• cancer vaccine;
  • glycolipid;
  • glycosylation;
  • immunotherapy;
  • monoclonal antibody


  1. Top of page
  2. Summary
  3. Introduction
  4. Function of GSLs
  5. Conclusions
  6. Acknowledgements
  7. Disclosure
  8. References


Metabolic Diseases, Host Responses, Allergies, Autoinflammatory Diseases, Type 1 diabetes and viruses.

Research into aberrant glycosylation and over-expression of glycolipids on the surface of the majority of cancers, coupled with a knowledge of glycolipids as functional molecules involved in a number of cellular physiological pathways, has provided a novel area of targets for cancer immunotherapy. This has resulted in the development of a number of vaccines and monoclonal antibodies that are showing promising results in recent clinical trials.


  1. Top of page
  2. Summary
  3. Introduction
  4. Function of GSLs
  5. Conclusions
  6. Acknowledgements
  7. Disclosure
  8. References

Membrane glycolipids represent an unexplored source of tumour-associated antigens. Many of these are over-expressed and altered in tumours compared to normal tissue. The glycome exceeds the complexity of the genome and proteome because of the large array of possible carbohydrate modifications and lipid backbones. There is evidence for the role of glycosylation in most cellular processes [1]. However, little attention has been directed at this area of study due to the difficulties in the structural and functional concepts of glycosylation. More recently, the expression mechanisms of these glycosyl epitopes, in terms of their respective glycosyltransferase genes [1] and their organization and function with membranes, has been elucidated. Antibodies which recognize tumour glycolipids are strong activators of complement and strong mediators of antibody-dependent cellular cytotoxcity (ADCC)/antibody-dependent cellular phagocytosis (ADCP). In addition, a large number of monoclonal antibodies (mAbs) targeting tumour glycolipids have the ability to induce direct cell death in target cells without the need for effector cells or complement [2–7].

Glycolipids consist of a lipid tail with a carbohydrate head and constitute about 3% of the outer monolayer of the plasma membrane [8]. They can be divided into three main groups, glycoglycerolipids, glycosylphosphatidylinositols (GPI) and glycosphingolipids (GSL), based on the type of lipid component. Of the three types, GSL are most important as targets for tumour immunotherapy, as they are widely up-regulated in cancers.

Synthesis and structure of GSL

GSL biosynthesis begins with ceramide (a sphingosine and a fatty acid). Most commonly, a glucose (Glc) is added to the ceramide by the Type I transmembrane protein glucosylceramide (GlcCer) synthase, forming GlcCer [9,10]. Galactose (Gal) is then added to GlcCer by β-1,4-galactosyltranferases forming lactosylceramide (LacCer) in the lumen of the Golgi apparatus (Fig. 1) [11,12].


Figure 1. The structure of LacCer. The core complex for most GSLs. Ceramide is produced through the sphingomyelin hydrolysis, de novo or salvage pathways and is made up of sphingosine and a fatty acid. To the ceramide Glc is added followed by Gal, producing LacCer. The addition of more monosaccharides can produce hundreds of different GSLs. Alternatively Gal can be added before Glc to the ceramide producing GalCer.

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LacCer is the acceptor for various transferases that generate three major classes of GSLs; Lacto(neo), globo series and gangliosides [13]. Alternatively, Gal can be added to the ceramide forming GalCer [12] which leads in turn to the synthesis of less common structures, including GM4 and sulphatide [12]. LacCer provides the basis of all glycosphingolipids, which are elongated with the addition of further sugars and sialic acid. A well-defined series of sialic acid and galactose addition leads to the synthesis of a number of gangliosides (e.g. GD3, GD2, GM3, GM2; Fig. 2). Lewis antigens can be added onto LacCer, which is mediated by a number of fucosyltransferases.


Figure 2. Schematic representation of the major pathways of ganglioside biosynthesis. The monosialoganglioside GM3, derived from lactosylceramide, is the common precursor for both ‘a’ and ‘b’ pathway gangliosides. Each ganglioside species consists of a ceramide backbone (CER), and a carbohydrate chain (Glc, glucose; Gal, galactose; GalNAc, N-acetylgalactosamine) containing one or more sialic acid (SA) residues. ‘a’ and ‘b’ pathway gangliosides downstream of GD1b/GM1a synthase were designated complex ‘a’ (CaG) and complex ‘b’ (CbG) gangliosides, respectively. Parallel steps in both pathways are catalysed by the same glycosyltransferases of the Golgi apparatus: GD3 synthase (α-2,8-sialyltransferase), GM2/GD2 synthase (β-1,4-N-acetylgalactosaminyltransferase); GD1b/GM1a synthase (β-1,3-galactosyltransferase); GT1b/GD1a synthase (β-2,3-sialyltransferase); GQ1b/GT1a synthase (α-2,8-sialyltransferase). Adapted from [13]. The letter refers to the number of sialic acids present in the glycan motif (M, mono-; D, di-; T, tri-; Q, quad-).

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Function of GSLs

  1. Top of page
  2. Summary
  3. Introduction
  4. Function of GSLs
  5. Conclusions
  6. Acknowledgements
  7. Disclosure
  8. References

GSLs perform many functions on the surface of cells. One main characteristic of GSLs that allows them to perform such versatile roles is their ability to form clusters. These clusters, termed ‘glycosynapses’, are able to interact with functional molecules on the cell surface [1]. Significantly, they have contrasting properties with lipid rafts (Table 1[14]). Glycosynapses rely on the ability of GSLs to bind via cis-carbohydrate to carbohydrate interactions, forming clusters in membrane and interacting with functional proteins.

Table 1.  Contrasting properties of lipid raft and glycosynapses. Adapted from Hakomori, 2002 [112].
Lipid raftGlycosynapse
1% Triton X-100 insoluble1% Triton X 100 soluble
0·5% Brij95 insoluble
Cholesterol-dependent (disrupted by cholesterol-binding reagent)Cholesterol-independent (resistant to cholesterol-binding reagent)
Tetraspanin-independentTetraspanin dependent
Diameter 10 nm to < 100 nmDiameter > 100 nm, usually 500–1000 nm
Highly mobileLess mobile – non-mobile
Not involved in cell adhesionInvolved in cell adhesion with concurrent signalling

Currently, three types of glycosynapses have been described (Fig. 3). ‘Glycosynapse 1’ represents a GSL–GSL or GSL–binding protein interaction between cells, mediated by the glycosyl epitope of the GSL [15]. The main GSL in these synapses mediates cell adhesion that leads in turn to activation of cytoplasmic signal transducers (e.g. TDa, TDb, TDc). This activation leads to downstream signalling and changes in transcription factor expression. This can lead to increased growth factor signalling, cell adhesion, spreading and enhanced cell motility [15,16]. ‘Glycosynapse 2’ is the term used for cell to cell adhesion based on O-linked mucin-type glycoproteins that are recognized by carbohydrate binding proteins [17]. ‘Glycosynapse 3’ refers to the adhesion of a cell to the extracellular membrane (ECM), which is mediated by N-glycosylated adhesion receptors complexed with tetraspanin and GSL [18–20]. GD2, GD3 and GM3 gangliosides have been shown to be associated with the tetraspanin CD51 in glycosynapses [18,21]. Alteration of this glycosylation affects interaction of integrin with tetraspanin, leading to significant inhibition or promotion of cell motility [22]. The activation of transmembrane receptors in type III glycosynapses also leads to intracellular signalling and changes in transcription factor expression [20].


