Monoclonal antibodies to human polymorphonuclear leucocyte granule antigens (original) (raw)
Biochimica Et Biophysica Acta-molecular Cell Research, 1989
The initiation of the respiratory burst and the degranulation of human polymorphonuclear leukocytes (PMN) in response to stimulation by uropathogenic strains off Escherichia coil is dependent on the expression of Type | fimhriae by those strains. These PMN responses correlate with an increasing tendency of the interacting E. coil strain to be retained on hydrophobic columns. The present work assessed the measurement of relative surface hydrophobicity in relation to PMN activation. Type | fimbr|ate organisms bound most readily to Octyl-Sepharose columns and were strongly agglutinated in the salt aggregation test. |n contrast, the same organisms partitioned to the dextran-rich (hydrophilic) phase of aqueous two-polymer phase systems. Electron microscopic observation of the organisms eluted from the Octyi-Sepharose columns and of the organisms recovered from both phases of the aqueous two-phase systems demonstrated, however, that both Type I and P-fimbriate organisms were retained on the columns and partitioned into the dextran-rlch phase as a consequence of their being fimbriate and failed to identify this as a major factor in the activation of PMN. lh addition electron microscopy demonstrated that e#ch P-fimbriate population had fewer organisms expressing fimbriae than did Type I fimbriate populations, confirming the importance of phase variation as a factor affecting the physicochemical characteristics of a bacterial population.
Infection and Immunity, 1983
In the present study, we assayed protein iodination in human granulocytes after interaction of the cells with mannose-specific (MS) type 1 fimbriated (MS+) and nonfimbriated (MS-) phenotypes of Escherichia coli pretreated with various amounts of anti-E. coli and antifimbrial antibodies. The MS+ phenotype stimulated protein iodination in granulocytes and possessed potent MS activity as measured by yeast aggregometry. In contrast, the MS- phenotype lacked all these activities. MS+ pretreated with moderate concentrations of antibodies, however, showed up to a 15-fold increase in granulocyte stimulation as compared to granulocyte stimulation induced by the non-antibody-treated MS+ phenotype or by the antibody-treated MS- phenotype. This marked antibody-mediated increase in stimulation of granulocytes was (i) dependent on the antibody concentrations, (ii) markedly reduced by methyl-alpha-D-mannoside, (iii) caused by immunoglobulin G as well as by F(ab')2, (iv) caused only by antifimb...
Blood, 1990
The ontogeny of a 57-Kd cationic antimicrobial protein (CAP57) that has substantial similarities to bactericidal permeability increasing protein (BPI) has been determined immunocytochemically. CAP57 was detected in the granules of mature peripheral blood neutrophils. However, it was absent from other cells of the peripheral blood: eosinophils, red blood cells (RBCs), and mononuclear cells. In human bone marrow, CAP57 was confined to the neutrophilic series. The earliest stage of development of the myeloid cells at which CAP57 was demonstrated was the promyelocyte. Double immunofluorescent labeling showed that CAP57 was detected in cells positive for myeloperoxidase. The absence of lactoferrin in certain cells (promyelocytes) containing CAP57 indicated that CAP57 was synthesized and packaged in a population of granules prior to the development of granules that contain lactoferrin. CAP57 could not be demonstrated in HL60 cells either by enzyme-linked immunosorbent assay (ELISA) or by ...
