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December 22, 2016
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June 28, 2011
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April 15, 1949
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Sanitized Copy Approved for Release 2011/06/28 :CIA-RDP80-00809A000600220201-4 CLASSIFI~'"''ION ~ CENTRAL INTELLIGENCE AGENCY~~ INFORMATION F~QM FOREIGN pOL;UR4ENT5 OR RADIO BROADCASTS COUNTRY ussR SUBJECT rlellosl ieeearah HOW PUBLISHED Periodical WHERE PUBLISHED Moeoov DATE PUBLISHED 1J4NCUAGE +~ REPOR DATE OF ENFORMAT14N 1947 DATE bIST. /~Apr 1949 SUPPLEMENT T4 REPORT N0. r. w: a, a aw ,uwwnw:..:..: m euww?lulow a n>t wnwulnow p nr eesntne.}w swY riwnw;9o aawuwrwowlaeo.nnoo Iw rwd Ngiww wr Y>. ' QrwwAwCnww Of-nn' row9 Iw ORMINTww.. Dogleg A a ga k E3 B TTol LI~C, No 4, 1946. , (877 Ber Abe T6~' 7`-- anelation ep6cifioal]y raquoated.) ANTIf~IPIC AND IAiMFTNOtFIsT]IC PROPERTIF3 OF NUCY.DDFROTETNS 0& DYSSl4TPi2TC BACTIItIA A. N. Belozerekip P. D. Cekker ~'hblee referred to herein are appealed) In their renearoh on the Dacteris ci ?ohe entero-typhoidal group up to now, *,he efforts of miorubiologiete and bioohemlete have been oentered ez- olueively on repeated saelyeee of bacterial antigens. AB for the nualeo- proieiae, whioh often oonetitute r.d r.~ish ae $0 perae~t of the dry weight of these baateria, they were igacred by the inveetigatere. Heeonroh en the eatigenio and immunogenio oharacterietioe of auoleo- protelne, hcwever, ie vital in the olnrification of the role of daoh oompene~t of th~.~ Daotarie7. oell in ima,~ro?cgy. Our laveetigatione were idltielly ooooarned with the nuoleoprotela of Flezner'e Bacillus {Shigelle paradyseateriae). The nuoleoprotein was obtained from the bacterial cell whioh waa pre- viouel~ tsented with triahloronaetlo aold to Seolate the whole antigen. The. nucleoprotein, obtaincfl by tY_:a method, proved to be e]Lghtl;? tonic is animals (a 3-milligram dose vas parfeotly tolerant for mioe). The antigenio propsrtiee of nuoleoprotein were olearly mnnifeated. Freolpitioe mad also agglntinlae of dyeent~rio bacteria of a low titer appeared in the earn of the launlzed animals. However, n detailed study of the preoipitative pro~pertiee of these sera produced very unempeuted re~ulta. These sera pro- dnoed positive preoipitation reaction when tested with nualeoproteiae and the while antigen. The titer of the preaipitiae in comparison to the whole antigen vaa often eigaifianntly ]figher than in the once of the nual~o- protei.n. Table 1 g*ves data '_t'_lcating the prop:.rtiee of the nucleoproteins that were obtained by various methodo. 1 - CLASSIFICATION ~!~~ Sanitized Copy Approved for Release 2011/06/28 :CIA-RDP80-00809A000600220201-4 Sanitized Copy Approved for Release 2011/06/28: CIA-RDP80-00809A000600220201-4 It aught bo propqundad that thle F::-:?.lpitation reaction occurs due to the `resenoe of similmr albuminous compua.nts Sn the nucleoprotein and in the ~ biuain content of the artlgend, flouo;?er,'thess t?et sera produced positive preglpitatioa reaction3 nct only with the antigens but also with pa~eeboharoee {haptea) from which the albumen had. bean removed. In addition, it vale possible to separate the antibody lntc antigens ami auol.eoproteins by the adabrptlon method. It to evident that these antieera contained two ono with respect to the antiigene, the other, to the albumen. the above Yaot merely proud that t~z. eu~~lboprotein preparations with whiah we worked also contalaed antigens. We nrdertook the tgN1K of ottaining a ~ur.i.eopxotein as free from the nntig?ne as possible. Taste were make with nucieoproteine that were obtained free a baoterial growth which vas washed three, ffve~. and eight times vtth triohloronoetio acid to ol2minate the antigens as much as possible. Ho!remery the results of this teat rer? identical to the previous tost as ~ ho seen from the data in Table 2. According to Table 2, the Aatieera, obtained