Figure 3. Schematic representations of glycosynapses. Type I glycosynapse with GSL clusters, proteolipid tetraspanin (PLtsp) and growth factor receptor. Type III glycosynapses contain N-glycosylated transmembrane adhesion receptors (usually integrin; ITR) complexed with tetraspanins (Tsp) and GSLs. Type III glycosynapse with integrin receptor (ITR) having α- and β-subunits and tetraspanin (Tsp.).

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Alterations in glycosylation in cancer

Glycolipids play an important role in many pathways of cellular physiology, and changes in the sugar head of the glycolipid can confer advantages to cancer cells. It has long been known that alterations of GSL, O- and N-linked oligosaccharides provide a survival advantage to tumours [23,24].

Incomplete synthesis.  Studies have shown that some cancers lack enzymes involved in the complete synthesis of some carbohydrate determinants due to silencing of the genes responsible. This is due to DNA methylation of transferases involved in glycan production and histone deacetylation [25]. Incomplete synthesis of the normally expressed glycans sialyl- 6-sulpho Lewis x and disialyl Lewis a leads to the increased expression of sialyl Lewis x and sialyl Lewis a [26,27]. Over-expression of sialyl Lewis a on either glycolipids or glycoproteins increases tumour cell adhesion and motility, resulting in increased metastasis [28,29].

Neo-synthesis.  As well as the incomplete synthesis of glycans on tumour cells, aberrant glycans can be expressed through neo-synthesis [30]. This is mediated by hypoxic regions of solid tumours, resulting in the enhanced transcription of glycosyltransferases [31,32]). The increase in these factors in response to hypoxia leads to the increase in glucose uptake (glucose-transporter type-I) leading to increased cell growth as well as an increase in glycan synthesis, leading to the increased expression of glycans including E-selectin ligands sialyl Lewis x and sialyl Lewis a [31].

Abnormal fucosylation.  Fucosylation is a common modification of glycoproteins. Up-regulation of enzymes involved in the addition of fucose residues to glycans can lead to an increase in fucosylation in cancer [33]. For example, α-fetoprotein (AFP) is increased in chronic hepatitis and liver diseases, but fucosylated AFP is over-expressed only in hepatocarcinomas (HCC). For this reason, it was approved as a tumour marker for HCC by the Food and Drug Administration (FDA) in 2005 [34].

Over-expression.  Gangliosides are sialylated glycolipids present on normal tissues, but have been shown to be over-expressed in malignant melanoma and tumours from neuroectodermal origin [35–37]. The glycolipid isoglobotetraosylceramide (IsoGb4) has been shown to be a marker of metastasis and over-expressed on tumours, with limited expression on normal tissue [38]. Sialyl Lewis a has been shown to be over-expressed in a range of cancers, including colorectal [39], breast [40] and ovarian [41,42] cancer. It is used as a serum marker in a range of cancers, including colorectal cancer to measure a patient's response to therapy [43]. GD2 is a disalylganglioside antigen expressed on the surface of tumours of neuroectodermal origin, including neuroblastoma and melanoma [44]. In addition, GD2 is also expressed on glioma and non-small cell lung cancer [44]. It is expressed abundantly on 100% of neuroblastoma tumours, regardless of stage [45]. Normal tissue expression is limited to neurones, skin melanocytes and peripheral fibres. Furthermore, GD2 has been shown to be involved in cell growth and differentiation and apoptosis [46]. The ganglioside GD3 has been described as a melanoma marker due to its over-expression [35] in this cancer and restricted normal expression. GM2 is over-expressed on a range of cancers, including malignant melanoma and neuroblastomas [47]. Enhanced expression of the Gb3Cer/CD77, a globotriaosylceramide, has been observed in most pancreatic and colon adenocarcinomas. As it is the receptor for Shiga toxin, the B-subunit of this toxin has been used a novel therapeutic strategy [48]. Studies have shown that some gangliosides can also be shed into the tumour microenvironment [49]. For example, GM2, GM3 and GD1a gangliosides have been shown to be shed by medulloblastoma cell lines [49]. Secreted gangliosides can be internalized by activated T cells triggering their apoptosis [50] and providing a mechanism for cancers to evade the immune system.

Abnormal sialylation.  Sialic acid is a common terminal addition to glycans in both normal and cancer tissues. As discussed previously, an increase in sialylated Lewis antigens can be observed on a range of cancers. The two most common forms of sialic acid expressed on mammalian cells are N-acetlyneuraminic acid (NeuAc) and N-glycolylneuraminic acid (NeuGc). The enzyme cytidine monophosphate (CMP)-N-acetylneuraminic acid converts CMP-NeuAc to NeuGc through hydrolysis. However, this enzyme is not expressed by humans due to a genetic mutation, resulting in the inability to synthesis NeuGc. However, it has been shown to be expressed by some cancers [7,51–53]. It has been shown that some cancers have a greater ability to incorporate the NeuGc from the diet into glycans through a scavenger pathway [54,55]. A possible advantage and explanation of tumour cells expressing NeuGc may be due to immunosuppression, with the ability of NeuGc to down-regulate CD4 on T cells in the tumour environment [56].

Glycolipids as targets for immunotherapy

The alteration of glycan structures on glycolipids suggests that they may be good targets for vaccines or mAbs.

Glycolipids as cancer vaccines.  Many issues complicate the use of glycan vaccines. First, isolation of the glycolipid from cancer cells is very difficult due to the heterogeneity of cell surface glycosylation. More recently, synthetic organic chemistry provides a solution to this problem [57]. Secondly, as glycolipids are often T cell-independent antigens (TI) they often elicit short-lived, low-affinity immunoglobulin (IgM) responses that lack memory. In addition, as most glycolipids are ‘self-antigens’ expressed at low levels on normal cells, they tolerize the immune system and their immunogenicity is low. Glycolipids are also shed into the blood by growing tumours, further reinforcing their immunotolerance. Studies have shown that TI antigens cross-link the B cell receptor (BCR) multi-valently, giving B cells a primary signal which results in proliferation [58,59]. However, Ig affinity maturation and production requires further signals, including ligation of CD40 and production of cytokines. This can be achieved by linking a synthetic glycan to a T cell carrier such as keyhole limpet haemocyanin (KLH), bovine serum albumin (BSA) or tetanus toxoid (TT) [60].

