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1 Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., Tokyo, Japan; 2 Department of Molecular Preventive Medicine, School of Medicine, The University of Tokyo, Tokyo, Japan; 3 Department of Internal Medicine and Molecular Science, Nagoya City University Graduate School of Medical Science, Nagoya, Japan; and 4 BioWa, Inc., Princeton, New Jersey
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ABSTRACT |
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 Top ABSTRACT INTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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INTRODUCTION |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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Rs) to the antibody constant region. ADCC is considered to be a major therapeutic function of antibodies, although there are others (e.g., antigen binding, induction of apoptosis, and complement-dependent cellular cytotoxicity; Refs. 1
, 2
).
FcRIIIa, the FcR mainly expressed on natural killer cells and responsible for ADCC activation, has two isoforms, 158Val and 158Phe. The FcRIIIa-158V allele shows higher binding capacity for IgG1 antibody compared with the FcRIIIa-158F isoform (3
, 4) . Importantly, Cartron et al. (5)
have reported recently that the anti-CD20 chimeric IgG1 antibody Rituxan was more effective for follicular non-Hodgkin lymphoma patients with FcRIIIa-158Val compared with patients with FcRIIIa-158Phe. Similar results have been reported by Anolik et al. (6)
of Phase I/II trials of Rituxan in the treatment of systemic lupus erythematosus. These reports underscore the importance of ADCC in the clinic.
One IgG molecule contains two asparagine N-linked oligosaccharide sites in its Fc region (7)
. The general structure of IgG N-linked oligosaccharides is complex-type, characterized by a mannosyl-chitobiose core with or without bisecting N-acetylglucosamine (GlcNAc)/L-fucose and other chain variants including the presence or absence of galactose and sialic acid. Several groups have reported that ADCC enhancement can be achieved by manipulating human IgG1 subclass antibody oligosaccharides. ADCC requires the presence of oligosaccharides in the Fc region and is sensitive to change in the oligosaccharide structure (8, 9, 10)
. Among all of the sugar components in the oligosaccharide, galactose (11
, 12)
, bisecting-GlcNAc (13
, 14)
, and fucose (15
, 16)
have been reported to affect ADCC. We clarified recently the greater importance of fucose among these sugar components; defucosylation of humanized anti-interleukin 5 receptor antibody or chimeric anti-CD20 antibody enhanced their ADCC >50-fold (16)
. Compared with fucose, the involvement of bisecting-GlcNAc in ADCC was minimal, and galactose did not contribute to ADCC (16)
. The influence of nonfucosylated oligosaccharide on ADCC has been also reported by Shields et al. (15)
using humanized anti-HER2 IgG1 and humanized anti-IgE IgG1. They showed that the improved ADCC of defucosylated IgG1 resulted from its improved binding to FcRIIIa. Defucosylation of human IgG1-type antibody is, thus far, one of the most powerful ways to improve antibody effector function. However, the superiority of defucosylated antibodies in vivo has yet to be proven.
Chinese hamster ovary (CHO) cell lines are one of the most widely used host cell lines for the production of recombinant pharmaceutical proteins. Many approved or developing therapeutic antibodies are produced by CHO cells, including Rituxan and the anti-HER2 IgG1, Herceptin, both of which are increasingly used in the treatment of non-Hodgkin’s lymphoma (17) and breast cancer (18) , respectively. Several in vivo and clinical studies indicate that ADCC is one of the essential therapeutic mechanisms of Rituxan and Herceptin (2 , 5 , 6) . However, the content of fucose in oligosaccharides of CHO-produced antibodies are relatively high, and their ADCC is much lower than that of defucosylated antibodies (15 , 16) .
Chemokine receptors mediate leukocyte migration through binding of soluble ligands. CC chemokine receptor 4 (CCR4) is a chemokine receptor that binds specifically to its ligands thymus and activation-regulated chemokine and macrophage-derived chemokine. CCR4 is expressed mainly on Th2-type CD4+ helper T cells in normal conditions (19, 20, 21) . Th2 cells regulate humoral immunity and are also thought to play a key role in immune disorders, such as allergies and autoimmune diseases (22) .
