consumption of an omelette meal consumed at lunch could increase satiety to a greater extent than a carbohydrate meal and may facilitate reduction of energy consumption between meals.

The effect of different protein hydrolysate/carbohydrate mixtures on postprandial glucagon and insulin responses in healthy subjects.

OBJECTIVES: To study the effect of four protein hydrolysates from vegetable (pea, gluten, rice and soy) and two protein hydrolysates from animal origin (whey and egg) on glucagon and insulin responses. SUBJECTS/METHODS: Eight healthy normal-weight male subjects participated in this study. The study employed a repeated-measures design with Latin square randomization and single-blind trials. Protein hydrolysates used in this study (pea, rice, soy, gluten, whey and egg protein hydrolysate) consisted of 0.2 g hydrolysate per kg body weight (bw) and 0.2 g maltodextrin per kg bw and were compared to maltodextrin alone. Postprandial plasma glucose, glucagon, insulin and amino acids were determined over 2 h. RESULTS: All protein hydrolysates induced an enhanced insulin secretion compared to maltodextrin alone and a correspondingly low plasma glucose response. A significant difference was observed in area under the curve (AUC) for plasma glucagon between protein hydrolysates and the maltodextrin control drink (P<0.05). Gluten protein hydrolysate induced the lowest glucagon response. CONCLUSIONS: High amino-acid-induced glucagon response does not necessarily go together with low insulin response. Protein hydrolysate source affects AUC for glucagon more profoundly than for insulin, although the protein load used in this study seemed to be at lower level for significant physiological effects.

Contribution of alveolar phagocytes to antibiotic efficacy in an experimental lung infection with Streptococcus pneumoniae.

OBJECTIVES: The effect of cyclophosphamide-induced leukocytopenia on the cellular defence and on the efficacy of penicillin treatment in a Streptococcus pneumoniae pneumonia model in mice was studied. METHODS: The number of alveolar phagocytes was determined in broncho-alveolar lavage (BAL) fluid as well as the number of bacteria in both BAL fluid and homogenized lung tissue. RESULTS: Eighteen and 21 h after infection, leukocytopenic animals had significantly lower numbers of alveolar phagocytes than controls, while the numbers of bacteria in both BAL fluid and lungs were significantly higher. The number of bacteria was inversely related to the dose of penicillin and the number of alveolar macrophages. The number of alveolar granulocytes was inversely related to the dose of penicillin. CONCLUSIONS: Leukocytopenia due to cyclophosphamide impairs the cellular defence in the lung against Streptococcus pneumoniae and the dose of penicillin must be increased to compensate for the higher outgrowth of bacteria in these leukocytopenic mice, compared to normal animals.

Imaging of bacterial infections with 99mTc-labeled human neutrophil peptide-1.

UNLABELLED: This study was undertaken to evaluate whether 99mTc-labeled human neutrophil peptide (HNP)-1 can be used as a tracer for rapid visualization of bacterial infections. METHODS: Mice were injected intramuscularly with 1 million Staphylococcus aureus or Klebsiella pneumoniae organisms and 5 min later were injected intravenously with 0.4 microg (0.8 MBq) 99mTc-HNP-1. At various intervals, detailed information about clearance and accumulation of this tracer at sites of infection and in various organs was obtained by scintigraphy. 99mTc-labeled immunoglobulin G (IgG), an established marker of infection and inflammation, was used for comparison. RESULTS: After injection into S. aureus- or K. pneumoniae-injected mice, 99mTC-HNP-1 was rapidly removed from the circulation, mainly through the kidneys and bladder, with half-lives of 170 and 55 min, respectively. Similar half-lives were observed for 99mTc-IgG in these animals. Visualization of foci with S. aureus or K. pneumoniae, as indicated by a ratio of 1.3 or higher between the targeted thigh muscle (containing bacteria) and the nontargeted (contralateral) thigh muscle (T/NT), was already achieved 5 min after injection of 99mTc-HNP-1. Similar T/NTs for 99mTc-IgG were obtained 4 h after injection of the tracer, indicating that imaging of foci of bacteria with 99mTc-HNP-1 is much faster than with 99mTc-IgG. To obtain insight into factors that contribute to accumulation of 99mTc-HNP-1 at sites of infection, the binding of this tracer to bacteria and leukocytes was assessed using a peritoneal infection model. Binding of 99mTC-HNP-1 to bacteria was approximately 1000 times higher than binding to leukocytes. Although the number of bacteria in the peritoneum was 1000-fold lower than the number of leukocytes, a significant correlation between binding of 99mTc-HNP-1 to bacteria on the one hand and accumulation of tracer on the other was still found, in contrast to 99mTc-IgG. CONCLUSION: 99mTc-HNP-1 allows rapid visualization of bacterial infections. Binding of this tracer to bacteria most likely contributes significantly to the accumulation of 99mTc-HNP-1 at sites of infection.

