84
ARTIFICIAL ANTIGENS WITH AGAR, GUM ACACIA AND
CHERRY GUM SPECIFICITY.
S. M. PARTRIDGE* AND W. T. J. MORGAN.
From the Biochemical Department of the Lister Institute, London.
Received for publication March 3, 1942.
THE conversion of a bacterial polysaccharide into an antigenic complex that
will induce the formation of immune-body complex specific for the polysaccharide component of the complex has been reported in earlier papers (Morgan
and Partridge, 1940a, -1941a, b; Partridge and Morgan, 1940). The method of
combining bacterial polysaccharide and protein components in formamide
solution has now been extended to include the formation of antigenic complexes that contain as the serological specific haptens polysaccharides- of
vegetable origin, and in the present paper the conversion of agar-agar, gum
acacia and cherry-gum into antigenic complexes is described. A preliminary
account of the work with the artificial agar antigen has already appeared
(Morgan and Partridge, 1940b).
EXPERIMENTAL.
The experiments were made with the polysaccharides, agar-agar, kanten,
gum acacia and cherry gum. In each instance the commercial material was
subjected to a simple purification procedure that removed the gross impurities
present and reduced considerably the nitrogen content of the polysaccharide
preparations.
The agar antigenic complex.
A commercial sample of agar was freed from gross impurities and inorganic
matter by solution in anhydrous formamide. The clear viscous solution was
fractionated by precipitation with alcohol, and the main fraction that separated
between the alcohol levels 50 per cent. and 60 per cent. was again dissolved
in formamide. The fraction was precipitated from solution by 60 per cent.
alcohol, and thoroughly washed with 60 per cent. alcoholic formamide and finally
with absolute alcohol. The agar was then dissolved in boiling water to make a
0-1 per cent. solution and after cooling, dialysed for several days against distilled water at 00. The purified agar was recovered by the addition of alcohol
to the dialysed solution, the material being subsequently washed with alcohol
and dried in vacuo. The substance contained 0-8 per cent. N and was tested
* Beit Memorial Research Fellow.
ARTIFICIAL ANTIGENS.
85
for antigenicity in rabbits (Table I, animals 140, 141, 170 and 171). Two
animals received eight doses of 0 05 mg. and two others ten doses of 02 mg.,
but in no instance were agar precipitins subsequently detected in the sera of
these animals. For convenience, kanten, a breakdown product of agar, was
used as hapten in the precipitation tests.
A portion of the agar (60 mg.) was dissolved in 3 ml. of anhydrous formamide (m.p. + 20) and a sample of conjugated protein (40 mg.) prepared from
the specific somatic antigen of Bact. shigae was added as a dry powder. The
conjugated protein was prepared according to the method described by Morgan
and Partridge (1940a, 1941a, b) and contained 11-4 per cent. N. The protein
slowly passed into solution. The mixture was then allowed to stand at 370
for 2 hours, and, after increasing the volume by the addition of 10 ml. of
formamide, was kept at 0-2° for 48 hours. The agar-protein complex was
recovered by the addition of alcohol to the formamide solution, the precipitate
was thoroughly washed with alcohol, was taken up in 10 ml. of boiling water
and, after cooling, was evaporated to dryness from the frozen state. Samples
of the material (10 mg.) were prepared for animal inoculation by suspension
in 10 ml. of sterile saline and boiling for a few secorfds to ensure solution or
fine dispersion. The opalescent, colloidal solution that was formed, however,
was unstable on cooling, and a sediment of fine particles appeared on standing
at room-temperature for several hours.
The whole suspension was given to rabbits Nos. 142 and 143 (Table I).
Three doses each of 0 05 mg. failed to induce the formation of specific precipitins,
but after a rest period of one week and a further course of five 0-2 mg. doses,
the serum of both animals gave precipitation when mixed with an equal volume
of 1 in 104 or 105 dilution of kanten. Definite, though slight, precipitation
was also given by the kanten preparation at a dilution of 1 in 106.
The gelatinous character of a solution of the antigenic complex suggested
that the preparation contained an excess of uncombined agar. The material
was therefore dissolved in boiling water, allowed to cool slowly and to stand
for some hours until the formation of the sparingly soluble floccules was complete. The suspension was then centrifuged and the deposit washed with
small quantities of boiling water. The residue was dehydrated with alcohol
and dried in vacuo. The material contained 8-9 per cent. N and 22 per cent.
agar, estimated according to the method of S0rensen and Haugaard (1933).