Several groups reported recently the selective expression of CCR4 on certain subsets of T-cell leukemia/lymphoma. Yoshie et al. (23) reported the frequent expression of CCR4 on adult T-cell leukemia (ATL) cells (22 of 24 cases were CCR4-positive by reverse transcription-PCR). ATL is the most aggressive and fatal leukemia, and there is currently no curative therapy. The very high rate of CCR4 positivity in ATL was also reported most recently by Ishida et al. (24) . Interestingly, their data demonstrated for the first time that the CCR4 expression correlates with poor prognosis. With respect to other T-cell malignancies, anaplastic large-cell lymphoma (25 , 26) , mycosis fungoides in transformation (25 , 27) , and cutaneous T-cell lymphoma (27) have been observed for the frequent expression of CCR4.
To verify the potential of defucosylated antibody with enhanced ADCC for anticancer drugs, we established the defucosylated chimeric anti-CCR4 IgG1 antibody KM2760. KM2760 has lower fucose content than an otherwise identical CHO-produced anti-CCR4 IgG1, KM3060, and exerts much higher ADCC when human peripheral blood mononuclear cells (PBMCs) are used as effector cells. We additionally established a T-cell leukemia mouse model engrafted with human PBMCs as effector cells. KM2760 showed significantly higher antitumor efficacy in this T-cell leukemia model compared with KM3060. The effectiveness of KM2760 against T-cell malignancy was also investigated in conventional syngeneic and xenograft mouse models.
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MATERIALS AND METHODS |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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Cell Lines.
CHO cell line DG44 (28)
was kindly provided by Dr. Lawrence Chasin (Columbia University, New York, NY). Rat hybridoma YB2/0 [American Type Culture Collection (ATCC) CRL-1662], mouse T-cell lymphoma cell line EL4 (ATCC TIB-39), human T-lymphoblastic leukemia cell line HSB-2 (ATCC CCL-120.1), human acute T-cell leukemia cell line Jurkat (ATCC TIB-152), human acute T-lymphoblastic leukemia cell line CCRF-CEM (ATCC CCL-119), and human cutaneous T-cell lymphoma cell line Hut78 (ATCC TIB-161) were purchased from ATCC. Human acute T-lymphoblastic leukemia cell line MOLT-4 (JCRB9031), TALL-1 (JCRB0086), and PEER (JCRB0830) were purchased from Japanese Collection of Research Bioresources (Tokyo, Japan). Human T-acute lymphoblastic leukemia cell line HPB-All was kindly provided by Dr. Ryuzo Ueda (Nagoya City University Medical School, Nagoya, Japan).
Establishment of Cells Producing Chimeric Anti-CCR4 Antibodies.
Hybridoma cells producing murine anti-CCR4 monoclonal antibody KM2160 was established by immunizing mice with a peptide corresponding to amino acid residues 2–29 of human CCR4 (21)
. KM2160 has been widely used as one of the most characterized antihuman CCR4 monoclonal antibodies (21
, 23
, 24
, 29, 30, 31)
. The heavy- and light-chain variable region cDNAs prepared from hybridoma cells by PCR were cloned into the pKANTEX93 chimeric IgG1 antibody expression vector (32)
to obtain the chimeric anti-CCR4 antibody expression vector pKANTEX2160. pKANTEX2160 was introduced into YB2/0 cells or DG44 cells via electroporation, and transfected cells were selected for gene amplification in methotrexate containing medium (32)
.
Production, Purification, and Oligosaccharide Profiling of Chimeric Anti-CCR4 IgG1s.
The chimeric anti-CCR4 IgG1s were produced by transfecting pKANTEX2160 into YB2/0 cells and CHO cells, and purified by the method described previously (16)
. YB2/0- and CHO-produced chimeric IgG1 were designated as KM2760 and KM3060, respectively. They were then subjected to the profiling analysis of oligosaccharides as described previously (16)
.
Construction of an EL4 Cell Line Stably Expressing CCR4.
A human CCR4 expression plasmid pcDNA3-CCR4 was kindly provided from Dr. Naofumi Mukaida (Kanazawa University, Ishikawa, Japan). A cytomegalovirus promoter was replaced with a CAG promoter in the BglII-HindIII site of plasmid pcDNA(CAG)-CCR3 (33)
to construct CAG-pcDNA-CCR4. EL4 cells were transfected with CAG-pcDNA-CCR4 by electroporation and grown in the presence of 0.5 mg/ml G418 sulfate to obtain G418-resistant clones. The CCR4 expression level of each clone was measured by flow cytometry as described below. The highest expressing clone was selected and designated CCR4/EL4.
Flow Cytometer Analysis.
