Number Of Neutrophilic Granulocytes Research Articles

DYNAMICS OF INDICATORS OF CELLULAR IMMUNITY IN PUPPIES DEPENDING ON THE MULTIPLICITY OF INTRODUCTION THE BIOLOGICAL STIMULUS

Study of immune-physiological indicators in puppies in critical periods of life, including and for the introduction of a biological stimulus, is an actual direction of modern researches. Actuality is related to a change in immunogenic properties of biological stimulus, a decrease in resistance of animal body, and lack of available methods for assessing the immune status of puppies after vaccination. The purpose of the research was to determine the dynamics of indicators of the cellular link of immunity in puppies depending on the frequency of a biological stimulus (BS) introduction. The experiment was conducted on 10 puppies from one brood. Puppies were divided into two groups: I – single vaccination at 51 days of life was done; II - two-time vaccination on the 28th and 51st days of life was done. Biocan DHPPi vaccine (Czech Republic) was used as a biological stimulus. For conduction the immunological studies, blood from puppies’ the jugular vein at 7, 14, 28, 51, 71, and 90 days of age was used. On days 28 and 51 of the study, blood was taken directly 5 minutes before biological stimulus administration. The absolute number of leukocytes, neutrophils, monocytes, lymphocytes and the phagocytic activity of neutrophils were determined in blood stabilized with heparin. During the first month of the postnatal period, the absolute number of neutrophils in the blood of puppies of both groups was almost the same, the difference between the groups was within 0.2–0.7 G/l. Starting from the 28th day, a significant increase in the number of neutrophils in the blood of puppies was recorded, similar to when determining the number of leukocytes. The analysis of the research results showed that the absolute number of lymphocytes, unlike other cells, had a clear tendency to decrease, starting from 14 to 70 days of age. Authors' research established that regardless of the frequency of biological stimulus introduction, a physiological crossover occurs in the body of puppies. However, it should be noted that the biological stimulus affects the quantitative composition of lymphocytes and neutrophils. This is expressed in the fact that in the group of puppies, where the biological stimulus was administered, the increase in the number of neutrophils was 2 times less than in the other group (where it was not administered on the 28th day). Regardless of frequency of the biological stimulus introduction on 70 days, the number of neutrophil granulocytes and lymphocytes in the blood of both groups did not differ significantly and maintained this trend during the next three weeks of observation. It was established that the administration of BS to puppies causes an immunosuppressive effect on the body, as evidenced by a decrease in the absolute number of neutrophils and lymphocytes in the blood. At the same time, administration of BS does not affect the phagocytic activity of blood neutrophil granulocytes. Physiological crossover, namely a decrease in the absolute number of lymphocytes and an increase in neutrophils in the blood of puppies occurs after 28 days of life without connection to administration of BS. Further research is needed to study the feasibility of a two-time administration of the vaccine as a biological stimulus in the studied terms.

Evaluation of the effects in the <i>in vitro</i> system of synthetic thymic hexapeptide on the expression levels of NF-κB, IFNα/βR and CD119 neutrophilic granulocytes in patients with chronic herpes viral co-infections

Background. Strategies used by herpes viruses with human cells are complex and multifaceted. On one hand, inborn defects in antiviral immune defense have been unveiled, which also affect interferon (IFN) system underlying development of chronic recalcitrant relapsing viral infections such as remittent respiratory viral infections, herpesvirus infections, and papillomavirus infections. On the other hand, numerous viruses are able to damage both immune system and IFN network. During inborn and acquired defects in IFN network, inborn or induced mutation in gene products involved in signaling cascades aimed at upregulating gene expression responsible for IFN production are observed. One of the strategies used by diverse viruses is altering some signaling pathways resulting in activated transcription factors including nuclear factor NF-kB. However, antiviral mechanisms executed by neutrophilic granulocytes (NGs), particularly affecting NF-kB expression have not been elucidated. Aim of the study: to study in vitro features of NF-kB expression and number of neutrophilic granulocytes (NG) expressing membrane IFN/R and IFNR in patients with atypical chronic active herpes virus infections (AChA-HVI), followed by assessing an effect of arginyl-alpha-aspartyl-lysyl-valyl-tyrosyl-arginine hexapeptide (HP), a synthetic analogue of the active center of the thymopoietin (active substance of drug Imunofan, Russia), on the expression of NG NF-kB and IFN/R and IFNR. Materials and methods. We observed 25 patients of both sexes aged 23 to 64 years with AChA-HVI, manifested by chronic fatigue syndrome and cognitive disorders. Study design: stage 1 clinical, ELISA, PCR methods, FC was used. Stage 2 the in vitro experiment: 32 blood samples from 8 healthy adults and 375 blood samples from 25 patients with AChA-HVI were analyzed: % NG expressing NF-kB, IFN/R, IFNR and the relevant MFI levels by using FC before and after incubation with HP. Results. Our study demonstrated low level (MFI) of NF-kB expression in 100% NG associated with decreased % of NG expressing IFN/R and IFNR in all patients with AChA-HVI and low serum level for IFN and IFN in comparison with healthy individuals. In the in vitro experiment there was shown that 100% of NG expressed NF-kB after exposure to HP. However, only 48% patients (SG 2) restored NF-kB expression level (MFI) to normal range and 52% of cases (SG 1) had no response. HP increased % of NG expressing IFN/R in SG 2 and increased % of NG expressing IFNR in SG 1. Conclusions. It was shown, that influence of HP in vitro has ambiguous effects on the expression of NF-kB, % of NG expressing IFN/R and IFNR in patients with AChA-HVI. We assume that different NF-kB response to HP is associated with inborn or secondary NF-kB deficiency.