Vaccination of GD2–KLH gave little antibody response, whereas an alternative molecule GD2–lactone–KLH gave an antibody response in 83% of patients. Interestingly, patients immunized with the highest dose of GD2–lactone–KLH gave a high antibody response with the ability to induce CDC in GD2-positive cell lines [61]. Patients immunized with synthetic fucosyl GM1–KLH in small cell lung cancer plus QS-21 showed IgM responses, but only one patient mounted an IgG response at low titre [62]. A GD3–KLH conjugate failed to raise an antibody response in humans [63]. However, melanoma patients injected with GD3–lactone–KLH plus QS-21 stimulated IgM responses. The most promising KLH conjugate was GM2. Early vaccines utilized GM2 alone plus bacillus Calmette–Guérin (BCG) or CX (cyclophosphamide), which gave a predominantly IgM response in Phase I trials, with some reports of an IgG response. Despite the lack of an affinity-maturated, class-switched antibody response, a highly significant increase in disease-free interval and overall survival in 17% patients was observed in patients treated with GM2/BCG [64]. In order to increase the immunogenicity of GM2, it was conjugated to the T cell carrier, KLH. This resulted in increased titres of both IgM and IgG responses in patients [65]. As well as increasing the immunogenicity of GM2 by adding KLH, other adjuvants were tested, including Dextox [monophosphoryl lipid A (MPL) and purified mycobacterial cell-wall skeleton-based adjuvant] and QS-21 (a saponin-based adjuvant) which out-performed other adjuvants [66]. The GM2–KLH vaccine showed promise in early clinical trials [67]. In the Phase III study, although there was a clear correlation between antibody response and survival, there was no clinical benefit from vaccination when correlated with placebo or no treatment groups [68].

In an attempt to raise an IgG response to this ganglioside, an anti-idiotypic monoclonal antibody that mimics GD3, BEC2 [69] was used as an immunogen. Despite early encouraging results in lung cancer patients, a Phase III trial failed to show any survival advantage for vaccinated patients [70].

NeuGc has been studied as a possible target for therapy [71–73] due to its absence from normal human tissue. There are two NeuGcGM3 ganglioside-based vaccines currently in Phase II clinical trials. The first racotumumab (known formerly as 1E10) is an anti-idiotype murine monoclonal antibody, which is a mirror image of an antibody that specifically recognizes NeuGcGM3. Thus, anti-idiotype antibodies can act as antigens inducing a response against the original antigen. The alternative vaccine is NeuGcGM3/VSSP, which results from conjugation of the ganglioside into very small-sized proteoliposomes (VSSP) derived from Neisseria meningitides. Both vaccines had acceptable safety outcomes and were able to induce specific humoral and cellular immune responses in patients. The response to vaccination was stronger in patients with lower tumour burden, better performance status and a good response to previous treatment [74–77]. The current Phase III trials will confirm any survival benefit.

Immunization with GM3 plus Salmonella minnesota mutant R595 vaccine did not induce an antibody response in mice [78]. However, incorporation of GM3 into very small-sized proteoliposomes produced by using anionic detergents to incorporate gangliosides into the outer membrane protein complex (OMPC) of N. meningitidis produced an IgG response in chickens, mice and monkeys [79]. Unfortunately, when this vaccine went into Phase 1 study in 26 patients with metastatic melanoma, it showed IgM responses in only some patients. There were, however, signs of tumour regression in two patients [80].

A sialyl Lewis a–KLH conjugate [81] and a Lewis y–KLH conjugate have also induced anti-carbohydrate antibodies. The latter was of interest, as several patients made responses to the Lewis y hapten that only recognized glycolipid and not glycoproteins [82]. Human antibodies recognizing tumour cells that could induce complement-mediated lysis were induced with all these vaccines; however, they were of low titre and predominantly of the IgM subtype, which have reduced ability to penetrate solid tumours compared to IgG.

To enhance further the immunogenicity of carbohydrate vaccines, new approaches are currently being tried. Attachment of a protein carrier to a glycan is often problematic, as the chemistry is difficult to control and gives wide batch variations [57]. Another major drawback is that the carrier proteins are highly immunogenic, leading to suppression of the anti-glycan response. A more potent vaccine targeting glycolipids is therefore required. Glycolipids can be processed by B cells and presented on CD1d [83], a major histocompatibility complex (MHC)-class 1-like molecule, to natural killer (NK) and NK T cells [84,85]. These antigens need to be multimerized and are often presented within liposomes [38,76,79]. Evidence from a study immunizing mice with bacterial glycolipids incorporated within liposomes and mixed with an anti-CD40 mAb displayed an increased antigen-specific antibody response to the pathogen with an increase in class-switching to IgG, showing that the anti-CD40 mAb could substitute for T cell help [86]. More recently, fully synthetic carbohydrate vaccines incorporating a glycan, the Toll-like receptor (TLR)-2 activator Pam2CysSK4, and a T cell epitope incorporated within liposomes, stimulated high IgG antibody titres [57].

However, whether these new approaches can overcome tolerance in humans and stimulate high titre, potent IgG antibody responses remains to be tested. It seems more logical to develop human monoclonal IgG antibodies, which can be administered repeatedly in high amounts.

Monoclonal antibody targeting of tumour-associated glycolipids.  In contrast to the thousands of mAbs recognizing protein antigens, only a limited number of anti-tumour glycolipid antigens have been identified. Of great interest, however, is that many of these mAbs induce direct killing by oncosis as well as mediating potent antibody-mediated cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC).

Lewis antigens.  A range of Lewis y antibodies have been identified, but a consistent problem with Lewis antibodies has been a degree of cross-reactivity with Lewis x and H type 2 structures, causing red blood cell agglutination and gastrointestinal toxicity [87–89]. More recent studies have shown that even this cross-reactivity of anti-glycan mAbs has been underestimated [90]. We have raised a new mAb, FG27, against Lewis y expressing glycolipids. In contrast to anti-Lewis y mAbs raised against cells, they are very specific and do not cross-react with other Lewis antigens such as Lewis-X (BR96 mab), Lewis b (SC101), B blood group (BR96), H blood group (BR55) or bi-antennary Lewis y antigens. FG27 failed to stain liver, lung, colon, jejunum, breast, kidney and the ileum, which contrasts with the other Lewis y cross-reactive mAbs. Indeed, its only cross-reactivity with normal tissues is against duodenum and stomach. Moreover, our monoclonal antibodies recognize Lewis y exposed at the cell surface and mediate oncosis, ADCC and CDC, which does not occur in the mAbs raised against Lewis y conjugated directly to the T cell carrier KLH [87].

