The effect of phagocytosis of Candida albicans blastospores on alkaline phosphatase levels in rat polymorphonuclear leucocytes

Summary The phagocytic function of young polymorphonuclear leucocytes with high levels of leucocyte alkaline phosphatase, which are present in the peripheral blood during an inflammatory response, was compared with that of normal polymorphonuclear leucocytes.Candida albicans blastospores were used as phagocytic targets. The phagocytic index of polymorphonuclear leucocytes with low levels of leucocyte alkaline phosphatase was higher than that of cells with high levels of enzyme.Monitoring of leucocyte alkaline phosphatase levels with increasing times of incubation of leucocytes with the blastospores showed a progressive decline in the level of the enzyme. Thus loss of the enzyme is linked to the phagocytic function, although the mechanism of this dynamic process is unclear.     

Fine-structural study of leucocytes in the goldfish, Carassius auratus

An electron microscopic study was performed on leucocytes from circulating blood of the goldfish. The leucocytes were divided into eight types: neutrophil, eosinophil, large granular leucocyte (LGL), medium-sized granular leucocyte (MGL), small granular leucocyte (SGL), fine granular leucocyte (FGL), lymphocyte, and monocyte. In this report the thrombocyte was excluded from leucocytes, and LGL, MGL, SGL and FGL were tentatively classified based on the size of intracytoplasmic granules possessed by each cell. The existence of goldfish monocytes was electron microscopically demonstrated for the first time in the present report.     

Expression of glutamate decarboxylase isoform, GAD65, in human mononuclear leucocytes: a possible implication of C-terminal end deletion by Western blot and RT-PCR study

Human peripheral blood leucocyte was examined for the expression of glutamate decarboxylase (GAD). Peripheral blood from healthy individuals was fractionated into polynuclear leucocytes and mononuclear leucocytes. Cell lysate from the mononuclear leucocytes was analysed by SDS-PAGE and Western blot. With antibody raised against unique C-terminal sequence for GAD65, two protein bands at 30 and 80 kDa were detected. However, with anti-GAD65/67 antibody recognizing very end of C-terminal, about 40 residues toward C-terminal, no protein bands were observed. Expression of mRNA coding for the epitope of these two antibodies was examined by using PCR technique. Results showed an evidence that mononuclear leucocytes express GAD65 with its C-terminal segment truncated. Our results have suggested an expression of GAD with the novel molecular weight may be caused by possible mononuclear leucocyte specific splicing errors.     

Response of common carp (Cyprinus carpio L.) leucocytes to hormone-induced stress

The influence of cortisone on leucocytes composition in the blood of common carp (Cyprinus carpio L.) is studied. Following the hormone injection, the relative number of leucocytes decreased and the number of neutrophils and blast-form cells increased in the leucocyte spectrum of experimental fish.     

Differences among the polyadenylated RNA sequences of human leucocyte populations: an approach to the objective classification of human leukaemias

We have constructed a complementary DNA (cDNA) library representing expressed sequences of the white blood cells from a patient with chronic granulocytic leukaemia. The library was screened by colony hybridization of 32P-labelled cDNAs synthesized from the polyadenylated RNAs of the white blood cells from patients with chronic granulocytic or chronic lymphocytic leukaemia. The autoradiographic patterns were compared and 70 recombinants were selected to comprise a panel which distinguished between these two types of leukaemia. Hybridization of this panel with complementary DNAs transcribed from the polyadenylated RNAs of a variety of normal and neoplastic leucocyte populations showed that the RNA sequences in high abundance in leucocytes from chronic granulocytic leukaemias differ quite radically from those in other leucocytes. The patterns of hybridization seen when this panel was challenged with cDNAs representing the RNAs of normal and leukaemic leucocyte populations were sufficiently different to distinguish clearly the peripheral blood leucocytes of chronic granulocytic leukaemias from other populations of white blood cells, both normal and leukaemic. We suggest that this approach might provide additional markers useful in the classification of the acute leukaemias, especially the undifferentiated leukaemias whose identification by conventional methods is uncertain.     