MiRNA-141-5p Affects the Levels of Neutrophil Elastase in Preeclampsia by Regulating MAPK1

Abstract Objective The objective of this study was to investigate the expression levels of microRNA-141-5p(miRNA-141-5p), MAPK1 and neutrophil elastase in patients with and without preeclampsia (PE), and the relationship between miRNA-141-5p and MAPK1 with respect to the secretion of elastase by neutrophils in patients with PE. Methods Thirty patients with PE and 30 healthy pregnant (HP) women were recruited from The Second Hospital of Shanxi Medical University, Taiyuan, China, between February 2017 and July 2018. Neutrophils were isolated from 8 mL peripheral blood samples and cultured. We recorded neutrophil count and morphology during culture. Apoptosis was detected by flow cytometry in different groups at 0, 24, and 48 h. The expression levels of elastase were detected in neutrophils by enzyme-linked immunosorbent assay, whereas the expression levels of miRNA-141-5p in peripheral blood neutrophils were detected by real-time polymerase chain reaction. We used TargetScanHuman Release 7.2 to analyze the target genes of miRNA-141-5p. The expression of MAPK1 in peripheral blood neutrophils was detected by western blotting. Data were analyzed by SPSS version 21.0 software, and comparisons between groups were carried out with the Student t test. Results There was no significant difference between the PE and HP groups (P > 0.050) with regard to age or body mass index. The weight of newborns in the PE group (2846.00 ± 600.00 g) was significantly lower than that in the HP group (3055.00 ± 230.68 g). The number of neutrophilic granulocytes(NGs) in blood samples from the PE group was significantly higher than that in the HP group (P = 0.003). There was no significant difference between the groups with regard to morphology. Apoptosis in the PE group was delayed when compared with the HP group at different time points. The P value of apoptosis in the PE and HP groups were respectively 0.790, < 0.001 and 0.030 at 0 h, 24 h and 48 h. The expression levels of miRNA-141-5p in the PE group were significantly lower than those in the HP group (P < 0.050). The expression levels of MAPK1 in neutrophils from the PE group were significantly higher than those in the HP group (P < 0.050) by western blot. The expression levels of elastase in neutrophils from the PE group were significantly higher than those in the HP group (P < 0.050). Furthermore, the number of NGs in peripheral blood from the PE group was higher than that of the HP group; however, the levels of apoptosis were lower. The expression levels of miRNA-141-5p in NGs decreased, the expression of MAPK1 increased, and the secretion of neutrophil elastase in the NG medium increased in the PE group than those in the HP group. Conclusion Collectively, our analysis suggested that miRNA-141-5p may be involved in the pathogenesis of PE by regulating the MAPK1 signaling pathway to activate neutrophils and increase the secretion of elastase.