CA19·9 is a mAb recognizing sialyl Lewis a [also known as carbohydrate antigen 19·9 (CA19·9)][42]. SC104 is a novel mAb inducing direct killing as well as ADCC/CDC. It recognizes sialyltetrasoyl ceramide [91] and is shortly to enter Phase I clinical trials. A human anti-sialyl Lewis a mAb was produced using peripheral blood mononuclear cells (PBMCs) isolated from a breast cancer patient undergoing sialyl Lewis a–keyhole limpet haemocyanin (KLH) treatment [92]. This mAb has shown specific binding to sialyl Lewis a alone, and promisingly induces ADCC and CDC of antigen-positive cell lines as well as anti-tumour activity in a xenograft model.

GD2.  A number of anti-GD2 mAbs have been produced, including 14.G2a, ch14·18 [93,94], ch.60C3 [5], 3F8 [95] and KM8138 [96], to target neuroblastoma, melanoma and non-small cell lung cancers. The benefit of targeting GD2 on the surface of cells is that, unlike other gangliosides studied, GD2 is not shed by the cells into the microenvironment [97]. 14·18 is an IgG3 murine mAb targeted to GD2 and 14.G2a is a class-switch variant developed to enhance the ADCC effect of the mAb. 14.G2a has also been shown to induce CDC in vitro. When administered with IL-2, 14.G2a showed minimal effectiveness and suffered similar human anti-mouse antibody (HAMA) responses to 3F8 [94]. To overcome the HAMA responses seen with the murine mAbs, chimeric mAbs ch14·18 and c.60C3 were produced. ch14·18 was shown to be more effective than its murine counterpart, with overall survival greater than with maintenance therapy [94]. The humanized mAb Hu18K322A was created from ch14·18 in order to increase the half-life of the mAb in vivo. As well as being humanized, Hu18K22A was engineered with an amino acid change in the Fc region at position 322 and produced in the YB2/0 cell line rather than Chinese hamster ovary (CHO) lines, which lacks fucosylation in the Fc region, with the purpose of increasing the efficacy of CDC and ADCC in vivo[98]. Currently, the use of anti-GD2 mAbs in clinical trials in conjunction with chemotherapy is the mainstay of neuroblastoma therapy [99].

GD3.  The ganglioside GD3 has been described as a valid target for mAb therapy due to anti-GD3 mAb-mediated melanoma cell lysis being observed with the mAb MB3·6 [100]. R24 is a mouse mAb that recognizes GD3 and has undergone numerous clinical trials [101]. It has been used to treat melanoma patients, and in one trial showed a complete response in one patient who lasted 2 years and a partial response in one patient who lasted 2 months [102]. It also displayed a human anti-mouse antibody response in patients, but the low level of efficacy in patients meant that the antibody was not humanized [103].

GM2.  DMF10·167·4 is a hamster mAb raised against a murine T cell lymphoma cell line and has been shown to induce apoptosis of that cell line in vitro[104]. Subsequently it was found to bind to GM2 and could bind to both melanoma and small cell lung cancer cell lines, but showed minimal binding to normal tissues [105]. Its ability to induce apoptosis in a range of cell lines in vitro also transferred to in vivo studies, where it was shown to inhibit the formation of tumours in murine models [105].

NeuGcGM3.  An anti-NeuGcGM3 mAb, 14F7, was produced in mouse and shown to bind to NeuGc GM3 and not NeuAc GM3 specific to cancer tissues [52,106]. It has been shown to kill NeuGc–GM3-positive cell lines by CDC, ADCC and directly, although high concentrations of mAb were required [107].

Galβ1-3GlcNAcβ1-3Gal.  RAV12 is a chimeric mAb that has been shown to bind a minimal epitope of Galβ1-3GlcNAcβ1-3Gal, which has been observed on 90% of intra-abdominal tumours, but also on mucosal and glandular/ductal epithelium [4,108]. RAV12 has been shown to directly kill the colorectal cancer cell, Colo 205, in vitro by oncosis [4]. A study has shown that RAAG12 is present on insulin-like growth factor-I receptor (IGF-IR) and binding of RAV12 leads to increased phosphorylation of IGF-IR in RAAG12-positive cell lines, leading to accelerated desensitization of the Akt/PKB pathway [109]. A recent Phase I study in 33 recurrent adenocarcinoma patients showed some anti-tumour activity of RAV12, although toxicity of the mAb precluded the delivery of maximal doses [110].

F77.  Interestingly, F77, a mAb targeted at an as-yet unidentified glycolipid target, is also able to induce direct cell death by oncosis. The mAb has been shown to bind to a large proportion of both primary and metastatic prostate cancer specimens by immunohistochemistry [3]. F77 is proposed to induce oncosis by recruiting antigen to lipid rafts and through the production of large membrane pores [3].


  1. Top of page
  2. Summary
  3. Introduction
  4. Function of GSLs
  5. Conclusions
  6. Acknowledgements
  7. Disclosure
  8. References

Carbohydrate vaccines have, in general, stimulated low-titre IgM responses that have failed to have a significant impact on tumour growth. It remains to be seen if new approaches can overcome tolerance in humans and stimulate high-titre, potent IgG antibody responses. It seems more logical to develop human monoclonal IgG antibodies. The only caveat to this assumption is the recent demonstration that glycolipid vaccines can also stimulate cellular immunity resulting in tumour regression mediated by CD8 cytotoxic T cells [111]. The combination of antibody and cellular immunity may prove to have strong anti-tumour efficacy. However, the precise mechanism of stimulation of cellular immunity by glycolipids needs further elucidation. The advantage of monoclonal antibodies is that they can be administered repeatedly in high amounts. The ganglioside mAbs targeting neurological tumours and melanoma have suffered from dose-limiting neurological toxicity. However, new mAbs targeting glycolipids expressed by epithelial tumours should not suffer from this problem. Glycolipids are poorly immunogenic but new methods to overcome this limitation are stimulating a new wave of mAbs, stimulating direct and immune-mediated tumour killing, which are just entering the clinic.


  1. Top of page
  2. Summary
  3. Introduction
  4. Function of GSLs
  5. Conclusions
  6. Acknowledgements
  7. Disclosure
  8. References

The authors are grateful for funding from the MRC and Lewis Trust.


