MEDICAL RESEARCH

• Document Type: CREST
• Collection: General CIA Records
• Document Number (FOIA) /ESDN (CREST): CIA-RDP80-00809A000600220201-4
• Release Decision: RIPPUB
• Original Classification: C
• Document Page Count: 5
• Document Creation Date: December 22, 2016
• Document Release Date: June 28, 2011
• Sequence Number: 201
• Publication Date: April 15, 1949
• Content Type: REPORT

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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.
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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 ~!~~
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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 antibedY.es: 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-
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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 poaiti.ve
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
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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. $rnohenko and a, x. re,ririn, max, ao ~, (194P~
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 9erled cf
e of ei[atigen i^t?eaatzat~on
ftuoleoprat.en
(etooo~~ng to
At~ount of Average titer of ~'?eigitiae
antigen Average regarding
in,7eated titer o?
(in milllgrame} agglntiai.ne ftuoleoproteia Whole aetiaen
ffisloserr3ki~ (1}}
4
5
1:6400
1:8000
1:12,000
- 1:64,000
BHCleiB9 8$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
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