CYTOLOGICAL CHARACTERISTICS OF REGENERATIVE PROCESSES IN PURULENT WOUND DEPENDING ON THE TIME OF SURGERY (MORNING EVENING) AND CHRONOTYPE OF PATIENTS (MORNING AND EVENING CHRONOTYPE)

Healing of postoperative wounds is a staged process. At the early stage of the wound process is characterized by the presence of a large number of unchanged and altered erythrocytes, the latter are characterized by shrinkage, change in shape, the formation of clusters. Hemorrhages in a wound and the initial phenomena of an inflammation are defined. In the period of regeneration of wound healing, inflammatory phenomena gradually disappear, and regenerative processes are increasingly developed. n the study of the dynamics of healing of postoperative wounds, the most informative is the method of "superficial wound biopsy". For cytological material is taken by lightly scraping the surface layer of the wound with a special spatula. This method usually manages to obtain elements not only of the exudate and the superficial (granular-fibrinous) layer of the wound, but also newly formed cells. The cytograms obtained in this way provide significant information about the course of regenerative processes in the wound. [1]. Using the method of "surface biopsy" of wounds to determine the features of the regenerative processes in the wound depending on the time of surgery (morning evening) and the chronotype of patients (morning and evening).
 Cytological examination was performed using the method of "surface biopsy" of the wound [2]. The material for the study was taken by lightly scraping the surface layer of the wound with a spatula on the 1st, 3rd, 5th and 7th day of the postoperative period. The resulting material was transferred to a glass slide, evenly distributed in a thin layer, fixed and stained by Romanowski-Gimza [3]. The obtained preparations were studied using a microscope Biorex-3 BM-500T with a digital photomultiplier DCM 900 with programs adapted for research data. Cytometric study was performed by the method of standard planes [4] at a magnification of x1000, determined the average number of erythrocytes, unaltered neutrophilic granulocytes, naked nuclear neutrophilic granulocytes, macrophages, lymphocytes, fibroblasts, fibrin threads and collagen fibers in 10. Microphotography of the selected areas for illustrations was performed using a microscope Biorex-3 BM-500T with digital photomultiplier DCM 900.
 It is established that the time of surgery, depending on the patient's chronotype, affects the course of the postoperative period and the wound healing process. Carrying out of operation according to the maximum activity of the patient reduces terms of realization of reparative process in a wound. Acceleration of hemostasis is accelerated and microcirculation and, accordingly, oxygenation of damaged tissues is restored faster. The number of unchanged and altered erythrocytes is probably lower in the superficial biopsies of wounds of patients who were operated with the chronotype in the most active time. In them, the dynamics of changes in the number of neutrophilic granulocytes indicates an earlier transition from the inflammatory stage of the wound process to regenerative. The obtained cytological data indicate the implementation of the reparative process 2 days earlier.

Особливості імунної відповіді організму піддослідних тварин на дію мийних засобів на основі аніонних поверхнево-активних речовин, призначених для догляду за дитячим посудом

Objective: We assessed the impact of the detergents, based on anionic surfactants, for the care of the children’s tableware on the indicators of the immune system of the experimental animals. Materials and methods: The effect of aqueous solutions of Alionka, Karapuz, Ushastyi Nian, Frosch Baby detergents on the indicators of the immune system was performed in the experiment with guinea pigs during 21 days. We determined the number of T- and B-lymphocytes, the number of neutrophilic granulocytes and active phagocytes; studied the reaction of the precipitation of circulating immune complexes with a solution of polyethylene glycol 6000. We used the methods of statistical processing of the results of medico-biological research. Results: The obtained results showed that the epicutaneous impact of all studied detergents for the care of children’s tableware could cause the immunomodulatory effects in the organism of guinea pigs. Changes in the largest number of the indicators of the immune system, which affected all links, were observed when exposed to Ushastyi Nian detergent. The least pronounced changes of the immunity were observed when exposed to the Frosch Baby detergent and they concerned only the indices of nonspecific resistance of the organism. The revealed inhibition of the functional activity of neutrophilic granulocytes, which was observed in animals of all experimental groups, may indicate a decrease in the protective functions of the organism of guinea pigs.

