Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub. 2008, 152(1):61–64.
© T. Pika, J. Minarik, P. Schneiderka, M. Budikova, K. Langova, P. Lochman, J. Bacovsky, V. Farbiakova, V. Scudla
THE CORRELATION OF SERUM IMMUNOGLOBULIN FREE LIGHT CHAIN
LEVELS AND SELECTED BIOLOGICAL MARKERS IN MULTIPLE MYELOMA
Tomas Pikaa*, Jiri Minarika, Petr Schneiderkab, Marie Budikovab, Katerina Langovac,
Pavel Lochmanb, Jaroslav Bacovskya, Vera Farbiakovad, Vlastimil Scudlaa
a 3rd Department of Internal Medicine, Teaching Hospital, 775 20 Olomouc, Czech Republic
b Department of Clinical Biochemistry, Teaching Hospital, 775 20 Olomouc
c Department of Statistics, Palacky University Olomouc, Czech Republic
d Department of Transfusion Service and Clinical Hematology, Třinec Hospital, Czech Republic
Received: March 23, 2008; Accepted: April 15, 2008
Key words: Multiple myeloma/Free light chain serum levels/Biological parameters
Aims: the presented study is aimed at the evaluation of correlation of free light chains serum levels – kappa, lambda
and their relation (K/L ratio) and serum levels of selected biological markers in a group of patients with multiple
myeloma examined at the time of the diagnosis.
Methods: 102 patients with multiple myeloma were included in this prospective study. Free light chains serum
levels were determined by FreeliteTM Binding Site system, for determination of serum levels of selected parameters the
following methods were used: REA thymidinekinase (TK), RIA (β2microglobulin (β2m), ICTP, PINP), enzymoimmu-
noassay (sIL-6R, sVCAM-1, sICAM-1, sOPG) and quantitative enzymatic immunoassay (sHGF, sVEGF, sSyndecan-1
(sSyn-1) a sFas).
Results: There was found a correlation in the kappa-group of the dominant kappa chain and serum readings of β2m
(r = 0.344, p = 0.005), TK (r = 0.263, p = 0.035), ICTP (r = 0.402, p = 0.001), PINP (r = 0.264, p = 0.039), sOPG (r =
0.328, p = 0.028), sSyn-1 (r = 0.255, p = 0.046) and sFas (r = 0.418, p = 0.001). In case of K/L ratio there was found a
statistically signifi cant association of levels of β2m (r = 0.316, p = 0.01), TK (r = 0.274, p = 0.027), ICTP (r = 0.346, p =
0.006), PINP (r = 0.261, p = 0.042), sSyn-1 (r = 0.283, p = 0.026) and sFas (r = 0.377, p = 0.002). In the lambda-group
the analysis confi rmed mutual dependence of the dominant lambda chain levels on β2m (r = 0.476, p = 0.003), ICTP
(r = 0.375, p = 0.022), sVCAM-1 (r = 0.383, p = 0.019), sHGF (r = 0.441, p = 0.006) and sFas (r = 0.334, p = 0.040).
In addition we ascertained a correlation of L/K ratio and levels of β2m (r = –0.473, p = 0.003), TK (r = –0.412, p =
0.011), ICTP (r = –0.331, p = 0.045), PINP (r = –0.409, p = 0.012), sHGF (r = –0.357, p = 0.028), sSyn-1 (r = –0.449,
p = 0.005) a sFas (r = – 0.371, p = 0.022).
Conclusions: The study confi rmed mutual correlation of FLC serum levels and the levels of several selected bio-
logical markers, in particular β2m, TK, ICTP, PINP, sSyn-1 a sFas at time of the diagnosis. It referred to the mutual
relation of bone marrow microenvironment, biological qualities of clonal plasmocytes and the intensity of the free
light chains production by the tumour cell population.
