ep?
ae
S
te been
‘a ehad
~
Stores.
*.
id high titres of
'g
frican
cronesians
comerous megakaryocytes in the bone-marrow and some in
Sveipheral blood, the possibility that the patient might have
a
ys.
was raised.
Ses been obtained.
‘three.
whe Ouchterl
ony
“ef
No convincing evidence that this is so
-@yhout transfusion the patient’s hemoglobin continued to
2, and in February, 1958, he was admitted to the Clinical
-_.
€$ Containing
Se .-<
‘f sodium
Dhos Rhea
ni
a:
m_ ethylene
ee
d:, sheet
Wirth a dic,
six we
‘Well of the
Same ts.
and Peripher
al y-
Center of the National Institutes of Health. During his time in
“Were plac
ed in
Fig. i
. Fig. 2
% I—Fhe precipitin reaction, seen in an agar-gel* Ouchterlony
by Oblique lum,
_ plate, between the serum of the patient in the centre well and
by the mictom
esdey
idegger (1955
Lusty
° 3-qeene, but not all, members of a panel of sera from normal
3 pervons. Unstained piate after 24 hours.
agar made up ..
_ Me 3—The precipitin reaction between the serum of the patient in
ody (or antigen
wr:
“€acting for 18 ha.
-.
Were visible.
Jz,
_,¥ te centre well against some, but not all, members of a panel of
+ The cwo degrees of positive reaction are shown: nos. 1; 3, and 5
+ ga 2+ ceactors and no. 2 is a weak + reactor.
vhich the Plates Wee-
Connaught Lahn
‘droxymeth! -2n-:+,.
Secpitel the patient had persistent anzmia withoutreticulocywas. The leucocyte count varied from 10,000 to 15,000 and
the platelet count was consistently above 500,000 per c. mm.
‘The differential count showed predominantly neutrophils with
tciztive increase of immature forms. Bone X-rays showed no
W906 80. 1507.
aste (Smithies po<Was Carried outs -
he components w::
iS componensys we
ities,
Refractory anemia
of unknown cause was
Gagnosed. The patient was given cells from 4 units of blood,
Wiech raised his hemoglobin to 11 g. per 100 ml., and was
ee
‘peration of 7S as:
Oacharged.
© density-cradie:
~ When the patient first received blood-transfusions in 1958
|» Seer were well tolerated. Since that time he has received 47
7) using 40, 30, >
wea of blood. The patient is of blood group AB, Rh+, and
bas never been evidence of red-cell incompatibility with
Mf Bozicevich cpa
", emer bloods. In the spring of 1960 the patient began to have
IE aS describe
tia)
4
. “Sxnrfusion reactions, Within about 1 hour of some, but notall,
famtfusions there was a fever, rising to 38-39°C (100-4—
* $022°F) accompanied by headache and muscle pain, but no
symptoms. The fever and symptomspersisted for about
of a patient wh.
t long and varie
A sc.end
positive subjects had rheumatoid factor, as determined
by bentonite-flocculation and latex-fixation tests. Some
panel subjects with rheumatoid factor pave reactions with
the antiserum, but others did not. A second sample of
serum from the patient, drawn 3 weeks after thefirst, ata
we andere
Fig. 4—Agar-gel
antibody.
. -%
i
immunocelectrophoresis
.
oo
to
_
characterise
the
The serum of the patient was placed in the circular wells and
submitted to electrophoresis, and a strongly positive serum concentrated three times was placed in the rectangular wells. A broad
precipitin line (arrow) is seen in the position of the y-globulin.
time when the patient had not received any medication
since discharge, gave identical reactions, as did a third
sample drawn 1 month Jater, and a fourth 3 monthslater.
Support for the interpretation that the precipitin in the
serum of this patient is an antibody came from experi-
ments showing that the active protein is a 7S-y-globulin.
the a-globulin antigen and Ag(a~) for sera that do not
contain the antigen. The precipitin in the serum of the
patient thus has the specificity anti-Ag(a-+-),
buminuria whit
nths.
unlike the diffuse reaction which slowly develops between ©
rheumatoid factor and y-globulin in some subjects
(Franklin 1960a).
Rheumatoid-factor reaction was
excluded because neither the patient nor most of the
We propose the designation Ag(a+) for sera containing
Ll retired Cecwrtiy
Budapest. he het
d paralysis of the
Precipitation Reactions
The serum of the patient gave well-defined precipitation reactions in agar with some, but not all, panel sera
(fig. 1). The reaction was clearly visible at 18 hours and
was intense after 2 days. The precipitation line was
convex toward the centre (precipitin) well, indicating that
the antigen is of relatively high molecular weight (Korn-
(fig. 1). The precipitation lines were clear-cut and quite
d at 18 houn
~
ally Visible}, 25
han
‘ RA-tess* i
5 hours, and then slowly subsided. The patient had been discharged from hospital before the precipitin in his blood was
discovered, so that it has not yet been possible to ascertain
whether only bloods containing the precipitating antigen give
transfusion reactions.
gold et al. 1959). Two intensities of reaction were defin-
tse
ral wells. Th
e pln
2rve
nd
635
able in most groups of sera studied (fig. 2). Some sera
lacked detectable antigen, the precipitation lines extending into the corresponding wells without deviation
* 88ar cores
frovith small volu - ta
ma -
tebe
THE LANCET
ORIGINAL ARTICLES
Ee ipved for a month and then ceased, and with the help of
sfusions the hamoglobin was raised to 14 g. per
ml. Because of the high white-cell count and the presence
debi
years. S$
:
€ Sera Were
«.
025, 1961
Characterisation of Antigen
E..
A panel serum giving a strongly positive precipitation
line in the gel-diffusion test was concentrated three times
by ultrafiltration and used for immunoelectrophoresis.
Waist, Weeknat
in the legs. Sod
spinal turd s.r:
per ml. Pycya2ain remiss
The antibody well wasfilled with serum from the patient.
A well-defined precipitation line developed between the
Iceration daticg
1938 and lhe
but svmpis:
antigen is a high-molecular-weight x,-component.
“he patiens war
he was repericevels, and tus’
ION suggesize
The eesics
%, region and the antibody (fig. 3). This result, along
with that in the Ouchterlony plates, suggested that the
i. t~Agar-gel immunoelectrophoresis to characterise antigen.
tenes beulti a strongly Positive reactor was concentrated three
ef the Patient wasn and submitted to electrophoresis. The serum
=ion of the as placed in the rectangular antibody well. The
Sobulin Abovern line (arrow) suggests that the antigen is an
.
Ove, unstained precipitin lines; below, protein stain.
en re
The
only a,-macroglobulin so far defined is the 19S-«, component described by Brown et aj. (1954) and Wallenius
et al. (1957), which appears to be equivalent to the’
slow-x, component found on starch-gel electrophoresis
(Poulik and Smithies 1958).
Purified «,-macroglobulin
(Behringwerke, Marburg) in a wide range of concentrations failed to give precipitation with the patient’s serum.
m2