Clinical use of Aglepristone for treatment of open-cervix pyometra in cats

The results of the clinical application of the integrated therapy of cats having an open-cervix of the pyometra are presented in this work. It has been proved that pathology affects mostly the animals in the age from 3 to 8 years. In the clinical study, it was found that in the open-cervix of the pyometra cats had also depression, anorexia, polydipsia, polyuria, increase in the abdomen, withdrawal of the purulent exudate from the vagina. In micropreparations taken from the vaginal mucosa, an increase in the number of neutrophilic granulocytes was observed, most of them with signs of apoptosis. Significant changes in functional reactivity of phagocytic cells were found. Using microbiological researches the polymicrobial association of pathogenic microorganisms have been identified in the exudate. Hematologic studies have shown decrease of hemoglobin content and signs of neutrophilic leukocytosis. In ultrasonography, patients with pyometra showed an increase in the body and horns of the uterus, which were stretched with accumulated fluid, thickening of the organ's wall, and a clear picture of the cystic endometrial hyperplasia of the endometrium was visualized. The research has tested a treatment regimen with the use of Aglepristone (Alizin® Virbac, France) in combination with Mastometrin and antibiotic therapy (Amoscillin 15%, INVESA, Spain). During the treatment the fever, vomiting and polydipsia have disappeared, the appetite hat restored. Laboratory studies have established a dynamic reduction in the number of leukocytes and fading reactive neutrophilia. The ultrasound has noted decreased diameter of the uterus. Major hematological and immunological parameters of homeostasis were normalized. The obtained clinical studies indicate that the complex scheme of therapy of cats for the open-cervix of the pyometra contributes to the restoration of the functional state of the uterus, the extinction of the pathological process and the normalization of the functions of all organs and systems.

SWI/SNF Protein SMARCD2 Orchestrates Transcriptional Networks Controlling Hematopoiesis and Neutrophil Granulocytes in Humans, Mice and Zebrafish

Differentiation of hematopoietic stem cells follows a hierarchical program of transcriptional-regulated events. We here identify SMARCD2 (Swi/Snf-related matrix associated actin dependent regulator of chromatin, subfamily D, member 2) as critical regulator of myelopoiesis in humans, mice, and zebrafish. We studied four patients from three unrelated pedigrees presenting with a novel syndromatic phenotype comprising congenital neutropenia, specific granule deficiency, susceptibility to myelodysplasia with excess of blasts, and various skeletal anomalies. All patients had homozygous loss-of-function mutations in SMARCD2. In contrast to wildtype alleles, the variant alleles did not give rise to proteins with capacity to interact with the SWI/SNF subunits BRG1, BAF170, BAF155, and BAF47, as shown by co-immunoprecipitation experiments. In vitro, knockdown of SMARCD2 in promyelocytic NB4 cells, differentiated in the presence of ATRA, led to decreased expression of genes encoding the primary granule proteins cathelicidin (CAMP) and alpha-1-antitrypsin (AAT) as well as specific granule proteins matrix metalloproteinase-8 (MMP8), transcobalamin (TCN1) and lactoferrin (LTF). This phenotype is reminiscent of patients with specific granule deficiency, characterized by mutations in CEBPE, a known transcription factor controlling terminal neutrophil development. We therefore hypothesized that SMARCD2 may act via CEBPe and performed immunoprecipitation studies in transfected cells. Upon pull-down of SMARCD2, CEPBE could be detected, and vice versa, suggesting that both proteins physically interact to control transcriptional networks. To interrogate effects of SMARCD2 deficiency on global chromatin accessibility we made use of ATAC sequencing of undifferentiated and ATRA-differentiated NB4 cells and compared this data with comprehensive RNA-sequencing results. A specific subset of genes was found deregulated in both assays, affecting vesicular trafficking, migration and signalling pathways. To validate a role for SMARCD2 in hematopoiesis in vivo, we generated murine and zebrafish model systems. We generated Smarcd2-/- mice by injection of Smarcd2+/- murine ES cells into blastocysts, transfer into pseudo-pregnant mice and interbreeding of heterozygous Smarcd2+/- offsprings. The mutant allele was inherited in a Mendelian fashion but no viable mice were born. 14.5dpc embryos were characterized by anemia and reduced size compared to their littermates. Analysis of fetal liver hematopoiesis revealed a complete absence of CD11b+Gr1+ and CD11b+Ly6c+ cells, whereas the number of LSK stem cells was not affected. Futhermore, Smarcd2-/- embryos showed aberrations in erythroid cells such as extensive anisocytosis, multinucleated cells, and perturbed mitosis. In cytokine-driven colony forming unit assays, GM-CSF, M-CSF, and G-CSF induced myeloid cell differentiation was decreased. Transcriptional profiling of LSK stem cells revealed a striking dysbalance affecting genes involved in signaling pathways and host defence, including CEBPE-dependent genes. Among a total of 12362 detected genes, we found 4290 to be differentially expressed (DESeq2, FDR<10%). Interestingly, the majority (79%) of the 605 genes with a relatively large difference (fold-change > 1.4, FDR<1%) were up- and not downregulated. Next, we generated three Smarcd2-deficient zebrafish models using a) morpholino-mediated knockdown in Tg(mpx:EGFP)i114 and Tg(lyz:dsRed)nz50 strains of the orthologous gene or b) Crisp/Cas9-mediated genomic engineering of this locus in Tg(mpx:EGFP)i114. In all models, the numbers of neutrophil granulocytes were significantly reduced. We conclude that SMARCD2 is a critical factor orchestrating transcriptional networks controlling hematopoiesis across species, in particular regulation and maintenance of neutrophil differentation and prevention of leukemogenesis. DisclosuresAbboud:Novartis: Honoraria; MAST Therapeutics: Research Funding; Eli Lilly and Company: Membership on an entity's Board of Directors or advisory committees, Research Funding; Pfizer: Membership on an entity's Board of Directors or advisory committees.