A specimen of the agar purified by formamide treatment was used for colorimetric comparison. The protein preparation gave only a trace of colour with
orcinol and sulphuric acid, under the conditions of estimation employed. If
one may judge from the limited number of animals used, the removal of the
excess agar enhanced the antigenicity of the agar protein complex. Three
intravenous doses of 0 05 mg. now gave rise to powerful kanten precipitins
in each of the 3 rabbits inoculated (Table I, animals 182, 183 and 184).
An antigenic complex was also prepared from agar and the conjugated
protein obtained from the " 0 " antigen of Bact. typhosum. The same method
for the formation of the complex was employed, and the resulting agar-protein
complex was very similar in general appearance to that derived from agar
and the " Shiga " protein component. The material again showed itself to
be antigenic, and although three doses of 0 05 mg. proved insufficient to elicit
7
86
S. M. PARTRIDGE AND W. T. J. MORGAN.
demonstrable kanten or agar precipitins, a further four similar doses gave rise
to strongly precipitating sera (Table I, rabbits 185 and 186).
In view of the claim made by Zozaya (1932) that agar could be rendered
antigenic by adsorption on collodion, the agar preparation employed in the
experiments described above was treated with a preparation of collodion
(Schering-Kahlbaum) particles prepared according to Zozaya's technique.
The particles were readily agglutinated with anti-agar immune serum, thus
indicating that an agar-collodion adsorption complex had been formed. The
agar-coated collodion particles were given intravenously to two rabbits (Table
I, animals 176 and 177). Four doses each of 0*2 mg. gave no demonstrable
anti-agar immune-body. After a week's rest a further course of four doses
of 0 5 mg. was given, but there was no evidence that even weak agar precipitins
had been formed.
Preparation of kanten for use as test hapten.
The anti-agar sera from rabbits immunized with the agar-protein complex
were found to give heavy precipitation with a preparation of partially degraded
agar called kanten. The preparation and properties of the substance have been
described in detail by Takahashi and Shirahama (1934) and by Pirie (1936).
Kanten lends itself more readily to purification than agar since it is readily
soluble in cold water or saline yielding clear mobile solutions, and for this
reason it has been found very convenient to use kanten in place of agar as
test hapten. A 1 per cent. solution of agar was heated at 1000 for 2 hours, set
aside to cool slowly and left overnight at 00. The liquid which separated from
the agar-gel was collected and after concentration was evaporated to dryness.
The crude kanten obtained in this manner contained 1-7 per cent. N, showed a
negative rotation, [o]M5461-747, and dissolved readily in cold water to yield a
clear solution. The material could be recovered from aqueous solution by
precipitation with acetone, but precipitation was not complete even at 80 per
cent. acetone level. A convenient method for the removal of nitrogenous
impurities, however, was found and was based on the insolubility of kanten
in 90 per cent. phenol solution. After three extractions with 90 per cent. phenol
the nitrogen remaining in the kanten fell to 0*3-0 4 per cent. This material
was used as test substance in place of agar in all experiments recorded in
Table I.
The gum acacia antigen.
A method of purification of the commercial gum was chosen which was
designed to avoid the use of strongly acid or alkaline reagents or heat treatment in order to reduce the risk of any degradation of the polysaccharide
taking place. A sample of commercial gum B.P. (1 gm.) was dissolved in
formamide (50 ml.), the small residue which remained undissolved was
removed and the clear solution was treated with alcohol. The bulk of the
material precipitated between the alcohol levels 40-50 per cent. Only a small
quantity of substance was thrown out of solution above 50 per cent. alcohol
and this was rejected. The main fraction was washed thoroughly with alcohol,
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S. M. PARTRIDGE AND W. T. J. MORGAN.
dissolved in water and dialysed against repeated changes of distilled water.
The acid polysaccharide was recovered by addition of an equal volume of
ethyl alcohol after acidification with 1 ml. glacial acetic acid. The substance
was redissolved in distilled water and again precipitated from solution by
addition of an equal volume of alcohol. The dried preparation contained
0-23 per cent. N and 0 4 per cent. ash.