  1. Top of page
  2. Summary
  3. Introduction
  4. Function of GSLs
  5. Conclusions
  6. Acknowledgements
  7. Disclosure
  8. References
  • 1
    Hakomori S. Glycosylation defining cancer malignancy: new wine in an old bottle. Proc Natl Acad Sci USA 2002; 99:102313.
  • 2
    Zhong LT, Manzi A, Skowronski E et al. A monoclonal antibody that induces neuronal apoptosis binds a metastasis marker. Cancer Res 2001; 61:57418.
  • 3
    Zhang G, Zhang H, Wang Q et al. Suppression of human prostate tumor growth by a unique prostate-specific monoclonal antibody F77 targeting a glycolipid marker. Proc Natl Acad Sci USA 2010; 107:7327.
  • 4
    Loo D, Pryer N, Young P et al. The glycotope-specific RAV12 monoclonal antibody induces oncosis in vitro and has antitumor activity against gastrointestinal adenocarcinoma tumor xenografts in vivo. Mol Cancer Ther 2007; 6:85665.
  • 5
    Alvarez-Rueda N, Leprieur S, Clemenceau B et al. Binding activities and antitumor properties of a new mouse/human chimeric antibody specific for GD2 ganglioside antigen. Clin Cancer Res 2007; 13:5613s20s.
  • 6
    Hellstrom I, Garrigues HJ, Garrigues U, Hellstrom KE. Highly tumor-reactive, internalizing, mouse monoclonal antibodies to Le(y)-related cell surface antigens. Cancer Res 1990; 50:218390.
  • 7
    Chou HH, Takematsu H, Diaz S et al. A mutation in human CMP-sialic acid hydroxylase occurred after the Homo–Pan divergence. Proc Natl Acad Sci USA 1998; 95:117516.
  • 8
    Alberts B, Johnson A, Lewis J, Raff M, Roberts K, Walter P. Molecular biology of the cell, 4th edn. New York: Garland Science, 2002.
  • 9
    Ichikawa S, Hirabayashi Y. Glucosylceramide synthase and glycosphingolipid synthesis. Trends Cell Biol 1998; 8:198202.
  • 10
    Sprong H, Kruithof B, Leijendekker R, Slot JW, van Meer G, van der Sluijs P. UDP-galactose: ceramide galactosyltransferase is a class I integral membrane protein of the endoplasmic reticulum. J Biol Chem 1998; 273:258808.
  • 11
    Lannert H, Bunning C, Jeckel D, Wieland FT. Lactosylceramide is synthesized in the lumen of the Golgi apparatus. FEBS Lett 1994; 342:916.
  • 12
    Won JS, Singh AK, Singh I. Lactosylceramide: a lipid second messenger in neuroinflammatory disease. J Neurochem 2007; 103 (Suppl. 1):18091.
  • 13
    Hettmer S, McCarter R, Ladisch S, Kaucic K. Alterations in neuroblastoma ganglioside synthesis by induction of GD1b synthase by retinoic acid. Br J Cancer 2004; 91:38997.
  • 14
    Lingwood D, Simons K. Lipid rafts as a membrane-organizing principle. Science 2010; 327:4650.
  • 15
    Iwabuchi K, Yamamura S, Prinetti A, Handa K, Hakomori S. GM3-enriched microdomain involved in cell adhesion and signal transduction through carbohydrate-carbohydrate interaction in mouse melanoma B16 cells. J Biol Chem 1998; 273:91308.
  • 16
    Kojima N, Hakomori S. Cell adhesion, spreading, and motility of GM3-expressing cells based on glycolipid–glycolipid interaction. J Biol Chem 1991; 266:175528.
  • 17
    Suzuki C, Kojima N. A cholesterol-independent membrane microdomain serves as a functional counter-receptor for E-selectin at the Colo201 cell surface and initiates signalling on E-selectin binding. J Biochem 2007; 142:5564.
  • 18
    Thorne RF, Mhaidat NM, Ralston KJ, Burns GF. Shed gangliosides provide detergent-independent evidence for type-3 glycosynapses. Biochem Biophys Res Commun 2007; 356:30611.
  • 19
    Mitsuzuka K, Handa K, Satoh M, Arai Y, Hakomori S. A specific microdomain (‘glycosynapse 3’) controls phenotypic conversion and reversion of bladder cancer cells through GM3-mediated interaction of alpha3beta1 integrin with CD9. J Biol Chem 2005; 280:3554553.
  • 20
    Zheng M, Fang H, Tsuruoka T, Tsuji T, Sasaki T, Hakomori S. Regulatory role of GM3 ganglioside in alpha 5 beta 1 integrin receptor for fibronectin-mediated adhesion of FUA169 cells. J Biol Chem 1993; 268:221722.
  • 21
    Zheng M, Fang H, Hakomori S. Functional role of N-glycosylation in alpha 5 beta 1 integrin receptor. De-N-glycosylation induces dissociation or altered association of alpha 5 and beta 1 subunits and concomitant loss of fibronectin binding activity. J Biol Chem 1994; 269:1232531.
  • 22
    Ono M, Handa K, Withers DA, Hakomori S. Glycosylation effect on membrane domain (GEM) involved in cell adhesion and motility: a preliminary note on functional alpha3, alpha5-CD82 glycosylation complex in ldlD 14 cells. Biochem Biophys Res Commun 2000; 279:74450.
  • 23
    Hakomori S. Aberrant glycosylation in cancer cell membranes as focused on glycolipids: overview and perspectives. Cancer Res 1985; 45:240514.
  • 24
    Reis CA, Osorio H, Silva L, Gomes C, David L. Alterations in glycosylation as biomarkers for cancer detection. J Clin Pathol 2010; 63:3229.
  • 25
    Kawamura YI, Toyota M, Kawashima R et al. DNA hypermethylation contributes to incomplete synthesis of carbohydrate determinants in gastrointestinal cancer. Gastroenterology 2008; 135:14251e3.
  • 26
    Itai S, Nishikata J, Yoneda T et al. Tissue distribution of 2-3 and 2-6 sialyl Lewis A antigens and significance of the ratio of two antigens for the differential diagnosis of malignant and benign disorders of the digestive tract. Cancer 1991; 67:157687.
  • 27
    Kannagi R. Molecular mechanism for cancer-associated induction of sialyl Lewis X and sialyl Lewis A expression – the Warburg effect revisited. Glycoconj J 2004; 20:35364.
  • 28
    Wei J, Cui L, Liu F et al. E-selectin and sialyl Lewis X expression is associated with lymph node metastasis of invasive micropapillary carcinoma of the breast. Int J Surg Pathol 2010; 18:193200.
