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Documente Cultură
DOI 10.1007/s00262-007-0431-z
O R I G I N A L A R T I CL E
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Received: 26 October 2007 / Accepted: 21 November 2007 / Published online: 6 December 2007
Springer-Verlag 2007
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N. Yamamoto N. Yamamoto
Division of Cancer Immunology and Molecular Immunology,
Socrates Institute for Therapeutic Immunology,
Philadelphia, PA 19126, USA
H. Suyama
Nagasaki Immunotherapy Research Group,
Yokoh, Nagasaki, Japan
H. Nakazato
Yokoyama Gastroenterology Hospital, Nagoya, Japan
Y. Koga
Nakagawa Hospital, Fukuoka City 811-1345, Japan
Introduction
N. Yamamoto (&)
Division of Cancer Immunology and Molecular Biology,
Socrates Institute for Therapeutic Immunology,
1040, 66th Ave, Philadelphia, PA 19126-3305, USA
e-mail: nobutoyama@verizon.net
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InXammation-derived macrophage activation is the principal macrophage activation process which requires serum
vitamin D3-binding protein (known as Gc protein) [8, 16,
25, 26, 27, 29] and participation of B and T lymphocytes [7,
18, 19, 22, 23, 24]. Gc protein carries one trisaccharide
composed of N-acetylgalactosamine with dibranched galactose and sialic acid termini at 420 threonine residue
[25, 33]. This oligosaccharide is hydrolyzed by the inducible membranous -galactosidase (Bgli) of inXammationprimed (or lyso-Pc-treated) B cells and by the membranous
Neu-1 sialidase of T cells to yield a macrophage-activating
factor, the protein with N-acetylgalactosamine as the
remaining sugar (MAF) [25, 28] (Fig. 1a). Thus, Gc protein
is the precursor for the principal MAF [25, 28]. However,
the MAF precursor activity of cancer patient Gc protein is
lost or reduced, because their serum Gc protein is deglycosylated by serum -N-acetylgalactosaminidase (Nagalase)
secreted from cancerous cells [34, 35, 37, 39] (Fig. 1b).
Deglycosylated Gc protein cannot be converted to MAF,
resulting in no macrophage activation. Macrophages are the
major phagocytic and antigen-presenting cells. Since macrophage activation for phagocytosis and antigen presentation to B and T lymphocytes are the Wrst indispensable
steps in the development of both humoral and cellular
immunities, lack of macrophage activation leads to immunosuppression [32, 34, 35, 37, 39, 42, 44, 45]. Advanced
cancer patients have high serum Nagalase activity, resulting
in no macrophage activation and severe immunosuppression that explain why cancer patients die with overwhelming infection (e.g., pneumonia) [37].
Stepwise treatment of highly puriWed Gc protein with
immobilized -galactosidase and sialidase generates
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administration and used for prognostic analysis. Assessment of patient response to each GcMAF administration
was performed by determining both the MAF precursor
activity of serum Gc protein and serum Nagalase activity.
Since serum Nagalase activity is proportional to tumor burden [11, 37, 46], assessment of curative response to
GcMAF therapy was performed by determining serum
Nagalase activity as a prognostic index.
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generates <0.85 nmol superoxide/min/106 cells, the precursor activity of patient serum Gc is considered to be lost.
Results
Therapeutic history and immunodiagnostic parameters
of metastatic colorectal cancer patients
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No.
Patient
Age
Sex
Therapeutic history
Surgery
Radiat./chemo.
Precursor activitya
superoxide (nmol)
NaGalaseb
(nmol/mg/min)
71
Colon resect.
Chemotherapy
1.88
2.72
63
Rectum resect.
Radiat./chemo.
2.01
2.56
68
Colon resect.
Chemotherapy
0.84
4.84
41
Colon resect.
Chemotherapy
0.72
6.71
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51
Colon resect
Chemotherapy
2.29
1.78
79
Rectum resect.
Radiat./chemo.
2.14
2.57
55
Colon resect.
Chemotherapy
1.68
3.28
82
Colon resect.
