Eyes with Ascaris Suum and Toxocara Cam's Larvae

In Vitro Interaction of Eosinophils from Ascarid-infected Eyes with Ascaris suum and Toxocara cam's Larvae

John H. Rockey,* Thomas John,* John J. Donnelly,*f Diane F. McKenzie,* Bert E. Stromberg4 and E. J. L. 5oulsbyf

The role of eosinophils in ocular ascarid infection is studied in an animal model. Primary intravitreal infection of guinea-pig eyes with ascarid (Toxocara cants, Ascaris suum) second-stage (L2) larvae resulted in an anterior chamber exudate containing 98% or more eosinophils. Anterior chamber aspirates were cultured in RPMI media 1640 with T. canis or A. suum L2 larvae at 37° C for 1-96 hours. The in vitro interaction of eosinophils with L2 larvae was studied by light, and scanning and transmission electron microscopy. Eosinophil interaction with L2 larvae in vitro was dependent upon soluble factors present in the anterior chamber aspirates from the infected eyes. Eosinophils adhered firmly to the surface of the L2 larvae, to a larval sheath, or to attached eosinophils. Eosinophils interacting with larvae displayed a range of granular matrix changes, core duplication, and reversal of the relative electron densities of the core and matrix, suggestive of eosinophil activation. An eosinophil secretory granule was seen to empty its contents onto a T. canis L2 larval surface. Electron-dense material was observed in the interphase between eosinophils and the larval cuticle or sheath. Large lipid droplets within muscle cells and ballooned-out epidermal cells were seen within larvae immediately beneath adherent eosinophils. Parasites were able to partially evade interaction with eosinophils in culture by shedding their sheaths. A similar phenomenon in vivo may allow the parasite to migrate from a site of inflammation. It is possible that a discarded sheath or membrane serves as an antigenic stimulus for a local intraocular granulomatous reaction free of parasite. Invest Ophthalmol Vis Sci 24:1346- 1357, 1983

Immunopathologic reactions of the eye in general immunopathologic reactions, but are especially nu- are the result of complex and incompletely understood merous in the intraocular and periocular granulo- interactions between any of a number of different cel- matous reactions caused by parasites.1"3 Experimental lular and/or humoral factors activated by extraneous intraocular infection with ascarid larvae produced eo- or abnormal chemical stimuli. The roles of distinct sinophilic granulomatous reactions in which eosino- ocular immunopathologic mechanisms may be difficult phils adhered firmly to larvae or a surrounding mem- to isolate in vivo, but may be studied to advantage in brane and appeared to be degranulating onto the larval tissue culture where specific cellular and humoral fac- surfaces. l>4"7 The mechanisms by which this adherence tors may be separated and manipulated independently. occurs, particularly the role of humoral factors in the Eosinophils are a prominent feature of several ocular eye (eg, locally produced antibodies), the extent of degranulation of adherent eosinophils, and the nature of the damage to the parasite, may be studied readily in vitro. The role of separable elements (eg, eosinophils) From the Department of Ophthalmology, Scheie Eye Institute, School of Medicine, University of Pennsylvania, Philadelphia, Penn- in ocular granulomatous reactions also may be inves- sylvania,* the Department of Clinical Veterinary Medicine, University tigated. In the present study, we examine the interaction of Cambridge, Cambridge, England,! and the Department of Vet- in culture of eosinophils and humoral factors from the erinary Pathobiology, College of Veterinary Medicine, University of ascarid-infected guinea pig eye with second-stage larvae Minnesota, St. Paul, Minnesota.^ of Toxocara canis and Ascaris suum. Supported by USPHS NEI Grants EYO3984 and EYO7041, a Fight For Sight Post-Doctoral Research Fellowship from Fight For Sight, Inc., New York City, grants from the UNDP/World Bank/ Materials and Methods WHO Special Program for Research and Training in Tropical Diseases and the Wellcome Trust, an unrestricted grant from Research to T. canis and A. suum Larvae Prevent Blindness, and by Gretel and Eugene Ormandy Teaching Second-stage larvae (L2) of T. canis and A. suum and Research Fund. 1 4 7 Submitted for publication July 9, 1982. were prepared as described previously. ' " Microscopic Reprint requests: John H. Rockey, M.D., Ph.D. Scheie Eye In- examination revealed that 95-98% of the larvae were stitute, Myrin Circle, 51 North 39th Street, Philadelphia, PA 19104. viable (motile). Freshly hatched A. suum U2 larvae in

