ON THE SEROLOGICAL CONSTITUTION OF FIN WHALE
III.

HUMAN B BLOOD GROUP SUBSTANCES INERYTHROCYTES AND SOME NOTES ON
ANTI-FIN Ju SPECIFIC ANTIBODIESKAZUO FUJINO

Many comparative studies have been undertaken on cell-antigens common
to human and various kinds of animals. Especially, reports on distribution of the human A, B, 0 blood group substances and Forssman's
antigen in animal kingdom, seem to be important and interesting fromthe standpoint of systematic serology. Terashima (1942) reports thathuman A blood group substance can be analyzed immunochemically into
four partial antigens, i. e., AI, An, Am and AIV, and shows that A typehuman cells possess all of these, A type dog cells An Am AIV, A type
pig cells Am Aiv and sheep cells Aiv respectively. Friedenreich andWith (1933) states that human B blood group substance may be divided
serologically into three components, i. e., B1i B2 and Ba and then theerythrocytes of B type human, rabbit and guinea pig contain B1B2B3 ,B,B 3 and Ba respectively. As regards 0 antigen system Inoue (1943)
describes that the erythrocytes of 0 type human, rat and rabbit possess
OIOnOm, 0110111 and Orn of the components of 0 blood group substances
respectively. Basing upon these results many analysis have been workedout on partial antigens being contained in erythrocytes, saliva and othersecretions or organ tissues of animals.
Symbols of Friedenreich and With have been used by some workers
ever since. But in the present paper, to avoiding the confusion withthose of B1 and B 2 of human blood B subgroups, abovestated three groups
are replaced with the symbols of BI, Bn and Bm respectively in accordance with the proposal of Furuhata (1957). Summarizing the reports
on B partial antigens in animal erythrocytes it is known that BIBnBm
exist in the cells of human (Friedenreich and With, 1933), orang-utan(Dahr, 1937), monkey and ape (Noda, 1949) (Ishii et al, 1954), tortoise
and frog (Sakuma, 1942), BnBm in the cells of rabbit (Friedenreich andWith, 1933), Kangaroo, tapir, weasel, cat (Dahr, 1937) (Ogura, 1953a) andgiraffe (Nakano, 1949) (Furuhata, Mori and Ro, 1949), Bm in the cells ofguinea pig (Friedenreich and With, 1933), capuchin monkey, elephant, dog,
badger, bison (Dahr, 1937), Japanese racoon-dog (Furuhata and Ro, 1948),
camel and whales (AZAMI, 1949). Furthermore, Ogura (1953b) reports thatthe B substance in the horse and sheep erythrocytes has a simpler structure than Bm of guinea pig cells. Accordingly to the classification of
171

172

K. FUJINO

Friendenreich and With (1933) the author detects the B substances inthe erythrocytes of whales, and confirmed the existence of some components of B antigen by adsorbing and immunizing experiments. The
author reports here on a new B partial antigen in fin whale cells which
has simpler structure than the already-known Bm in guinea pig cells,
and simultaneously notes some observations on the anti-fin Ju normal
antibodies in the sera of immune animals and the appearance of immune
antibody into egg-white of a fowl.
HUMAN B BLOOD GROUP SUBSTANCES IN FIN WHALEERYTHROCYTESMATERIALS AND METHODS

Anti-BI, anti-En and anti-Em sera were prepared by the method of
Yamaguchi (1943). Details of those and other items on materials andmethods are as follows.
Anti-B1 agglutinin. When a rabbit is immunized with human B(BIBnBm) blood group erythrocytes, anti-BI antibody is produced alone
in relation to B antigen, because the cells of rabbit possess Bn and Bm
components. This phenomenon is termed as the intravital filtration(Furuhata, 1957, p. 156). Then species-specific and anti-C agglutinins
which are produced simultaneously should be adsorbed away from thisserum by the A type human erythrocytes. These A type cells are takenfrom five or more individuals and are mixed together for use of adsorption. After these procedures may be obtained the anti-BI agglutinin
available for the experiments.
Anti-BII agglutinin. Being adsorbed the anti-C and the anti-0 (Eisler'sor Kagaya's anti-human cell heterogeneous antibody, (Eisler, 1930; Kagaya,
1940) agglutinins with A and 0 types human cells and anti-Em agglutininwith guinea pig cells from the serum of fowl immunized with rabbit
erythrocytes (BnBm), available anti-En agglutinin can be obtained alone.
A type cells which are used for adsorption are taken from several individuals similary to the previous case.
Anti-Bm agglutinin. If the anti-C and the anti-0 agglutinins areadsorbed away from serum of a fowl immunized with the erythrocytes ofguinea pig (Bm), the anti-Em agglutinin will be taken out alone.
Erythrocytes of human and fin whales. Collecting, preserving andother treatments of these erythrocytes are just the same as those inprevious report (Fujino, 1953).
Erythrocytes of rabbits and guinea pigs. Bloods were taken from ear
vein in rabbit and from heart with syringe in guinea pig. After separation
and washing with salt solution, these erythrocytes were used to various