Antibacterial activity of human neutrophil defensins in experimental infections in mice is accompanied by increased leukocyte accumulation.

Neutrophil defensins (or human neutrophil peptides-HNP) are major constituents of the azurophilic granules of human neutrophils and have been shown to display broad-spectrum antimicrobial activity. Other activities of these defensins, which are released from stimulated neutrophils, include cytotoxic, stimulatory, and chemotactic activities toward a variety of target cells. We studied the potential use of HNP-1 for antibacterial therapy of experimental bacterial infections in mice. In experimental peritoneal Klebsiella pneumoniae infections in mice, HNP-1 injection was shown to markedly reduce bacterial numbers in the infected peritoneal cavity 24 h after infection. This antibacterial effect was found to be associated with an increased influx of macrophages, granulocytes, and lymphocytes into the peritoneal cavity. These leukocytes appeared to be a requirement for the antibacterial effect, since in leukocytopenic mice administration of HNP-1 did not display antibacterial activity. HNP-1 treatment also reduced bacterial numbers in experimental K. pneumoniae or Staphylococcus aureus thigh muscle infections. In this model, radiolabeled HNP-1 was found to accumulate at the site of infection, whereas most of the injected HNP-1 was rapidly removed from the circulation via renal excretion. These results demonstrate that neutrophil defensins display marked in vivo antibacterial activity in experimental infections in mice and that this activity appears to be mediated, at least in part, by local leukocyte accumulation.

Radiolabelled antimicrobial peptides for imaging of infections: a review.

At present, it is difficult to distinguish between bacterial infections and sterile inflammatory processes using radiopharmaceuticals. This is so for a variety of reasons, including binding to bacteria with low affinity (e.g. infection) and binding to a specific micro-organism (e.g. radiolabelled monoclonal antibodies or F(ab)2 fragments thereof against micro-organisms). In this review, we propose that radiolabelled antimicrobial peptides should be the first choice in the development of new radiopharmaceuticals for imaging of bacterial infections. Antimicrobial peptides are a recently discovered component of the innate defence system of plants, animals and humans. These peptides, which now number more than 100, with proven microbicidal activity against a variety of micro-organisms, share certain properties, such as their small size and cationic charge. The latter allows them to bind preferentially to a broad spectrum of micro-organisms. We have recently demonstrated that radiolabelled human defensins allow the rapid visualization of bacterial infections in mice. Furthermore, binding of this antimicrobial peptide to bacteria is the major factor contributing to the accumulation of this tracer in bacterial infections. Based on these considerations, we believe that radiolabelled antimicrobial peptides will be an important asset in the imaging of infections in patients.

Detection of experimental infections with 99mTc-labeled monoclonal antibodies against TNF-alpha and interleukin-8.