0146-0404/83/1000/1346/$ 1.40 © Association for Research in Vision and Ophthalmology 1346

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sterile saline were used immediately for intravitreal larva was counted and tabulated in five cumulative injections. A. suum and T. canis L2 for in vitro studies groups: 0, s^l—3, ^4-6, ^7-10, and ^10 eosinophils were incubated in sterile saline containing 0.03-0.08 per larva. mg/ml gentamicin sulfate for 12-24 hours at 37° C Electron Microscopy in a 95% air-5% CO2 atmosphere. Suspensions containing larvae with adherent eosin- Eosinophils ophils were fixed in 2.5% glutaraldehyde-4 mM CaCl2- Female, Hartley guinea pigs (500-600 g, Skippack 64 mM Na cacodylate buffer, pH 7.4, at 4° C for 14- Farms, Skippack, PA) were used for eosinophil pro- 16 hours. Specimens were washed three times and re- duction by intravitreal infection with second-stage as- suspended in fresh Na cacodylate buffer containing carid larvae.14"7 Intravitreal injections were performed 131 mM sucrose. Larvae-eosinophil preparations for with ether inhalation anesthesia. A. suum larvae transmission electron microscopy (TEM) were post- (3,000-3,500 L2 in 0.1 ml sterile physiologic saline) fixed with 1% OsO4 in 0.12 M Na phosphate buffer, were injected through the pars plana and the motile pH 7.4, at room temperature for 2 hours, and dehy- larvae were viewed by direct ophthalmoscopic ex- drated sequentially with 70%, 95%, and three changes amination in the vitreous. Anterior chamber paracen- of absolute ethanol (10 min each), infiltrated for 16 tesis was performed under ether inhalation anesthesia hours with Spurr resin diluted 1:1 with propylene oxide, 10 days after primary intravitreal infection, using a and for 48 hours in two changes of undiluted resin, tuberculin syringe and a 30-gauge needle. Each guinea embedded in fresh resin, and polymerized at 70° C pig eye was tapped only once, and 0.01-0.05 ml of for 38-72 hours. anterior chamber fluid was obtained from each eye. Thin sections for light microscopy were stained with toluidine blue. Ultrathin sections for electronmicros- copy were stained with uranyl acetate and lead citrate, Eosinophil-Parasite Interaction In Vitro and examined in a Hitachi® HS-8 or a Philips® EM Anterior chamber aspirates from infected guinea pig 201 electron microscope. eyes were added to 0.5 ml of synthetic culture medium For scanning electron microscopy (SEM), fixedlar - (10.4 g RPMI 1640, Grand Island Biological Co., vae-eosinophil suspensions were dehydrated through Grand Island, NY, and 0.3 g gentamicin sulfate per graded ethanol (25%, 50%, 70%, 95%, and 100%), L HEPES buffer, pH 7.2). Fibrinous material was re- washed three times in amyl acetate, placed on a glass moved mechanically with a sterile wire inoculating coverslip, dried at room temperature over silica gel, loop. T. canis or A. suum L2 larvae (1,000 larvae in sputter coated with 100 A gold, and examined with 0.1 ml sterile saline; viability 95% or greater) were an AMR 1000 scanning electron microscope. added to the eosinophil cultures and incubated in a 95% air-5% CO2 atmosphere at 37° C for 1-96 hours. Results Eosinophil-parasite interaction was examined at 10- min to 24-hour intervals in 50- to 100-/il samples by The anterior chamber exudates present 10 days after dark-field, phase contrast, or differential interference a primary intravitreal infection with ascarid larvae were microscopy, and in air-dried smears stained with composed almost entirely of eosinophils. Figure 1 Wright's or Luna's eosinophil granule stain. shows a histologic section through an anterior chamber exudate, and the adjacent cornea stained with Luna's eosinophil granule stain. Except for a few macrophages Aqueous Humor Factors and Eosinophil- almost all of which had phagocytized one or more Parasite Interactions eosinophils, all of the cells were eosinophils. Light mi- Selected anterior chamber paracentesis samples were croscopy of smears from anterior chamber aspirates, divided equally between two culture tubes, each con- stained with Wright's or Luna's eosinophil granule taining 0.25 ml RPMI medium. To remove aqueous stain, showed that 98% or more of the cells were eo- humor antibodies and/or other soluble factors, the eosinophils. No larva was seen in the stained smears or sinophils in one tube were centrifuged, the supernatant upon dark-field examination of the anterior chamber was removed, and the cells were washed and resus- aspirates in culture medium. pended in fresh RPMI medium. Eosinophils in the The interaction of eosinophils with viable (motile) corresponding second tube were left unwashed. Equal ascarid larvae in tissue culture was followed readily by numbers of ascarid L2 larvae (1,000-1,200 larvae in dark-field light microscopy because the large brightly 0.1 ml sterile saline) were added to each tube and the refractile granules of the eosinophils were identified cultures were incubated in the standard manner. The easily (Fig. 2). Rapid oscillatory motion of the granules total number of eosinophils attached to each ascarid of many, but not all, eosinophils was seen in eosinophils