A group of 6 rabbits was inoculated with varying doses of the purified gum
acacia as set out in Table II (animals 178, 179, 199, 200, 205 and 206), but no
precipitins specific for gum acacia were detected in any, of the sera examined.
Uhlenhuth and Remy (1933, 1934) showed that certain commercial preparations of gum arabic are antigenic, but that after purification to remove a
nitrogenous contaminant, the resulting gum no longer gave rise to immunebody. Similarly, Seideman (1940) showed that the serum of animals immunized with gum arabic preparations that gave positive protein reactions
responded with the formation of immune-bodies reactive against a purified
non-antigenic gum arabic.
The sample of " Shiga " conjugated protein used to prepare the gum acacia
antigen was obtained from the bacterial antigen isolated by the phenol method
(Morgan and Partridge, 1941b). The purification of the conjugated protein consisted of a single precipitation with acetic acid from solution in cold, dilute
NaOH solution. The sample contained 112 per cent. N, 1-07 per cent. P, and
showed [cx]546l-48 ± 3°. The finely powdered protein (20 mg.) was mixed with
the purified gum acacia (30 mg.) and formamide, m.p. + 20, (1 ml.) was added.
The mixture was thoroughly stirred and allowed to stand at 0-20 overnight.
Solution was not quite complet9; the small residue was rejected, however, and
the clear supernatant was transferred to another centrifuge tube and 4 ml.
alcohol added. The precipitate of protein-gum acacia complex was washed
three times with alcohol to effect the complete removal of formamide, and the
substance was suspended in 10 ml. of 0 9 per cent. NaCl solution. An opalescent suspension of the complex was obtained which slowly settled on standing.
The suspension was centrifuged and the clear supematant discarded. The
insoluble material contained 8-9 per cent. polysaccharide. It was again
taken up in 10 ml. of 0-9 per cent. NaCl solution and finely dispersed by means
of a glass tissue grinder. The protein content of the suspension was estimated
and the volume adjusted so that the suspension contained 1 mg. of protein
per ml. The dose given to the animals (Table II, animals 217, 221 and 227)
was O 5 ml.
A sample of the conjugated protein component of the " 0 " antigen of
Bact. typhoaum was also used in place of the " Shiga " conjugated protein as
the second component in the gum acacia antigenic complex. The protein
was prepared from a diethyleneglycol primary extraction product of a culture
of the " 0 " strain " 901 " and contained 11.5 per cent. N. Details of its
preparation and properties will be given in a later paper. The technique used
to accomplish the combination was the same as that already described. The
complex proved to be a powerful antigen, and three intravenous doses of
0 05 mg. (calculated as polysaccharide) only were sufficient to induce the
formation of strong gum acacia precipitins in the two animals immunized
(Table II, animals 197 and 198).
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Cherry-gum antigen.
A specimen of cherry-gum (0-75 mg.) kindly supplied by Prof. J. R. Marrack
was dissolved in water (25 ml.). A small insoluble residue was removed, and
the gum was precipitated from solution by the addition of 50 ml. alcohol containing a few drops of glacial acetic acid. The material was thoroughly
washed with absolute alcohol and dried in vaCuo. The gum contained 0-52
per cent. N and was further purified by extraction with 90 per cent. phenol
solution. The powder swelled to a clear gelatinous mass but did not dissolve.
After three extractions the phenol was removed from the gum by trituration
with alcohol. The material after drying in vacuo weighed 0-46 g. and was
free from N. The antigenic complex was prepared exactly as described above
for the gum acacia antigen. The complex after precipitation from formamide
with alcohol was taken up in saline, vigorously shaken and- centrifuged. The
deposit was again shaken with saline and finely dispersed with the aid of a
glass tissue grinder. The resulting suspension was not stable, but slowly
settled out on standing overnight in the cold room. Four rabbits were given
six intravenous doses of the purified cherry-gum (Table III, animals 235, 243,
257 and 272), and three animals (Nos. 232, 262 and 263) were given the equivalent amount of gum in the form of cherry-gum protein complex. It will
be seen from the results given in Table II1 that the hapten alone failed to
induce the formation of specific cherry-gum precipitins, whereas the complex
caused the production of specific immune-body after three doses only.
Cross Precipitation Tests.
