J. Nihon
Univ.
Sch.
Dent.,
Natural
Vol. 34, 237-239,
1992
Antibody against Thomsen-Friedenreich Antigen in Bovine Milk
Masao KUSAMA1, Kaoru KUSAMA2 and Itaru MORO2 (Received21 November1991and accepted3 February 1992) Key words:
natural antibody, Thomsen-Friedenreich antigen, bovine milk, fetal bovine serum, enzyme-linked immunosorbent assay Abstract
The presence of natural antibody against Thomsen-Friedenreich (TF) antigen in fresh bovine milk was demonstrated by an enzyme-linked immunosorbent assay (ELISA). The amount of the antibody against TF antigen was decreased after heat treatment, and the antibody was not detectable by ELISA in the milk after pasteurization. A small amount of the antibody was detected in fetal bovine serum by ELISA, suggesting the transfer of passive immunity to the calf. Introduction The presence of Thomsen-Friedenreich antigen (TF antigen, /3 Gal(1-3) a GalNAc-) has been detected in several kinds of Gram-negative bacteria such as Escherichia coli and in their lipopolysaccharides (LPS)111.The presence of natural antibody against TF antigen (anti-TF antibody) is detectable in human serum as well as in sera of various animals{11.Decreased levels of anti-TF antibody in sera of cancer Bovine and
in
protect we
patients
milk
is
compared
a very
cooking. the
Fresh
living
the
in normal
nutritious
drink
bovine
body,
demonstrated
enzyme-linked
to those
popular
such
milk as
presence
contains
lactoferrin of
immunosorbent
anti-TF assay
Fresh
was
1). also
to
coated
was
by as
10 ƒÊg/ml
in serum
草 間 正 夫1,草
PBS
saline
(Cappel) 間 薫2,茂
were
of
citrate
Each
as
(pH to
a standard
in
5 milch
fat
well
asialoglycophorin
(OA-PBS)
active
reported[1-3]. various
fresh
foods
components In
bovine
that this
milk
study,
using
an
Methods
from
addition
phosphate-buffered
in
immunoglobulins[4,5].
antibody
and
and
follows:
been
used
(ELISA).
collected
components
ELISA
with
ovalbumin bovine
milk
precipitated
subjected
with
bovine Cellular
have
is also
various
and
Materials
(Table
serum and
cows
removed
of
7.4)
(PBS),
solution
The
96-well
whey
microplates
Chemical each
was
and
casein
obtained
After
incubated
binding. diluted
parturition
was
(NUNC)
Co.).
well
non-specific and
after
centrifugation
solution.
(Sigma
block
4 months by
samples
Diluted were
was washing with
1%
normal added
to
呂 周2
1 Faculty of Home Economics, Tokyo Kasei University, 1-18-1, Kaga, Itabashi-ku, Tokyo 173. 2 Department of Pathology, Nihon University School of Dentistry, 1-18-13 Kanda-Surugadai, Chiyoda-ku, Tokyo 101. To whom all correspondence should be addressed: Dr. Kaoru KUSAMA, Department of Pathology, Nihon University School of Dentistry, 1-8-13 Kanda-Surugadai, Chiyoda-ku, Tokyo 101, JAPAN.
238
Table
the
wells
and
containing
incubated
0.05%
ature
wells
20
min.
Optical
density
Electric
Co.
(Gibco)
were
effect
of
Ltd.,
solution was
room
(H
measured
on the
the
1%
fresh
milk
the
after
of
to
be
Figure
Fig.
1 shows
bovine
1
Calibration a standard
serum.
the
calibration The
amount
curve of the enzyme solution.
microplate
at
63•Ž
antigen
of
temper-
2 M
bovine To
bovine
was
H2SO4.
serum
examine
milk,
used
400-times-diluted
5.0)
(Corona
5,10,15,20,25
serum
in
at room pl
fetal
ELISA.
fresh
for
bovine
and
to in
25
for
(pH
photometer
whey
antigen
normal
H202 of
PBS with
X10,000) buffer
30%
subjected
anti-TF
anti-TF 1,000
a
with
incubated
(KPL,
addition
serum,
and
heating
Diluted
amount
estimated
of
pl/ml by
by
was
antibody
0.3
bovine
OA-PBS
activity
ELISA.
nm
washing
well
citrate-phosphate
stopped
492
Normal
with
M
and
at
each
rabbit
0.1
was
the
the
whey
and
30
a
standard
as normal
min
bovine
units/ml.
Results
normal
+ L)
milk
After
10 times,
with
reaction
in bovine
temperature.
Tween) IgG
Tokyo).
to
and
serum
the
diluted
subjected
antibody
o-phenylenediamine
was
from
was
at
(PBS
incubated
Then
heating
obtained
1 h
20
were
1 mg/ml
for
of anti-TF
anti-bovine
Washed
containing
for
Tween
peroxidase-labelled 1 h.