This study was designed to assess monoclonal antibodies (MAbs) directed against tumor necrosis factor-alpha (TNF-alpha) (anti-TNF) or interleukin-8 (anti-IL-8) as radioactive agents for the detection of Staphylococcus aureus-or Klebsiella pneumoniae-infected thighs in mice. At 5 min (acute infection) or 20 h (established) post-infection, 20 micrograms of the 99mTc-labeled MAbs were injected. At various time intervals, the accumulation of the radiotracer in the infected thighs was assessed and expressed as a target-to-nontarget (T/NT) ratio. The binding of 99mTc-labeled MAbs to circulating mononuclear cells and granulocytes was quantitated 20 h after injection. The pharmacokinetics of the MAbs, in relation to the control agents 99mTc-labeled polyclonal human immunoglobulin (IgG) and a 99mTc-labeled nonspecific IgG1 MAb, were also studied. In acute infections, 99mTc-anti-TNF accumulated to a higher extent (p < 0.05) in S. aureus-infected thighs in mice until 4 h after the injection than 99mTc-IgG and was higher at 0.25 h in K. pneumoniae-infected mice (p < 0.03) compared with 99mTc-IgG. In established S. aureus and K. pneumoniae infections, 99mTc-anti-IL-8 detected the infection more intensely than 99mTc-IgG until 1 h after injection. In both S. aureus and K. pneumoniae infections, localization of sites of infection correlates (p < 0.05) with increased binding of the 99mTc-labeled MAbs to granulocytes and mononuclear cells in both acute and established infections. It was concluded that 99mTc-labeled MAbs, directed against TNF-alpha and IL-8, accumulate in bacterial infections in mice to a higher extent than does 99mTc-IgG after infection and is related to the binding of the antibodies to blood leukocytes. With these 99mTc-labeled MAbs, information might be gained about the development of an infection.

Ontogeny of milky spots in the human greater omentum: an immunochemical study.

BACKGROUND: Milky spots in the human greater omentum are preformed specific accumulations of primarily macrophages within the stroma of the greater omentum. To obtain a better understanding of milky spots in the human greater omentum, the development and the earliest forms of milky spots in the human greater omentum were studied, with special attention to the macrophage population. METHODS: Specimens of human greater omentum were obtained from fetuses of 20 to 40 weeks gestation and one newborn three days old (n = 6). Using mature macrophages (RFD 7), activated macrophages (RFD 1), B-lymphocytes (CD 22), and T-lymphocytes (CD 2), and immunoperoxydase labeling, the percentage of these cells in developing milky spots and the development of milky spots were studied by light microscopy. A time-dependent increase in the percentage of positive staining cells and the size of clusters was analyzed using the non-parametric Spearman rank correlation test. RESULTS: Small accumulations of cells with about 50% monocytes/macrophages were present at 20 weeks of gestation. With increasing gestational age the number of clusters of cells increased significantly (P < 0.01) as well as their size (P < 0.01). Starting at 29 weeks, vascularized clusters of cells were seen; true milky spots were present at 35 weeks. A significant (P < 0.05) increase in the percentage of mature macrophages was found in developing milky spots, whereas no activated macrophages were seen. The percentage of B-lymphocytes and T-lymphocytes found in the clusters of cells and milky spots increased significantly (P < 0.05) but did not exceed 10% of the total number of cells. CONCLUSIONS: From our data it can be concluded that milky spots are specific structures in the greater omentum formed between the 20th and 35th week of gestation. Further, we concluded that immature cells (promonocytes) mature locally in developing milky spots.

Localization of a bacterial infection with 99Tcm-labelled human IgG: further improvement with enriched IgG subclass preparations.

The aim of this study was to determine the contribution of various IgG subclasses to the scintigraphic detection of a staphylococcal infection. An experimental thigh infection in mice was used to determine the accumulation of the various 99Tcm-labelled IgG preparations with enriched IgG1, IgG2 or IgG4 subclass. Multiple-regression analysis was used to investigate a relationship between the IgG subclasses and the time-dependent accumulation in infected sites. Eighteen hours after infection with Staphylococcus aureus bacteria 20 micrograms of 99Tcm-labelled IgG preparations enriched with one of the IgG1, IgG2 or IgG4 subclasses by thiophilic absorption were administered intravenously and target-to-nontarget (T/NT) ratios were determined at 15 min, 1 h, 4 h and 24 h after injection of the tracer. Moreover, the binding of these preparations to S. aureus was assessed using an in vitro bacterial pellet model as an indication for the potency of detecting infections. As a control agent, 99Tcm-labelled polyclonal IgG (HIG) was used. In vivo, the T/NT ratios were significantly (P < 0.05) higher for the IgG1-enriched preparation at all time points, and for the IgG2-enriched preparation at 4 h and 24 h after injection, compared with HIG. In contrast, IgG4 did not yield higher T/NT ratios at any time. Using multiple-regression analysis, it became evident that IgG3 at all time intervals, IgG1 for early scans (up to 4 h) and IgG2 for late scans (24 h) contribute significantly (P < 0.05) to the accumulation. The abundance of IgG subclasses in the various preparations appeared to influence the accumulation of tracer at infected sites. The percentage of binding to S. aureus in vitro was significantly (P < 0.05) higher for enriched IgG subclass preparations than for HIG. We conclude that specific subclass enrichment of 99Tcm-labelled IgG preparations improves the scintigraphic detection of staphylococcal infections at various time intervals post-injection.

