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  • Bech Von Magnus 1959

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    Anca Elena Dulcinatu
    15 Ansichten11 Seiten
    Bech von magnus 1959

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    Bech von Magnus 1959

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    Bech von magnus 1959

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    © All Rights Reserved
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    15 Ansichten11 Seiten

    Bech Von Magnus 1959

    Hochgeladen von

    Anca Elena Dulcinatu
    Bech von magnus 1959

    Copyright:

    © All Rights Reserved
    Als PDF herunterladen oder online auf Scribd lesen
    Markieren Sie unangemessene Inhalte
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    [PROM STATENS SERUMINSTITUT, COPENHAGEN, DIRECTOR: J. ORSKOY, 3. STUDIES ON MEASLES VIRUS IN MONKEY KIDNEY ‘TISSUE CULTURES 1, Ivolalion of Virus from 5 Patients with Measles. by ‘Yioco Brcrt and Paar yor Maones Recsivet 209187 In 1994 Enders & Peebles (8) reported the successful isolation of virus ike agents from blood and throat washings of patients with measles. These agents multiplied in cultures of human and simian renal cells and produced characteristic eytopathie changes. ‘The mi ‘rient fluid from infected cultures was found to contain antigen which fixed complement in the presence of measles convalescent sera. In ad- dition such sera were shown to inbibit the peopagaton of the isolated agents in tissue cultures. These original observations by Enders & Peebles were later confirmed hy Cahen el al. (8). These investigators ‘isolated several virus strains and studied the intracellular localization of the virus in tissue cultures by means of the fluorescent antibody. technique. Recently Dekking & McCarthy (4) reported that they had been able to propagate measles virus in the KB strain of humen carcinoma cells AU about the same time Black, Reissig & Melnick (2) sueceoded in adapting measles virus to another human cancer cell steain, UEP 2 ‘The present paper reports the isolation of five strains of measies virus in cultures of trypsinized monkey kidney cells. The identification ‘ofthe strains by serological tests and by transmission of the disease 10 monkeys is also described, MATERIALS AxD METHODS Tiesne cultures: Calls of trgpsolznd Lines from shore monkers, csnomolgous ranks or habeons were wie rutnel. The technique of Younger (19) forthe ure ‘aration of thew tien eaitares wus Wied ith some moafeatons es previously [caruea (10): Dus the outgrowth of the calls the growth median eanolted of lnctalarain hjdeoyaate 08 per ceat (1) in ents 2 clots seeding with vires the medium was changed, Each tube received 1.4 mi of slther synthetissuelumn 198 (Ia) or bovine smaitic Hid @) containing peel ‘ed-as an Inilestor {Cina dilution 02 per cent). all medis contained penelia "and steeptompein (01 mua. fe (8) with 2 per eost cy 6 Collection of specimens: Ie and blood ion of views was atlemplet Grom throat washings se elther with 15 maf 2 ining I per emt Saco {iptose ‘OT mg‘per mt wee ded to the thu. From the throats of very Soune chilten apeehbeas were oh ine hy en each wer egg nr otf on a te {to fulds Jus dsc, In ll Intancestheapecmens vere immediately froneh In Gbasce a then stored tn an lcetriel ecptrese (60°C) Reve intaltion tet clare he fan matte wer thawed py 3° mal ‘hood in caler experiments heparin was added to the blood (2 ml of a9 048 ‘er cont solstion ek heparin per 18 ma of Sood) In ate experiments the lot ‘Sas allowed to coagulate Tle celle which had not become tached tothe Sat fod'n the sum, sid tie mitre was sed Inoculum tor sae od samples were sored att" C unl the tne of Inoculation. of trae caller: Tsu callarer were tartated wit 09 sl of al'aoont of fond fs {raon (1 ration pes mite) ‘Sobeutures from thers passage were carved out betien the Gh and the 16k ig on te Sth dar titer inoculation. "Fhe nutrient madiom wat 8 ule fin the courts af pateageSabaltres from the Tater pease were fen the Std the 1h day ical ln Tatas ata red fz stapenson of et inc by loosen icteThe amount Sooclated varied, but it wan gaually U2 a folume of fin the imcalatd tabes war about ? mi Sera ‘Complement ficaion tates Complement lation Tete were made scoring to a method af Fulton & Dumbell (3) as mold hy Seetinge, Enders & Holloway Gt ke Sntien the undilsted suspension of cells snd el debs nthe citer esi in eare asa wa ae eel ater arent te ‘shorter or longer poet of storage at 20°C Controls with antigen consisting fultare fields wih'cclls tron uafsoeulated tue ealtares were fneaded in each ‘Aperinent Ail antigens hod heen inactivated at 56°C: for hil am how "Phe antlerum consist of a pool of serum from about) measor patents Bed act weeks afer the