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Fig. 1. Histologic section of an anterior chamber exudate in a guinea pig eye 10 days after intravitreal infection with ascarid larvae. All of the cells are eosinophils except for a rare raacrophage which has phagocytized one or more eosinophils (ME). Corneal endothelium (A). (Luna's eosinophil granule stain.)

attached to parasites. The eosinophils were attached the two points of adherence through a long, taut elastic either directly to the larval surface, to a refractile cytoplasmic band drawn out from one extreme of the striated sheath, with surface corrugations complemen- eosinophil by the extension of the larval loop. Larvae tary to those of the parasite, that encased the larvae often were covered by several layers of eosinophils. (Fig. 3A), or to sheaths from which the larvae had Some elongated masses of eosinophils were identified escaped (Fig. 3B). Some A. suum larvae were seen to as containing ascarid larva only by the active flexing be freely mobile within their sheath. On occasion, a and extending motion of the enclosed larva. Dense larva was observed to slide in and out of a ruptured collections of eosinophils about ascarid larvae, com- sheath to which were attached many eosinophils (Fig. parable to the central collections of eosinophils in in- 3A). The larger T. cams second-stage larvae were more traocular eosinophil granulomas,1'4"7 also were seen in active than the A. suum larvae, and rapidly lost their tissue culture (Fig. 4). sheaths which, with attached eosinophils, were found Examination of smears stained with Wright's or free in the culture medium. The sheaths were very Luna's reagents revealed that almost all of the cells thin and were observed to collapse against larvae be- attached to the parasite larvae or sheaths were eosin- cause of thermally induced currents in the culture me- ophils. Only a very rare attached macrophage or poly- dium on the microscope slide. The adherence of eo- morphonuclear neutrophil was seen. Eosinophil gran- sinophils to the larvae was very firm. Eosinophils at- ules were dispersed over the larval surfaces (Fig. 5). tached at two opposing points across a loop of an Interactions occurred not only between eosinophils and actively flexed T. canis larva were seen to maintain larval surfaces, but also between larval-attached eo-

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Fig. 2. Dark-field microscopy of anterior chamber eosinophils interacting in culture with a second-stage larva of T. canis.