Phagocytosis and killing of suspended and adhered bacteria by peritoneal cells after dialysis.

OBJECTIVE: To determine the effect of dialysis fluid containing various glucose concentrations on the phagocytosis and killing of Staphylococcus aureus by rat peritoneal cells under conditions mimicking the in vivo situation. DESIGN: Phagocytosis and killing were evaluated by quantitation of the killing capacity of macrophages after in vivo phagocytosis of the bacteria as well as by an in vitro flow cytometric assay of the phagocytosis and killing of adhered bacteria by peritoneal cells. ANIMALS: Male Wistar rats. MAIN OUTCOME MEASURE: It was expected that the intraperitoneal administration of dialysis fluid would impair the capacity of peritoneal cells to eliminate bacteria. RESULTS: The first test revealed no effects of glucose concentration or dwell time on the killing of phagocytosed bacteria by macrophages, median percentages ranging between 29% and 64%. In the second series of experiments no effect of glucose concentration on the phagocytosis and killing of adhered bacteria was found either; however, longer dwell times significantly enhanced both the phagocytosis (at a dwell time of 1 hour, under 20%; at dwell times of 4 or 18 hours, above 20%, p < 0.02) and the killing (at a dwell time of 1 hour, under 53%; at dwell times of 4 and 18 hours, above 70%, p < 0.01). CONCLUSIONS: Glucose concentration has no effect on the phagocytosis and killing of Staphylococcus aureus, whereas the dwell time significantly enhances both of these functional capacities of peritoneal cells if the bacteria are adhered to surfaces.

Novel isolation and purification method permitting functional cytotoxicity studies of macrophages from milky spots in the greater omentum.

Milky spots in the greater omentum are well organized perivascular infiltrates of leukocytes which are probably involved in the clearance of tumor cells from the peritoneal cavity. In milky spots, macrophages are the predominant cell type forming a distinct population of cells. To investigate whether these macrophages have a function in the control of metastatic spread in the peritoneal cavity, a novel isolation and purification method was developed in order to study the functional cytotoxicity of macrophages from milky spots in the greater omentum against tumor cells in vitro. In order to obtain a cell suspension, greater omenta of unstimulated healthy male WAG/RIJ rats were incubated in collagenase/DNase suspension and filtered. Subsequently, macrophages were isolated and purified using flow cytometry by sorting unstained cells on the basis of size and internal complexity. Macrophages and other cells were identified by routine May-Grünwald-Giemsa staining and by immunophenotyping with the specific macrophage monoclonal antibody ED 1. Furthermore, macrophage subtypes were characterized by ultrastructural analysis. Functional cytotoxicity of the isolated macrophages was assayed against the syngeneic CC 531 tumor cell line in a colorimetric MTT assay. From three greater omenta of healthy rats 1.16 +/- 0.16 x 10(6) macrophages were isolated with a purity of 83 +/- 2% and a viability of > or = 96%. The macrophages were of the exudate (monocytic), exudate-resident and resident cell type and were in equal proportions. The contaminating cells were mainly mesothelial. A maximum cytotoxicity of approximately 30% was reached with the macrophage fraction at an effector-to-target ratio of 10. Furthermore, it was established that the mesothelial cells did not exhibit cytotoxicity.

Contribution of phagocytic cells and bacteria to the accumulation of technetium-99m labelled polyclonal human immunoglobulin at sites of inflammation.