Blood system peculiarities in the bank vole (Clethrionomys glareolus) under chronic environmental pollution

The parameters of peripheral blood and hemopoietic organs in mature and immature bank voles inhabiting a chemically polluted area were studied. Variability of the blood system parameters depending on the level of toxic load and the animals’ reproductive status was determined. Alteration of the cell composition of erythrocytes and leucocytes, the structure of erythrocytes, and the hemoglobin fractions and leucocyte functions describe the adaptive response to the factors of a changed environment more than the concentration of leucocytes, erythrocytes, and blood hemoglobin.     

Leucocytes of anadromous and landlocked strains of Atlantic salmon (Salmo salar L.) in the smolting period

Using flow cytometry and leucocyte specific monoclonal antibodies, neutrophils and B-cells were studied in blood and head kidney from wild strains of Atlantic salmon. The strains were Vosso and Blege, being an anadromous and landlocked strain, respectively. Smoltification was induced using a simulated natural photoperiod and sampling was performed monthly for 6 months and ended for the Blege strain at the time of seawater transfer while samples were collected from the Vosso strain after 4 weeks in seawater. Throughout the observation period, the mean proportions of neutrophils in both head kidney leucocytes (HKL) and peripheral blood leucocytes (PBL), were highest for the Vosso strain. The opposite was observed for B-cells where the Blege strain had higher or similar mean proportions compared to the Vosso strain. There were some differences between HKL and PBL. The mean proportion of neutrophils was always higher in HKL than in PBL and the mean proportion of B-cells was higher in PBL than in HKL. The fluctuations during the observation period, in the proportions of B-cells and neutrophils of the analysed cell population, showed mainly the same pattern in both strains. Differences between the strains were observed at various times in the mean of total number of leucocytes per gram head kidney and per millilitre of blood. The fluctuations throughout the experimental period in total numbers of leucocytes in head kidney and blood followed mainly the same pattern in both strains. The results of the leucocyte analyses suggest that there are differences between the anadromous and landlocked strains with respect to what cell type is present in highest proportion in the leucocyte samples from head kidney and blood. The striking similarity between the strains is the profiles of proportions of B-cells and neutrophils in HKL and PBL during the smoltification period.     

Changes in blood leucocyte populations induced by acute maximal and chronic submaximal exercise

Absolute (x 10(3).mm-3) or relative (%) numbers of blood leucocyte types (monocytes, lymphocytes, neutrophils) and lymphocyte subsets (T11+, T4+, T8+, B1+, and NKH1+) reacting with specific monoclonal antibodies were determined at rest, immediately after maximal exercise on a treadmill, in six controls (C), and in six young cyclists before training (BT) and after 5 months of training (AT). Maximal exercise significantly increased the absolute number (mobilization) of virtually all the types of leucocytes and subsets of lymphocytes in C, BT and AT subjects. In these subjects mobilization of natural killer cells (NKH1+) and cytotoxic/suppressor T lymphocytes (T8+) was greater than mobilization of the other leucocyte types and lymphocyte subsets; however, maximal exercise induced no significant changes in the relative numbers of any leucocyte types and lymphocyte subsets, except in the case of T4+ lymphocytes in At cyclists. Chronic submaximal exercise induced increased mobilization of neutrophils and decreased mobilization of lymphocytes during maximal exercise, except in the case of B lymphocytes (B1+) and NKH1+ cells, and decreases in the absolute and relative number of neutrophils at rest. It remains to be seen how these results can explain the modifications of leucocyte activities noted in vitro after isolated or chronic exercise.     

The effect of triisopropyl phosphate on the mobility of surface concanavalin a receptors and on the locomotion of polymorphonuclear leucocytes

The non-phosphorylating organophosphorus compound triisopropyl phosphate, which is known to inhibit rabbit leucocyte locomotion, can stimulate the locomotion of guinea pig leucocytes under certain conditions. Different methods of preparing guinea pig leucocyte monolayers can give preparations with different proportions of motile cells. With preparations that contain relatively slowly moving cells triisopropyl phosphate increases the number of stationary cells without significantly affecting the speed of the cells that remain motile. Most rabbit leucocytes labelled with fluorescein-labelled concanavalin A form caps within 5–10 min at 37 °C. In contrast the rate of cap formation in guinea pig leucocytes is much slower and after 20 min many cells have only random patches. Triisopropyl phosphate accelerates cap formation in guinea pig leucocytes but not in rabbit leucocytes. The local anaesthetic nupercaine inhibits cap and patch formation in rabbit and guinea pig leucocytes. Inhibition of rabbit leucocyte locomotion is induced by concanavalin A at 1 μg/ml. These results are briefly related to the known effects of triisopropyl phosphate on the isolated leucocyte plasma membrane.     

