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From www.bloodjournal.org by guest on June 14, 2017. For personal use only. Phagocytosis By E by J. MARn OSINOPHILS over 100 Mesnil in eases CuE, were years remains semble other granulocytic locomotion, response to fer in the specific levels of peroxidase and precise role general, cells chemotactic activities of certain and aryl sulfatase is available cent knowledge of phagocytosis studies of Archer and Hirsch, peritoneal exudates.5 with a 3 year the Cancer Medical submitted MARTIN Associate Francisco, Medical J. Research Center, This investigation (GM-01791) from San ) for and the in tissues Eosinophils . normal also dif- and functions of the difficulty of obtaining elements. Most of our re- has been provided separated from horse period in example, they have high low levels of hexokinase metabolism we had eosinophilia AND and Francisco, the of by the blood elegant or rat opportunity the of peripheral ob- blood. studies of the metaboWhenever possible, we that of eosinophils from METHODS March 5, 1968; M.D.: a research Service. for Associate Professor Research Institute, Cancer California. accepted JoN the Department of Medicine, either with drug reaction MT.). They University of California. was supported by the U. S. Public Health CLIME, Director, institute San Center, San Francisco, University of California Medical Requests for reprints should University of California Medical 922 re- of Leukocytes California First dis- mammals Leukocytes were obtained from 3 patients with eosinophilia, associated parasitic disease (patient V.P.), with asthmatic bronchitis and possible (patient B.C.) or with a syndrome resembling eosinophilic leukemia (patient From and various subjects. MATERIALS Source in higher distribution infections These 3 patients provided the material for detailed lism and phagocytic functions of human eosinophils. have tried to compare the activity of these cells with healthy morphology and physiologic behavior, including and particle ingestion.3 They on pronounced granulocyte of phagocytosis the from stems from other cellular by eosinophils using cells During 3 patients on man enzymes; activity human eosinophils. This lack no doubt large numbers of these cells free from of capable literature their stimuli LEH.RER type be in normal in their pattern of states ( e.g., parasitic alkaline phosphatase.4 Only limited information serving to eosinophils in I. ROBERT a distinctive observed a voluminous In differ from neutrophils and in certain disease AND as were their unclear. Eosinophils HANIFIN recognized and Despite cells, Human JON first ago,’ 1895.2 of these functions by grant (CA-07723) publication July of Medicine, University 11, and grant 1968. Department of a training of Medicine, California Medical and Center, M.D.: Fellow in Medicine, University of California California. ROBERT I. LEHRER, M.D.: Fellow in Medicine, Center, San Francisco, California. be addressed to Dr. M. J. Cline, Cancer Research Institute, Center, San Francisco, Calif. 94122. HANIFIN, BLOOD, VOL. 32, No. 6 (DECEMBER), 1968 From www.bloodjournal.org by guest on June 14, 2017. For personal use only. PHAGOCYTOSIS were the also obtained peripheral matory from blood an otherwise (R.I.L.) and normal from subject with 8 to 10 per cent with neutrophilia associated 7 patients eosinophils with in inflam- or polycythemia. diseases Preparation of Leukocytes Leukocytes were isolated by dextran sedimentation, washed in Hanks’ balanced salt solution containing 1/10 volume fetal calf serum, and suspended at 2 X 10 or 2 X lOVrnl. in Eagle’s minimal essential medium (MEM) containing either 3 per cent human albumin, 10 per cent fresh or fresh frozen human serum, or 20 per cent fetal calf serum.6 In studies involving glucose Leukocyte per cent phils utilization, populations contaminating designated eosinophils at or beyond at least 90 per cent cells were removed by hypotonic lysis.7 at least 80 and usually 95 to 100 state. Populations designated neutrometamyelocytes and myelocytes. contained the metamyelocyte mature granulocytes, contained Preparation red eosinophils of Particles Polystyrene particles,* 1.3 p in diameter, were suspended in MEM at a final concentration of 0.5 to 1.0 mg./ml. Candida albicans grown in Sabouraud’s broth was washed with saline and killed by heating at 100 C. for 10 minutes. The ratio of yeast to leukocytes in