The purpose of this study was to assess the contribution of phagocytic cells and bacteria to the accumulation of technetium-99m labelled polyclonal human immunoglobulin (HIG) at sites of inflammation. Mice were intraperitoneally injected with Staphylococcus aureus (SA animals), with heat-inactivated newborn calf serum (NBCS, to mimic a non-bacterial inflammation) or with physiological saline (controls); 1 h thereafter they received HIG. At various intervals after the administration of HIG the mice were killed, and the percentages of radioactivity in the peritoneal effluent and attached to the cellular and bacterial fraction thereof were established. Furthermore, the total number of cells and that of bacteria in the fluid were quantitated. The percentage of activity in the effluent in the SA animals was (P < 0.02) higher than those in the NBCS-injected animals and controls from 4 h onwards. In all groups of mice this percentage was highest at 4 h and decreased (P < 0.01) afterwards. The percentage of cell-bound activity and the total number of cells remained fairly constant or increased with time in the SA animals (P < 0.01). The bacteria-bound activity remained rather constant throughout the experiment and ranged between 4% and 6%. In the SA-infected animals the percentage of cell-bound activity was correlated with the total number of cells (macrophages but especially neutrophils) but even more strongly with the number of cell-associated bacteria. In the NBCS-injected animals a correlation was demonstrated between the cell-bound activity and the total number of cells (only neutrophils).(ABSTRACT TRUNCATED AT 250 WORDS)

A new 99mTc labelling method for leucocytes: in vitro and in vivo comparison with 99mTc-HMPAO.

A new method for the labelling of mixed leucocytes with 99mTc-tropolone was optimized and compared with a 99mTc-HMPAO leucocyte labelling procedure in vitro and in vivo. In the present study, leucocytes obtained from patients suffering from Crohn's disease, were isolated and labelled with 99mTc-HMPAO or labelled according the new 99mTc-tropolone procedure using 9.8 mM tropolone, 1 microM stannous chloride and 0.8 mM potassium borohydride (KBH4) at pH 5.5-6. Labelling efficiency with 99mTc-tropolone yielded 92 +/- 3%, which is higher compared to the 99mTc-HMPAO labelling procedure (64 +/- 13%) using 10(8) of leucocytes. In vitro stability and viability of both the tropolone and the HMPAO labelled cells was investigated. The viability test of the 99mTc-labelled leucocytes was performed in autologous plasma at 37 degrees C and compared with unlabelled leucocytes. After 18 hours of incubation a significant (P < 0.05) higher stability was observed for 99mTc-tropolone labelled leucocytes (84 +/- 5%) compared with that of 99mTc-HMPAO labelled leucocytes (73 +/- 5%). The viability of the 99mTc-labelled leucocytes observed for both labelling procedures was similar to unlabelled leucocytes. In vivo experiments were performed in mice. 99mTc-tropolone or 99mTc-HMPAO labelled murine mixed leucocytes were injected in mice, with a Staphylococcus aureus ATCC 25923 thigh infection. Analysis of scintigraphic images yielded a faster clearance of the 99mTc-tropolone labelled leucocytes. This was most likely due to a significant (P < 0.02) higher liver uptake at 4 hours after administration of the 99mTc-tropolone labelled leucocytes (19%) in comparison with 99mTc-HMPAO labelled cells (9%). Faster and significant (P < 0.02) higher accumulation of the 99mTc-tropolone labelled leucocytes was observed at the site of infection compared with 99mTc-HMPAO labelled leucocytes at all time-intervals after the administration of the 99mTc-labelled leucocytes. The new 99mTc-tropolone leucocyte labelling procedure, offers an attractive low-cost agent for research purposes.

Effect of glucose in dialysis fluid on antibacterial defence in the peritoneal cavity.

To study the effect of glucose concentration and dwell time of dialysis fluid on peritoneal antibacterial defence, an experimental infection with Staphylococcus aureus was induced in rats. For this purpose rats were inoculated intraperitoneally with Staphylococcus aureus at different intervals after the administration of various dialysis fluids. Twenty-four hours later the numbers of bacteria and cells in the peritoneal cavity were determined. The number of bacteria was correlated positively with the glucose concentration. Furthermore, an inverse correlation between dwell time and the number of bacteria was observed. Neither finding could be attributed to a glucose-dependent growth of the bacteria or disruption of the killing capacity of peritoneal cells in vitro. A glucose-dependent increase in the volume of the peritoneal fluid could partially explain the differences found in vivo. It is concluded that the glucose in dialysis fluid impairs antibacterial defence in the peritoneal cavity and that longer dwell times enhance this defence.