IMMUNOLOGICAL AND HEMATOLOGICAL PARAMETERS IN CAPTIVE HARBOR SEALS (PHOCA VITULINA)

Peripheral blood was collected from 13 captive seals (12 harbor seals and 1 gray seal) of various ages to study different immunological and hematological parameters. In vitro mitogenic reactivity of blood lymphocytes was measured by means of a microculture lymphocyte transformation test (LTT). After stimulation with different doses of the mitogens concanavalin A (Con A) and pokeweed mitogen (PWM), all examined seals showed significant proliferative responses to each mitogen. Furthermore, mitogenic reactivity significantly decreased with animal age suggesting this parameter of seal lymphocyte function is age-related. The present experiments support that the LTT is a suitable tool to monitor the functional capacity of seal lymphocytes. By means of the erythrocyte-rosette (E:-rosette) test it was demonstrated that a subpopulation of mononuclear seal bloosd leucocytes formed rosettes with sheep red blood cells (Srbc). This observation indicates that the phenomenon probably represents a marker for T lymphocytes in the seal as in several other mammalian species. Furthermore, the percentage of Srbc rosette-forming cells decreased with the age of the animals. Total blood leucocyte counts and differential leucocyte counts were determined by light microscopy. The number of leucocytes varied considerably among individual animals. Parallel to a significant decline of the percentage of lymphocytes with age, a corresponding increase in the percentage of neutrophils was demonstrated. In all of the seals, the percentages of monocytes and eosinophils were found to be low with only minor individual variation. The LTT and the E-rosette test are suggested as potential tools to elucidate immunological disorders in the seal.     

Immunology and growth faltering of Anga children,Papua New Guinea: Preliminary work

Nutrition–infection interactions among poor children of the less-developed world influence growth and development. However, the relative importance of each is difficult to determine, because the relationship is mediated by immunological status. In this analysis, relationships between immunological measures and anthropometry were sought among 41 Anga, Papua New Guinea children aged 0–7 years. These had elevated serum total leucocyte and leucocyte subset counts relative to western reference values. Although there was no correlation between anthropometric nutritional status and total leucocytes and leucocyte subsets for this group, the small group (n = 8) with very high total leucocyte count (greater than 15,000/mm²) had significantly lower mean Z score of stature for age (−3.78), and weight for stature (−1.35) than those with leucocyte counts lower than this cut-off (weight for stature Z score: −0.59; stature for age Z score: −2.68, respectively. Low stature-for-age Z score was associated with lower total lymphocyte count and increasing age, against a background of elevated lymphocyte levels relative to western reference values among the older children; low weight-for-stature Z score was associated with lower neutrophil count, against a background of normal neutrophil levels across all age groups. The pattern of weight and stature growth seen in the Anga may reflect extended nutritional deficits which result in stunting of a degree to which the most growth-compromised children die, leaving those above a threshold associated with high mortality alive. Thus, the anthropometric and immunological characteristics of the older children in this small sample may reflect the biology of survival under severe ecological conditions, where poor linear growth and elevated leucocyte status relative to normative values are characteristics of survivorship. Am J Phys Anthropol 106:515–520, 1998. © 1998 Wiley-Liss, Inc.     

Tyrosine hydroxylase of the blood leukocytes

Tyrosine hydroxylase activity has been established in blood plasma leucocytes of rat, cat and man. Tyrosine precursors and some nuclear erythroid cells. GFU-GM did hydroxylase activity in leucocytes shows the Km for tyrosine inhibited by high concentrations of L6 tyrosine (substrate inhibition), alpha-methyl-para-tyrosine dopamine. The kinetic properties of leucocyte tyrosine hydroxylase are qualitatively similar to the properties of brain tyrosine hydroxylase.     