sinophils and outer-layer eosinophils where eosinophils from the dark electron-dense outer cuticular lamina; were interconnected by bridges of varying width (Fig. similar granular material also is found within the sur- 5). This type of eosinophil-eosinophil interaction gen- face invaginations. There is periodic condensation of erally was not seen between eosinophils free of parasite electron-dense material within the eosinophil cyto- interactions. plasm immediately beneath the eosinophil plasma Scanning electron microscopic observations of larva- membrane over the parasite cuticular annular striae eosinophil interactions occurring in tissue culture are (Fig. 7A). In Figure 7B, an eosinophil has engulfed shown in Figure 6. The ascarid larva shown here is fragments and ends of the larval sheath and also has covered almost completely by eosinophils which in penetrated beneath the sheath to interact closely with many instances have flattened out over the parasite the larval cuticular surface. In Figure 8A an eosinophil surface. The annular striae and the intervening trans- has penetrated beneath a larval sheath and has become versely arranged surface invaginations or indentations attached both to the larval cuticle and the inner surface of the parasite surface are seen in the few regions not of the larval sheath. Long thin pseudopodia of other covered by eosinophils. eosinophils have interacted with the outer surface of Transmission electron microscopy of eosinophil- the parasite sheath. The parasite sheath is seen to be parasite interactions occurring in tissue culture showed composed of two electron-dense layers separated by an intimate interaction between the eosinophils and an electron-lucent layer. In Figure 8B a uniform gran- the T, canis or A, suum L2 larval cuticle or sheath. ular membrane located between the eosinophil plasma In Figure 7A, the eosinophil plasma membrane shows membrane and the larval cuticular surface has pulled a high degree of complementarity with the outer sur- away from the cuticular surface but has retained small faces of the annular striae of the parasite cuticle, but spike-like projections that show a high degree of com- bridges over the transversely arranged surface invag- plementarity to the surface invaginations of the larval inations or indentations between the annular striae. cuticle annular striae. A thin layer of moderately electron-dense granular Eosinophils displayed a wide range of granular material separates the eosinophil plasma membrane changes both in the granule matrix and in the crys-

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Fig. 3A. Dark-field microscopy of anterior chamber eosinophils interacting in culture with a second-stage larva of A. suum. Eosinophils are adherent to a sheath (A) that partially encases the larva. During observation this parasite was seen to move in and out of the sheath through a rupture. 3B. Dark-field microscopy of anterior chamber eosinophils interacting with an isolated ascarid sheath.

talloid core. A range of early eosinophil granule matrix looned-out epidermal cells were seen within the ascarid changes and core duplications is illustrated in Figure larvae immediately beneath areas of firm attachment 8B. In Figure 8C an eosinophil secretory granule is of eosinophils to the surface of the parasite (Figs. 7 A, seen emptying its contents onto a T. canis Ul lateral 8A). Early parasite cuticle disruption also was noted. ala. The contents of the secretory granule are closely Vacuolization was seen subcuticularly in muscle, epi- similar to the altered matrix materials of some of the dermal and gut cells. eosinophil granules of Figure 8B. Other eosinophils The interaction of eosinophils with ascarid larvae displayed reversal of core-matrix patterns with electron- in vitro was dependent upon soluble aqueous humor lucent crystalloid cores and electron-dense granule factors present in the aspirates from the infected eyes. matrices (Fig. 7C). Some eosinophils that were attached Whereas 94% of the ascarid larvae had one to three to ascarid larvae showed extensive degranulation and or more, and 50% more than ten, attached eosinophils cytoplasmic vacuolization (Figs. 7, 8). Microfilaments per larva when both soluble and cellular anterior and centrioles were seen in some eosinophils (Fig. 8A). chamber aspirate components were present, only 11% Large lipid droplets within muscle cells and bal- had one to three or more, and none more than seven

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Fig. 4. Dark-field microscopy showing a dense collection of eosinophils adherent to ascarid larvae.

Fig. 5. Eosinophil-ascarid larva interaction seen in a Wright-stained smear of culture fluid. Eosinophil granules are dispersed over the larval surfaces (A). Eosinophil-eosinophil bridges are seen in the layers of eosinophils attached to the parasite (#).

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Fig. 6A. Scanning electron microscopy of eosinophils interacting with an ascarid larva. 6B. Segment of the ascarid larva with adherent eosinophils at higher magnification.