Chemotherapy
1.29
3.89
5.12
0.39d
C
a
b
c
d
Healthy humanc
Precursor activity <0.85 is unable to support activation of macrophages and that is considered to be loss of activity
Nagalase assayed before entering GcMAF therapy
Average of six healthy humans
This activity level is enzyme activity of -galactosidase and not of Nagalase
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Table 2 Correlation of the MAF precursor activity of colorectal cancer patients with their serum -N-acetylgalactosaminidase activity
2.15
2.02
2.21
1.89
2.75
1.55
20
26
1.15
3.18
1.03
3.20
1.01
3.21
0.99
3.25
0.97
3.45
0.82
4.08
0.68
2.01
2.56
2.08
2.41
2.25
2.36
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2.43
17
2.72
2.01
14
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1.88
11
NaGalase
(nmol/min/mg)
2.31
2.28
2.42
1.97
10
2.51
1.85
11
2.91
1.62
15
3.21
1.47
19
3.69
1.03
25
3.80
0.88
36
3.93
0.80
41
4.01
0.53
45
4.11
0.44
0.84
4.84
1.18
4.48
1.24
4.02
1.33
3.77
1.44
3.42
Precursor activity
superoxide (nmol)
Time assay
week
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2.12
3.38
10
2.22
3.02
16
2.31
2.38
25
2.44
1.92
32
2.55
1.75
40
3.98
0.99
46
4.01
0.62
65
4.21
0.40
0.72
6.71
1.06
5.48
1.31
4.92
1.34
4.82
1.35
4.77
1.37
4.68
11
1.34
4.40
16
1.41
4.09
19
1.49
3.86
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Precursor activity
superoxide (nmol)
NaGalase
(nmol/min/mg)
23
1.52
3.32
27
1.55
3.68
30
2.11
2.34
34
2.34
1.76
43
3.10
1.25
52
3.62
0.63
64
3.83
0.50
2.29
1.78
2.41
1.62
2.40
1.55
3.21
1.33
3.32
0.85
13
3.42
0.78
15
3.81
0.66
25
4.29
0.55
Time assay
week
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Patient no.
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Table 2 continued
Discussion
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As we reported previously, GcMAF has a potent antiangiogenesis activity [9]. Thus, GcMAF has another antitumor
activity via antiangiogenesis. These Wndings were conWrmed by the group of Folkman [10]. This antiangiogeneic
eVect of GcMAF on malignant tumors is growth inhibition
of the tumors. In contrast, GcMAF is the most potent macrophage-activating factor that can greatly activate macrophages to eradicate a variety of cancers [46]. For the
assessment of therapeutic eYcacy of GcMAF for metastatic
colorectal cancer patients, GcMAF was used as a single
remedy modality. Therefore, anemic cancer patients were
excluded from GcMAF therapy.
Approximately 85% of patients diagnosed with colorectal cancer can have an operation intended for cure [2].
However, micrometastasis and recurrent tumors are commonly noted. Micrometastatic cells in bone marrow have
been shown to correlate with a poor clinical outcome [2].
Micrometastasis in bone marrow lead to the recurrence and
cause anemia. Tumor resection of anemic patients aborts
anemia. Then, they became eligible for GcMAF therapy.
Even patients deemed to have incurable cancer should have
palliative resection of tumors to abort anemia, prevent
obstruction, and reduce the risk of invasion to adjacent
organs such as bladder [4]. As soon as they became nonanemic, they should be eligible for GcMAF therapy.
Administration of GcMAF to cancer patients is a very
eVective procedure for cancer therapy in spite of the presence of high Nagalase activity in patient sera [46].
Although Gc protein is eYciently deglycosylated by serum
Nagalase secreted from cancerous cells [11, 34, 36, 37]
(Fig. 1b), serum Nagalase has no eVect on GcMAF. Thus,
more abnormality, the activated macrophages kill undiVerentiated cells more rapidly than diVerentiated cells [46].
Thus, the rapidly decreasing serum Nagalase activities
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References
1. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of
protein-dye binding. Anal Biochem 172:248254
2. Braun S, Pantel K (1999) Micrometastatic bone marrows involvement detection and prognostic signiWcance. Med Oncol 16:154
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