AMERICAN JOURNAL OF CLINICAL PATHOLOGV
Vol. 32, No. 3, September, 1959, pp. 257-260
Printed in U.S.A.
A CASE OF MULTIPLE MYELOMA WITH TWO ABNORMAL SERUM GLOBULIN
FRACTIONS DETECTED BY MEANS OF PAPER ELECTROPHORESIS
ARYEH SZEINBERG, P H . D . , S. L. WEISSMAN, M.D., AND GISELLA TAUMAN, Men. CHBM.
Government Hospital, Tel Hashomer, Israel
Modern technics of study have considerably broadened our knowledge of the abnormal proteins formed in multiple myeloma.
Examinations of large series of patients reveal that a diversity of pathologic proteins
may be formed in this condition. 1317 ' 20
When analyses of serum were performed by
means of moving-boundary electrophoresis
or starch-gel electrophoresis, cases with 2 or
more pathologic fractions were detected. 3 ' s '
12, 13, i7,20 rpjie differentiation ability of paper
electrophoresis seems to be much lower, inasmuch as, even in large surveys of cases of
multiple myeloma by means of this technic,
only .1. abnormal protein fraction has been
observed by several investigators. 4 '"• 7 The
only reports known to us, describing cases of
myeloma with 2 abnormal protein fractions
detected by paper electrophoresis, are those
of Tsevrenis and Samios18 (2 cases with 2
protein bands between beta and gamma
globulin) and Szeinberg and his associates16
(a case with a pre-albumin fraction and a
fraction moving to the cathodic side of
gamma globulin). In the present communication, we describe the findings in an additional case with 2 abnormal serum proteins
detected by means of paper electrophoresis
of specimens from 45 patients with myeloma.
METHODS
Determinations of total protein, albumin,
and globulin were performed by the method
of Wolfson and his associates.19 Paper electrophoresis was performed with a Servall
apparatus Type LKB 3276 in a barbiturate
buffer, pH 8.6, ionic strength 0.125. 110 v.,
0.8 ma. per strip for 20 hr. The strips were
stained with bromphenol blue and the relative concentrations of protein were estimated
with the aid of a photoelectric densitometer.
Received, March 30, 1959; accepted for publication May 12.
Dr. Szeinberg is Laboratory Director, Dr.
Wcissman is Head of Orthopedic Department,
and Mrs. Tauman is Biochemist.
Salting out of protein fractions was performed by adding 0.4 ml. of serum to 10 ml.
of sodium sulfate solutions of various concentrations (13 to 15 per cent). After 4 hr. of
incubation at 37 C , the tubes were eentrifuged and the precipitates washed twice
with solutions of sodium sulfate of the concentration originally used. After the last
washing, the precipitates were dissolved in
0.4 ml. of physiologic saline solution and
examined by means of paper electrophoresis.
Heal precipitation. Samples of serum were
incubated at 45 C , 50 C , and 56 C. for 2
hr., and then observed for the formation of a
precipitate or gelatification.
CASE R E P O R T
A man, 36 years old, was admitted to the
orthopedic department with complaints of
severe pains in the right iliac crest. A year
previously, he had started to limp suddenly,
without any known cause. Three months
prior to his admission to the hospital he
began to feel severe pains in the right hip.
Upon admission, the blood sedimentation
rate (Westergren) was 115 mm. after 1 hr.
Serum total protein was 7.6 Gm. per 100
ml., with 3.2 Gm. of albumin and 4.4 Gm. of
globulin. No protein was detected in the
urine. Radiologic examinations revealed a
large osteolytic focus in the right ilium. No
abnormalities were observed in any of the
other bones. The patient was operated and a
large cavity filled with soft tissue was observed in the right ilium. A complete
curettage of the focus was performed and
bone chips were inserted into the cavity.
Upon histologic examination of the removed
tissue, plasma cell myeloma was detected.
Approximately 2 weeks later, a sternal puncture was performed, and examination of the
bone marrow revealed typical findings of
plasma cell myeloma. The diagnosis of
multiple myeloma was thus established, although only a solitary tumor was seen in the
radiologic examination of the bones.
257
•25S
SZEINBEKG ET
Protein Investigations
AL.
Vol. 32
synthesis of protein by the abnormal cells.