Interaction of growth with social behaviour in Tilapia zilli raised in three different temperatures

Three thermal levels were tested on 10-day old T. zilli fry obtained from one spawning to study the effects of temperature on the occurrence of growth depensation in the population reared under controlled environmental conditions. Results showed highest individual growth rate, mortality rates and growth depensation at 30 °C whilst the greatest biomass increase was obtained at an intermediate temperature (27 °C). The enhanced depensation and high mortality rates at the upper limit of the thermal gradient for growth of T. zilli were attributed to an alteration in the behaviour of fish which favoured a few, large, dominant and socially aggressive individuals.     

MOBILIZATION OF B AND T LYMPHOCYTES AND HAEMOPOIETIC STEM CELLS BY POLYMETHACRYLIC ACID AND DEXTRAN SULPHATE

The leucocytosis which can be evoked by the polyanions dextran sulphate (DS), polymethacrylic acid (PMAA) and the copolymer of PMAA and styrene (PMAA—STYR) was studied in mice. After intravenous administration of these polyanions peak numbers of leucocytes were found in the peripheral blood 3 hr after injection. All three types of polyanions increased the number of lymphocytes, granulocytes and monocytes. Dose—response studies revealed that the nature of the polyanion determined the degree of leucocyte mobilization. The most potent mobilizer was found to be DS. This polyanion could evoke a six-fold increase of the number of peripheral blood leucocytes. By means of the membrane fluorescence technique it could be demonstrated that optimal doses of DS, PMAA and PMAA—STYR mobilized both B and T lymphocytes. The ratio between the number of B and T cells mobilized was greater for DS than for the other two polyanions. Intravenous injection of DS, PMAA and PMAA—STYR also increased the number of circulating haemopoietic stem cells (CFU-S). The most potent stem cell mobilizer appeared to be PMAA—STYR. This polyanion evoked a twenty-five-fold increase in the number of CFU-S.     

Mobilization of B and T lymphocytes and haemopoietic stem cells by polymethacrylic acid and dextran sulphate.

The leucocytosis which can be evoked by the polyanions dextran sulphate (DS), polymethacrylic acid (PMAA) and the copolymer of PMAA and styrene (PMAA--STYR) was studied in mice. After intravenous administration of these polyanions peak numbers of leucocytes were found in the peripheral blood 3 hr after injection. All three types of polyanions increased the number of lymphocytes, granulocytes and monocytes. Dose--response studies revealed that the nature of the polyanion determined the degree of leucocyte mobilization. The most potent mobilizer was found to be DS. This polyanion could evoke a six-fold increase of the number of peripheral blood leucocytes. By means of the membrane fluorescence technique it could be demonstrated that optimal doses of DS, PMAA and PMAA--STYR mobilized both B and T lymphocytes. The ratio between the number of B and T cells mobilized was greater for DS than for the other two polyanions. Intravenous injection of DS, PMAA and PMAA--STYR also increased the number of circulating haemopoietic stem cells (CFU-S). The most potent stem cell mobilizer appeared to be PMAA--STYR. This polyanion evoked a twenty-five-fold increase in the number of CFU-S.     

Anti-HLA immunization following transfusions of leukocyte-poor blood

We have followed the evolution of leucocyte antibodies in patients receiving multiple blood transfusions of leucocyte-poor blood. The number of leucocytes present in one unit of transfused blood was always less than 1 X 10(8). We observed that such preparations were able to re-stimulate antibodies which had disappeared, but especially they were able to provoke a first immunisation in some patients never immunized in the past. The specificities of these antibodies corresponded to one or several antigens present in the donor of the transfused unit. This study shows that leucocyte immunisation cannot always be avoided in spite of the selection of blood products containing very few white cells.     