Multiple myeloma (MM) is a malignant hematological
disease characterised by the uncontrolled proliferation
and accumulation of neoplastically transformed plasma
cells, production of monoclonal immunoglobulin (MIG)
detectable in the serum and/or urine and various degrees
of organ involvement (hypercalcemia, renal insuffi ciency,
osteolytic bone lesions or osteoporosis, anemia and im-
munodefi ciency)1, 2. Apart from the complete molecule
of monoclonal immunoglobulin, the plasma cells also
produce single free light chains (FLC) which are not
fi xed to the MIG molecule3. In the last few years, the as-
sessment of free light chain serum levels has become an
irreplaceable part of the examination algorithm in diag-
nostics, screening and evaluation of treatment response
not only in patients with multiple myeloma but also in
other malignant or potentially malignant monoclonal
gammopathies4-6. The bone marrow microenvironment
in relation to the pathological plasmacellular infi ltration
is characterised by a very large system of intercellular and
cytokine interactions among the stromal cells of bone
marrow, bone elements and tumour cells themselves, in
close relation to their own biological qualities of the dis-
ease7. Some unusual biochemical parameters appear to
be very promising as possible future markers not only
in the neoplastic transformation of potential malignant
monoclonal gammopathy of undetermined signifi cance
(MGUS) and its diff erentiation from the early stage MM
but also as markers of advanced stage, progression and
monitoring of treatment response in patients with multi-
ple myeloma8-10. The aim of this study was to determine
the relation of FLC serum levels to 12 selected less usual
biological parameters in patients with multiple myeloma
who were examined at the time of diagnosis.
T. Pika, J. Minarik, P. Schneiderka, M. Budikova, K. Langova, P. Lochman, J. Bacovsky, V. Farbiakova, V. Scudla
MATERIALS AND METHODS
The sample studied included 102 patients examined
at the time of diagnosis and who fulfi led the SWOG and
IMWG criteria for multiple myeloma diagnosis11. The age
range was 43-82, median 63 years, men and women ra-
tio 1:1.04. Immunochemical type IgG was found in 58 %
(n = 60), IgA 22 % (n = 22), Bence – Jones 17 % (n = 17),
IgD 1 % (n = 1) and nonsecretoric type 2 % (n = 2). The
kappa light chain secretion was found in 68 % (n=69) and
lambda secretion in 32 % (n = 33) of patients. With the
use of Durie-Salmon staging system there 12 % (n = 12)
of patients were in stage I, 28 % (n = 29) in stage II and
60 % (n = 61) in stage III. The A substage (creatinine un-
der 177 μmol/l) was found in 75 % of patients12. Using the
International Prognostic Index (IPI) staging system 12 %
(n = 12) of patients were in the 1st stage, 22 % (n = 22)
in the 2nd stage and 66 % (n = 68) in the 3rd stage13.
Serum levels of free light chains were set by the FreeliteTM
Binding Site system (Immunotech, standard serum inter-
val: kappa 3.3-19.4 mg/l, lambda 5.7-26.3 mg/l, lambda
/kappa (K/L ratio) 0.26-1.65) (ref.3). Serum levels were
determined by the following methods: β2microglobulin
(β2m) by the radioimmunoanalysis method (RIA, stand-
ard range 0-2.5 mg/l), tymidine kinase (TK) by radioen-
zymatic method (REA, 0-6 IU/l), intercellular C-terminal
telopeptide collagen-I (ICTP, m: 2.1-5.0, w: 2.1-5.6 μg/l)
and N-terminal peptide procollagen-I (PINP, m: 21-78, w:
19-102 μg/l) by RIA method; vascular cell adhesive mol-
ecule-1 (sVCAM, 395-714 ng/ml), intercellular cell adhe-
sive molecule-1 (sICAM, 269-691 ng/ml), solubile form of
IL-6R (sIL-6R, 10-90 ng/ml) and solubile osteoprotegerine
(sOPG, 3.7-4.4 μmol/l) by enzymeimmunoassay method;
hepatocyte growth factor (sHGF, 671-1992 pg/ml), vascu-
lar endothelial growth factor (sVEGF, 62-707 pg/ml), syn-
decan-1/sCD138 (sSyn-1, 37-123 ng/ml) and Fas antigen
(sFas 4792-1750 pg/ml) were determined by a quantitative
sandwich enzymatic immunoassay method. For statistical
evaluation the Spearman Rank Correlation Coeffi cient
test was used. Kappa and lambda secretion, respectively
kappa and lambda groups were assessed separately due
to the reciprocal relation of serum levels and K/L ratio
of free chains.