phagocytosis experiments was 1 : 1 or 5:1. Escherichia coli or Staphylococcus aureus 502A, grown for 18 hours in Trypticase soy agar and washed in Hanks’ solution, were added to leukocyte suspensions in ratios of 8:1 or 20:1. Blood 0 Rh-positive group red cells were with a serum containing a high were washed and added to leukocytes E. coli endotoxinf was used. 2 Pliagocytosis washed with buffered saline titer (1:2000) of anti-D in a final concentration hours and incubated antibody. of 0.5 The per for red cells cent (v/v). of Particles Leukocytes the presence were incubated in 95 per cent 02-5 per cent CO2 at 37 C. for 60 minutes in or absence of metabolic inhibitors before the addition of the test particles. Two types of leukocyte preparations were used: 1) suspensions, consisting of 1 ml. of cells (2 x 107/ml.) which had been rotated at 30 r.p.m. in 5-mi. plastic tubes in an incubator or of 2 ml. of cells shaken in 50-ml. siliconized Erlenmeyer flasks in a Dubnoff metabolic shaker (2 (3 cycles/second), x 107/ml.) capacity) for warm 60 MEM. minutes. was with or Ciemsa were counted per cent. In studies in were they added by of duplicate on pour-plates and the for added of a phase samples hypotonic of lysis contrast of with fixed in coefficient white of cell of nonadherent was continued Four of of leukocytes cells and the methanol microscope. mixture the 1 ml. in Leighton along smears the the free incubation were of coverslips washed preparations; killing, minutes were examination with consisting to glass radioisotopes observation each 30 variation and colony suspension tubes (1 ml. cells with for 5 to 120 test particles. and stained hundred cells was bacteria counts 6 ± were of the of agar.8 Studies Incorporation phosphatase The direct to adhere which quantitated of bacterial microorganism and production except after particles by preparations, allowed experiments, then every 2) glass-fixed been minutes, some Phagocytosis Metabolic had Test In removed and which that of H3-uridine into lysozyme activities of C1402 Hyamine flask as CO2-absorber. *Dow Chemical fDifco, Detroit, was from glucose-i-C14 added Michigan; RNA, production by previously of lactate and acid described methods.6’9 was determined as described previously,9 to the center well of a modified Erlenmeyer of variation for radioactive counts was ± 7 per cent. dfrectly The coefficient Co., Midlands, acid-precipitable were measured Michigan. lot 491-028. From www.bloodjournal.org by guest on June 14, 2017. For personal use only. 924 CLINE, Isolation of Leukocyte suspended granule LEHBER Granules were isolated from eosinophils or neutrophils by the method in phosphate-buffered saline at pH 7.4. Additions were suspension, and the change in optical density at 450 mt with time Granules HANIFIN, CASE of Williams’0 and then made to the was determined. SUMMARIES a 47 year old woman from El Salvador was admitted to the University of California Medical Center because of progressive loss of hearing. She had a history of previous infestation with Trichuris, Strongyloides and Ascaris. Examination of the stools revealed the larvae of Strongyloides stercoralis and the eggs of Trichuris trichiura. A routine WBC count showed 12,000 cells/cu. mm., with 58 to 78 per cent eosinophils. She was treated for parasitic disease, and the eosinophilia eventually disappeared. B.C., a 45 year old white housewife, entered the University of California Medical Center with a complaint of hay fever and asthma of 3 years’ duration. She had been hospitalized on several occasions previously because of pulmonary and sinus infections, symptoms of depression and suicide attempts. She had been treated with a variety of medications: V.P., antibiotics, antihistasnines, “antidepressants,” bronchodilators and expectorants, including iodide. Two weeks before the present admission cough and sputum production increased. X-ray examination showed infiltration at the base of both lungs and the right middle lobe. A sputum culture grew E. coli, and treatment with ampicillin was begun. The WBC count then and at the time of admission was 42,000 celLs/cu. mm., with 68 to 72 per cent eosinophils. Physical examination was unremarkable except for evidence of respiratory distress. During the patient’s hospital stay pronounced eosinophilia of the peripheral blood persisted, and repeated bone marrow aspirations revealed eosinophiic hyperplasia. A marrow chromosome