Effect of interferon-gamma in dialysis fluid on peritoneal defence in rats.

BACKGROUND: A major drawback of continuous ambulatory peritoneal dialysis (CAPD) is the occurrence of peritoneal infection. This might be explained by a non-optimal phagocytic capacity of peritoneal cells which can be improved by stimulating factors. AIM: To investigate the effect of addition of interferon-gamma (IFN) to dialysis fluid with various glucose concentrations or to saline (as control) on the peritoneal defence against Staphylococcus aureus in an experimental dialysis model in rats. METHODS: Twenty-four hours after the administration of either dialysis fluid containing various glucose concentrations or saline with or without IFN, bacteria were injected intraperitoneally. At the time of the bacterial infection and 24 h later cellular and bacterial parameters were studied. RESULTS: The addition of IFN to dialysis fluid or saline resulted in a significant (P < 0.01) increase in the number of peritoneal macrophages at the time of infection; this was accompanied by a significant increase in both the number of Ia-positive peritoneal macrophages (P < 0.01) and the production of nitrite by macrophages (P < 0.05) at the time. IFN in dialysis fluid as well as in saline significantly (P < 0.01) reduced the recovery of bacteria from the peritoneal cavity 24 h after infection. Only the absence of IFN glucose increased the recovery of bacteria from the peritoneal cavity at the same time. CONCLUSION: In this experimental model the addition of IFN to dialysis fluid lowered the recovery of staphylococci from the peritoneal cavity by means of activation of an increased number of macrophages.

Optimized localization of bacterial infections with technetium-99m labelled human immunoglobulin after protein charge selection.

To improve the scintigraphic detection of bacterial infections a protein charge-purified fraction of polyclonal human immunoglobulin was applied as a radiopharmaceutical. This purification was achieved by attaching the immunoglobulin to an anion-exchanger column and by obtaining the column-bound fraction with buffer. The binding to bacteria in vitro and the target to non-target ratios of an experimental thigh infection with Staphylococcus aureus or Klebsiella pneumoniae in mice were evaluated to compare the purified and the unpurified immunoglobulin. The percentage of binding to all gram-positive and gram-negative bacteria used in this study was significantly (P < 0.03) higher for the purified than for the unpurified immunoglobulin. For the in vivo study, mice were infected in the thigh muscle with Staph. aureus or K. pneumoniae. After 18 h 0.1 mg of technetium-99m labelled polyclonal immunoglobulin or 99mTc-labelled protein charge-purified polyclonal human immunoglobulin was administered intravenously. At all time intervals the target (infected thighs) to non-target (non-infected thighs) ratios for both infections were significantly higher (P < 0.03) for protein charge-purified polyclonal immunoglobulin than for unpurified polyclonal human immunoglobulin. Already within 1 h the infected tissues could be detected by the purified immunoglobulin. It is concluded that 99mTc-labelled protein charge-purified immunoglobulin localizes both a gram-positive and a gram-negative thigh infection more intensely and faster than 99mTc-labelled unpurified immunoglobulin.

Rat macrophage lysosomal membrane antigen recognized by monoclonal antibody ED1.

The monoclonal antibody (mAb) ED1 is being used widely as a marker for rat macrophages. The distribution of the recognized antigen in tissues and isolated cells strongly supports this use as a macrophage marker, since the majority of macrophages are recognized and only seldomly are other cell types stained by mAb ED1. In the present study we further characterized the recognized antigen by a detailed description of the localization of the antigen and by determining biochemical and functional properties. We show that the antigen is expressed on the membranes of cytoplasmic granules, like phagolysosomes, as well as on the cell surface. The amount of ED1 expression in a single cell can be correlated to phagocytic activity of the respective cell type, but the mAb ED1 is not able to block latex phagocytosis or bacterial killing. The mAb ED1 appears to recognize a heavily glycosylated protein of 90,000-110,000 MW, depending on the cell type used as antigen source. A possible relation with other known lysosomal glycoproteins with a similar molecular weight is discussed.