In view of the overlapping specificity shown by the partial hydrolysis
products of certain vegetable gums with anti-pneumococcal serum (Type II)
(Marrack and Carpenter, 1938), it was considered of interest to investigate
the reactions of the anti-sera described above with the pneumococcal specific
polysaccharides (Types I and II) and with the heterologous gums. The
undiluted immune serum was mixed with an equal quantity of a dilution of
the gum or polysaccharide preparation, incubated for 2 hours at 370 and
examined after standing at 0° for 18 hours. The results of these tests are
given in Table IV. Gum acacia failed to give any precipitate when mixed
TABLE IV.-Showing the Specificity of the Anti-polysaccharide Immune Sera.
.
Kanten
Gum aoaoia.
Precipitation test withAnti-gum acacia serum. Anti-cherry gum serum.
Dilution of test hapten. Dilution of test hapten.
Anti-agar serum.
Dilution of test hapten.
Test hapten.
.
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Gum tragacanth.
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ARTIFICIAL ANTIGENS.
91
with an anti-cherry-gum serum, whereas cherry-gum gave distinct, although
weak, precipitation with anti-gum acacia serum. Preparations of kanten,
gum tragacanth, hyaluronic acid, the specific polysaccharides of pneumococcus
(Types I and II) and the blood group " A " polysaccharide isolated from pig
gastric mucin failed to give any precipitation. The slight precipitation given
by the Type I pneumococcus hapten with the anti-agar serum was probably
due to agar substance in the preparation of the specific substance.
Complement-Fixation Test8.
Complement-fixation tests were carried out with anti-sera prepared against
agar, gum acacia and cherry gum and the corresponding polysaccharide
haptens. The soluble degradation product of agar, kanten, was employed
as hapten in the tests with anti-agar serum, but owing to its strong anti-complementary action up to a dilution of 1 : 100,000 the tests have been considered
untrustworthy. Similarly, with the cherry-gum-anti-cherry gum system the
tests were rendered unsatisfactory owing to the anticomplementary action, up
to a dilution of 1: 200, shown by anti-cherry gum serum. On the other hand
neither the gum acacia at 1: 1000 dilution nor its homologous serum at 1: 20
dilution showed any anticomplementary action, and the complement-fixation
tests showed that 1 M.H.D. of complement (0*2 ml.) was fixed by as little as
1 part of gum in 1: 10,000,000 parts of saline (0x2 ml.) in the presence of a
1: 20 dilution of the immune-body (0 2 ml.). Anti-sheep cell serum, 3-5
M.H.D., was used as amboceptor throughout the tests. Owing to the unsatisfactory nature of the agar and cherry gum complement-fixation tests no crossreactions within the three systems were investigated.
Anaphylaxis Experiments.
Groups of guinea-pigs (300-450 g.) were passively immunized with antiagar, anti-gum acacia and anti-cherry gum rabbit serum by receiving an intraperitoneal injection of 2-3 ml. of the immune serum. After 24-36 hours each
animal was given an intravenous dose of the homologous hapten and observed
for symptoms of anaphylaxis. Three animals immunized with anti-agar
serum showed severe anaphylaxis after a dose of 1 mg. kanten; one animal
died after 2 minutes. Two other animals which received 0.1 mg. of kanten
showed definite symptoms of anaphylaxis, but subsequently recovered.
Five guinea-pigs passively sensitized with 2 ml. of anti-gum acacia serum and
shocked with 1 mg. of the homologous gum showed severe anaphylaxis. Three
-of the animals died in 2-3 minutes after receiving the injection; the remaining
two animals slowly recovered from shock during the following 2-3 hours.
Similarly, a group of guinea-pigs, each of which had received an intraperitoneal injection of 3 ml. of anti-cherry gum rabbit serum likewise showed all
the usual symptoms of shock when given an intravenous dose of the homologous gum 24 hours after the serum. Owing to the limited amounts of
immune sera that were available no anaphylaxis tests that involved the use
of heterologous haptens to induce shock were made.
92
S. M. PARTRIDGE AND W. T. J. MORGAN.
DISCUSSION.
The method of combining bacterial polysaccharide and protein components
in formamide solution to form antigenic complexes has now been extended to
include the formation of antigens in which polysaccharides of vegetable origin
function as haptens. In this manner the polysaccharides agar-agar, gum
acacia and cherry gum have been combined with the conjugated protein components of the " 0 " somatic antigens of Bact. shigae and Bact. typhosum and
powerful antigenic complexes have been formed.