from rabbits Smmanised with the euoleoproteins which were isolated after repeated washing with tri~? :aoidt, ooatainod preoipitlne with relation to the albumen, and to the rhale antt~en. Eere also the tSter of the ahtigena vas considerably On the strength of thane results, ve decided to fractionate the nucleo- protein since it vns quite evident that the nucleoprotein isolated direot]y from the baoterial growth by means of a weak alksline aulution was not an ioseparabls oomyonent. The fractionation vas performed by A. ft. Beloserekiy's method (1 ). As a result of this fraotioaetion we obtained a serles of produota oorrespouding to the ouolear and oy*oplaematic elements of the baoterial cell. according to their ohemlaal properties. Moat aetonlehiug foot 4as that this admiztars of baelo slemeate vas found in ell the frnotioaeted products sad it appeared especially clearly in all of than products which corresponded to the anolear elements o_? the cell, Iimunlestlon of rabbits with nnoleo- protsiw~ ?,.e,a with mxolooproteine containing nlbumea nod tbymonuoleio acid, prodaoed satisern the,t precipitated a given nucleoprotein in titi~li -~ of 1:8,000 and the antigens is titers of 1;32,000 - 1a128,00U. lYperiments made with nuoleopro(:eins isolated from strains of Flezner's. Iiaoillna sad from ~~her typ tpidal bacilli gave completely analogone results proving that a common oharaoterietio eziste in these phenomena. the foot that Doth the xapeat~dly washed aaoterial eubstnnan obtained from 8Glbrd m.larobss nod the complicated chenioal treatment of the nualea- protein in its fraotionntion process completed +ailod in ieolatin~ the nuoAsopratsln from the besib elements auggestrt that these Daeio elements enter into an 1udlssoluble argnnio union with the baoterial nucleoprotein. its turned to the investigation of the uuoleoproteia of B-form dysenteric bautsrin. It vea ee~abliehed that in the process of immunization, the anoleoproteln of the F-farm bacilli produced pxeoipitine only io titers of 1:4,000 - 1s8~00f1. The latter fully cenforme with our data on tae antigenic strnotare of H-foam bacilli. Oar enbssgnent work involved the in4eeti?stion of the immunogenic properties of the onoleoprottlin. The results proved that the basic antigen rsapoaelble for the imunization effect le the baoterial antigen. As for the anoleoprotsln, Sts ismaizntion effect i9 either negligible or oomplete~~ lacking. Only when Smmunlzatien ie made with large doses is the nuoleo- Sanitized Copy Approved for Release 2011/06/28: CIA-RDP80-00809A000600220201-4 Sanitized Copy Approved for Release 2011/06/28: CIA-RDP80-00809A000600220201-4 rotein a able of rodG?__n~a some im.^,ir_....atio triat tkiis ninor ~mmuniz '?lor_ e`feet 1~ dui ton effnrt. It is quite probable ? ~` the remnaxiia of tkie whole antigen. The great amount of ezperimertal material. which has been at our disposal 1n the course of several "ea'rs work has convinced ue that the 3-form nucleo- proteins contain a neg~.iglb',y small amount of the apeci4lc antigenic propsrty. This basic element ^iay ho elthsr s apeci.fie na'yasccharide ccnnected'vith the nucleoprotteir. molecule or a whole antigen. We attempt;,d:'o salts this problem by treating the nucleoprotein 0.1 iQ by 'coiling in acetic acid, to decoetpoae the whole antigsn 1f present. The study of the antigenic prap6.?tiea treat o3 by this method showed that the preparation lost its basic a"ement. The resulting antisera produced reactions only against nueiecproteins, and ^egative reactions against the whole antigen and nolyaaccharids. These testa prove that the basic element of nucleoprotein le moot likely the whole ~,nLlgen which, by the above-mentioned treatment, turned into hapten, tte pol?oeeaoxaride deprived of all antigenie properties. The assumption that the whale antigen is the basic element united with the ~rcleoprotein 1c~ strengthened by the data oP Boivin (3) who demonstrated that the i+hole antigen can be divided in