Expression of cell surface CCR4 molecules was determined by flow cytometry. Leukemia T cells (2 x 106) were simultaneously stained with 10 µg/ml of biotin-labeled KM2760 and 250 µg/ml of human IgG (Welfide, Osaka, Japan) for blocking Fc receptors. Biotinylated KM2760 was prepared using EZ-Link Sulfo-NHS-LC-Biotin (Pierce, Rockford, IL) as described by the manufacturer. Phycoerythrin-conjugated streptavidin (Becton Dickinson Japan, Tokyo, Japan) was used as the secondary reagent. The stained cells were analyzed using a EPICS XL-MCL flow cytometer (Beckman Coulter, Tokyo, Japan).
Preparation of Murine Effector Cells.
Splenocytes were taken from C57BL/6 mice and stimulated in 50 ng/ml (>500 units/ml) recombinant human interleukin 2 (Peprotech EC, London, England) for 7 days. Thioglycollate-induced macrophages were taken from i.p. cavities of mice 4 days after the i.p. injection (2 ml/mouse) of 3% Brewer thioglycollate medium containing 0.3 mM thioglycollate (Difco, Detroit, MI).
ADCC Assay.
ADCC was determined by 4 h 51Cr release assay using PBMCs from healthy volunteers or interleukin 2-activated murine effector cells as effector cells as described previously (16)
. For macrophage-mediated ADCC, the incubation time was extended to 18 h.
Construction of Human PBMC-Engrafted Mice.
Human PBMCs were isolated from the peripheral blood of a healthy donor using Lymphoprep (Axis Shield, Dundee, United Kingdom). PBMCs (5 x 106) were injected i.p. in a volume of 0.2 ml suspended in PBS into SCID mice under sterile conditions. The mice were pretreated with a combination of irradiation using a 137Cesium irradiator (2.5 Gy, 3 days before PBMC injection) and 50 µl antiasialo GM1 antiserum (Wako, Richmond, VA) given i.p. 1 day before PBMC injection.
Assessment of Antitumor Activity in Hu-PBMC-SCID Mice.
CCR4/EL4 cells (5 x 104) were injected i.p. into SCID mice engrafted with human PBMCs, 4 days after PBMC injection. One, 3, and 5 days later, anti-CCR4 IgG1s were injected i.p. Mice were observed daily to monitor the death due to ascites tumor development.
Experiments in a Syngeneic Disseminated Tumor Model.
CCR4/EL4 cells (5 x 104) were injected i.v. into C57BL/6 mice on day 0. On days 1, 3, and 5, anti-CCR4 IgG1s were injected i.v. Twelve days after tumor injection, mice were anesthetized and sacrificed by exsanguination. Livers and kidneys were taken and their weight were measured as an indication of metastases. The organ weights were normalized by dividing by body weights. Treated versus control value was also used as an indication of metastasis, which was calculated as follows: treated versus control (%) = [(Organ weight of treated group) - (Organ weight of naïve group)]/[(Organ weight of control group) - (Organ weight of naïve group)] x 100. For in vivo depletion of natural killer (NK) cells, 50 µl antiasialo GM1 antisera were injected i.p. on days -3, -1, and 1, and metastases were evaluated on day 11. Almost complete elimination of NK cells in spleens of the treated mice on day 6 was confirmed using flow cytometry.
Experiments in a Human T-Cell Leukemia Xenograft Model.
BALB/c-nu mice were injected s.c. with CCRF-CEM cells (2 x 107). Three h, 3 and 6 days later, antibodies were injected i.v. Tumor volume was calculated by the following equation:
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Statistical Analysis.
Statistical significance of differential findings between experimental groups of animals was determined by two-tailed unpaired t test.
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RESULTS |
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; Table 1
). Although the difference in the contents of fucose was most prominent, there were also some changes in other sugar components, such as galactose or GlcNAc, their differences were not considered sufficient to contribute to the difference in ADCC (16)
. The CCR4-binding activities of KM2760 and KM3060 were measured by ELISA, and the two antibodies showed identical binding to CCR4 (data not shown).