Abnormal serum levels of free light chains were re-
corded in 92 % of patients and, a pathological K/L ra-
tio was found in 91 % cases. Abnormal β2m levels were
recorded in 95.1 %, in TK in 58.8 %, sIL-6R in 68.6 %,
sVCAM in 74 %, sICAM in 32.6 %, ICTP in 72.7 %, PINP
in 21 %; abnormal levels (under and above standard in-
terval) were recorded in case of sOPG in 97.3 %, sHGF
in 39.3 %, sVEGF in 7.7 %, sSyn-1 in 69.4 % and in case
of sFas antigen in 3.8 %. A signifi cant correlation of se-
rum levels of the kappa dominant chain and serum levels
of β2m (r = 0.344, p=0.005), TK (r = 0.263, p = 0.035),
ICTP (r = 0.402, p = 0.001), PINP (r = 0.264, p = 0.039),
sOPG (r = 0.328, p = 0.028), sSyn-1 (r = 0.255, p = 0.046)
and sFas (r = 0.418, p = 0.001) were found. In the case
of the K/L ratio and particular parameter assessment,
a correlation of the levels of β2m (r = 0.316, p = 0.01),
TK (r = 0.274, p = 0.027), ICTP (r = 0.346, p = 0.006),
PINP (r = 0.261, p = 0.042), sSyn-1 (r = 0.283, p = 0.026)
and sFas (r = 0.377, p = 0.002). In the lambda group the
correlation analysis confi rmed mutual dependence of the
dominant lambda chain level and levels of β2m (r = 0.476,
p = 0.003), ICTP (r = 0.375, p = 0.022), sVCAM-1
(r = 0.383, p = 0.019), sHGF (r = 0.441, p = 0.006) and
sFas (r = 0.334, p = 0.040). Likewise, an association of
K/L ratio and the levels of β2m (r = –0.473, p = 0.003),
TK (r = –0.412, p = 0.011), ICTP (r = –0.331, p = 0.045),
PINP (r = –0.409, p = 0.012), sHGF (r = –0.357,
p = 0.028), sSyn-1 (r = –0.449, p = 0.005) and sFas (r =
– 0.371, p = 0.022) was revealed. No signifi cant correla-
tion of alternative chain levels and monitored parameters
levels was found for any of the groups.
The results show the mutual relation of serum levels
of FLC, K/L ratio and selected untraditional markers of
multiple myeloma. The abnormal percentage of serum
free light chain levels and K/L ratio for the analysed group
approximately corresponds to previously published stud-
ies which have reported comparable pathological percent-
ages of FLC even if there was a higher percentage in the
case of the K/L ratio than FLC levels themselves in most
studies4, 14, 15. The correlation of FLC serum levels, K/L
ratio and β2m levels not only results from the biological
qualities of both parameters (level increase in case of renal
insufi ciency, polymerization), but also both parameters
are markers of the size of the tumour mass. The interval
levels of β2m, the most important prognostic factor in
multiple myeloma, are the new staging systems compo-
nents – International Prognostic Index (IPI), South-West
Oncology Group (SWOG) (ref.12, 16); there was a correla-
tion of these system stages and FLC levels, also the com-
bination of IPI system and serum levels of FLC and K/L
ratio appears to be very promising prognostic marker17-20.
The relation of FLC and serum tymidinekinase as prolif-
eration markers and as tumour mass markers is explain-
able as well; an association was found of FLC serum levels
and OLOMOUC staging system phases in which β2m and
TK serum levels were used21-23. The detected relation of
the bone metabolism parameters ICTP, PINP and FLC
levels can also be explained by the association of these
markers to the stage of bone involvement and also to the
progress and prognosis of the disease24, 25. Detected re-
lation of FLC, K/L ratio serum levels and Syndecan-1
confi rms the unique position of this less usual parameter.
Syndecan-1 is a proteoglycan released from the surface
of viable malignant plasma cells, mainly concerned with
the proliferation process, apoptosis of tumour and also
bone cells and relating bone involvement; infl uences the
cytokine net in direct and indirect way in conditions of
The correlation of serum immunoglobulin free light chain levels and selected biological markers
in multiple myeloma
the bone marrow microenvironment. It is considered to
be one of the most important molecules in the multiple
myeloma pathobiology26. Syndecan-1 appears to be a new
important prognostic marker, also a possible parameter
for discernment of monoclonal gammopathy of undeter-
mined signifi cance and multiple myeloma8, 9, 27-29.