SEROLOGICAL CONSTITUTION OF FIN WHALE

173

experiments.
Immune animal. Mature rabbits and fowls, weighing more than 2.0 kg
in the former and 1.5 kg in the latter, were used as immune animal. Prior to
immunization, anti-human A and anti-human B normal antibodies, thatis the '' Serum type '', were detected on the sera of these animals.
Immune sera. Each immune animal was given intravenously on alternatedays a series of (seven times') injections of 10% erythrocytes suspens10ns. A dose of injection of these immune antigens is 5 ml. in a rabbit and 3 ml. in a fowl. Seven days after last injection, animals weresacrificed for bleeding by puncture of cervical artery. After separation andinactivation these immune sera (1) were added with 1/10 volume of 5%
carbolic acid and preserved in refrigerator at the temperature of 0° to
5°C, or (2) were kept in freezer at the temperature of - 5° to -10°C
after mixing with 1/10 volume of lo/o sodium azide (NaN 3).
Adsorption tests and immunizing experiments. For the purpose of
detection of B antigen in the fin whale erythrocytes, adsorption tests
and immunizing experiments must be made on their cells. Firstly thestated three partial antibodies were used in such adsorption tests. After
thorough adsorption with fin whale cells, descendings of their titers
against human B cells were examined. If the descent occurs, it is assumed that the fin whale cells used for adsorption possess the corresponding or simpler B antigen. When no descent occurs, existence ofcorresponding B antigen will be denied. Secondarily the existence of B
antigen should be confirmed with immunizing experiments. Prior to immunization normal anti-human B antibody of immune animal (fowl) was
examined. If acendings of titers of anti-human B agglutinins take place
after immunization with fin cells, B antigen exists in their cells. In
this case immunizing and post-immunizing procedures are just the sameas those in preparation of anti-Biii agglutinin.
In case of fowls' immune sera, the cells of antigen show a tendency
to cling on bottom wall of hole-glass before formation of agglutinating
lumps, and therefore the more frequent shaking or mixing seem to benecessary during reacting period than in the case of rabbit sera.
EXPERIMENTS AND RESULTS

Preparation of anti-Bu anti-En and anti-Em sera. These partial
antibodies were prepared with the method used by Yamaguchi (1943).
Their titers did not generally reach up to so high, and are showed in table
1, i. e., 1: 256 in anti-BI, 1: 16 in anti-Bu, and 1: 256, 1: 64 and 1: 32
in anti-Bm agglutinins respectively. But these sera were useful enoughto analysis of partial antigens.

No. 10 rabbit
male (o')4l

Anti-Bn No. 18 fow)
no. 45 female (a'fJ)
serum

Anti-BI
no. 13serum

Name of Immune
serum
animalNormal sera of imm. animal/

Ads. byhuman B
cells

JA
l~

n
-tit*+-----+±------

Ads. byhuman Acells

-tit**+----JA * + - - - - - l~G

No ads.

No ads.

Rabbitcells

Human
B typecellsl~Ads. 3>by human
A cells.

l~{~Ads. byhuman
A cells
Further3l
ads. byguineapig cells

l~

No 2)
ads.

JA

l~

fA

No1l
ads.