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, respectively). The CCR4/EL4 cells were murine thymoma EL4 cells expressing high levels of exogenous human CCR4, established by transfecting the EL4 cells with an expression vector encoding CCR4. KM2760 exhibited much higher ADCC than KM3060 in all three of the experiments. The difference in potency was prominent especially for donor A (Fig. 2A
, the maximum cytotoxicity of KM2760 and KM3060 were 51% and 8%, respectively) and donor B (Fig. 2B
, 100% and 4%, respectively). KM2760 also showed higher potency for donor C; however, the maximum cytotoxicity of KM3060 was relatively high in this case (Fig. 2C
, 100% and 71%, respectively). These results suggest that defucosylated IgG1 antibodies exhibit much more potent ADCC than highly fucosylated CHO-produced IgG1s. This is true despite the considerable variation in ADCC of CHO-produced IgG1s among individuals.
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. A similar advantage of defucosylated IgG1 in E:T ratio-based observation has also been reported for humanized anti-HER2 IgG1 (15)
. We also confirmed by flow cytometry that KM2760 or KM3060 did not bind to parental EL4 cells not expressing human CCR4. Likewise, KM2760 and KM3060 did not exert any ADCC on the parental EL4 cells (data not shown), indicating that the potent cytotoxicity of KM2760 depends on the presence of CCR4.
T-Cell Lymphoma and Leukemia Cell Lines Frequently Expressing CCR4 Are Lysed by KM2760-Mediated ADCC.
To confirm the reactivity of KM2760 to neoplastic T cells, we used biotinylated KM2760 to stain eight randomly selected human T-cell leukemia/lymphoma cell lines. CCR4/EL4 cells were stained as the positive control. As shown in Fig. 3
, five of eight cell lines were significantly KM2760-positive (Fig. 3, A–E)
, indicating that T-cell leukemia/lymphoma frequently express CCR4. This finding coincides well with recent reports (23
, 25, 26, 27)
.
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, all of the target cell lines were lysed in the presence of KM2760, although the extent of antibody-dependent and independent cytotoxicities were varied among cell lines.
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. Our preliminary study showed that the engrafted PBMCs survive for at least 2 weeks in the peritoneal cavity, and human PBMCs obtained from peritoneal cavities of engrafted mice ex vivo induced potent ADCC by KM2760 and weak ADCC by KM3060 (data not shown), in the same manner as when fresh PBMCs were used as effector cells.
The antitumor activity of antibodies in human PBMC-engrafted mice is shown in Fig. 5A
. When CCR4/EL4 cells were injected i.p. 4 days after the engraftment of human PBMCs, without additional treatment the mice survived only for an additional 15.2 days on average because of the ascites tumor development. Significant survival prolongation was observed in mice that received either KM2760 or KM3060. However, both 1 µg and 10 µg of KM2760 were significantly more potent (the mean survival time, 22.7 days and 23.2 days, respectively) than the same and larger doses (10 µg and 100 µg) KM3060 (17.7 days and 18.0 days, respectively). In contrast, 10 µg KM2760 showed no significant survival prolongation in mice without human PBMCs (Fig. 5B)
, indicating that the antitumor efficacy shown in Fig. 5A
entirely depended on human PBMCs.
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). When splenocytes were used as effector cells, they showed increased natural cytotoxicity in the absence of IgG1 primarily induced by lymphokine-activated killer cells. However, regarding IgG1-dependent cytotoxicity, the activated splenocytes or macrophages could only show marginal activities, and no significant differences were found between the two IgG1s. Taken together, these data indicate that murine effector cells do not reproduce the enhanced ADCC of KM2760; consequently, murine effector system might differ from that of human regarding the mechanism of action of defucosylated IgG1.
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. Unexpectedly, KM2760 showed more potent efficacy than KM3060. The difference was more profound for kidney metastases, i.e., the weight of treated organs were almost the same as that of naïve animals, whereas the difference for liver metastases were found to be modest. Depletion of NK cells by administration of anti-asialo GM1 antisera did not significantly affect the treated versus control value of KM2760- and KM3060-treated organs (Fig. 6C)
. These results suggest that there is an unidentified mechanism of the superiority of KM2760 in murine effector system that differs from human effector mechanism in which enhanced ADCC of defucosylated IgG1 is due to the strong binding to FcRIIIa on NK cells (15)
. We also performed the same experiment with FcR chain-deficient mice of C57BL/6 background. In contrast with wild-type mice, the antitumor efficacy of both KM2760 and KM3060 were completely abrogated in FcR-deficient mice (data not shown), indicating that the antitumor efficacy observed in the syngeneic tumor model using wild-type mice depended on the activating signals via murine FcRs.
In Vivo KM2760 Antitumor Activity of KM2760 in Mice Bearing Human T-Cell Leukemia.