1 Levels
and curve of
Discussion of
antibody
immunoassay
the
ELISA
for
anti-TF
antibody
against
TF
antigen
in
for anti-TF
antibody
in normal
using
fresh
bovine
bovine
serum
as
239
milk
was
2,140•}1,005
globulin
isotype
Further When to
40%
further TF
studies fresh of
in
that
antigen
the
on
was of
the
bovine
In
Fig.
of
fresh
detected
fact,
milk
we
were
of ELISA,
Decrease
at
bovine
the
unable
in level
in
for
the
of anti-TF
5 min, The in
ELISA
detect
of
any
antibody
are
the
antibody
milk
heat
of
antibody pasteuriza-
detectable
immunity
after
heat
heat
reactivity
anti-TF of
were
passive
The
and
because
was against
pasteurization.
of
needed. decreased
antibody
antigenicity
units/ml)
in bovine
immuno-
was
The
reactivity
of
milk
of 2.
the
IgA and IgM[4].
reactivity
amount Fig.
probably
transfer
of
with bovine
after
a loss
(180•}20 the
amount
antibody
examined,
suggesting
large
shown
by in
to
antibody
A
in comparison
milk. as
milk
resulted
milk
the
63•Ž
10 min
in
bovine
1).
is IgG,
anti-TF
heated
heating
available
by
2
was
for
amounts
serum
isotypes
by
commercially Small
milk
non-heated
not
fresh
(Table
bovine
milk
in
antibody.
tion.
the
bovine
decreased
treatment
units/ml
in fresh
in to
the
fetal calf.
treatment.
References [1] SPRINGER, G. F., DESAI,P. R., MURTHY, M. S., TEGTMEYER, H. and SCANLON, E. F.: Human carcinoma-associated precursor antigens of the blood group MN system and the host's immune responses to them, Prog. Allergy, 26, 42-96, 1979 [2] SPRINGER, G. F. and DESAI,P. R.: Detection of lung and breast carcinoma by quantitating serum anti-T IgM levels with a sensitive, solid-phase immunoassay, Naturwissenshaften, 69, 346-348, 1982 [3] BRAY,J., MACLEAN,G. D., DUSEL,F. J. and MCPHERSON, T. A.: Decreased levels of circulating lytic anti-T in the serum of patients with metastatic gastrointestinal cancer, Clin. Exp. Immunol., 47, 176-182, 1982 [4] BUTLER, J. E.: Bovine immunoglobulin: An augmented review, Vet. Immunol. Immunopathol., 4, 43-152, 1983 [5] RAINARD, P. and CAFFIN,J. P.: Sequential changes in serum albumin, immunoglobulin (IgGi, IgG2, IgM) and lactoferrin concentrations in milk following infusion of Escherichia coli into the udder of immunized and unimmunized cows, Ann. Rech. Vet., 14, 271-279, 1983
Univ.
Sch.
Dent.,
Natural
Vol. 34, 237-239,
1992
Antibody against Thomsen-Friedenreich Antigen in Bovine Milk
Masao KUSAMA1, Kaoru KUSAMA2 and Itaru MORO2 (Received21 November1991and accepted3 February 1992) Key words:
natural antibody, Thomsen-Friedenreich antigen, bovine milk, fetal bovine serum, enzyme-linked immunosorbent assay Abstract
The presence of natural antibody against Thomsen-Friedenreich (TF) antigen in fresh bovine milk was demonstrated by an enzyme-linked immunosorbent assay (ELISA). The amount of the antibody against TF antigen was decreased after heat treatment, and the antibody was not detectable by ELISA in the milk after pasteurization. A small amount of the antibody was detected in fetal bovine serum by ELISA, suggesting the transfer of passive immunity to the calf. Introduction The presence of Thomsen-Friedenreich antigen (TF antigen, /3 Gal(1-3) a GalNAc-) has been detected in several kinds of Gram-negative bacteria such as Escherichia coli and in their lipopolysaccharides (LPS)111.The presence of natural antibody against TF antigen (anti-TF antibody) is detectable in human serum as well as in sera of various animals{11.Decreased levels of anti-TF antibody in sera of cancer Bovine and
in
protect we
patients
milk
is
compared
a very
cooking. the
Fresh
living
the
in normal
nutritious
drink
bovine
body,
demonstrated
enzyme-linked
to those
popular
such
milk as
presence
contains
lactoferrin of
immunosorbent
anti-TF assay
Fresh
was
1). also
to
coated
was
by as
10 ƒÊg/ml
in serum
草 間 正 夫1,草
PBS
saline
(Cappel) 間 薫2,茂
were
of
citrate
Each
as
(pH to
a standard
in
5 milch
fat
well
asialoglycophorin
(OA-PBS)
active
reported[1-3]. various
fresh
foods
components In
bovine
that this
milk
study,
using
an
Methods
from
addition
phosphate-buffered
in
immunoglobulins[4,5].
antibody
and
and
follows:
been
used
(ELISA).
collected
components
ELISA
with
ovalbumin bovine
milk
precipitated
subjected
with
bovine Cellular
have
is also
various
and
Materials
(Table
serum and
cows
removed
of
7.4)
(PBS),
solution
The
96-well
whey
microplates
Chemical each
was
and
casein
obtained
After
incubated
binding. diluted
parturition
was
(NUNC)
Co.).
well
non-specific and
after
centrifugation
solution.