2
4

8

*-tit*++ - -+-tit-tit*+--**+---++-tit-tit*+-tit-tit*+----tit -tit -tit -tit -tit -tit*+-tit -tit -tit -tit -tit-tit**-tit -tit -tit -tit -tit -tit**-tit -tit -tit -tit -tit**+

1

"

+

+

+

16 32 64 128 256 512

Dilution,,.-- - - - - -Immune sera

Treatment Human
Dilution
Immune Treatment Humancellcell ,,------- ~antigen
1 2 4 8 16 32 64 128/

TABLE 1. AGGLUTININ TITERS OF IMMUNE ANTI-BI, ANTI-Bn AND ANTI-Bm SERAPREPARED FOR ADSORPTION TESTS

6

§

'Tj

0

~

.....

q

q

l~

[A

{~

No ads.

Ads. byhuman
A cells

(~

l~

( A

No ads.

No ads.

Treatment Humancell-tit*------

1 2 4 8 16 32 64 128

DilutionqqGuineapig
cellAds. by3)human
A cellsNo
ads.

Ads. by3)human
A cellsNo
ads.

Ads. by3)human
A cells

No
ads.

(~
(~

(~
(~

(~
{~

Immune Treatment Humanantigen
cell
2 4

8

+1t-ttt+H-1t+---tit**±---+ * +H- +H-1t + - ++1t+----**+H-+H-*++--ttt-ttt-ttt-ttt-l!lt*+**+H-+H-+H-**+-!tt-ttt-ttt-ttt * + - --tit -tit -tit -tit -tit -tit**++

16 32 64 128 256 512-tit -tit -tit -tit**+ -tit -tit -tit -tit -tit -tit -tit*-ttt+H-+H-**+--

1

Dilution

Immune sera

1) Even before adsorption, no anti-B11 or anti-Bm antibody is not involved in this serum. (see Material 1))2) This serum contains anti·Bm antibody besides anti-Bu one.
3) These sera are used for adsorption test with fin whale erythrocytes later on.
4) Serum type.

No. 17 fowl
female (a')

No. 8 fowl
male (o')

No. 4 fowl
male (o')

Remark:no. 44serumq

no. 20serumqAnti-BIII
no. 17serum

Name of Immune
serum
animalNormal sera of imm. animal

TABLE 1. AGGLUTININ TITERS OF IMMUNE ANTI-BI, ANTI-Bn AND ANTI-Biii SERAPREPARED FOR ADSORPTION TESTS (cont.)

.....
-..:i
01

~

t'"'

~

~

z~

0

"Ij

z

0

>-l

c:::

::i

~

&:t'"'
z8

s8

~

gJ

176

K.FUJINO

Adsorption tests of partial antibodies with fin whale erythrocytes.
After purifying procedures each partial antibody was used for adsorption
test with fin whale erythrocytes. Comparisons of their titers against
TABLE 2. ADSORPTION TESTS OF ANTI-BI, ANTI-BI! AND ANTI-BUIAGGLUTININS WITH THE ERYTHROCYTES OF FIN WHALES
Dilution of serum3J
Name ofserum

RemarkTreatment
2

4

8 16 32 64

128 256512* + + No descent* + + of titer
After
ads.
* ++
by
* ++
"
Control+* -Ht * +Anti-Brr
No descent+-no. 45
After ( no. A29 Jui cell
* -Ht * +of titerserum
ads.
no. A31 Ju 2
+
+-*
-Ht *
by
no. A45 Jui Ju2
++-"
*
-Ht *
Control-Ht -Ht -Ht -Ht -Ht -Ht *
+
*
cell
+
+
-Ht -Ht -Ht -Ht * *no. 285286
Anti-BUI After-Ht -Ht -Ht -Ht * * + +"Titer- ads.
no. 17no. 287-Ht -Ht -Ht -Ht -Ht * + +descend
"
byiJ
serum
-Ht -Ht -Ht * * ++"no. 289-Ht -Ht -Ht -Ht * * + +"no. 293-Ht -Ht -Ht -Ht * * + +"Control+*****+
Titer
cell
"
"
+
+--Afterno. 20descend
*
*
ads.
no. 3
serum
"
by2J ("°'** ++--no. 4-Ht * ++--"
Control+****+Titer+-" 44" After {"'' A29 Ju, cell
-Ht -Ht * *descend
no.
ads.
A31
Ju2
"
no.
serum
****+-by
no. A45 Jui Ju2 "****+-Remark: 1) and 2), Blood types of fin whale cells were not examined in these case.
Control