In vivo antitumor activity of KM2760 against human neoplastic T cells was also investigated in a xenograft model bearing murine effector system. We chose human CCRF-CEM cells for this study, because CCRF-CEM was found to be the only human cell line among all five of the selected CCR4-positive cell lines shown in Fig. 3, A–E
, which reproducibly formed a s.c. solid tumor in nude mice (data not shown). As shown in Fig. 7
, KM2760 showed a significant tumor formation delay (3–4 weeks later than the control group).
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DISCUSSION |
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TopABSTRACTINTRODUCTIONMATERIALS AND METHODSRESULTSDISCUSSIONREFERENCES |
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The high frequency of CCR4 expression on neoplastic T cells has been reported (29
, 31, 32, 33)
and is especially high in ATL (80–90%). Our results also showed that CCR4 is frequently expressed on neoplastic T-cell lines (5 of 8) and that all of these CCR4-expressing cell lines were lysed by KM2760-mediated ADCC. KM2760 is also expected to recognize and lyse ATL cells in patients, because Ishida et al. (24)
have shown that neoplastic cells from 
90% of ATL patients can be stained by KM2160, the original murine antibody of chimeric KM2760 sharing the same variable region with KM2760. Because CCR4 expression is mostly localized to Th2 cells (19, 20, 21)
, we expect low toxicity of CCR4-targeted therapy. There is currently no curative therapy for ATL, and the 4-year survival rate is only 5–6% for acute or lymphoma type patients (35)
. CCR4-targeted antibody therapy with potent effector function is, therefore, a promising candidate to overcome this difficulty.
KM2760 with low fucose-containing oligosaccharides (93% defucosylated) exerted potent ADCC with human PBMCs as effector cells. In this study, we observed that KM2760 ADCC with PBMCs from three donors was much higher than that of KM3060 (9% defucosylated oligosaccharides). Interestingly, KM3060 ADCC was relatively high in the experiment with donor C (Fig. 2C)
. We did not investigate the FcRIIIa genotypes of PBMC donors in this study; however, it may be that one of three donors used in this study carries the FcRIIIa-158Val genotype, which corresponds to an isoform capable of stronger binding to IgG1. In addition, KM2760 can exert potent ADCC in the presence of fewer effector cells (Fig. 2D)
. This feature could be therapeutically beneficial, because the number of effector cells able to penetrate into a large tumor mass is much less than available for circulating tumors.
In this report, the therapeutic efficacies of KM2760 and KM3060 were compared in a CCR4-positive T-cell leukemia model with human PBMC-engrafted mice. Human PBMCs can be engrafted into the peritoneal cavity of SCID mice for a limited duration (34) . We adapted this mouse model to reflect therapeutic efficacy of ADCC by human effector cells. In this mouse model, KM2760 was shown to be significantly more effective in survival prolongation than larger amounts of KM3060. This result suggested that the high potency of KM2760 was retained in vivo. In support of the clinical relevance of the model, the antitumor activity of KM2760 entirely depended on the presence of engrafted PBMCs. This is the first report to demonstrate the superiority of defucosylated IgG1 as a therapeutic agent in vivo. The potent in vivo efficacy of defucosylated IgG1 achieved with a very low dose could ameliorate the problem of cost, which is one of the major defects of antibody drugs.