Detected correlation of FLC, K/L ratio and sFas levels
is now more of the academical meaning which would be
observed. Solubile Fas antigen is a part of antiapoptotic
mechanisms participating in the immortality of tumour
The results confi rm the relation of serum levels of free
light chain kappa, resp. lambda and K/L ratio and several
less usual parameters in multiple myeloma. The study con-
fi rmed the association with tumour mass, biolocal quali-
ties of myeloma cells, parameters of bone metabolism,
myeloma bone disease and the bone marrow microenvi-
ronment itself. The conclusions of the study give source
materials for use of less usual markers for distinguishing
monoclonal gammopathy of undetermined signifi cance
from multiple myeloma and for evaluation of progress and
treatment response in patients with MM. Kappa/Lambda
ratio particularly seems to be more sensitive marker than
free light chains levels.
Supported by IGA MZ CR NR/9500-3 and VVZ MSM
1. Dispenzieri A, Lacy M Q, Greipp PR. Multiple Myeloma, p. 54.
In: Gertz MA, Greipp PR: Multiple myeloma and related plasma
cell disorders. Springer, Heidelberg, 2004.
2. Adam Z, Scudla V, Neubauer J. Mnohocetny myelom. In: Adam
Z, Vorlicek J et al. Hematologie II. Prehled malignich hematolo-
gickych nemoci. Praha, Grada publishing 2001: p. 461-502.
3. Bradwell AR. Serum free light chain analysis, 4th edition, p. 12-21.
The Binding site, Birmingham, 2006.
4. Scudla V, Minarik M, Schneiderka P, Kouril M, Kapustova M,
Vytrasova M et al. Vyznam stanoveni serovych hladin volnych le-
hkych retezcu imunoglobulinu v diagnostice a hodnoceni aktivity
mnohocetneho myelomu a vybranych monoklonalnich gamapatii.
Vnitr Lek 2005; 51(11): 249-1259.
5. Bradwell AR. Serum free light chain measurements move to center
stage. Clin Chem 2005; 51(5):805-807.
6. Mayo MM, Johns GS. Serum free light chains in the diagnosis
and monitoring of patients with plasma cell dyscrasias. Contrib
Nephrol 2007; 153:44-65.
7. Bacovsky J, Scudla V. Cytokinova sit u mnohotneho myelomu.
Vnitr Lek l994; 40 (8):517-520.
8. Englis M. Stanoveni volnych lehkych retezcu imunoglobulinu v seru
v diagnostice a monitorovani monoklonalnich gamapatii. http://
9. Scudla V, Pika T, Budikova M, Ordeltova M, Minarik J, Zemanova
M et al. Prispevek k hodnoceni vztahu angiogennich cytokinu a
vybranych biologickych ukazatelu k prognostickym faktorum mno-
hocetneho myelomu. Cas Lek Ces 2006; 145:929-935.
10. Scudla V, Pika T, Budikova M, Ordeltova M, Minarik J, Zemanova
M et al. Evaluation of selected biological parameters in stages of
multiple myeloma examined according to Durie-Salmon (D-S)
and International Prognostic Index (IPI). Haematologica 2007;
11. International myeloma working group criteria for the classifi ca-
tion of monoclonal gammapathies, multiple myeloma and related
disorders: a report of the International Myeloma Working Group.
Br J Hemat 2003; 121:749-757.
12. Durie BGM, Salmon SE. A clinical staging system for multiple
myeloma. Cancer 1975; 36:824-854.
13. Greipp PR, San Miguel J, Durie BGM, Crowley JJ, Barlogie B,
Blade J et al. International staging system for multiple myeloma. J
Clin Oncol 2005; 23:3412-3420.
14. Mead GP, Carr-Smith HD, Drayson MT, Bradwell AR. Serum free
light chain levels in patients with intact immunoglobulin myeloma.
Clin Chem 2003; 49:A107, D-61.