Interleukin-17 enhances the removal of respiratory syncytial virus in mice by promoting neutrophil migration and reducing interferon-gamma expression.

The aim of this study was to observe the effect of interleukin (IL)-17 on early immune response and inflammation in the lungs of respiratory syncytial virus (RSV)-infected mice. Specific pathogen-free BALB/c mice were randomly assigned to control, RSV-infected, RSV-infected with phosphate-buffered saline, and RSV-infected + IL-17 intervention groups. The RSV infection model was set up by nasal mucosa immunization. The intervention group was provided with restructured IL-17 (intranasal). The viral load and cytokine concentrations in the lung tissues and broncho-alveolar lavage fluid (BALF) were determined by real-time-polymerase chain reaction and enzyme-linked immunosorbent assay. RSV caused acute lung inflammation in mice with a significantly higher number of neutrophils and cytokines such as interferon-gamma (IFN-γ), IL-1β, IL-6, and G-CSF in the BALF than that in the control group. IL-17 intervention led to a significant increase in the number of neutrophilic granulocytes in the BALF. Alternately, IL-17 intervention led to a significant decrease in the IFN-γ concentration and a significant increase in the IL-1β, IL-6 and G-CSF levels in the BALF. IL-17 induced a reduction in the viral load and an increase in the survival rate of mice on the third day of infection. IL- 17 mucosal immunity enhances the removal of RSV and strengthens the immune defense by promoting neutrophil migration and reducing the IFN-γ levels in mouse lungs.

Revision arthroplasty : Histopathological diagnostics in periprosthetic joint infections

Histopathological differences in synovia and synovial-like interface membrane (SLIM) patterns can be used to differentiate periprosthetic particle-induced reactions, bacterial infections (bacterial synovitis and osteomyelitis), mechanical-induced tissue alterations, adverse reactions to implant material, and arthrofibrosis (SLIM consensus classification). Because of differences in treatment the diagnosis of a bacterial implant infection is very important. Histopathological tests and scoring systems are important diagnostic tools in identifying deep implant infections in patients with unclear clinical history as well as radiographic and laboratory studies. Modern enzyme PCR-based methods, histochemical- and immune-histopathological techniques (CD3,CD15, CD68) are useful in identifying specific and nonspecific infections, as well as differentiating postsurgical changes from recurrent infections in patients with a spacer. In all histopathological scoring systems for bacterial infection, quantifying the number of neutrophil granulocytes in a defined number of high power fields is crucial. Neutrophil granulocytes can be detected through histochemical methods and more specifically by immune-histopathological techniques and by various quantification systems (histopathological scores) leading to the diagnosis of bacterial peri-implant infection. One important function of histopathology, apart from diagnosing infection, is to rule out other mechanisms of implant failure, such as tumor infiltrations, particle-induced reactions, and adverse reactions to implant materials.

Effect of praziquantel prolonged administration on granuloma formation around Schistosoma japonicum eggs in lung of sensitized mice