Anti-BI
no. 13
serum{no. A29 Jui cell
no. A31 Ju2 11
no. A45 Jui Ju2-Ht -tttttt -Ht -Ht -Ht-Ht -Ht -Ht -Ht -Ht -Ht-Ht -Ht -Ht -Ht -Ht -Ht-Ht -Ht -Ht -Ht -Ht -Ht

11

r

r

288

3) Agglutinin titers against human B type erythrocytes.

human B cells between before and after adsorption are shown in table
2. Fin whale cells used for adsorption were examined previously on
their blood groups (Ju-system only), but in cases of Nos. 17 and 20 sera
these examinations were not made because of the lack of available reagent for blood grouping.
In this table agglutinin titers in the column of "control" show the
pre-adsorbing titers against human B cells and the others show thethe post-adsorbing those. As no descent of the titers is seen in the

SEROLOGICAL CONSTITUTION OF FIN WHALE

177

anti-BI and anti-Bn sera, it may be assumed that the corresponding
B antigens, i. e. BI and Bn, are not contained in the fin whale cells.
While in the case of anti-Bm agglutinin descents of titers can be seen
independently to blood groups of adsorbing cells at the degrees of
1/2 to 1/4 in no. 17 serum, 1/8 in no. 20 serum and 1/2 in no. 4 serum
respectively. However, the anti-Bm agglutinin is not adsorbed thoroughlyaway in any case. From these facts it can be thought that the fin whale
cells possess a portion of Bm antigen independently to their blood types,
at least to Ju system. To confirm the existence of B antigen which
seems to have simpler structure than Bm was worked out the following
immunizing experiment.
Immunization of fowl with fin whale cells. In this experiment, prior
to immunization the serum type and its titers of fowl's serum was examined, and after immunization the ascent of titer against human B
cells was examined. As shown in table 3, such ascent of this titer wasrecognized at the degrees from 0 up to 8 times in no. 38 serum, from0 up to 16 times in no. 47 serum and from 2 up to 16 times in no. 46
serum respectively. According to these results the existence of B partial
antigen was immunologically confirmed.
All these anti-B agglutinins produced by the immunization was able
to be completely adsorbed with both guinea pig cells and any other fin
whale cells than immune antigens belonging to Jul type (nos. 47 and 48),
Ju2 type (no. 59) and ·Ju1Ju2 type (nos. 45 and 48).
According to the results of adsorbing and immunizing tests in previous
items 2) and 3), it was established that the B antigen in fin whale cells
had the simpler structure than Bm in guinea pig cells. Though the comparative survey between this antigen and BIV in sheep and horse cells
which was described by Ogura (1953b) has not been worked out, theauthor should like to use " BIV' " provisionally as the symbols of thissubstance. By using of this symbols the erythrocytes of B type human,
rabbit, guinea pig and fin whale are shown to have following antigenic
structures in relation to B antigen system.
BI Bn Bm BIV'
B type humanrabbitBn Bm BIV'
guinea pig
Bm BIV'fin whaleBIV'
DISCUSSIONAs already stated BIV' antigen is contained in fin whale (Balaenoptera
physalus) cells independently to Ju blood group system. And so far as
author's survey reaches it can be assumed that this BIV' partial antigen

46

No. 20Remark:+-~---+-----

0B

Ads. by j Human A
human 0 l
"
B'l
cells
,,
0

q
0
. No. 49 fin Ju,

1"

ffi -It+ - - -It+----

ffi ffi -It+± ffi - I t + - - + ± ----[

Human ANo
ads,

-It+----

No. 40 fin Jui

I.

-It++± - -

B

•
,,

Human· A[

Ads. by [ Human A· human 0 \
•
B
cells
\
,,
0

lr

·.

16

it+-~--

32

-----

4 8

..-------

Dilution

_,..

Ju,
type
cells

No. 49

Ju,
type
cells

No. 40

cells

t~'ie

No. 144

f

No
ads.

f

No. 40 fin Ju,

Human A
n
B
•
0

•
B
•O

No. 144 fin Ju,
Human A

Human A•B•
0

Cells foragglutinogen

Ads. byhuman A
cells

adsorp.