preparation showed no specific abnormalities. A liver biopsy revealed normal potassium hepatic architecture without for ova and parasites were The patient’s symptoms was costeroids patient was instituted. discharged, evidence of infiltration. Numerous examinations of the stool negative; and no evidence of lymphoma was found. the eosinophilia continued until treatment with cortiThereafter, the WBC count decreased rapidly. At the time the the WBC count was 16,000 cells/cu. mm., with 1 to 7 per cent eosinophils. man, was transferred to the University of California Medical of fatigability, anorexia and persistent cough of 11 months’ duration of 30 pounds, he had been admitted to another hospital 5 months and spleen were enlarged, and the WBC count was 26,000 cells/cu. cent eosinophils. The patient underwent splenectomy and biopsy nodes at that time. Histologic examination revealed diffuse infiltration the spleen with mature eosinophils. Infiltration by eosinophils was also seen in periarteriolar sheaths and within the subendothelium of trabecular veins. The lymph nodes showed reticular and lymphoid hyperplasia and diffuse infiltration of the cortical and medullaiy areas by mature eosinophils. M.T., Center In a 25 year old white for evaluation. Because and a weight loss previously. The liver mm., with 47 per of mesenteric lymph of the red pulp of studies at the Medical and by biopsy, showed granulocytes. An attempt Center, specimens of bone marrow, obtained by aspiration solidly cellular marrow with a predominance of eosinophulic to prepare marrow chromosomes was unsuccessful. Numerous attempts to demonstrate parasitic disease failed to reveal evidence of infestation. The patient received 60 mg. of prednisone daily for 1 week, but the hematologic findings remained unchanged. Unfortunately, he left the hospital before a definitive diagnosis could be established. a REsULTh Particle Ingestion Eosinophils polystyrene in suspension ingested spherules, gram-positive a variety of particles and gram-negative in vitro, bacteria, including and living From www.bloodjournal.org by guest on June 14, 2017. For personal use only. 925 PHAGOCYTOSIS Table 1.-Phagocytosis Ratio Test l)lVSt Human to Eosinophils and Phago . incubation TVeutrophils cvtic . lndex* Eosinophils Time Leukocvtes (minutes) Neutrophils from B.G. V.P. MT. R.I.L. vrene spherules ( of Particles Particles by 1 mg. :1 x l0 30 70 ± 9 29 54 33 I :1 60 93 ± 3 - - - S :1 1 20 69 ± 17 - - 40 96 ± 0.5 - - - (i(lid(l ‘U (ilblCa7lS E.seheric/j coli ‘1(1/)/i 20:1 index 1)olystyrene tlw phagocytic the for mean ± of dul)licate and dead particles one S.D. samples in at particularly evident containing a large corresponding 7 0 less of in Table apparent nor glass-fixed The In in detail Using ficiency glass minutes 50 contrast, conditions and elsewhere.’2 cells isolated of ingestion and in after free the particles to 70 per of the from patient of heat-killed suspension. addition results mean cells ± ingested in (Table S.D. was was of was considerably at ingesting a was employed. eosinophils example, these 1 ). This percentage but inability from only less than eosinophils less than the 20:1 ratio of 12 per the of preparations cent cent ingested eosinophils to smears of time ingest the of eosinophils times large incubation phagocytized a 2 hour period. Under similar condineutrophils showed no erythrophagof human experiments monocytes are on monocytes M.T., we studied C. albicans by eosinophils yeast had at various sampling from representative stained Phagocytosis of The the the For cells over of human results as to leukocytes and 1. The was ocytosis. or neutrophils studied phagocytic phagocytic, particles human red preparations 5 more per cent of neutrophils were phagocytic and or more organisms. Under similar conditions, mixture (Fig. 1). Neither suspensions antibody-coated tions glass-fixed than fungi neutrophils. 20 populations. the of particles of ingested were which efficient and ratio 100 ± 6 or cell. For C. albicans, Results are expressed 1er samples Similar results were obtained periods up to 120 minutes. Data shown of with conditions. bacteria of neutrophil of duplicate neutrophils, number anreus leukocytes. 