Schistosomiasis remains a major public health problem and it is an immune disease. The schistosome egg is the primary parasite factor responsible for the overt disease. The eggs release the soluble antigen, which induces intensive tissue reaction, a granulomatous reaction to the eggs. If granuloma formation could be suppressed, overt disease might not develop. Praziquantel is an effective antischistosomal drug especially for adult worms. However, whether praziquantel has a suppressing effect on granuloma formation around schistosome eggs directly remains unclear. The purpose of the present study was to investigate the effect of praziquantel, especially administered persistently, on granuloma formation around Schistosoma japonicum eggs in the lung of sensitized mice. Thirty-six mice were divided into three groups averagely. Group A was a control group. First, the mice were injected with schistosomal eggs hypodermically in abdomen, and 10 days later injected with schistosomal eggs intravenously via a tail vein. Group B was a praziquantel short administration group. In addition to the injections of schistosomal eggs as the same of Group A, the mice were administered with praziquantel in a daily dose of 300 mg/kg for 3 days, from 1 day before the intravenous injection of the eggs. Group C was a praziquantel prolonged administration group. In addition to the injections of schistosomal eggs as the same of Group A, the mice were administered with praziquantel in a daily dose of 150 mg/kg for 5 days weekly until the mice were sacrificed. Three mice of each group were sacrificed on days 7, 14, 28, and 56, respectively after the intravenous injection of the eggs, and the lung tissues were fixed with formalin and the slices were HE stained. The granulomas containing eggs in their centers were selected, and 25-30 granulomas from the animals of each group were measured at each time period. The mean areas of egg granulomas of each group were calculated, and the neutrophilic granulocytes, eosinocytes, lymphocytes, fibroblasts, and macrophages within the egg granulomas were counted. The mean numbers of them of each group were calculated. All the data of each group were analyzed and compared statistically. On day 56 after the intravenous injection of the eggs, the mean area of schistosomal egg granulomas in group B was (227.4 ± 728.0) × 10(3) μm(2), less than that of [(297.9 ± 153.3) × 10(3) μm(2)] in group A, and the suppression rate was 23.7% (P < 0.05). On days 7, 14, 28, and 56, the mean areas of schistosomal egg granulomas in group C were (575.8 ± 155.6) × 10(3) μm(2), (310.5 ± 854.0) × 10(3) μm(2), (267.7 ± 513.3) × 10(3) μm(2), and (214.9 ± 446.4) × 10(3) μm(2), respectively, significantly less than those of [(692.7 ± 232.6) × 10(3) μm(2), (439.4 ± 165.0) × 10(3) μm(2), (385.7 ± 129.3) × 10(3) μm(2), and (297.9 ± 153.3) × 10(3) μm(2)] in group A. The suppression rates were 16.9%, 29.3%, 30.6%, and 27.9%, respectively (P values <0.05). On day 56, the mean numbers of neutrophilic granulocytes were 11.4 ± 5.0 in group A and 5.2 ± 3.1 in group C, respectively, with the suppression rate of 54.4% in group C (P < 0.05). On day 56, the mean numbers of eosinocytes within the egg granulomas were 2.3 ± 2.0, 0.1 ± 0.3, and 0.3 ± 0.6 in groups A, B, and C, respectively, with the suppression rate of 95.7% in group B and 87.0% in group C (P values <0.05). On day 56, the mean numbers of macrophages within egg granulomas were 14.3 ± 6.9 in group C, compared with 18.6 ± 8.2 in group A, the suppression rate was 23.1% (P < 0.05). On day 56, the mean numbers of fibroblasts within the egg granulomas were 6.6 ± 4.4 and 5.8 ± 2.6 in groups B and C, respectively, and compared with 14.3 ± 7.8 in group A, the increasing extents decreased by 53.8% and 59.4%, respectively (P values <0.05). Therefore, the administration of praziquantel, especially the prolonged administration, can suppress the formation of schistosomal egg granulomas, including reduction in the areas of granulomas and suppression of the inflammatory cells and the hyperplasia of fibroblasts within granulomas.

Reduced phagocytic activity of polymorphonuclear leukocytes in alpha(1,3) fucosyltransferase VII-deficient mice.

Deficiencies in adhesion molecules or their counter-receptors in humans may have severe consequences as exemplified by leukocyte adhesion deficiency (LAD) I or II syndromes. Because such diseases occur with great rarity, animal models are valuable for studying the role of particular adhesion molecules and their natural ligands in immunity. We studied selected immune parameters and general health in mice with a defect in the sialyl-Lewis X antigen (selectin ligand) caused by disruption of the gene encoding alpha(1,3)fucosyltransferase VII (Fuc-TVII). Leukocytes from Fuc-TVII -/- and control mice were tested for adherence to cellophane membranes or polymer particles in vivo and phagocytic activity in vitro. While no difference in adherence was found, the number of neutrophil granulocytes in exudate induced by intraperitoneal injection of polymer beads was reduced in knock-out mice. Moreover, the phagocytic activity in Fuc-TVII -/- mice was significantly reduced. These animals have splenomegaly due to increased hematopoiesis and reduced weight but do not exhibit clinical signs of immunodeficiency. In conclusion, the lack of Fuc-TVII activity leads to several morphological and functional abnormalities without an impact on survival rate.