No.

0B

l "" B0

JHuman A

No. 49 fin Ju,q•

Human AAds byHuman A
human A)
•
B
cells
l
• 0
\

I

1

Ads. byhuman Al·cells

r

l
No
ads.

Immune Treatment
antigen
DilutiontH- tH- tH- -It+ -ffitH-tH--lt+---ffitH-tH-tH-ffi-lt+-tH-iffftH-tH--lt+--tH- tH- tH- tH- tH- tH- tH- tH- tH- -IttH- tH- -It -It+ -ffiffiffiffi-lt-lt-lt+ffi tH- -It -It -It+tH- tH- tH- -It++
ffi tH- tH- tH- tH- tH- tH- tH- tH- tH-

ffiffi-lt+-

+

-It

Titer
ascendTiter
ascendTiter
ascendRemarks

Therefore this+16 32 64 128 256 512 1024 2048 4096

*ffi ffi- Iffit *++--- ------+-It ffi ffi ffi ffi * ffi ffi -It +2

ffi ffi - I t + - - - - -I_,..--~~~~~~~~~~~~~~~~

Immune sera

1) This normal agglutinin was not a\lsorbed completely by human A type cells, but was adsorbed by guinea pig (Bm) cells.
agglutinin may be assumed to be one corresponding to B III or simpler structural B antigen than B Ill.
2) Serum type.

male (er'$')No. 19female (a')

47

No
ads.

No
ads.

38

Human AB
0
No. 144 fin Jui

No. 16
male («')'lCells_ ~oragglutmogen~

Serum . Fow!,
TreatmentNo.
immunized...___ANTI-B AGGLUTININS PRODUCED BY FOWLS·-IMMUNIZED WITH FIN WHALE ERYTHROCYTESNormal sera of ·fowl

TABLE 3.

,_.

~

8

>zj

r

CX1

SEROLOGICAL CONSTITUTION OF FIN WHALE

179

occurs in the cells of all fin whale individual independently to any otherblood group systems of this species and constitutes a component ofspecies-specific antigen of their erythrocytes. At present it has been
immunochemically ascertained that the erythrocytes of sperm whale(Physeter catodon) and dolphin (Prodelphinus caeruleoalbus) possess somekind of more complicated structural B antigen than Brv', likely to be Bm(Fujino, unpublished data). In past days such analysis on B substancein whale cells has been reported by AZAMI (1949) alone. After investigating on two whales, he recognized the existence of Bm, still the
specification on these whale species was not noted. However, such
structural differences of B substance by whale species seem to offer tothe author a prospect of discussion upon the evolutional relationshipamong various whale species, particularly between whalebone and toothedwhales. But any discussions on such problem should be taken up after
obtaining the comprehensive data on the distribution of A, B and 0
blood group substances among cetaceans.
As previously cited, Ogura (1953b) reports that the erythrocytes of
sheep and horse have a simpler structural B substance than Bm, and are
to be named as Brv. Comparative survey between Brv and Brv' will be
discussed in future issue.
NORMAL AGGLUTININ IN IMMUNE ANIMAL AND THEIRPRODUCING ABILITY OF IMMUNE AGGLUTININ

The author is working at immunogenetical researches on identification ofbreeding populations of whales in concerning to fishery problems (Fujino,1956). To carrying out such extensive investigations it is very important to obtain immune antibodies, which have high specificities andhigh titers, for blood grouping.
After examining the normal agglutinins of animals specific to human
A and B blood group antigens, Hibino (1935) introduces a term of '' Serumtype of animals ", and classifies the animal sera into four groups, i. e.
a'(1', a',(1' and o', accordingly as the existence of anti-A or anti-B heterohemagglutinins. And then he illustrates that serum type of animal is aserological constitution, which is closely related to the producing abilityof corresponding immune antibodies. While basing upon the quantity ofhuman A-like substance in the saliva, Koshino (1938, 1939) divides rabbits serologically into two groups of secretor and nonsecretor, and confirmes that the nonsecretor is more excellent producer of anti-human A
immune antibody than the secretor. Then he concludes that serum type andsecretor-nonsecretor type of rabbit are important serological characters tobe previously tested in case of preparation of immune anti-A antibody.