  • 29
    St Hill CA, Baharo-Hassan D, Farooqui M. C2-O-sLe glycoproteins are E-selectin ligands that regulate invasion of human colon and hepatic carcinoma cells. PLoS ONE 2011; 6:e16281.
  • 30
    Kannagi R, Yin J, Miyazaki K, Izawa M. Current relevance of incomplete synthesis and neo-synthesis for cancer-associated alteration of carbohydrate determinants – Hakomori's concepts revisited. Biochim Biophys Acta 2008; 1780:52531.
  • 31
    Koike T, Kimura N, Miyazaki K et al. Hypoxia induces adhesion molecules on cancer cells: a missing link between Warburg effect and induction of selectin-ligand carbohydrates. Proc Natl Acad Sci USA 2004; 101:81327.
  • 32
    Ogawa J, Inoue H, Koide S. Glucose-transporter-type-I-gene amplification correlates with sialyl-Lewis-X synthesis and proliferation in lung cancer. Int J Cancer 1997; 74:18992.
  • 33
    Miyoshi E, Moriwaki K, Nakagawa T. Biological function of fucosylation in cancer biology. J Biochem 2008; 143:7259.
  • 34
    Moriwaki K, Miyoshi E. Fucosylation and gastrointestinal cancer. World J Hepatol 2010; 2:15161.
  • 35
    Tsuchida T, Saxton RE, Morton DL, Irie RF. Gangliosides of human melanoma. Cancer 1989; 63:116674.
  • 36
    Dubois C, Magnani JL, Grunwald GB et al. Monoclonal antibody 18B8, which detects synapse-associated antigens, binds to ganglioside GT3 (II3 (NeuAc)3LacCer). J Biol Chem 1986; 261:382630.
  • 37
    Hamilton WB, Helling F, Lloyd KO, Livingston PO. Ganglioside expression on human malignant melanoma assessed by quantitative immune thin-layer chromatography. Int J Cancer 1993; 53:56673.
  • 38
    Brodin T, Thurin J, Stromberg N, Karlsson KA, Sjogren HO. Production of oligosaccharide-binding monoclonal antibodies of diverse specificities by immunization with purified tumor-associated glycolipids inserted into liposomes with lipid A. Eur J Immunol 1986; 16:9516.
  • 39
    Yamada N, Chung YS, Takatsuka S et al. Increased sialyl Lewis A expression and fucosyltransferase activity with acquisition of a high metastatic capacity in a colon cancer cell line. Br J Cancer 1997; 76:5827.
  • 40
    Jeschke U, Mylonas I, Shabani N et al. Expression of sialyl lewis X, sialyl Lewis A, E-cadherin and cathepsin-D in human breast cancer: immunohistochemical analysis in mammary carcinoma in situ, invasive carcinomas and their lymph node metastasis. Anticancer Res 2005; 25:161522.
  • 41
    Charpin C, Bhan AK, Zurawski VR Jr, Scully RE. Carcinoembryonic antigen (CEA) and carbohydrate determinant 19-9 (CA 19-9) localization in 121 primary and metastatic ovarian tumors: an immunohistochemical study with the use of monoclonal antibodies. Int J Gynecol Pathol 1982; 1:23145.
  • 42
    Magnani JL, Nilsson B, Brockhaus M et al. A monoclonal antibody-defined antigen associated with gastrointestinal cancer is a ganglioside containing sialylated lacto-N-fucopentaose II. J Biol Chem 1982; 257:143659.
  • 43
    Nakagoe T, Sawai T, Tsuji T et al. Circulating sialyl Lewis(x), sialyl Lewis(a), and sialyl Tn antigens in colorectal cancer patients: multivariate analysis of predictive factors for serum antigen levels. J Gastroenterol 2001; 36:16672.
  • 44
    Mujoo K, Cheresh DA, Yang HM, Reisfeld RA. Disialoganglioside GD2 on human neuroblastoma cells: target antigen for monoclonal antibody-mediated cytolysis and suppression of tumor growth. Cancer Res 1987; 47:1098104.
  • 45
    Wu ZL, Schwartz E, Seeger R, Ladisch S. Expression of GD2 ganglioside by untreated primary human neuroblastomas. Cancer Res 1986; 46:4403.
  • 46
    Yoshida S, Kawaguchi H, Sato S, Ueda R, Furukawa K. An anti-GD2 monoclonal antibody enhances apoptotic effects of anti-cancer drugs against small cell lung cancer cells via JNK (c-Jun terminal kinase) activation. Jpn J Cancer Res 2002; 93:81624.
  • 47
    Portoukalian J, Zwingelstein G, Dore JF. Lipid composition of human malignant melanoma tumors at various levels of malignant growth. Eur J Biochem 1979; 94:1923.
  • 48
    Distler U, Souady J, Hulsewig M et al. Shiga toxin receptor Gb3Cer/CD77: tumor-association and promising therapeutic target in pancreas and colon cancer. PLoS ONE 2009; 4:e6813.
  • 49
    Chang F, Li R, Ladisch S. Shedding of gangliosides by human medulloblastoma cells. Exp Cell Res 1997; 234:3416.
  • 50
    Sa G, Das T, Moon C et al. GD3, an overexpressed tumor-derived ganglioside, mediates the apoptosis of activated but not resting T cells. Cancer Res 2009; 69:3095104.
  • 51
    Devine PL, Clark BA, Birrell GW et al. The breast tumor-associated epitope defined by monoclonal antibody 3E1·2 is an O-linked mucin carbohydrate containing N-glycolylneuraminic acid. Cancer Res 1991; 51:582636.
  • 52
    Oliva JP, Valdes Z, Casaco A et al. Clinical evidences of GM3 (NeuGc) ganglioside expression in human breast cancer using the 14F7 monoclonal antibody labelled with (99m)Tc. Breast Cancer Res Treat 2006; 96:11521.
  • 53
    Higashi H, Sasabe T, Fukui Y, Maru M, Kato S. Detection of gangliosides as N-glycolylneuraminic acid-specific tumor-associated Hanganutziu–Deicher antigen in human retinoblastoma cells. Jpn J Cancer Res 1988; 79:9526.
  • 54
    Bardor M, Nguyen DH, Diaz S, Varki A. Mechanism of uptake and incorporation of the non-human sialic acid N-glycolylneuraminic acid into human cells. J Biol Chem 2005; 280:422837.
  • 55
    Gabri MR, Otero LL, Gomez DE, Alonso DF. Exogenous incorporation of neugc-rich mucin augments n-glycolyl sialic acid content and promotes malignant phenotype in mouse tumor cell lines. J Exp Clin Cancer Res 2009; 28:14654.
  • 56