Biochimica Et Biophysica Acta-molecular Cell Research, 1990
The generation of the 5-1ipoxygenase product, leukotriene B 4 (LTB4) by human mononuclear phagocytes (monocytes) following incubation with 25 different uropathogenic strains of Escherichia coli correlated with the haemolytic activity of the strains (r--0.572, P < 0.01). LTB 4 generation by human neutrophils (PMN), however, was unrelated to this haemolytic potential (r= 0.164). In contrast, both prelabelled monocytes and PMN were stimulated by haemolytic strains of E. coli and by haemolytic culture supernatants to release significant amounts of [3Hlarachidonic acid. There was a significant correlation between haemolytic activity and [3H]arachidonic acid release generated by individual strains from monocytes (r--0.804, P < 0.001) and PMN (r--0.888, P < 0.001). In addition, nonhaemolytic strains but not their culture supernatants were capable of causing slow release of both [3H]arachidonic acid and LTB 4 from PMN and mononuclear cells. These results suggest that both the possession of haemolytic activity, and the direct interaction of bacteria with the leukocyte surface are mechanisms by which uropathogenic strains of E. coli may cause the release and metabolism of araehidonic acid. In addition, there was synergistic augmentation by nonhaemolytic bacteria of the PMN LTB 4 response to haemolytic culture supernatants or to low doses of the calcium ionophore A23187. These results support an ionophore-like mechanism for the activation of the cell by haemolysin. LIB 4 generation by PMN incubated with haemolytic supernatants was also augmented by particulate zymosan in a manner dependent on the dose of zymosan, suggesting that the direct interaction of E. coli with PMN may involve an activation mechanism similar to that for zymosan. These results demonstrate differing responses of peripheral mononuclear cells and PMN from the same donors to identical strains of E. coli and suggest that the generation of the potent chemotactic agent LIB 4 in response to E. coli infection in vivo need not depend solely on the elaboration of cytotoxic haemolysins by individual strains.
Clinical and diagnostic laboratory immunology, 1994
Peritoneal dialysis effluent from patients with end-stage renal failure contains a low-molecular-weight solute that inhibits the killing of phagocytosed Staphylococcus epidermidis by polymorphonuclear leukocytes (PMN). This observation has been investigated by using luciginen-enhanced chemiluminescence to measure PMN NADPH oxidase activity, CD1lb/CD18 expression and lactoferrin release to measure secondary granule discharge, and cellular levels of l-glucuronidase (EC 3.2.1.31) to measure changes in primary granules. Peritoneal dialysis effluent had no effect on the loss of intracellular 0-glucuronidase from normal unstimulated PMN or from PMN stimulated with S. epidermidis. It did, however, cause a concentration-dependent (0 to 70%; vol/vol) increase in expression of CD11b/CD18 and NADPH oxidase activity. CD11b/CD18 expression increased over 20 min before starting to plateau. Release of lactoferrin by the same cells demonstrated a strong positive correlation with integrin expression (P < 0.001, Spearman's rank correlation coefficient). When dialysis effluent-treated PMN were stimulated with formyl-methionylleucylphenylalanine, integrin expression, release of lactoferrin, and NADPH oxidase activity were greater than in PMN treated with formylmethionylleucylphenylalanine alone. Under these conditions, a concentration-dependent increase in CDllb/ CD18 and lactoferrin release were observed only at a concentration between 0 and 30% (vol/vol) dialysis effluent, while a concentration-dependent increase in oxidase activity was seen at a concentration between 0 and 70% (vol/vol). The results suggest that dialysis effluent does not affect PMN primary granule release but does cause increased release of secondary granules and an increase in NADPH oxidase activity in both unstimulated and stimulated PMN.
A New Microbicidal Pathway for Polymorphonuclear Leukocytes
2016
In anucleate, granule-poor, motile fragments from human blood neutrophils (cytokineplasts; CKP), the nitric oxide synthase inhibitor N`-monomethyl-L-arginine (NMMA) produced a modest decrease in uptake of staphylococci from supernatants (P < 0.02, n = 7), and a marked decrease in the killing of cytoplast-associated bacteria (P < 0.001, n = 7). After 60 min of incubation with bacteria, NMMA-treated cytoplasts had a mean of over 3.5 times as many live, CKP-associated staphylo-cocci as did controls (51 % of the inocula versus 14%), despite having taken up fewer. Effects on both uptake and killing were reversible by L-arginine but not by D-arginine. Results were the same with other granule-poor cytoplasts (U-cytoplasts, U-CYT), which, unlike CKP, retain activatable oxidase activity. Killing by intact PMN, including those from a patient with chronic granulomatous disease, was not inhibited by NMMA. Thus, the ability to discern effects of NMMA correlated with the paucity of granules...