Additional evidence of the effectiveness against T-cell malignancy was shown in a syngeneic or a xenograft mouse model that does not bear human effector cells. Using FcR chain-deficient mice, Clynes et al. (2)
have shown that the mechanisms of action in clinically effective Rituxan and Herceptin in such mouse models were FcR mediated, suggesting that antibodies with a human IgG1 constant region could exert antitumor activity in vivo via murine FcR on murine leukocytes. However, unlike human PBMC-mediated ADCC, the ADCC of the two anti-CCR4 IgG1s on CCR4/EL4 cells mediated by mouse effector cells were very weak and indistinguishable from each other in this study. On the other hand, we unexpectedly found that KM2760 was more efficacious than KM3060 in the syngeneic tumor model, especially for kidney metastases in a FcR-dependent way. Taken together, our results suggested that KM2760 functions in mice more efficiently than KM3060 via an unidentified mechanism, which differs from that in humans, and which may vary with the tumor microenvironment. A similar phenomenon has been reported for the case of anti-gp75 antibody, in which the antibody was capable of suppressing syngeneic melanoma in mice in a FcR-dependent manner, whereas the ADCC by murine effector cells could not be detected in vitro (36
, 37)
, although it was not a comparative study of fucose variants. This inconsistency between in vivo and in vitro activity profiles observed in the present study and in the case of anti-gp75 antibody lead us to postulate the two possible mechanisms: (a) murine ADCC effective in vivo sometimes might be too weak to be detected in vitro; or (b) the antibodies can attack tumor not in a direct cytolytic way in vivo. Because KM2760 and KM3060 does not exhibit complement-dependent cellular cytotoxicity nor direct cytotoxicity in vitro (data not shown), these effector functions are not likely to be important for anti-CCR4 IgG1s in mouse model. In addition, we also found the incapability of murine NK cells in in vivo efficacy of KM2760 and KM3060, implying the involvement of FcR-expressing effector cells other than NK cells, such as macrophages. The importance of macrophages in FcR-mediated IgG1 therapy in the mouse model has also been pointed out by Clynes et al. (2)
, as they found that the therapeutic efficacies of Rituxan and Herceptin were potentiated in mice that lack inhibitory receptor FcRIIb, which are expressed on macrophages but not on NK cells. In contrast to NK cells, which selectively express FcRIII, macrophages express all of the three subtypes of FcRs both in human and mouse systems. One possible explanation is that the in vivo superiority of KM2760 in mouse system may be due to the differential binding of the two fucose variant IgG1s to at least one of the FcRs on macrophages. At this point, we have no additional clues to how KM2760 functions in murine effector system, and it requires additional investigation. Considering the clinical relevance of the model, our results suggest that the improved ADCC of defucosylated human IgG1 could be evaluated only in a human effector system, such as human PBMC-engrafted mice.
The antitumor efficacy of KM2760 was also confirmed in the xenograft model bearing human leukemia CCRF-CEM cells. The efficacy in a xenograft model seemed to be somewhat modest, because even 400 µg of administration did not lead to complete suppression. A possible explanation for this is that the two mouse models differ in the accessibility of effector cells to tumor cells, i.e., tumor cells disseminated by blood flow in the syngeneic model can be easily accessed by mouse effector cells, and, therefore, more sensitive to antibody therapy compared with s.c. tumor model. In addition, a little lower CCR4 expression level in CCRF-CEM cells compared with CCR4/EL4 cells (Fig. 3)
may also contribute to the lower responsiveness of the xenograft model to the KM2760 therapy.
Many therapeutic antibodies currently approved or under clinical development are produced by CHO cells. Compared with YB2/0 cells, CHO cells express higher levels of 
1,6-fucosyltransferase, which catalyzes the fucosylation of complex-type oligosaccharides (16)
, and in consequence, CHO-produced antibodies are likely to have highly fucosylated oligosaccharides. In support of this, we recently analyzed the oligosaccharide profile of Rituxan and demonstrated that the content of nonfucosylated oligosaccharide was only 6% and the ADCC of Rituxan was much lower than that of YB2/0-produced defucosylated anti-CD20 antibody (16)
. Defucosylated antibody with enhanced ADCC and potent antitumor activity gives the additional value to the next generation of therapeutic antibodies.
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ACKNOWLEDGMENTS |
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FOOTNOTES |
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Requests for reprints: Kenya Shitara, Tokyo Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., 3-6-6 Asahi-machi, Machida-shi, Tokyo 194-8533, Japan. Phone: 81-42-725-0857; Fax: 81-42-725-2689; E-mail: kshitara{at}kyowa.co.jp
Received 7/11/03. Revised 1/ 9/04. Accepted 1/15/04.
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S. Iida, H. Misaka, M. Inoue, M. Shibata, R. Nakano, N. Yamane-Ohnuki, M. Wakitani, K. Yano, K. Shitara, and M. Satoh Nonfucosylated Therapeutic IgG1 Antibody Can Evade the Inhibitory Effect of Serum Immunoglobulin G on Antibody-Dependent Cellular Cytotoxicity through its High Binding to Fc{gamma}RIIIa. Clin. Cancer Res., May 1, 2006; 12(9): 2879 - 2887. [Abstract] [Full Text] [PDF]
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L. Sturla, F. Fruscione, K. Noda, E. Miyoshi, N. Taniguchi, P. Contini, and M. Tonetti Core fucosylation of N-linked glycans in leukocyte adhesion deficiency/congenital disorder of glycosylation IIc fibroblasts Glycobiology, October 1, 2005; 15(10): 924 - 934. [Abstract] [Full Text] [PDF]
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