    de Leon J, Fernandez A, Mesa C, Clavel M, Fernandez LE. Role of tumour-associated N-glycolylated variant of GM3 ganglioside in cancer progression: effect over CD4 expression on T cells. Cancer Immunol Immunother 2006; 55:44350.
  • 57
    Buskas T, Thompson P, Boons GJ. Immunotherapy for cancer: synthetic carbohydrate-based vaccines. Chem Commun (Camb) 2009; 36:533549.
  • 58
    Mond JJ, Vos Q, Lees A, Snapper CM. T cell independent antigens. Curr Opin Immunol 1995; 7:34954.
  • 59
    Tolar P, Hanna J, Krueger PD, Pierce SK. The constant region of the membrane immunoglobulin mediates B cell-receptor clustering and signaling in response to membrane antigens. Immunity 2009; 30:4455.
  • 60
    Cipolla L, Peri F, Airoldi C. Glycoconjugates in cancer therapy. Anticancer Agents Med Chem 2008; 8:92121.
  • 61
    Ragupathi G, Livingston PO, Hood C et al. Consistent antibody response against ganglioside GD2 induced in patients with melanoma by a GD2 lactone–keyhole limpet hemocyanin conjugate vaccine plus immunological adjuvant QS-21. Clin Cancer Res 2003; 9:521420.
  • 62
    Krug LM, Ragupathi G, Hood C et al. Vaccination of patients with small-cell lung cancer with synthetic fucosyl GM-1 conjugated to keyhole limpet hemocyanin. Clin Cancer Res 2004; 10:6094100.
  • 63
    Livingston PO. Approaches to augmenting the immunogenicity of melanoma gangliosides: from whole melanoma cells to ganglioside–KLH conjugate vaccines. Immunol Rev 1995; 145:14766.
  • 64
    Livingston PO, Wong GY, Adluri S et al. Improved survival in stage III melanoma patients with GM2 antibodies: a randomized trial of adjuvant vaccination with GM2 ganglioside. J Clin Oncol 1994; 12:103644.
  • 65
    Kitamura K, Livingston PO, Fortunato SR et al. Serological response patterns of melanoma patients immunized with a GM2 ganglioside conjugate vaccine. Proc Natl Acad Sci USA 1995; 92:28059.
  • 66
    Helling F, Zhang S, Shang A et al. GM2–KLH conjugate vaccine: increased immunogenicity in melanoma patients after administration with immunological adjuvant QS-21. Cancer Res 1995; 55:27838.
  • 67
    Slovin SF, Keding SJ, Ragupathi G. Carbohydrate vaccines as immunotherapy for cancer. Immunol Cell Biol 2005; 83:41828.
  • 68
    Eggermont AM, Suciu S, Rutkowski PM et al. EOTRC Melanoma Group, randomized phase III trial comparing postoperative adjuvant ganglioside GM2–KLH/QS-21 vaccination versus observation in stage II (T3-T4N0M0) melanoma: final results of study EORTC 18961. J Clin Oncol 2010; 28:201218.
  • 69
    Chapman PB, Wu D, Ragupathi G et al. Sequential immunization of melanoma patients with GD3 ganglioside vaccine and anti-idiotypic monoclonal antibody that mimics GD3 ganglioside. Clin Cancer Res 2004; 10:471723.
  • 70
    Giaccone G, Debruyne C, Felip E et al. Phase III study of adjuvant vaccination with Bec2/bacille Calmette–Guérin in responding patients with limited-disease small-cell lung cancer (European Organisation for Research and Treatment of Cancer 08971-08971B; Silva Study). J Clin Oncol 2005; 23:685464.
  • 71
    Hernandez AM, Rodriguez N, Gonzalez JE et al. Anti-NeuGcGM3 antibodies, actively elicited by idiotypic vaccination in nonsmall cell lung cancer patients, induce tumor cell death by an oncosis-like mechanism. J Immunol 2011; 186:373544.
  • 72
    Fernandez LE, Gabri MR, Guthmann MD et al. NGcGM3 ganglioside: a privileged target for cancer vaccines. Clin Dev Immunol 2010; 2010:814397405.
  • 73
    Fuentes D, Avellanet J, Garcia A et al. Combined therapeutic effect of a monoclonal anti-idiotype tumor vaccine against NeuGc-containing gangliosides with chemotherapy in a breast carcinoma model. Breast Cancer Res Treat 2010; 120:37989.
  • 74
    Alfonso S, Diaz RM, de la Torre A et al. 1E10 anti-idiotype vaccine in non-small cell lung cancer: experience in stage IIIb/IV patients. Cancer Biol Ther 2007; 6:184752.
  • 75
    Guthmann MD, Castro MA, Cinat G et al. Cellular and humoral immune response to N-Glycolyl-GM3 elicited by prolonged immunotherapy with an anti-idiotypic vaccine in high-risk and metastatic breast cancer patients. J Immunother 2006; 29:21523.
  • 76
    Hernandez AM, Toledo D, Martinez D et al. Characterization of the antibody response against NeuGcGM3 ganglioside elicited in non-small cell lung cancer patients immunized with an anti-idiotype antibody. J Immunol 2008; 181:662534.
  • 77
    Osorio M, Gracia E, Rodriguez E et al. Heterophilic NeuGcGM3 ganglioside cancer vaccine in advanced melanoma patients: results of a Phase Ib/IIa study. Cancer Biol Ther 2008; 7:48895.
  • 78
    Livingston PO, Ritter G, Calves MJ. Antibody response after immunization with the gangliosides GM1, GM2, GM3, GD2 and GD3 in the mouse. Cancer Immunol Immunother 1989; 29:17984.
  • 79
    Estevez F, Carr A, Solorzano L et al. Enhancement of the immune response to poorly immunogenic gangliosides after incorporation into very small size proteoliposomes (VSSP). Vaccine 1999; 18:1907.
  • 80
    Guthmann MD, Bitton RJ, Carnero AJ et al. Active specific immunotherapy of melanoma with a GM3 ganglioside-based vaccine: a report on safety and immunogenicity. J Immunother 2004; 27:44251.
  • 81
    Ragupathi G, Damani P, Srivastava G et al. Synthesis of sialyl Lewis(a) (sLe (a), CA19-9) and construction of an immunogenic sLe(a) vaccine. Cancer Immunol Immunother 2009; 58:1397405.
  • 82
    Sabbatini PJ, Kudryashov V, Ragupathi G et al. Immunization of ovarian cancer patients with a synthetic Lewis(y)-protein conjugate vaccine: a phase 1 trial. Int J Cancer 2000; 87:7985.
  • 83