Agar, gum acacia and cherry gum do not possess the property of forming
soluble complexes with the conjugated protein in aqueous solution in the same
way as certain other bacterial and tissue haptens (Morgan and Partridge,
1941b; Morgan, 1941). It is now shown that when these gums are mixed
in formamide solution with a sample of the bacterial protein, the protein
firmly retains part of the polysaccharide, yielding a complex that cannot be
altered greatly in composition by repeated washing with water or dilute salt
solution. Although the polysaccharide content of the association complex
is in each instance low compared with that of the natural bacterial antigens,
the complexes are nevertheless found to induce the formation of powerful
anti-gum immune sera on injection into rabbits. It is now well known that
proteins when combined with mono- and di-saccharides, polysaccharides and
many simple organic compounds (see Landsteiner, 1936; Marrack, 1938)
acquire a new immunological specificity, and when the complexes are subsequently injected into animals they induce the formation of immune body
that is largely specific for the attached group. It is, however, frequently
difficult to secure a chemical linking of protein with the more complex and
labile polysaccharides such as the specific acetyl polysaccharide of the pneumococcus (Type I) or the " A " specific blood group polysaccharide of gastric
mucin or saliva, without damaging the polysaccharide molecule and thus
modifying its immunological specificity. For this reason a method of forming
an antigenic polysaccharide-protein complex that involves a procedure no
more violent than solution of the components in anhydrous formamide at
room temperature is likely to prove of considerable value in future investigations.
Some years ago Sordelli and'Mayer (1931) found that the immune-sera of
horses that had been immunized with bacteria grown on agar medium developed
agar precipitins. Utilizing this observation, Morgan (1936) showed that it
was possible to detect, by means of anti-agar horse serum, agar and agar
breakdown products in a. number of specific polysaccharide preparations..
The potency of these anti-agar horse sera, however, is not high even after the
horses have received a prolonged course of immunization lasting over a period
of several months. Furthermore, such sera arise only occasionally, and must
be considered as accidental by-products in the manufacture of therapeutic
sera for clinical use. The potent anti-agar rabbit serum, on the other hand,
produced by means of the artificial agar-protein complex has proved of considerable value in following the removal of agar and kanten from antigenic
material isolated from agar grown bacteria. Indeed, the precipitation reaction
ARTIFICIAL ANTIGENS.
93
of agar or kanten with its homologous immune-body is so sensitive that material
which fails to yield any of the colour reactions in the tests elaborated by Pirie
(1936) for the detection of agar will nevertheless give definite precipitation
when tested with anti-agar serum. The strong anti-complementary properties of kanten suggest that the anti-complementary nature of certain
bacteria grown on an agar medium may, in part, be due to a surface coating
of kanten. Since the original observation of Sordelli and Mayer no explanation for the conversion of agar into an antigenic complex has been forthcoming,
but in view of our observations it seems likely that agar or its breakdown
products combines with certain protein compon'ents of the bacterial cell and
in this manner is transformed into a full antigen.
The anti-gum acacia and anti-cherry gum immune sera were considered
suitable for studying the serological relationships that might exist between
these sera and other heterologous haptens, such as the specific polysaccharides
of pneumococcus (Types II and III), thus contributing to the valuable observations of Marrack (1937) and of Marrack and Carpenter (1938) on the cross
reactions shown by the acid hydrolysis products of vegetable gums and pneumococcus (Types II and III) immune-body (see also Morgan, 1937). Heidelberger,
Avery and Goebel (1929) showed that the native gum acacia reacted only
occasionally, and then very weakly, with Type II and III anti-pneumococcus
serum, and that it was necessary to partially hydrolyse the gum before it
showed any pronounced cross-reactions with the pneumococcus Types II and
III immune-body. These workers suggested that even the slight activity of
the original gum might be satisfactorily accounted for on the assumption that
traces of the specifically reacting material were formed during a period of
exposure in the process of refining the commercial product or by enzyme
action. Indeed, it is clear from our experiments that there is no cross precipitation reaction between the specific polysaccharide hapten of the pneumococcus (Type II) and gum acacia and gum cherry immune-body. It seems
improbable therefore that the aldobionic acid structure, which plays such an
important part in the cross reactions shown by the partially hydrolysed gums
and the pneumococcus (Type II) immune-body, exists as a serologically
important configuration in the native gum-acacia and cherry-gum. This
conclusion is supported by the observation that glucuronic acid fails to inhibit
either of the homologous gum-anti-gum precipitation reactions. Galacturonic
acid is also inactive in similar inhibition tests. Furthermore, virulent cultures
of pneumococcus Types II and III are not agglutinated by the gum acacia
and cherry gum immune-body.