Clinical and Immunological Characteristics of Cats Affected by Feline Infectious Peritonitis

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The delayed-type hypersensitivity reaction is dependent on IL-8. Inhibition of a tuberculin skin reaction by an anti-IL-8 monoclonal antibody.

Cell-mediated immune reactions are essential to our immune response toward foreign organisms such as microorganisms, or in the response toward foreign tissue Ags, as seen in the rejection of allogeneic transplanted organs. Similar reactions form the basis for the development and the progression of delayed-type hypersensitivity (DTH) reactions. We found that the alpha-chemokine IL-8 plays an important pathophysiologic role for the development of a DTH reaction because infusion of a neutralizing anti-IL-8 mAb (WS-4) was able to suppress the development of a tuberculin skin reaction in rabbits, as judged by histologic, biochemical, and clinical examinations. Thus, the number of neutrophil granulocytes and lymphocytes at the site of tuberculin injection was decreased considerably, and the clinical signs of inflammation were suppressed almost completely at 24 h after intracutaneous injection of tuberculin, as judged by the size of the infiltrates. In contrast, we did not see any effect on the visible erythema of the skin. We found that the tissue content of myeloperoxidase (MPO), reflecting the number of infiltrating neutrophils, was lowered significantly. Furthermore, immunohistochemical analysis confirmed that IL-8 immunoreactivity is actually enhanced in the skin of positive tuberculin reactions. The results indicate that IL-8 plays an important role for the early accumulation of leukocytes in the skin and for the clinical signs of a DTH reaction.

Reduced variance of bone‐marrow transit time of granulopoiesis—a possible pathomechanism of human cyclic neutropenia

Abstract. Human cyclic neutropenia (CN) is a haematological disorder characterized by oscillations in the numbers of neutrophilic granulocytes and other blood cells with a stable period of approximately 21 days. In most cases the neutrophils oscillate well below normal values such that these patients are chronically neutropenic. A comprehensive concept of the origin of CN is proposed. It assumes an abnormally small variance of the transit time of bone marrow cells (compared to normal human granulopoiesis) for the origin of the characteristic cycles. Furthermore, a reduced responsiveness of the immature granulopoietic bone marrow cells to the mitotic feedback stimuli is assumed to account for the subnormal neutrophil peaks. Together with feedback control provided in a simulation model of normal human granulopoiesis these two abnormalities can explain experimental and clinical cell kinetic data for bone marrow and blood in CN.

Hematological, blood biochemical, and cytological bronchoalveolar lavage studies in prepubertal gilts after endotoxin inhalation and ingestion.

The effect of oral intake of endotoxin (ET) combined with inhalation of ET-contaminated feed-dust was studied in 6 prepubertal gilts. The animals were given 100 mg of ET each in their ordinary feed ration and meanwhile stayed for 4 h in a continuous flow exposure chamber where the endotoxin concentration was approximately 9 micrograms/m3. Blood samples were collected during the exposure and for 8 h afterwards. Bronchoalveolar lavage (BAL) was performed 24-26 h after the exposure. Six additional animals received ordinary feed and were exposed to feed-dust without added ET. Blood samples were analysed for total white blood cell count (B-WBC) and differential count: polymorphonuclear (PMN) and mononuclear (MN); total red blood cell count (B-RBC); platelets (B-PLT); total serum bile acids (S-BA); glutamate dehydrogenase (S-GLDH); and a blood plasma prostaglandin F2 alpha metabolite (15-ketodihydro-PGF2 alpha; P-PG). The major findings were decreased numbers of MN, PMN and B-RBC 3 to 6 h after the start of the ET exposure and increased numbers of neutrophilic granulocytes in the BAL fluid of the ET-exposed animals. It is concluded that a combined oral and airborne ET challenge induces hematological and cytological changes that may contribute to the development of respiratory disorders.