100 7 containing experiments. percentage of eosinophils are five least independent phagocytic 20 or more bacteria. during incubation at mean 12 leukocytes Staph. per of the a high to leukocytes, had ingested cent numbers as were percentage staphylococci 96 per cent periods a series the the of of fungi The efficiency experimental when of percentage number three and experiments, percentage 78 per least the coli or 20 or more of eosinophils particles such as either albicams. with the polystyrene In (1 1 ) for expressed Suspensions (Iefifle(1 10 or more E. is defined as the are C. varied 60 here 15 parti(les, ifl(Iex eosinophils the ± ylo(oeeos (IU1CU$ as 15 i(i was to the the complete cells. After phagocytic. are reported course time and ef- 5 and 15 adherent between 60 minutes, to fungi From www.bloodjournal.org by guest on June 14, 2017. For personal use only. 926 CLINE, . . ‘ LEHRER . .. . S. \ HANIFIN, h4. .. “S p . . ., . . _(> *.. ..* ,.. 1 Fig. 1.-A smear of suspension A central neutrophil has aureus. rounding were eosinophils found in suspended effects were containing either serum cent or no bacterial of the fixed phagocytosis. eosinophils 5 to the close 30 cells ingested when serum ratio in at pelleted 37 between yeast was determine at a variety when present, the of Staphylococcus but the Giemsa and 63.5 stain, per C. in the medium suspended serum x cent sur1000. of the heated the presence and eosinophils, 17 to 25 per of pH, of pH’s between serum in centrifugation incubating effect or to cent or were and which at 56 C. for g. of than lacked 2); for eosinophils 150 absence less medium eosinophils 5.5 on phagoin MEM required native serum (Table heated serum. The requirement experiments, by yeast of the suspending from M.T. were yeast or in additional were minutes contact autologous ingestion in albumin confirmed a 1:3 component Eosinophils albumin, Maximal was less was in for MEM per of the protein then studied. 30 minutes. phagocytosis To 91 little eosinophils and of microorganisms, cells. The cytosis yeast show of neutrophils, ingested a number , ,. ...j and and incubated serum. 1 per Despite cent serum; of the however, phagocytic. were 9 and suspended then incubated in serum with and heat- From www.bloodjournal.org by guest on June 14, 2017. For personal use only. 927 PHAGOCYTOSIS Table 2.---Effects of of Candida Suspending Protein Serum albicans and by pH the Phagocytosis (Patient M.T.)5 Medium Phagocytic Content 3% albumin 10% serum 10% serum 20% serum on Eosinophils PH heated at 56 C. for 30 mm. Index 7.4 7.4 7.4 5.5 1.0 60.5 22.0 82.5 20% serum 6.5 90.5 20% 20% serum serum 7.4 8.0 91.0 20% serum 9.0 70.0 *The results killed yeast. proached 15 are the As 7.4; minutes, means the of three incubation which experiments. proceeded, consequently, at independent the phagocytosis time there 72.0 was was no pH of all recorded significant suspensions only during deviation from pH. As shown in Table range, and only above 2, particle pH 8 does ingestion phagocytosis takes place over decline slightly. sults polystyrene as the particle. for Ingestion were obtained Metabolic As Requirements an approach ingestion by metabolic to . of these the eosinophils 3.-Effect Particle study of these cells before In all eosinophils affected Table the eosinophils, inhibitors ) spherules rations had with were the of Metabolic of as to cause on Human the test Phagocytosis of the Conditions Co ntaini Potassium cyanide 73.0 80.0 -- 1 x iO 69. 73.2 21 33 1 x 10 Sodium 2 x 10 fluoride lodoacetate 1 x 10 ng More Than indicates (‘ont. Exp. Exp. 54 40 23 24 54 45 72.9 12.4 21 3 54 it; 1d 75.1 25.3 21 0 54 6 M - Cont. - M Exp. - - :34 69 :i 0 M 73(1 67.3 21 11 54 23 :34 77.0 72.0 21 7 54 26 - 25 M 7 x I0 M 77.0 73. 54 45 34 28 control and exp. indicates experimental - cell - suspensions. cubated in 5 per cent C02-95 per cent O with saline as the only addition. Results independent experiments performed at different times on different populations population MT. Coat. 4 x 10 (‘ortisol *Cont i Exp. 5 Particles from 2.5 x 10 Puromcin Colchicine i to Particles V.P. (‘ont. 1)initrophenol prepainhibitors population Po1ystrene B.C. Neutrophils Concentration Anaerobic of Leukocytes Eosinophils Experimental particle (polystyrene glass-fixed If the of pH re- a variety particles than reason. initial for with a fraction (7 of Le ukocytes Inhibitor the first a broad Similar requirements preincubated suspensions rather for the following Inhibitors by metabolic were addition studies, used so the test apthe of eosinophils. Control cell suspensions are the means of neutrophils - were in- of at least three or on the same From www.bloodjournal.org by guest on June 14, 2017. For personal use only. 928 cu, Table 4.