Influence of cytostatic agents on the pulmonary defence of mice infected with Klebsiella pneumoniae and on the efficacy of treatment with ceftriaxone.

The effect of cytostatic treatment on the cellular defence and the efficacy of treatment with ceftriaxone in Klebsiella pneumoniae pneumonia was studied. Mice, made monocytopenic and granulocytopenic by cyclophosphamide or monocytopenic by etoposide, were infected intratracheally with K. pneumoniae (approximately 10(4) CFU) and then treated with ceftriaxone. At various intervals, the numbers of bacteria in the broncho-alveolar lavage (BAL) fluid and in lungs homogenised after lavage were determined. Cyclophosphamide reduced the numbers of granulocytes in the BAL fluid significantly but reduced only slightly the number of alveolar macrophages at the time of inoculation, 12 and 15 h later. The number of CFU in cyclophosphamide-treated mice was higher than that in controls, being significant in the homogenised lungs at 15 h after infection. In etoposide-treated mice, the numbers of alveolar phagocytes in BAL did not differ from those in control mice, whereas the number of bacteria was lower (only significantly in BAL fluid at 15 h after infection) than that in the controls. In this short experimental infection cytostatic treatment did not affect the outgrowth of Klebsiella pneumoniae substantially or the efficacy of treatment with ceftriaxone.

Improved detection of a staphylococcal infection by monomeric and protein A-purified polyclonal human immunoglobulin.

The present study was undertaken to compare the technetium-99m labelled non-specific polyclonal human immunoglobulin (Ig) with 99mTc-labelled monomeric human immunoglobulin (m-Ig), 99mTc-labelled, protein A-purified, human immunoglobulin (A-Ig) and 99mTc-labelled monomeric, protein A-purified, human immunoglobulin (mA-Ig) as tracer agents for the detection of a thigh infection with Staphylococcus aureus. In vitro the binding of the various tracer agents to bacteria at various intervals was determined. For the in vivo evaluation, mice were infected and received one of the various labelled proteins. Scintigrams were made 0.25, 1, 4 and 24 h later. All 99mTc-labelled Igs bound to bacteria in vitro: the percentages of binding for the m-Ig (from 1 h onwards) and A-Ig and mA-Ig (from 3 h onwards) were significantly higher than that for Ig. The in vivo target-to-non-target (T/NT) ratios were significantly higher from 4 h onwards for all purified Igs than for Ig. Protein A-purified Igs yielded higher T/NT ratios than m-Ig. Furthermore, the amount of activity in the liver was significantly lower 24 h after administration of m-Ig, A-Ig and mA-Ig than after administration of Ig. It is concluded that in this experimental infection 99mTc-labelled monomeric Ig localizes a staphylococcal thigh infection better and faster than 99mTc-labelled unpurified Ig. However, the accumulation obtained with protein A-purified Ig or protein A-purified monomeric Ig was the highest of all tracer agents tested.

Separation of alveolar macrophages and dendritic cells via autofluorescence: phenotypical and functional characterization.

In a previous study we demonstrated that an increase of monocytes and dendritic cells (MDC) was In the current study, the bright autofluorescence of alveolar macrophages (AMs) was used to separate them efficiently from the MDC. Sorting of freshly isolated BAL cells resulted in a high-autofluorescent fraction, consisting predominantly of AMs, and a low-autofluorescent fraction containing the MDC, lymphocytes, and granulocytes. Thus, a clear separation between suppressive (AM) and stimulating (MDC) activity was obtained as shown in antigen-specific T cell responses. Flow cytometric parameters, density fractionation, and a series of ED monoclonal antibodies raised against rat macrophage antigens showed that both AMs and MDC were diverse populations. After overnight culture, more than 80% of an MDC population with a density range of 1.065-1.079 changed to a lower density (< 1.056) and morphologically developed into DCs with many processes. Concomitantly, monoclonal antibody ED1 expression changed from a granular pattern to a discrete juxtanuclear spot localization.