Hyperventilation-induced changes of blood cell counts depend on hypocapnia

Voluntary hyperventilation for 20 min causes haemoconcentration and an increase of white blood cell and thrombocyte numbers. In this study, we investigated whether these changes depend on the changes of blood gases or on the muscle work of breathing. A group of 12 healthy medical students breathed 36 l.min-1 of air, or air with 5% CO2 for a period of 20 min. The partial pressure of CO2 decreased by 21.4 mmHg (2.85 kPa; P < 0.001) with air and by 4.1 mmHg (0.55 kPa; P < 0.005) with CO2 enriched air. This was accompanied by haemoconcentration of 8.9% with air (P < 0.01) and of 1.6% with CO2 enriched air (P < 0.05), an increase in the lymphocyte count of 42% with air (P < 0.001) and no change with CO2 enriched air, and an increase of the platelet number of 8.4% with air (P < 0.01) and no change with CO2 enriched air. The number of neutrophil granulocytes did not change during the experiments, but 75 min after deep breathing of air, band-formed neutrophils had increased by 82% (P < 0.025), whereas they were unchanged 75 min after the experiment with CO2 enriched air. Adrenaline and noradrenaline increased by 360% and 151% during the experiment with air, but remained unchanged with CO2 enriched air. It was concluded that the changes in the white blood cell and platelet counts and of the plasma catecholamine concentrations during and after voluntary hyperventilation for 20 min were consequences of marked hypocapnic alkalosis.(ABSTRACT TRUNCATED AT 250 WORDS)

Correlative histologic and arthroscopic evaluation in rheumatoid knee joints

The correlation between arthroscopic observations and histologic changes in rheumatoid arthritis is still controversial. Synovial samples of 21 knee joints in rheumatoid arthritis patients were comparatively investigated by endoscopy and histology. Biopsies were scored by an endoscopist and subsequently dissected. Different histochemical and immunocytochemical staining techniques were used to define inflammatory activity. Arthroscopic and histological values were compared by rating scales and variance analysis. Our study indicates that synovial biopsy is of diagnostic value in rheumatoid arthritis. However, its usefulness depends on the histochemical methods used. The results revealed highly significant correlations of endoscopic features with the number of neutrophilic granulocytes, intravascular leukocytes, and peroxidase-positive macrophages. However, no relationship was found between the detection of lymphocytes or resident macrophages and inflammatory scores. The close correlation between endoscopic and histological findings suggests that arthroscopic evaluation allows a valuable classification of the inflammatory activity in rheumatoid synovitis.

The Effect of Glucocorticosteroids on the Neonatal Blood Count

To investigate the influence of glucocorticosteroid therapy on the neonatal blood count, the haematologic data of 68 preterm and term infants, who had received a single dose of 1 mg dexamethasone i.v., were reviewed. White blood cell (WBC) count and platelet count increased after steroid therapy. The increase in WBCs was associated with an increase in the number of neutrophilic granulocytes, whereas the number of eosinophils decreased. We conclude that glucocorticosteroids after the neonatal blood count and influence its value as a diagnostic marker for bacterial infections.

Histopathology of initial gingivitis in humans. A pilot study.

The present study concerns the inflammatory alterations in the gingival margin during initial gingivitis in 11-13 year old human subjects. At day 0 of the experiment, all participants had clean teeth and healthy gingiva. All active oral hygiene measures were excluded for 4 days. From upper and lower premolars, which were extracted for orthodontic reasons, contralateral gingival biopsies, including the tooth and the adjacent gingiva, were obtained on days 0 and 4. The presence of inflammatory cells in the junctional epithelium and the adjacent connective tissue was determined quantitatively in semi-thin sections. The collagen content of the gingival margin was also determined. From day 0 to day 4 there was only a slight increase in the number of neutrophilic granulocytes in the junctional epithelium and adjacent connective tissue, while a more pronounced increase was found in the number of mononuclear leukocytes. A loss of collagen was noticed in 4 of the subjects, while 2 did not show any changes in collagen content. The inflammatory reaction seen in the present study differs somewhat from that observed in adult humans and adult dogs. The results correspond more to the reaction seen in juvenile dogs.

Untersuchungen zum weissen Blutbild von Manikern, endogen Depressiven, Schizophrenen und Neurotikern

Manic, endogenous depressive, schizophrenic and neurotic patients, 80 (40 females, 40 males) in each diagnostic group, are compared with regard to the white blood count with one another and with normal controls. Significant differences of means were found (1) between some groups of patients concerning the total number of leucocytes (µM ≠ µS, µD), the number of neutrophil granulocytes (µM ≠ µS, µN), the number of eosinophil granulocytes (µM ≠ µS, µD; µN ≠ µS, µD) and the number of lymphocytes (µN ≠ µD), and (2) between all groups of patients and normal controls concerning the number of monocytes (µD, µN, µM, µS ≠ µK). The findings are discussed.