Acta Veterinaria Brno, 1989
It i s 0 v, Dan i e la,' J. L 0 k aj. : ~ tl'SIIblllhlb#1OIy _1""1-. bllOtY CoItmrlnlliOlU qf Alltlmlcrobllll .."", 011 1M Clwmlhmrl1liM:ell« of PIttIgo. cyt08l11g Gltlllllloc:yta. Ada vet. Brno, sa, 1989: 353-362." " This Study w,as aimed' at determinina the etrect of commonlyadminiitered antj.; bac:teriaI drupon bumanpolymorPboouc1ear leukocytes (PMNL). 1'be functiOl~a). ~ty of DeutroPbllic ~uloc:ytes wu assayed by measurina"luminol«pendent chemiJumiDesCenc:e (CL)usina luminon1eter LKB 12Sl.Rice starch and Opsonized, E. coil were used in testinl plJaaoCytosis' by isolated lfIIIlulocytes. The cells were iric:ubated with antibiOtics in subinluDitory' and inhibitory concentrations with a ,special attention to their therapeutic levels. The, testedanbDiotics prOduc:ed variOUs "J ' effects on pbagoc:yte' CL i'eprdless their chemical relatiOnships. Those anb"biotics which can penetrate into the pbaaoc:yte were especially effective. Lincomycin, ampicillin, sentamicin, cblorampbenicol and rolitetracycline bad no effect, clindamycin, oxacillin and polymyxin B increased CL in therapeUtical concentrations; doXycycline. rifampicin, ~ycin, adriamycin and trim~pbametboxazole in therilpbeutical criilcentrations reduc:ed CL activity c(~inllfIIIlulocytes. Clindamycin and trimetboprim-sulpbametbOJl'aZolepoteQtiated CLactivityinsu~~' concentrations. ' " " ' , .4nlimlcmb/~1 ag6Us. im1ll1lnomodUiallon, PoIYmorphonucl«u I~a" .~ihuni nacence During the last 20 years, a growins interest bas arisen in the study ofinteracuons betWeen antimi~ial.nts and host defence systems. Since polymQJPbOnuc1ear ncutrophi4 (PMNL) act as the first ,line, of defena; against bacteria, functional alterations of these cells ,by anb"biotics could explain, at least in ~ som~ of the ~cies, obs~ betweeJl in vitroanb"bacteri!al activity and clinical efticacy (L a b r 0 ,et ale 1986). ' , " , , ' , , : When })MNL interact with soluble and particulate' stimuli, ~ cells respOnd with a, bu~ in oxidative metabolism which generates chemic::ally reactive molecules, e. g., superoxide lQlions (<h) and bydrpgen peroxtde (H~2).Thi~activation, which is an essential step, i~ bostdefence against invading microorpnisms, is accompanied byJi&bt emission or chemiluminescence by the ce~ (AI I e,p et ale 1972). The addition ofluminolto a CL system ,bas been sbown to amplify the response and to simplify the measuring proc:edure (A I len $ld ,~o 0 s e ,1976). , "The luminol-enbanc:ed s~tem obviates the need for workios under darlt~ted ,conditions CD e Cb a t eJ e t an~ S b.id e yl~2),~tStbe'* of Yely.few cells (8 t e v ~ n,s et ale 1978) and can in fact be used to measure CL in samples of whole blood (KatQ et ale :1981). For this reason, the luminol-enbanc:ed CL bas been used i~y as a means
Infection and immunity, 1999
The effects of pathologically relevant concentrations (0.38 to 12.5 microM) of the proinflammatory, Pseudomonas aeruginosa-derived pigment 1-hydroxyphenazine (1-hp) on Ca2+ metabolism and intracellular cyclic AMP (cAMP) in N-formyl-L-methionyl-L-leucyl-L-phenylalanine (FMLP; 1 microM)-activated human neutrophils, as well as on the release of myeloperoxidase (MPO) and elastase from these cells, have been investigated in vitro. Ca2+ fluxes were measured by the combination of a fura-2/AM-based spectrofluorimetric method and radiometric procedures, which together enable distinction between net efflux and influx of the cation, while radioimmunoassay and colorimetric methods were used to measure cAMP and granule enzymes, respectively. Coincubation of neutrophils with 1-hp did not affect intracellular cAMP levels or the FMLP-activated release of Ca2+ from intracellular stores but did retard the subsequent decline in the chemoattractant-induced increase in the concentration of cytosolic fre...