180

K. FUJINO

Same idea as abovestated points must be considered in preparation of
the reagent for blood grouping on fin whales. The author notes here somedata on the anti-Ju normal agglutinin in animal sera and the results ofproduction of anti-Ju immune agglutinins by these animals. These data
are not obtained under the intentional survey, but may serve to someextent for future work. As showing in table 4, the heterohemagglutinin
specific to Jul antigen occurs in the normal serum of a fowl among three andthe heterohemagglutinin specific to Ju2 antigen occurs in the normal sera of
the three rabbits among five and a fowl among three. It can be seen fromtable 4 that the titers of anti-Ju2 immune agglutinins reach up to higher
in rabbits (nos. 34, 35 and 36 sera) than in fowl (no. 46 serum), but incontrary to this tendency the titers of anti-Jul immune agglutininsreach up to higher in fowls (nos. 38 and 47 sera) than in rabbits (nos.
33 and 37 sera). In table 5 are shown some examples of agglutinin
titers to explain the degree of specificity of these immune sera.
TABLE 4. AGGLUTININ TITERS OF NORMAL AND IMMUNE SERAPRODUCED BY RABBITS AND FOWLS IMMUNIZED WITH THE
FIN WHALE ERYTHROCYTES
Immune animal

Normal agglutinin
After immuniz.

type
Species No. Sex Serum&A+ or ARabbit 33 male
II
37
II
Fowl
16
20 female"
Rabbit 35 male
II
II
34
II
II
36
II
Fowl
19
II

Remark:0 1 -A+a'-A+a'a'a 1 -A+0 1 -A+0 1 -Aa'fl'

Immune1)antigen

Serum
Blood No.
group
Exist. Titer Exist. Titer Anti-Ju 1 Anti-Ju 2
111 Jui33+ 1: 8 1: 160 1: 8
anti-Ju2