The haemoflagellate Trypanoplasma borreli induces the production of nitric oxide,which is associated with modulation of carp (Cyprinus carpio L.) leucocyte functions

In an attempt to characterise the role of nitric oxide (NO) in immune responses of carp, carp leucocytes obtained during an acute T. borreli infection were examined, for their capacity to generate NO. In a second set of experiments the impact NO on viability of the parasite and on the modulation of functional carp leucocyte responses were tested in vitro. Both in carp head-kidneys and in the peripheral blood, the fractions of lymphoblasts among separated leucocytes were increased. However, the relative proportions of granulocytes among head-kidney leucocytes (HKL) significantly decreased during infection, whereas granulocytes appeared among peripheral blood leucocytes (PBL). The cellular dynamics of HKL and PBL of infected carp were paralleled by an enhanced spontaneous NO release in vitro. NO production was further increased after addition of viable parasites to these cultures. The hypothesis that NO had a possible role in granulocyte activation and lymphocyte proliferation in carp was supported by the reduction of mitogen-induced proliferative responses of PBL from healthy carp in the presence of NO donor substances. The negative effects of NO on lymphocyte proliferation were contrasted by enhancing effects on granulocyte functions: the inhibition of NO generation in T. borreli-stimulated HKL cultures by the l-arginine analogue L-NMMA reduced the viability of granulocytes and their phagocytic activity. Even massive amounts of nitric oxide produced by donor substances (up to 600 micromol l(-1) NO(-)(2)) caused no reduction in the numbers of viable T. borreli flagellates in vitro. Thus, in carp, T. borreli seems to induce high amounts of NO in vivo which are apparently not harmful for the parasite but which may interfere with co-ordinated interactions of activated cells aiming at the defence of the parasite.     

Snapper (Pagrus auratus) leucocyte proliferation is synergistically enhanced by simultaneous stimulation with LPS and PHA

Important channels of communication between mammalian leucocytes have long been recognised. Here, data are reported that suggest similar integrations may occur between snapper leucocytes upon mitogen stimulation. Cell surface immunoglobulin (IgM) expression was used in conjunction with intracellular fluorescence staining and flow cytometry to differentiate proliferating peripheral blood leucocyte subsets (PBLs). Independent activation using phytohaemagglutinin (PHA) or lipopolysaccharide (LPS) drove both mIg(-)and mIg(+)cells into cycle. It is not known if the proliferation of mIg(+)cells was mediated by a mutually exclusive effect of the mitogen on each cell population, cognate cellular interaction or a soluble growth factor. Simultaneous activation of PBLs with PHA and LPS consistently induced significantly more cells to proliferate than the sum of proliferating cells stimulated solely with PHA or LPS. Together, the results suggest that different leucocyte subsets have the capability to influence their respective responses to mitogenic stimulation. Therefore, like in the mammalian immune system, communication may occur between snapper leucocyte subsets.     

Immunological aspects of benign melanocytic tumours and melanoma in situ

Summary Extracts were prepared from 15 malignant melanomas, 10 naevi, 8 pieces of skin containing atypical melanocytes and 4 samples of normal skin. Peripheral blood leucocytes from 52 patients with benign naevi that had altered sufficiently to cause the patient to seek attention, 57 patients with malignant melanomas, and 98 normal individuals were tested against these extracts in a capillary leucocyte migration test. Melanoma extracts preferentially inhibited the leucocytes of melanoma patients and to a lesser extent those of naevus patients. Melanoma patients' leucocytes were more frequently inhibited by naevus extracts (whether from intradermal or from compound naevi) than were the leucocytes of naevus patients and control donors. Extracts of skin containing atypical melanocytes (perimelanomatous skin and lentigenous skin) selectively inhibited melanoma patients' leucocytes.     

Immunostimulatory effect of azadirachtin in Oreochromis mossambicus (Peters)

The effect of azadirachtin, a triterpenoid derived from Azadirachta indica on the immune response was studied in the freshwater teleost, O. mossambicus. Bovine serum albumin (BSA) and sheep erythrocytes (SRBC) were used as antigens to evoke immune response. The immune responses in fish were measured by quantifying antibodies produced and counting the peripheral blood leucocytes in control and experimental fish. In general, azadirachtin significantly enhanced the antibody response and leucocyte count in a dose dependent manner. An inverse relationship was observed between the dose of azadirachtin and the degree of immunostimulation. Timing of azadirachtin administration in relation to immunization revealed that the maximum enhancement of antibody response was observed when the stimulant was given two days prior to immunization. The observed immunostimulatory property of azadirachtin has an implication in the maintenance of finfish health in freshwater intensive aquaculture practices.