-Effect of Metabolic RTVA Inhibitors Human by H1-Uridi,ie on Leukocytes HANIFIN, Incorporation Conditions I nhihitor N(Lit 98 ± 5 95 1 12 i 03 ± S 74 1 00 I x H) \l 96 ± 10 I 06 57 2 x 10 \I 6 ± 4 5 \l 7 1 ± 12 71 \l 51 ± 16 85 ± 13 1)initrophenol IO(loac(tat( 1 x 10 2.5 (‘olchicine *l.t.siil1 s ts leave (Xl)I(’SS((1 tie I li(’ ire the reflected hibition surface, staining exceed 8 per itors of on and the The results an that intact of these neither poration nitrogen and eosinophils, terfered with particle uptake of uridine incorporation, H3-uridine vital experiments are nor 32 79 85 . rol (OUt unIv 1 leukocyte s. 1(l(lit . ion would not have To differentiate between cells, all suspensions as nonviable into RNA given in of the judged cells control, not effect was inwere experiments. by supravital did the affect of In the the inhib- in control measured glycolytic In particle ingestion these studies was fluoride iodoacetate and or (fluoride there 3 or included a both neuin- effect on inhibited slightly ap- incor- iodoacetate) a significant only is uridine tested With and It conditions for profoundly was 4. aerobic metabolism dinitrophenol. inhibitors before whereas Tables require of oxidative cyanide and _____ 60 111 t he t exl)eIini(llt eosinophils for ingestion dyes. - preparations. The inhibitors potassium trophils In of of 100 as rol 2 eosinophils of additional pathway into RNA. atmosphere, (liffel(’n in at the conclusion nonviable cells fraction an neutrophils mitochondrial us adherent blue of As leukocyte ti total population. simple killing This data. incorporation experimental parent cent. the inrurl)orat h salimu’ ( ) it of (cait 152 ((‘nt remaining the and ( I 1-mimidiiu e1 let erniiii;i 2 per cent trypan did the percentage interpretation of i ( 2-5 I limee of ag( ni ( the the behavior of phagocytosis stained with no instance mir ((‘lit if :i t least mmo:iiis glass 5 1)(1 ifl sI x 11) t lie LS 111(111 )tt ((l 5iiS1)flSli)fl5 I(’ill x I 0- 1 liromiiyein :i rol ) #{188}1 ‘1’. \I I x 10 fluoride \‘.P. (‘ont (Vttfli(l( SO(liLIIII from SiIR)j)hils B.(. of nu)sphere it Potassiuni rophils rat lou (‘omuent Nitrogen into :5 Eo I’x%)eriml1(’ntal LEHRER the H3- inhibitory. Phag- ‘I 0 _-*-_._. ln_J 40- 00 I”,. 0.0 5 ‘S.’ 5- viz:5 5%’ o 5-.-.- b- 20- .-.. -----#{149} .-- SW Neutrophils m Eosinoph0s 5-55 I 0 I 20 I I 40 I I 60 I I 20 1 I 40 I and phagocytic TIME, MINUTES Fig. 2.-Cellular (#{149} #{149}) leukocytes. acid phosphatase activity control (0 0) I 60 From www.bloodjournal.org by guest on June 14, 2017. For personal use only. 929 PHACOCYTOSLS Table 5.-Effect of Phagocytosis on Incorporation Metabolisni of of Production 113-Uridine Cell Control from Phagocvtic Type Neutrophils 696 124 ± Lactate (‘ontrol Production Phagocytic 1342 194 ± 163 5 ± 2534 124 65 ± 4 ± 148 12 ± from: 435 676 2505 -- .- 291 :3782 572 1284 MT. 172 *The methods of measuring trophils differed from the counting rate, 408 H3-uridine used employed mycin a slight 60 x effect less the 80 per cent studies ingestion, transferred particles titate fraction behaved determine however, often intracellular and than the magnitude and V.P.) of are from for neutrophils (13G. or three in- into the assessed the increase gives in Figure particle other precise suspending free by their lost at contrast 37 C. nonphagocytic dense an of were loss granule by residual of of activity data eosinophils were the in on the acid and measured cellular fraction granparticle and difficult in residual total the medium, of and to quanthe granule is most cellular although enzyme neither degranulation. phosphatase activity As occurred neutrophils. in are of polystyrene enzymes into the degranulation extent The size granule After enzyme Therefore, the occasional It is technically of in cellular events microscopy Suspensions of performed. the cytoplasm. measurement in enzyme 30 to col5M ) had neighbors. particles that from medium. in the quantitative ingestion phase period observations measurements 2, a decrease metabolic by vacuoles. When large numbers there is some leakage of granule by within PhagocytosLi activity are synthesis eosinophils. suggesting phosphatase enzymes inhibition of protein The dose of puro- protein by by granules, acid or of enzyme shown Three of degranulation method following of neu- similarly. The