'Repeated examinations of several speci- No final conclusions have yet been reached
mens of serum by means of paper electro- with regard to the nature of these proteins.
phoresis revealed a constant finding of 2 According to Deutsch and16 his associates- and
prominent, compact, and homogenous pro- Smith and his associates, these proteins are
tein bands between beta and gamma globu- probably normal components of serum, but
lin (the position of M globulin). In the- are present in abnormal amounts. Slater
position for normal gamma globulin, only a and his associates suggested that they are
related to constituents of
small quantity of protein was seen (Fig. 1, not normal, but
14
normal
serum.
According
to Putnam it is
lino 2). The quantitative evaluation of the
electrophoretic strip revealed the following more credible to regard these globulins as
composition of the serum proteins: albumin, truly abnormal proteins (differing struc38 per cent; alpha-1 globulin, 5.5 per cent; turally from the normal proteins in various
alpha-2 globulin, 12.5 per cent; beta globu- loci such as the N terminal residue), the
lin, 5.5 per cent; abnormal Mi fraction, 15 product of an abnormal cell with a perverted
resembling
per cent; abnormal M 2 fraction, 18 per cent; mechanism of protein synthesis,
9 10
'
in
this
respect
a
mutation.
gamma globulin, 5.5 per cent.
Recent studies of other conditions with
Differential salting-out with solutions of
sodium sulfate yielded the following results: synthesis of an abnormal protein (for exat the concentration of 13 per cent sodium ample, the hemoglobinopathies) suggest
sulfate, no precipitate was formed; at 13.5 that the synthesis of each individual protein
per cent sodium sulfate, a part of the ab- is governed by a single gene. If the producnormal fraction M 2 was precipitated (Fig. 1, tion of the myeloma proteins is regarded as a
line 4); at 13.7 per cent and 13.8 per cent mutation, then a constant appearance of 2
sodium sulfate, all of the fraction M 2 and a or more abnormal globulins in a single
part of Mi were precipitated, whereas, at 14 patient might represent an alteration in
per cent, both abnormal fractions precipi- several genes.
Another possible explanation of this findtated completely. The electrophoretic exing
is a supposition that 1 or several such
amination of the precipitate revealed that
globulins
represent an unfinished molecule,
both fractions were separated, and in quantities comparable to those obtained with or an abortive product of the synthesis of
intact serum (Fig. 1, line 3). The gamma protein. Such possibilities have been raised
globulin was not precipitated with this con- in connection with the relation between1, 5the
myeloma proteins in serum and urine. - "
centration of salt.
Inasmuch
as the abnormal pattern is conIncubation of the samples of serum at 45
stant
in
each
instance, however, such an
to 56 C. for 2 hr. did not result in formation
explanation
of
the appearance of multiple
of any precipitate or gelatification.
pathologic fractions in serum would entail
supposition that the ability of releasing
DISCUSSION
unfinished or abortive molecules into the
Multiple myeloma is of great biochemical
blood circulation is also a constant and
interest because of the profuse synthesis and
characteristic property of the myeloma cell
the diverse nature of proteins elaborated in
(in some persons), controlled by an unknown
this condition. The abnormal proteins
mechanism.
formed by various patients are considerably
different in their electrophoretic mobility,
SUMMARY
solubility in solutions of salts, molecular
weight, amino end-groups, and antigenicity,
A case of multiple myeloma is described,
but remain qualitatively peculiarly constant and the patient had 2 abnormal serum
throughout the course of the disease. 9 ' 10 ' n globulins that were detected by means of
It seems, therefore, justifiable to presume paper electrophoresis. Both fractions had
that in each instance their structure is fixed electrophoretic mobility between beta and
by a specific- inherent abnormality of the gamma globulin. The abnormal globulins
Sept. 1959
ABNORMAL, G L O B U L I N
IN MYELOMA
259
F I G . 1. Paper electrophoresis of proteins i n : line 1—normal
serum; line8—serum of the patient; line S—precipitate
formed
by a 14 per cent solution of sodium sulfate; and line 4—precipitate formed by a 13.5 per cent solution of sodium sulfate.
could be precipitated from the serum by
means of a 14. per cent solution of sodium
sulfate, and they retained their distinct
mobility.
SUMMAKIO
IN INTEKL1NGUA
Es describite un caso de myeloma multiple. Le patiente habeva 2 fractiones anormal de globulina serai le quales esseva detegite per medio de electrophorese a papiro.
Am be iste fractiones habeva un mobilitate
electrophoretic inter illo de globulina beta e
globulina gamma. Illos poteva esser precipitate ab le sero per medio de un solution de
.1.4. pro cento de sulfate de natrium. Lor
distincte mobilitate esseva retenite post lor
precipitation.
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ET
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