attached eosinophils when soluble factors were elim- peroxide and superoxide anions.8 l4 Eosinophils have inated from the cultures of larvae and eosinophils (Ta- surface membrane receptors for the Fc fragment of ble 1). IgG and IgE heavy chains and for complement components (C3b, C3d, C4).15-17 Discussion All soluble factors required for eosinophil adherence and damage to the ascarid larvae seen in the present The interaction of anterior chamber eosinophils system were contained in the anterior chamber exu- from the ascarid-infected guinea pig eyes with second- dates. The ascarid-infected eyes have been shown to stage larvae in culture required soluble factors absent have uveal infiltrates of plasma cells locally producing from washed cell preparations. A number of soluble immunoglobulins, detected by immunofluorescence factors have been shown to be important for eosinophils with anti-guinea pig IgG (H- and L-chains) antibodies to adhere to parasite surface membranes and for the within ten days after a primary intravitreal infection.1 cytotoxicity and killing of parasites by eosinophils in The soluble factors lacking in the washed eosinophil tissue culture. These include noncytophilic (eg, human preparations may include noncytophilic (eg, IgG2) anti- IgG) and cytophilic (eg, rat IgG2a, IgE) antiparasite ascarid antibodies directed against surface antigens of antibodies, complement, the lymphokine Eosinophil the A. suum and T. canis second-stage larvae and larval Stimulation Promoter (ESP), mast cell mediators such sheaths. The presence of IgE antibody in anterior as Eosinophil Chemotactic Factor of Anaphylaxis chamber aspirates from guinea pig eyes after intravit- (ECF-A) tetrapeptides and histamine, and hydrogen real infection with A. suum and T. canis larvae has

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Fig. 7A. Transmission electron microscopy (TEM) of eosinophil-ascarid larva interaction. Periodic condensations beneath eosinophil plasma membrane over parasite cuticular striae (A) (X7.20O). 7B. TEM of eosinophil-ascarid larva interaction. Fragments of a ruptured larval sheath engulfed by the eosinophils (A). Lipid droplet in ascarid muscle (L) (X6,2OO). 7C. TEM of an eosinophil in culture with ascarid larva. The granules show reversal of the normal electron densities of the central core and the matrix (X6,6OO).

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Fig. 8A. TEM showing eosinophils that have penetrated beneath the sheath of an ascarid larva and have interacted with both the sheath and the parasite cuticle. Pseudopodia of eosinophils interacting with the sheath (A). Lipid droplets in ascarid muscle cell (L). The cuticle is separated from the underlying muscle cells by epidermal cells that have become ballooned-out (A) (X4.500). 8B. TEM of eosinophil-parasite interaction in culture. The eosinophil has separated partly artifactitiously from the parasite. A uniform membrane (A) with projections closely corresponding to the indentations of the parasite surface is seen between the eosinophil plasma membrane and the parasite cuticular surface. The eosinophil granules show a range of matrix changes. The epidermal cells (A) have a normal appearance with the cuticle very closely approximated to the underlying muscle cell (X7.200). 8C. TEM of a vacuolated, partially degranulated eosinophil adherent to a T. canis larva in culture. Secretory granule (SG) opening out onto the parasite surface and lateral ala (X4,800).