fate of the individual because of the motion of the cells and the phagocytic eosinophils clearly had surrounded into phagocytic are phagocytized, conveniently activity or affected The data studies Accompanying granules were lysosomal cytoplasm - tested were sensitive of cortisol (7 x 10 uptake be extruded intact from the cell. order to support the morphologic ulation, - by populations differences single of yeast were observed over a 15- to 30-minute prominent cells of production values. are from of eosinophils concentrations Changes ingesting granules phagocytic to on particle Morphologic and glass-fixed preparations granules was difficult to their cytoplasmic contents; shape these 94 -- appears to be more sensitive to than phagocytosis by neutrophils. inhibitory Eosinophils lose their and C’402 to control for eosinophils populations 10M. High two and fewer and Although of phagocytic Those inhibited The at 2.5 Metabolic I{NA populations. the ratios populations. into 83 (MT.). by eosinophils by puromycin minutes. chicine incorporation for eosinophil did not affect different studies ocytosis synthesis those they of seven dependent could In Phagocytic (Mg.) 220 to Control (c.p.m.) B.G. only Eosinophils of C’O V.P. studies and Glucose-I-C1tm c.p.m. Eosinophils Neutrophils phagocytic eosinophils: From www.bloodjournal.org by guest on June 14, 2017. For personal use only. CUNE, 930 Table 6.-Effect of Endotoxin Eosinophils on the Metabolis,mi in Autologous of Seruv,a Incorporation (2 Human x 1O No. of Neutrophils of Neutrophils the from MT. of (a shunt), in Table All three was metabolic The more eosinophils. tion the of the higher Fig. dotoxin, ± 200 3532 rise into pronounced This events study, the was were Endotoxin after than does the not incubation matter of in both types ingestion of of the ingested 30 minutes a vacuole 866 ± C1402 from hexose-monophos- of lactic in neutrophils, for within of the acid. eosinophils necessarily showing 6434 6524 production in two shunt 162 ± 3424 by stimulated of neutrophils 3.-An eosinophil after demonstrating particulate 3280 production exhibited production hexose-monophosphate percentage RNA, in neutrophils observation 388 measurement pattern in C1402 ± 748 H-uridine in a single same (‘ontrol (c.p.m./hr.) 817 semiquantitative and, 5, the kocytes. 2006 I incorporation glucose-i-C’4 phate Endotoxin of Ci402 Glucose-1-C (c.p.m.’hr.) 7 F:osinophils and Production from (‘ontrol Studies LEHRER cells/mi.) I13-Uridine (‘eli Type HANIFIN, of As phagocytic polystyrene three reflect leuparticles populations a greater but shown neutrophils. arid of stimula- is consistent with particles. with Escherichia (arrow). coil en- From www.bloodjournal.org by guest on June 14, 2017. For personal use only. 931 PHAGOCYTOSIS 40 - 20 - 0 , Granules Neutrophil 8 Eosinophil 310 210 Granules 510 40 TIME, MINUTES Fig. 4.-The ules suspended change in optical in buffered saline absorpancy at pH 7.4 -s). S)orO.1NHCi( (S Effects at 450 m of isolated leukocyte gran0), 0.8 per cent deoxycholate (0 of Endotoxin We have changes in shown previously that E. coli endotoxin produces human neutrophils resembling those associated sis.13 The changes include increased production of C’402 and of lactate, increased incorporation of H3-uridine into tide and RNA, panied ides. and by the These which particles contaminate the than of the present studies the almost certainly cells effect 0.i of endotoxin indicate of the on same those walls eosinophils seen 105,000 was produces RNA by effect examinaat on endotoxin-treated to part- cell microscopic preparations in diameter. accom- small bacterial that endotoxin H3-uridine into Occasional similar of Electron such to i lysosomal are containing fragments of certain events of vacuoles approximately particles of metabolic preparations. the has activity These . centrifugation approximately glucose-i-C’4. containing cellular ) within (Table 6) incorporation but from vacuoles by the lysozyme endotoxin results in the neutrophils, C1402 are obtained measuring similarly. The matic changes in and appearance tion of pellets reveals particles In a decrease ( a-glucosidase enzymes metabolic with phagocytofrom glucose-i-C14 acid-soluble nucleo- the tested less draeosinophils production eosinophils in g had ( Fig. neutrophils 3). Degranulation Is the integrity influenced be by obtained In two tration and that granule factors? of