15. Katzmann JA, Abraham RS, Dispenzieri A, Lust JA, Kyle RA.
Diagnostic performance of quantitative kappa and lambda free
light chain assays in clinical practice. Clin Chem 2005; 51:5,878-
16. Jacobson JL, Hussein MA, Barlogie B, Durie BGM, Crowley JJ.
A new staging system for multiple myeloma pacients based on
the Southwest Oncology Group experience. Brit J Hemat 2003;
17. Pika T, Zemanova M, Minarik J, Schneiderka P, Bacovsky J, Scudla
V. A relationship of serum imunoglobulin free light chain levels
measurement to multiple myeloma staging. Trans Hemat dnes
18. Nowrousian MR, Brathorst D, Sammet C, Kellert M, Daniels R,
Schuett P et al. Serum free light chain analysis and urine immun-
ofi xation electrophoresis in patients with multiple myeloma . Clin
Cancer Res 2005; 11(24):8706-8714.
19. Kyrtsonis MC, Vassilakopoulos TP, Kafasi N, Anagnostopoulos
A, Delimpasi S, Terpos E et al. Prognostic implifi cation of the
combination SFLCR and ISS in MM. Haematologica 2007; 92
20. Kyrtsonis MC, Vassilakopoulos TP, Kafasi N, Maltesas D,
Anagnostopoulos A, Terpos E et al. The addition of sFLCR
improves ISS prognostication in multiple myeloma. ASH 2007,
Abstract No 1490.
21. Scudla V, Bacovsky J, Vytrasova M, Ordeltova M, Budikova M,
Papajik T et al. Prognosticke faktory a klinicke stazovaci systemy
u mnohocetneho myelomu v souboru 237 nemocnych lecenych
v obdobi 1991-2002 konvencni chemoterapii. Klin Onkol 2002;
22. Pika T, Zemanova M, Minarik J, Langova K, Scudla V. Vztah se-
rovych hladin volnych lehkych retezcu imunoglobulinu ke stupni
progrese mnohocetneho myelomu. Vnitr Lek 2006; 52(11):1120,
Abstract No 56.
23. Scudla V, Bacovsky J, Papajik T, Budikova M, Srovnalik K.
Tymidinkinaza sera u mnohotneho myelomu. Vztah k vybranym
laboratornim ukazatelum nemoci. Vnitr Lek 1994; 40:151-156.
24. Abildgaard N, Bentzen SM, Nielsen JL, Heickendorff L. Serum
markers of bone metabolism in multiple myeloma: Prognostic
value of the carboxy-terminal telopeptide of type I collagen. Brit.
J. Hematol. 1997; 96:103-110.
25. Bacovsky J, Scudla V, Vytrasova M, Budikova M, Myslivecek M.
Monitoring of bone resorption and bone formation in multiple
myeloma. Biomed Papers 2002; 146(2):59-61.
26. Dhodapkar MV, Sanderson RD. Syndecan-1 (CD 138) in myeloma
and lymphoid malignancies: A multifunctional regulator of cell
behavior within the tumor microenviroment. Leuk Lymph 1999;
27. Kyrtsonis MC, Vassilakopoulos TP, Siakantaris MP, Kokoris SI,
Gribabis DA, Dimopoulou MK et al. Serum syndecan-1, basic fi -
broblast growth factor and osteoprotegerin in myeloma patients
at diagnosis and during the course of the disease. Eur J Haematol
T. Pika, J. Minarik, P. Schneiderka, M. Budikova, K. Langova, P. Lochman, J. Bacovsky, V. Farbiakova, V. Scudla
28. Schaar CG, Vermeer HJ, Wijermans PW, Huisman W, Cessie S,
Kluin-Nelemans HC. Serum syndecan-1 in patients with newly di-
agnosed monoclonal proteinemia. Haematologica 2005; 90:1437-
29. Seidel C, Sundan A, Hjorth M, Turesson I, Dahl IMS, Abildgaard
N et al. Serum syndecan-1: a new independent prognostic marker
in multiple myeloma. Blood 2000; 95:388-392.
30. Shima Y, Nishimoto N, Ogata A, Fujii Y, Yoshizaki K, Kishimoto
T. Myeloma cell express Fas antigen/APO-1 (CD95) but only some
are sensitive to anti-Fas antibody resulting in apoptosis. Blood