been reported previously.1*4"7 The time of appearance origin. A temperature-dependent coating of T. canis of aqueous humor IgE antibody corresponded to a second-stage larva, detected by fluorescent antibody rapid increase in the intraocular eosinophil infiltrate in vivo, recently was described by Smith et al.41 following a single intraocular infection with second- Eosinophils also were attached to a parasite sheath stage ascarid larvae.5 Intraocular IgE antibody also composed of two electron-dense layers separated by could play a role in the eosinophil-induced adherence an electron-lucent layer. A similar sheath that had sep- and damage to second-stage larvae of A. suum and T. arated from an ascarid larvae lying between the retina canis observed in the present experiments.8 and pigment epithelium of an intravitreally infected Cuticle disruption, lipid droplet formation within guinea pig eye is seen in Figure 2 of reference 1. In the muscle cells, and ballooned-out epidermal cells culture, the A. suum and T. canis larvae were able to immediately beneath the area of eosinophil attachment move freely within or to discard these sheaths with to the parasite surface were evidence of eosinophil- attached eosinophils. induced larval damage in the present studies. Eosin- The larval sheaths, or a surface coating of parasite ophils contain a number of enzymes and mediators origin, from which larvae may escape, offer a mech- (Table 2) that may contribute to eosinophil-mediated anism whereby the ascarid parasites may evade the parasite damage. host's immune response and destruction by eosino- The granules of the mature eosinophil leukocyte, phils. A discarded sheath or membrane, left behind when examined with an electron microscope, are seen by an escaping larva, could serve as an antigenic focus as typically ellipsoidal, membrane-bound structures for a local intraocular eosinophil granulomatous re- consisting of an electron-dense core and an electron- action while the viable larva is free at a distant site. 33 lucent matrix. The crystalloid core contains the eo- In human ocular toxocariasis with eosinophil cho- sinophil major basic protein (MBP) and eosinophil rioretinal granuloma, even on serial sectioning, it is 1819 cationic proteins (ECP). Eosinophils attached to common not to find the parasite.2 This may be ex- ascarid larvae in culture showed a wide range of granule plained by the parasite having left behind its membrane changes or depletion of eosinophil granules (degran- or sheath which provokes the chorioretinal lesion, while ulation). These eosinophil granular changes may be the viable larva has migrated elsewhere in the eye or the result of "activation" secondary to eosinophil-par- out of the globe. This mechanism also may explain asite interactions. Eosinophils that had radiolucent the observation of Glickman (personal communica- crystalloid cores and more dense granule matrices were tion) that in monkeys injected intravitreally with T. present in cultures with larvae. Such eosinophil granule canis second-stage larvae, the granulomatous lesions reversal patterns have been reported before to indicate eosinophil "activation."3435 The discharge of granule content onto larval surfaces also was seen from eosin- Table 1. Dependence of eosinophil adherence to ophils attached to ascarid larvae. ascarid L2 larva in culture on soluble anterior Electron-dense material was observed between the chamber factors attached eosinophils and the cuticular surface of the A. suum and T. canis larvae. Similar electron-dense Number of attached eosinophils per ascarid larva material between parasites and attached eosinophils Soluble factors 0 ^4-6 ^7-10 has been observed by others.3436"40 In contrast, in adherence studies between neutrophils and schistosomula 6* 94 71 59 50 of Schistosoma mansoni, electron-dense material was 89 11 1 0 0 38 absent. The electron-dense material between the as- * Percentage. carid larval cuticular surface and the eosinophil plasma Anterior chamber aspirates from L2-infected eyes of three animals were added separately to culture medium and subdivided. The effect of removing membrane may represent host material (eg, specific soluble factors by washing one-half of the cells in culture medium on eosinophil- antibody, complement) bound to the parasite surface. parasite adherence was examined in Luna's stained smears: 400-500 parasites were counted per culture tube. Percentage of the total larvae with a given Alternately, it may represent a larval coating of parasite number of attached eosinophils is tabulated.