separate the the the experimental of Only on the absorbancy granules at 450 of eosinophils conditions. the eosinophils a limited difficulty experiments, of deoxycholate by measuring these the same because phils. strate of the in the mp. acquiring effects integrity The and and answer large of a high of eosinophil results, neutrophils of the to this shown neutrophils question numbers of could eosino- hydrogen ion concen- granules was studied in Figure behave similarly 4, demonunder From www.bloodjournal.org by guest on June 14, 2017. For personal use only. 932 CLINE, HANIFIN, LEHRER DiscussioN The that the precise cells function of the are phagocytic, teria and bacterial toxins, that are in contact with gest a role in host and Certainly, other phages, are abundant The present are phils. and The The role as yet sequestration and the processing in that hydrogen ion as efficient serum Watson in is than of that occur eosinophils in these are efforts are of subsequent antibody as the free late suspension.15 in an infectious studies resemble ingestion Their oxidative metabolism, slightly reduced study on in and by In limited ), and enzymatic studies, sensitive into the glass range are at suspension, suggested least provided that surface phago- antibodies, whereas phagothe high levels of antibodies from are occurs the granules consequences be unaffected reduced protein by of by particle inhibitors of and production and phagocytosis both ingestion and particle of glycolysis Lactate shunt eosinophils deoxycholate. known inhibitors observed eosino- for of phagocytosis. by synthesis. all stimulated following with metabolic Degranulation, tests, patients requirements3’#{176}”6 to of RNA and to free hexose-monophosphate neutrophils. to acid resemble a broad metabolic is strikingly inhibitors eosinophils of H3-uridine eosinophils by of the appears but the neutrophils, Wood, Smith and may be a more efficient process eosinophils their some capacity The their physioeosinophil or over adherent They that in in at least and in Eosinophils ingestion cells possible phagocytosis process. indicate neutrophils phagocytic by production. phagocytes. same allergic by antigenprocesses.14 Among may be important are unique to of particle fungi various is unknown. Eosinophils yeast eosinophils certain in aggregates eosin- as neutro- are attracted in immunologic of immune other capable with normal abundant they processes for patients that macrodefensive of bacteria shown or in is un- and in phagocytizing From studies of rabbit that surface phagocytosis in sug- Whether efficient from strains have disarming the ingesting available. concluded present philia equally eosinophils may require only low levels of opsonizing by freely suspended cells may require The tion more unreported, concentrations. phagocytosis cytosis cytosis neutrophils instances the eosinophil these functions they flora microorganisms. including of certain antigens of duplications neutrophils cell of membranes bacterial some evidence that their involvement and phagocytic capacities logic functions-whether its cells, that that of the are of indicate observation roles merely in phagocytes and experimental aggregates suggest specific these less effective than neutrophils ingested microorganisms. the clinical conditions antibody and studies, in mucous and against phagocytic as efficient Additional considerably in killing are abundant environment The observations inactivating bac- cells are present in the circulating blood to be critical in such defense reactions studies not is unknown. capable of particularly reactions known. more eosinophil enzymes external defense numbers of these active inflammation ophiia are the sufficient sites of units. human possess neutrophils the (one incorpora- in both human morphologically in both cell appear types. to be From www.bloodjournal.org by guest on June 14, 2017. For personal use only. 933 PHAGOCYTOSIS In in conclusion, the human range appear but to be not eosinophils in similar the in the of phagocytosis. We of physiologic functions appear efficiency of requirements may thus for add that to particle these these resemble human ingestion. The and the aspects two cell types consequences metabolic of of types neutrophils two phagocytosis human to white cells the list have in common. SUMMARY Eosinophils tested were phagocytic for Escherichia coli, so less red cells. Metabolic over lysosomal had