its entirety poly after the ingoetion of tho baaterinl grovbh by proteolytic ferments. How mllpuld re treat the materiel ve have obtained through ezperimenteY Above a1L, we mast decisively disagree with the propoa pion that the aorre? spondiag antigenic prcpertriae of nnoleoproteine which we have studied are inflneaoed by their compounding with the whole antigen. Against such a ooa- ceptiou, there ezieta the fact that the whole antigsn may be isolated in its entirety frcm the nucleoprotein only by the action of proteolytic fer.neate which completely destroy the nucleoprotein molecule. On the other harxl. vhnt- ever the method used (fractionation, multiple washing with triohlo:0eoetio acid, etc.,) ae long ae as intact nucleoprotein molecule ezieta it will elvaye pro- dace reactions eimulatiog she presence of a whole antigen. Oar esperimental data vas ooafirmed by the vork of A. G. Kravchenko and A. I. Inrkla (4) 3rhlch has ,lust been puhI,iehed? 6n the Daeie of sl'_ *he ezperimentnl data we have at our disposal, we =ome to the oonoluaion that the main co:ponarte of the whole antigen localize themaelver on the ezterior of the cell sad can be easily extracted. Another ineiguifioant component of the whale antigen i~ organirally united with elements of the protoplasm of the bacterial cell aad,in particular, with the unclear elements. This component of the whole antigen, ~rganioel],y united with the bavt;erial nuoleopreteire, bee s antigenic eigni?ioance which has been pointed out earlier by Peshko~ and Belozerekly. The study of the chemical composition of the protoplasm of the R-form oY Flezner'o Aaaillue and the study of tho sttigenic an3 Smciunogenlc properties of the H-form nueleoprateia proved very important.- Ho qulntitative ~r qualitative change wsa noted Sn the composition of the basic substances of the protoplasm in the R-firm of the bacteria i^ comparison pith thy S-form. The sole change observed in the '?-form vas the complete ~tisappearanae cf the whole antigen: The transition In the R-form ie attended not only by i.ha loe~ of the superficial~r distributed whole antigen, but ales b~ the lose of that comFansnt ~:hic:t is cr~.nlcnl~y united with the nuclear elements of the bacterial yell. The cztremely closeralattnn between the yhole antigen and the nucleo- proteids and, is particular, bet-+een the vhcl~ antigen and the atomic nncleo- psoteids ie extremely a,-ident an3 has Brest bYOlog'_cal s*_gnificance. - 3 - ~FIt1AL Sanitized Copy Approved for Release 2011/06/28: CIA-RDP80-00809A000600220201-4 Sanitized Copy Approved for Release 2011/06/28 :CIA-RDP80-00809A000600220201-4 Rhi?, eaidenb2~y, is the sautes of the function of the eugerfioial~,y looallzed whole aatig~n. The tranait4on into the R-form ie acocapanied by the lose of this reproductive poker. BIBLIQC$;AYHY A. S. B?ioterek4y, Microbiology, 10, 185 (194'1) A. P. Sonni:l[oe`, Zh1~46I, P? ~ -2 { 1942 ) A. Bo4vin eE Z. Meerobeanu, 1tew. d'immnnologie, 1, 553 (1935) e-. a. $rnohenko and a, x. re,ririn, max, ao ~, (194P~ Sanitized Copy Approved for Release 2011/06/28 :CIA-RDP80-00809A000600220201-4 9erled cf e of ei[atigen i^t?eaatzat~on ftuoleoprat.en (etooo~~ng to At~ount of Average titer of ~'?eigitiae antigen Average regarding in,7eated titer o? (in milllgrame} ftuoleoproteia Whole aetiaen ffisloserr3ki~ (1}} 4 5 1:6400 1:8000 1:12,000 - 1:64,000 BHCleiB9 8$Ode 3 1.5 1:2400 1:8000 1:12,000 - 1:64,000 ftualeopzotei4 (aooor~iag to $acakrDV (2}} 4 2.5 1:6400 1:8000 1:16,000 ?'atla 2. Af6'E'If~d1C PRO~kt OF NTICLF~QPitOTE11PS OBTAIfF:ED by R,a"PEAT~D kTASiiIIiCB OF FIrB HACZLYIPS Titer produoing preoipitntion raaotiona For the whole antigen ~'or Lhe tauoleoprotein 1 1:32,000 1:4,000 i 1:64,000 1:8,000 1 1132,000 1:8,000 3 1:64,000 1:16,000 5 1:128,000 1:8,000 5 1:64,000 1:8,00^ 5 1:64,00 1:8,ooa 5 1:8,coo 1:2,00 5 1:64.ooe 1:4,000 8 1.7.2,000 1:1b,000 P 1:128,000 1:16,000 Sanitized Copy Approved for Release 2011/06/28: CIA-RDP80-00809A000600220201-4