Cretcher and Butler (1928) and Butler and Cretcher (1929) showed that
by mild acid hydrolysis gum acacia loses, its arabinose and rhamnose components very readily. Under these conditions of hydrolysis it is known from
the work of Heidelberger, Avery and Goebel (1929) that a new serological
specificity develops and that the resulting partially degraded gum now reacts
with pneumococcus (Types II and III) antibody. These observations suggest
that the serological specificity of the native gum acacia might be due in part
to the specific configuration of the arabinose and rhamnose molecules, especially if the ring structure of the complex acidic nucleus brought forward by
Norman (1937) is accepted. In such a structure for the native gum the
94
S. M. PARTRIDGE AND W. T. J. MORGAN.
arabinose and rhamnose units are joined to the nucleus by glucosidic linkage,
and thus, by masking the reactive surface of the aldobionic acid molecule they
might be expected to play a dominant role in determining the serological specificity of the native gum. It has been found, however, that d-arabinose,
I-rhamnose and d-fucose fail to inhibit the acacia-anti-acacia or the cherrygum-anti-cherry gum precipitation reactions. There is, therefore, no evidence
based on specific inhibition that the pentose and methyl pentose components
of the natural gums play any dominant part in determining their immunological specificity. We have had no opportunity of investigating further many
points of considerable interest arising out of this work.
SUMMARY.
1. The formation of antigenic complexes from the non-antigenic polysaccharides, agar, gum acacia and cherry gum and the conjugated protein
component of the specific somatic antigens of Bact. sIhgae and Bact. typlhosum
are described.
2. The artificial antigens induce the formation of immune-body specific for
the polysaccharide components of the artificial antigenic complexes.
3. The homologous gum-anti-gum precipitation reactions are not inhibited
by glucuronic or galacturonic acid and the anti-gum sera fall to agglutinate
virulent pneumococci (Types II or III).
REFERENCES.
AND
C.
BUTLER, L.,
CRETCHER, L. H.-(1929) J. Amer. chem. Soc., 51, 1519.
CRETCHER, L. H., AND BuTLER, C. L.-(1928) Science, 68, 116.
HEIDELBERGER, M., AVERY, 0. T., AND GOEBEL, W. F.-(1929) J. exp. Med., 49, 847.
LANDSTEINER, K.-(1936) 'The Specificity of Serological Reactions.' Baltimore.
MARRACK, J. R.-(1937) ' Proc. 2nd Intemat. Congr. for Microbiol.' London.-(1938)
'The Chemistry of Antigens and Antibodies.' London.
Idem AND CARPENTER.-(1938) Brit. J. exp. Path., 19, 53.
MORGAN, W. T. J.-(1936) Biochem. J., 30, 909.-(1937) J. Hyg., 37, 372.-(1941)
Chem. Ind., 60, 722.
Idem AND PARTRIDGE, S. M.-(1940a) Biochem. J., 34, 169.-(1940b) Chem. Ind.,
59, 849.-(1941a) Ibid., 60, 722.-(1941b) Biochem. J., 35, 1140.
NORMAN, A. G.-(1937) 'The Biochemistry of Cellulose, the Polyuronides, etc.'
London.
PARTRIDGE, S. M., AND MORGAN, W. T. J.-(1940) Brit. J. exp. Path., 21, 180.
P1E, N. W.-(1936) Ibid., 17, 269.
SEIDEMAN, R. M.-(1940) J. Immunol., 38, 237.
SORDELLi, A., AND MAYER, E.-(1931) C. R. Soc. Biol., Pari8, 107, 736.
TAKAILSHI, E., AND SHIRAHAMA, K.-(1934) J. Fac. Agric. Hokkaido Univ., 35, 101.
UHLEKNHUTH, P., AND REMY, E.-(1933) Z. ImmunForsch., 79, 318.-(1934) Ibid.,
82, 229.
ZOZAYA, J.-(1932) J. exp. Med., 55, 325.
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