development of the ash Serum hd boon inatiated x 56°C for bait se ose RESULTS Virus leolation In the spring of 1955 and of 1956 isolation experiments were carried out on 18 patients with elinical symptoms of meas'es (Table 1). From 9 patients material was collected during the first 24 hours after the onset of the rash. Virus was recovered from the throat of five of these patients, in 2 instances from cotton swabs (Nos. I and 11) and in two cases from throat washings (Nos. 4 and 10). From one patient (No, 5) virus was recovered both from cotton swabs and from throat washings. ‘The throat washings were kept frozen at —60° C for varying lengths of time before inoculation into tissue cultures. As ean be seen in Table 1, virus could be recovered even if 4 days had elapsed before inocula- tion took place (patient No. 5) ‘Virus was isolated from heparinized blood in osly one out of three TABLE 1 otation of Measles Virus in Monkey Kidney Tseae Caltuces. inte war Recovered from ® out of 13 Palen ° elt tlottt cote! biel ma 1. ‘Onterowth of ermal epithelia cele fn am uninewlatel monkey Kees smanifetations in money Wdney tgue eultaesfaoealated : sith vactolen. fig 3 hater stage showing ‘us, $ and 5. Uatnoenated monkey kidney ites cultures showing vaewlatl syn- ‘al siruetanes resembling those caused hy measles tus Fg. 4) and Of rome: ‘what diferent se and appearance (ig. 80 attempts (No. 4), and in no instance from serum (none out of 5 pa- tients bled wilhin 24 hours after onset of the rash). ‘From 4 patients virus isolation was attempted when more than 24 hhours had elapsed after the appearance of the exanthem. Virus was not recovered from any of these patients. Cytopathic changes: The cytopathic changes observed in the cultures on isolation of measles vicus were similar to those deseribed by Enders & Peebles (8): Atter period of one to several days syneytium-like areas develop in the tissue. Initially, these changes are most predo- nant at the margin of the tissue. The syneytia seem to arise from the melting together of cells whose boundaries gradually obliterate, With ‘continued incubation the syncytia increase in siza, and new arise in fther places, Very soon large numbers of small vnetoles appear ven trally in the affected areas giving rise to foamy- or lace-like formations (Big. 2) As the degeneration continues the vacuoles disappear, and the chang- cd areas now appear uniformly gritty and darker than the surrounding tells, The cell debris may loosen from the glass and float into the eul- ture medium leaving an area without tissue. Usually, however, the residue of the destroyed tissue is found to be arranged like dark bor ‘ers along the margin of the remaining tissue ‘At this time there appear in the cultures rather large granular for- tations, more of less cireular in outline and showing clther a quite smooth or a erenated margin (Fig. 3). These formations are probably condensation products of cell debris from the specifically degenerated areas. They are soon irrespective of whether the cultures have been Kept stationary or rotating. They may be fixed to the normal tissue. but frequently they Float in the eulture medium. ‘The destruction of the tissue continues us the individual syncytia {increase in size and new ones appear. In monkey kidney tissue the speed of destruction and the extent of cytopathle changes may vary considerably from one passage to another. Now and ten, larger or simaller areas of apparently normal tissue will remain even after 2} ‘reeks. Mostly, however, the remaining tissue shows a loosening of the Structure and granulation of the cells. This general oversall degenera- tion of the tissue may oceasionally be observed already after a few ays? incubation. ‘As described by Enders & Peebles (@), and later by Rustigian et al (18) and by Cohen et al. (3) eytopathie changes sinilar to those caused bby measles viras may be observed also in uninoculated cultures of monkey kidney tissic (Figs. 4-5). Theso changes are probably caused by virus-like agents, so called “foamy agents”, which seem to be fre- quently preseot in kidney cells from apparently healthy monkeys. Specific measles antigen is, however, produced only in cultures infected ‘with measles virus, In the present study the ability of tissue culture Tmaterial to fix complement im the preseces of eonvalescent- oo phase measles serum was therefore used as & eriterion for the presence fof measles virus. Serological Studies. Complement fixation teste: Table 2 shows the results of complement tests on sera from four patients with measles. Blood was taken le after the onset of the rash, and again from 9 to 37 days Inter during convalescence. TABLE 2 Complement Feation Titers of Sera from Cases of Meaates. 1 nine | SEARS ‘

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