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Table 2. Biochemical constituents of eosinophils References 1. Rockey JH, Donnelly JJ, Stromberg BE, Laties AM, and Soulsby Substance Reference EJL: Immunopathology of ascarid infection of the eye: role of IgE antibodies and mast cells. Arch Ophthalmol 99:1831, 1981. Major basic protein (MBP) (granule crystalloid core) 18 2. Wilder HC: Nematode endophthalmitis. Trans Am Acad Eosinophil cationic proteins (ECP) Ophthalmol Otolaryngol 55:99, 1950. (granule crystalloid core) 19 3. Ashton N: Larval granulomatosis of the retina due to Toxocara. Peroxidase 20-22 Br J Ophthalmol 44:129, 1960. /3-Glucuronidase 20-22 4. Donnelly JJ, Rockey JH, and Soulsby EJL: Intraocular IgE an- Arylsulfatase B 20,23 tibody induced in guinea pigs with Ascaris suum larvae. Invest Ribonuclease 20 Ophthalmol Vis Sci 16:976, 1977. Cathepsin 20 5. Rockey JH, Donnelly JJ, Stromberg BE, and Soulsby EJL: Im- Acid phosphatase 20,21 munopathology of Toxocara canis and Ascaris suum infection Alkaline phosphatase 20,21 j8-Galactosidase 21 of the eye: the role of the eosinophil. Invest Ophthalmol Vis Sci a-Mannosidase 21 18:1172, 1979. Collagenase 24 6. Soulsby EJL, Stromberg BE, Donnelly JJ, and Rockey JH: In- Plasma membrane adenosine triphosphatase 25 traocular immunoglobulin E induced by intravitreal infection /3-Glycerophosphatase 22 with Ascaris and Toxocara spp. larvae. Ophthalmic Res 12:45, Lactate dehydrogenase 26 1980. Histaminase 27 7. John T, McKenzie D, Rockey JH, Donnelly JJ, Soulsby EJL, Phospholipase D 28 and Stromberg BE: In vitro interaction of eosinophils from As- Phospholipase B 29, 30 carid-infected eyes with larvae of Toxocara canis and Ascaris Plasminogen 30 suum. ARVO Abstracts. Invest Ophthalmol Vis Sci 20(Suppl):3, Eosinophil-derived neurotoxin 31 Neurotensin (NT) 32 1981. 8. Capron M, Bazin H, Joseph M, and Capron A: Evidence for IgE-dependent cytotoxicity by rat eosinophils. J Immunol 126:1764, 1981. usually do not contain a larva, while larva with limited 9. Butterworth AE, Remold HG, Houba V, David JR, Franks D, or no surrounding inflammatory reactions may be David PH, and Sturrock RF: Antibody-dependent eosinophil- 5l found nearby in the eye. mediated damage to Cr-labeled schistosomula of Schistosoma mansoni: mediation by IgG, and inhibition by antigen-antibody The anterior chamber exudates of guinea pig eyes, complexes. J Immunol 118:2230, 1977. aspirated 10 days after a primary intravitreal infection 10. Capron M, Rousseaux J, Mazinque C, Bazin H, and Capron with ascarid second-stage larvae, were composed almost A: Rat mast cell-eosinophil interaction in antibody-dependent entirely of eosinophils. In contrast, eosinophil prep- eosinophil cytotoxicity to Schistosoma mansoni schistosomula. arations from humans and experimental animals often J Immunol 121:2518, 1978. are impure (eg, contain neutrophils) even after en- 11. Anwar ARE, McKean JR, Smithers SR, and Kay AB: Human eosinophil- and neutrophil-mediated killing of schistosomula of richment. For example, eosinophils from rat peritoneal Schistosoma mansoni in vitro. I. Enhancement of complement- cavities contained only 47-60% eosinophils after en- dependent damage by mast cell-derived mediators and formyl 1042 richment by repeated plating in Petri dishes. The methionyl peptides. J Immunol 124:1122, 1980. ascarid-infected guinea pig eye, therefore, also offers 12. Kazura JW, Mahmoud AAF, Karb KS, and Warren KS: The a ready source of eosinophils for in vitro and in vivo lymphokine eosinophil stimulation promoter and human schis- studies of eosinophil functions in general, and the spe- tosomiasis mansoni. J Infect Dis 132:702, 1975. 13. Kazura JW, Blumer J, and Mahmoud AAF: Parasite-stimulated cific roles of eosinophils and isolated eosinophil con- production of H2O2 from human eosinophils and neutrophils. stituents in ocular and periocular immunopathology. Abstract. Clin Res 27:515A, 1979. The eosinophil probably plays a complex role in 14. Tauber AI, Goetzl EJ, and Babior BM: The production of su- acute and chronic ocular immunopathologic reac- peroxide (O~2) by human eosinophils. Abstract. Blood 48:968, 1 1976. tions. The present in vivo and in vitro models should 15. Capron M, Capron A, Dessaint J-P, Torpier G, Johansson SGO, prove useful in further separating and defining eosin- and Prin L: Fc receptors for IgE on human rat eosinophils. J ophil and other interacting ocular immunopathologic Immunol 126:2087, 1981. mechanisms that are of importance in ocular parasite 16. Butterworth AE, Coombs RRA, Gurner BW, and Wilson AB: infections and other granulomatous or nongranulo- Receptors for antibody-opsonic adherence on the eosinophils of matous inflammatory diseases of the eye. guinea pigs. Int Arch Allergy Appl Immunol 51:368, 1976. 17. Anwar ARE and Kay AB: Membrane receptors for IgG and Key words: Toxocara canis, Ascaris suum, eosinophil, an- complement (C4, C3b and C3d) on human eosinophils and tibody, culture, eosinophil granuloma neutrophils and their relation to eosinophilia. J Immunol 119:976, 1977. Acknowledgment 18. Lewis DM, Lewis JC, Loegering DA, and Gleich GJ: Localization of the guinea pig eosinophil major basic protein to the core of The authors wish to thank Mr. R. C. Patterson for expert the granule. J Cell Biol 77:702, 1978. technical assistance. 19. Olsson I, Venge P, Spitznagel JK, and Lehrer PI: Arginine-rich

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