    Cox D, Fox L, Tian R et al. Determination of cellular lipids bound to human CD1d molecules. PLoS ONE 2009; 4:e5325.
  • 84
    Blanca IR, Bere EW, Young HA, Ortaldo JR. Human B cell activation by autologous NK cells is regulated by CD40–CD40 ligand interaction: role of memory B cells and CD5+ B cells. J Immunol 2001; 167:61329.
  • 85
    Leadbetter EA, Brigl M, Illarionov P et al. NK T cells provide lipid antigen-specific cognate help for B cells. Proc Natl Acad Sci USA 2008; 105:833944.
  • 86
    Dullforce P, Sutton DC, Heath AW. Enhancement of T cell-independent immune responses in vivo by CD40 antibodies. Nat Med 1998; 4:8891.
  • 87
    Kitamura K, Stockert E, Garin-Chesa P et al. Specificity analysis of blood group Lewis-y (Ley) antibodies generated against synthetic and natural Ley determinants. Proc Natl Acad Sci USA 1994; 91:1295761.
  • 88
    Tolcher A, Sugarman S, Gelmon K et al. Randomized phase II study of BR96-doxorubicin conjugate in patients with metastatic breast cancer. J Clin Oncol 1999; 17:47884.
  • 89
    Pai LH, Wittes R, Setser A, Willingham MC, Pastan I. Treatment of advanced solid tumors with immunotoxin LMB-1: an antibody linked to Pseudomonas exotoxin. Nat Med 1996; 2:3503.
  • 90
    Manimala JC, Roach TA, Li Z, Gildersleeve JC. High-throughput carbohydrate microarray profiling of 27 antibodies demonstrates widespread specificity problems. Glycobiology 2007; 17:17C23C.
  • 91
    Durrant LG, Harding SJ, Green NH, Buckberry LD, Parsons T. A new anticancer glycolipid monoclonal antibody, SC104, which directly induces tumor cell apoptosis. Cancer Res 2006; 66:59019.
  • 92
    Sawada R, Sun SM, Wu X et al. Human monoclonal antibodies to sialyl-Lewisa (CA19·9) with potent CDC, ADCC, and antitumor activity. Clin Cancer Res 2011; 17:102432.
  • 93
    Barker E, Mueller BM, Handgretinger R, Herter M, Yu AL, Reisfeld RA. Effect of a chimeric anti-ganglioside GD2 antibody on cell-mediated lysis of human neuroblastoma cells. Cancer Res 1991; 51:1449.
  • 94
    Frost JD, Hank JA, Reaman GH et al. A phase I/IB trial of murine monoclonal anti-GD2 antibody 14.G2a plus interleukin-2 in children with refractory neuroblastoma: a report of the Children's Cancer Group. Cancer 1997; 80:31733.
  • 95
    Kramer K, Humm JL, Souweidane MM et al. Phase I study of targeted radioimmunotherapy for leptomeningeal cancers using intra-Ommaya 131-I-3F8. J Clin Oncol 2007; 25:546570.
  • 96
    Nakamura K, Tanaka Y, Shitara K, Hanai N. Construction of humanized anti-ganglioside monoclonal antibodies with potent immune effector functions. Cancer Immunol Immunother 2001; 50:27584.
  • 97
    Kramer K, Gerald WL, Kushner BH, Larson SM, Hameed M, Cheung NK. Disialoganglioside G(D2) loss following monoclonal antibody therapy is rare in neuroblastoma. Clin Cancer Res 1998; 4:21359.
  • 98
    Yang RK, Sondel PM. Anti-GD2 Strategy in the Treatment of Neuroblastoma. Drugs Future 2010; 35:66580.
  • 99
    Modak S, Cheung NK. Disialoganglioside directed immunotherapy of neuroblastoma. Cancer Invest 2007; 25:6777.
  • 100
    Cheresh DA, Honsik CJ, Staffileno LK, Jung G, Reisfeld RA. Disialoganglioside GD3 on human melanoma serves as a relevant target antigen for monoclonal antibody-mediated tumor cytolysis. Proc Natl Acad Sci USA 1985; 82:51559.
  • 101
    Chapman PB, Yuasa H, Houghton AN. Homophilic binding of mouse monoclonal antibodies against GD3 ganglioside. J Immunol 1990; 145:8918.
  • 102
    Kirkwood JM, Mascari RA, Edington HD et al. Analysis of therapeutic and immunologic effects of R(24) anti-GD3 monoclonal antibody in 37 patients with metastatic melanoma. Cancer 2000; 88:2693702.
  • 103
    Kaminski MJ, MacKenzie CR, Mooibroek MJ et al. The role of homophilic binding in anti-tumor antibody R24 recognition of molecular surfaces. Demonstration of an intermolecular beta-sheet interaction between vh domains. J Biol Chem 1999; 274:5597604.
  • 104
    Fernandes DM, Baird AM, Berg LJ, Rock KL. A monoclonal antibody reactive with a 40-kDa molecule on fetal thymocytes and tumor cells blocks proliferation and stimulates aggregation and apoptosis. J Immunol 1999; 163:130614.
  • 105
    Retter MW, Johnson JC, Peckham DW et al. Characterization of a proapoptotic antiganglioside GM2 monoclonal antibody and evaluation of its therapeutic effect on melanoma and small cell lung carcinoma xenografts. Cancer Res 2005; 65:642534.
  • 106
    Carr A, Mullet A, Mazorra Z et al. A mouse IgG1 monoclonal antibody specific for N-glycolyl GM3 ganglioside recognized breast and melanoma tumors. Hybridoma 2000; 19:2417.
  • 107
    Carr A, Mesa C, Arango M, Vazquez AM, Fernandez LE. In vivo and in vitro anti-tumor effect of 14F7 monoclonal antibody. Hybrid Hybridomics 2002; 21:4638.
  • 108
    Coberly SK, Chen FZ, Armanini MP et al. The RAV12 monoclonal antibody recognizes the N-linked glycotope RAAG12: expression in human normal and tumor tissues. Arch Pathol Lab Med 2009; 133:140312.
  • 109
    Li JC, Li R. RAV12 accelerates the desensitization of Akt/PKB pathway of insulin-like growth factor I receptor signaling in COLO205. Cancer Res 2007; 67:885664.
  • 110
    Burris HA III, Rosen LS, Rocha-Lima CM et al. Phase 1 experience with an anti-glycotope monoclonal antibody, RAV12, in recurrent adenocarcinoma. Clin Cancer Res 2010; 16:167381.
  • 111
    Mazorra Z, Mesa C, Fernandez A, Fernandez L. Immunization with GM3 ganglioside nanoparticulated vaccine confers an effector CD8+ T cells-mediated protection against melanoma B16 challenge. Cancer Immunol Immunother 2008; 57:177180.
  • 112
    Hakomori S. The glycosynapse. Proc Natl Acad Sci USA 2002; 99:22532.