(Sigma
block
4 months by
samples
Diluted were
was washing with
1%
normal added
to
呂 周2
1 Faculty of Home Economics, Tokyo Kasei University, 1-18-1, Kaga, Itabashi-ku, Tokyo 173. 2 Department of Pathology, Nihon University School of Dentistry, 1-18-13 Kanda-Surugadai, Chiyoda-ku, Tokyo 101. To whom all correspondence should be addressed: Dr. Kaoru KUSAMA, Department of Pathology, Nihon University School of Dentistry, 1-8-13 Kanda-Surugadai, Chiyoda-ku, Tokyo 101, JAPAN.
238
Table
the
wells
and
containing
incubated
0.05%
ature
wells
20
min.
Optical
density
Electric
Co.
(Gibco)
were
effect
of
Ltd.,
solution was
room
(H
measured
on the
the
1%
fresh
milk
the
after
of
to
be
Figure
Fig.
1 shows
bovine
1
Calibration a standard
serum.
the
calibration The
amount
curve of the enzyme solution.
microplate
at
63•Ž
antigen
of
temper-
2 M
bovine To
bovine
was
H2SO4.
serum
examine
milk,
used
400-times-diluted
5.0)
(Corona
5,10,15,20,25
serum
in
at room pl
fetal
ELISA.
fresh
for
bovine
and
to in
25
for
(pH
photometer
whey
antigen
normal
H202 of
PBS with
X10,000) buffer
30%
subjected
anti-TF
anti-TF 1,000
a
with
incubated
(KPL,
addition
serum,
and
heating
Diluted
amount
estimated
of
pl/ml by
by
was
antibody
0.3
bovine
OA-PBS
activity
ELISA.
nm
washing
well
citrate-phosphate
stopped
492
Normal
with
M
and
at
each
rabbit
0.1
was
the
the
whey
and
30
a
standard
as normal
min
bovine
units/ml.
Results
normal
+ L)
milk
After
10 times,
with
reaction
in bovine
temperature.
Tween) IgG
Tokyo).
to
and
serum
the
diluted
subjected
antibody
o-phenylenediamine
was
from
was
at
(PBS
incubated
Then
heating
obtained
1 h
20
were
1 mg/ml
for
of anti-TF
anti-bovine
Washed
containing
for
Tween
peroxidase-labelled 1 h.
1 Levels
and curve of
Discussion of
antibody
immunoassay
the
ELISA
for
anti-TF
antibody
against
TF
antigen
in
for anti-TF
antibody
in normal
using
fresh
bovine
bovine
serum
as
239
milk
was
2,140•}1,005
globulin
isotype
Further When to
40%
further TF
studies fresh of
in
that
antigen
the
on
was of
the
bovine
In
Fig.
of
fresh
detected
fact,
milk
we
were
of ELISA,
Decrease
at
bovine
the
unable
in level
in
for
the
of anti-TF
5 min, The in
ELISA
detect
of
any
antibody
are
the
antibody
milk
heat
of
antibody pasteuriza-
detectable
immunity
after
heat
heat
reactivity
anti-TF of
were
passive
The
and
because
was against
pasteurization.
of
needed. decreased
antibody
antigenicity
units/ml)
in bovine
immuno-
was
The
reactivity
of
milk
of 2.
the
IgA and IgM[4].
reactivity
amount Fig.
probably
transfer
of
with bovine
after
a loss
(180•}20 the
amount
antibody
examined,
suggesting
large
shown
by in
to
antibody
A
in comparison
milk. as
milk
resulted
milk
the
63•Ž
10 min
in
bovine
1).
is IgG,
anti-TF
heated
heating
available
by
2
was
for
amounts
serum
isotypes
by
commercially Small
milk
non-heated
not
fresh
(Table
bovine
milk
in
antibody.
tion.
the
bovine
decreased
treatment
units/ml
in fresh
in to
the
fetal calf.
treatment.
References [1] SPRINGER, G. F., DESAI,P. R., MURTHY, M. S., TEGTMEYER, H. and SCANLON, E. F.: Human carcinoma-associated precursor antigens of the blood group MN system and the host's immune responses to them, Prog. Allergy, 26, 42-96, 1979 [2] SPRINGER, G. F. and DESAI,P. R.: Detection of lung and breast carcinoma by quantitating serum anti-T IgM levels with a sensitive, solid-phase immunoassay, Naturwissenshaften, 69, 346-348, 1982 [3] BRAY,J., MACLEAN,G. D., DUSEL,F. J. and MCPHERSON, T. A.: Decreased levels of circulating lytic anti-T in the serum of patients with metastatic gastrointestinal cancer, Clin. Exp. Immunol., 47, 176-182, 1982 [4] BUTLER, J. E.: Bovine immunoglobulin: An augmented review, Vet. Immunol. Immunopathol., 4, 43-152, 1983 [5] RAINARD, P. and CAFFIN,J. P.: Sequential changes in serum albumin, immunoglobulin (IgGi, IgG2, IgM) and lactoferrin concentrations in milk following infusion of Escherichia coli into the udder of immunized and unimmunized cows, Ann. Rech. Vet., 14, 271-279, 1983