of RNA, enzymes similar a broad on glucose and in glycolysis, extent. and neutrophils oxidation presence and incorporation degranulation and by the did puromycin enhanced oxidation in but antibody-coated range of including similar albicans, and eosinophils. eosinophils En- and neutro- in vitro. SUMMARIO Eosinophulos testate esseva pro styreno, br minus coli, del Concomitantes in enzymas del le stimulation lysosomal-esseva effectos super le in un phagocytose-incluse del oxydation oxydation de in e Candida area esseva de sed erythrocytos in poly- isto ilos revestite de pH glycolyse, illos incite albicans, de valores de e particulas phagocytava large le de glucosa, neutrophilos glucosa eosinophiia, de e in ie presentia puromycina e colchicina, grados. del simile non Inhibitores a vane con ingereva 520A, Illos oxydative. particulas patientes eosinophilos aureus occurreva metabolismo INTERLINGUA de Le neutrophilos. metabolic ARN, IN sanguine Staphylococcus que le ingestion le phagocytic. efficace inhibitores ad ab Le phagocytose anticorpore. inhibiva isolate capacitate Escherichia esseva de pH Inhibitors to a variable of glucose were effects did phagocytosis, stimulation with eosinophilia and polystyrene particles, and Candida not phagocytize 502A They occurred concomitants into dotoxin aureus neutrophils. of oxidative metabolism. inhibited particle ingestion of uridine phils than Phagocytosis of from the blood of patients Eosinophils ingested inert Staphylococcus efficiently of inhibitors colchicine loss isolated ability. per promovite e le e eosinophilos. eosinophilos de uridina incorporation disgranulation e le Endotoxina e neutrophilos perdita habeva in studios de simile effectuate vitro. in ACKNOWLEDGMENT The electron were micrographs Medical California Center, kindly prepared by Dr. William Davis, University of San Francisco. REFERENCES 1. Schultze, M.: Em tisch und seine Verwendung ungen des Blutes. Arch. 1856. 2. des Mesnil, A.: Sur le vert#{233}br#{233}s inf#{233}rieurs biennes de artificielles; l’immunit#{233}. heizbarer Objectbei UntersuchMikr. Anat. 1:1, mode aux de Inst. micro- a contribution Ann. r#{233}sistance invasions Pasteur l’#{233}tude 9:301, 1895. 3. phil Hirsch, leukocytes. J. C.: Neutrophil In: Zweifach, and eosinoB. W., Grant, L. and McCluskey, R. T. (Eds): The Inflammatory Process. New York, Academic Press, 1965, p. 245. 4. Cline, M. J.: Metabolism of the circulating leukocyte. Physiol. Rev. 45:674, 1965. 5. lation cytes Archer, of C. granules T., and Hirsch, from and study of their J. Exp. Med. 118:277, 1963. 6. Cline, M. J.: Isolation eosinophil enzyme and J. G.: Isoleuco- content. character- From www.bloodjournal.org by guest on June 14, 2017. For personal use only. 934 ization J. Lab. CLINE, of RNA Clin. Med. from 68:33, J., human 1966. Ltd., leukocytes. 1953, 4th ed. Clime, M. J., and Lehrer, R. I.: Phagomonocytes. Blood in press. 13. Clime, M. J., Melinon, K. L., Davis, w. C., and Williams, H. E.: Mechanism of endotoxin interaction with human leucocytes. Brit. J. Haemat. in press. 12. III, Davidson, J. S., and Burk, D.: Leukocyte preparations from human blood: Evaluation of their morphologic and metabolic state. J. Lab. Clin. Med. 59:779, 1962. 8. Meynell, C. G., and Meynell, E.: Theory and Practice in Experimental Bac- cytosis eosinophil: Phagocytosis teriology. complexes. Proc. 7. Fallon, J. D., Trier, H. Cambridge, Frei, E., Cambridge University 1965, 9. Cline, p. 15. M. J.: Phagocytosis and synthesis of ribonucleic acid in human granulocytes. Nature 212:1431, 1966. 10. Williams, H. E.: a-Glucosidase acPress, tivity ophys. 11. in human leucocytes. Biochim. Bi- Acta 124:34, 1966. Fisher, R. A., and Yates, F.: Statistical Tables for Biological, Agricultural and Mcdical Research. Edinburgh, Oliver and Boyd HANIFIN, by 14. human Sabesin, S. M.: Soc. A function of antigen-antibody Biol. Exp. of Med. the 112: 1963. 667, 15. Wood, Watson, B.: W. B., Jr., Smith, Studies on the M. R. and mechanism of recovery in pneumococcai pneumonia. IV. The mechanism of phagocytosis in the absence of antibody. J. Exp. Med. 84:387, 1946. 16. Karnovsky, of phagocytic 143, 1962. M. activity. L.: Metabolic Physiol. basis Rev. 42: From www.bloodjournal.org by guest on June 14, 2017. For personal use only. 1968 32: 922-934 Phagocytosis by Human Eosinophils MARTIN J. CLINE, JON HANIFIN and ROBERT I. 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