anti-Jui

~~~

+

1: 2

+
+
+

~~

~~No.

1: 40
155
"II
144
1: 800II
1: 64040
1: 20 1: 320 107 Ju1JU2
1: 64
1: 12800 113
Ju2
1: 64
II
1: 640 139
f/
1: 4 1: 2 1: 160
49

37
38
47
35
34
36
46

1) Erythrocytes of fin whale.

According to this table it may be assumed that the degree of specificity of anti-Jul is rather low both in fowls and rabbits, and that thedegree of anti-Ju2 specificity is low in fowl but rather high in rabbits,
especially higher in those with normal anti-Ju2 agglitinin. After summarizing the abovestated points, followings seem to be concluded on
preparation of anti-Jul and anti-Ju2 reagents.
1) To use rabbit that has corresponding normal agglutinin as theanti-Ju2 immune agglutinin producer.
2) To use fowl as the anti-Jul agglutinin producer.
By accomplishing the more extensive detection of normal anti-Jul agglutinin in fowl serum and of Ju blood group substances in various or-

-flt
-flt
-flt

Jui
Ju2

Jui
Ju2

Jui
Ju2

Jui
Ju2

Jui
Ju2

{111
139

{159
158

{165
139

{1E8
139

{ 40
49

38

33

34

35

46

40

80

-flt -flt -flt
-flt -flt -flt

-tt

-tt

+
+

10240

20480
81920
40960

-----------

5120-------** -------++

160 320 640 1280 2550

-flt -flt -flt -flt -flt
-flt -flt -flt -flt -flt

20

Dilution of antiserum

*
-flt
-

-

___,

**
+ ---------* -flt* -flt -flt -flt -flt -flt -flt
+ + -flt -flt**
-flt -flt -flt
* -flt+ --flt -flt --flt - + -- - -- --flt -flt -flt -flt*- - - - -flt -flt -flt -flt -flt -flt+
- - - -flt -flt -flt -flt -flt -flt * * + **10~

Blood group

No.

Serum
no.

Cell antigen

fowl

"

{Anti-Jui= 20
Anti-Ju2=320
Anti-Ju2= 160"

rabbitfowl

ImmuneanimalAnti-Ju2= 12 ,800

Anti-Ju1=160

Anti-Ju1=800

Titers of bloodgroup specific aggl.

TABLE 5. SOME EXAMPLES OF AGGLUTINATION REACTIONS OF ANTI·Jul AND ANTl-Ju2
IMMUNE (UNADSORBED) SERA AGAINST FIN WHALE ERYTHROCYTES
tTl-......
00......

tTl

>
t""

::et

"rj

"Ij

0

0

c:::

'"'3
'"'3

-..,
-z
z:a
Ul

z

0

(')

( ')

>
t""

C'l

0
t""
0

:;;:I

Ul

K. FUJINO

182

gans or glands of immune animals, however, more excellent reagents
must be obtained.
APPEARANCE OF IMMUNE ANTIBODIES INTO EGG-WHITE OFA FOWL INJECTED WITH FIN WHALE ERYTHROCYTES

No. 20 female fowl in table 3 (producer of no. 47 serum) was in a stateof egg laying during the period of immunizing experiment. So the author
Injection

1

Il

]][

v

N

VI

W

10

0

j

Oi

..----.-A

5.,,.,.. . . . . . --------.- c
.,;"

')(

,"'"•
--------- D,',.,1..--,,,,/ ,,,,-<.
- -"'..,.,,." .... """"

3

5

7

9

.,;

II

Day's

13 15 17 19 t
Rabbitsacrificed

Fig. 1. Production of anti-Jui and anti-B agglutinins in serum and egg-white of no. 20
fowl (Din Log2 D=limiting dilution titers) A: anti-Jui in serum, B: anti-B in serum
C: anti-Jui in egg white, D: anti-B in egg white.

tried to trace the appearance of immune antibody into egg-white. In
the following are reported some notes on such observation. As alreadystated the no. 20 female fowl was given intravenously on alternate days
a series of (seven-times') injections of 10% suspension of Jul type fin
whale erythrocytes, and sacrified for bleeding 7 days after last injection.
During the period of experiment this fowl laid a egg respectively ineach 4th, 7th, 12th, 14th and 20th day from the start of injection, be-SEROLOGICAL CONSTITUTION OF FIN WHALE

183

ing up to 5 eggs in all. The author traced ascendings of anti-Jul andanti-human B agglutinin titers in these egg-whites and sera obtained bytest bleedings. After centrifuging and inactivating, these agglutinins
were titrated with hole-glass (slide glass) method. Limiting dilution titersof these agglutinins are plotted in figure 1.
It may be assumed from this figure that both anti-Jul and anti-B
agglutinins in the egg-white begin to appear few days later than those in
the serum and don't reach up to so high titers as those of the latter. As
this observation was performed at the base of coastal whaling, no quantitative analysis have been carried out on globulins in egg-white and serum.
SUMMARY

1. Basing upon the classification of Friedenreich and With (1933) the
author carried out the analysis of the human B blood group substancein the erythrocytes of cetaceans from the view points of systematic
serology. A new partial antigen which has simpler structure than Bm in
the guinea pig cells were detected immunologically in the fin whale erythrocytes independently to their blood groups. This antigen was named provisionally as Brv'. By this symbol the B blood group substances in theerythrocytes of human, rabbit, guinea pig and fin whale may be
expressed as B1B11BmB1v', BnB111B1v', BmBrv', and Brv' respectively.
Comparative survey between Brv' and Brv in the erythrocytes of sheepor horse (Ogura, 1953b) will be discussed in the future issue.
2. Normal agglutinins specific to Ju antigens of fin whale erythrocytes
were detected in the sera of rabbits and fowls which were used as immune animal. In case of preparation of reagents for blood grouping on
Ju system of fin whales, it is desirable that fowl is used as the immune
animal for anti-Jul agglutinin producer and rabbit which has normal antiJu2 agglutinin is used as the anti-Ju2 agglitinin producer.
3. Appearance of immune antibodies into egg-white of a fowl wasobserved on the way of preparation of the anti-Jul agglutinin. It may
be assumed from the results of this observation that both anti-Jul and antihuman B agglutinins begin to appear into egg-white few days later thaninto serum and don't reach up to so high titers as those of the latter.
ACKNOWLEDGEMENT

The author should express his hearty thanks to Prof. Tanemoto Furuhata
and the late Dr. Katsumi Yamaguchi for suggestion on this study.

184

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