Bioengineering of Surface GD3 Ganglioside for Immunotargeting Human Melanoma Cells

Zou, Wei; Borrelli, Silvia; Gilbert, Michel; Liu, Tianmin; Pon, Robert A.; Jennings, Harold J.

doi:10.1074/jbc.m402787200

Cited by 51 publications

(34 citation statements)

References 47 publications

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“…However, anti-LPS titers obtained with (MDWNMHAA) 50 -TT appeared lower than with (MDWNMHAA) 32 -TT in week 8, despite the higher anti-peptide titers (G1, week 8). This may be the result of a carrier-induced suppression effect, which can decrease the production of antigen-specific antibodies, as observed earlier in a similar immunological study with a strongly immunogenic PS-mimetic peptide conjugate [(DRPVPY) 39 -TT] (6). No clear difference in the LPS-cross-reactive antibody response was observed when (MDWNMHAA) 50 -TT was administered to mice three times (G2) instead of four (G1), suggesting that when a more immunogenic mimetic peptide conjugate is used, fewer injections are required.…”

Section: Cross-reactivity: Anti-ps (Ps-tt) Antibodies Bind To Mdw Nmhmentioning

confidence: 70%

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Immunological Evidence for Functional Rather than Structural Mimicry by aShigella flexneriY Polysaccharide-Mimetic Peptide

Borrelli

Hossany

Pinto

2008

Clin Vaccine Immunol

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Shigellosis, caused by Shigella species (gram negative), is a prominent, and the most infectious, diarrheal disease. Shigella flexneri, the species responsible for the highest mortality rate, is endemic in most developing countries (23). The 13 serotypes of S. flexneri, with the exception of serotype 6, result from structural modifications of the O-antigen polysaccharide (PS), the outer portion of the lipopolysaccharide (LPS) on the bacterial surface, which is both an essential virulence factor and a protective antigen (10). The basic O-antigen PS repeat, common to the 12 serotypes, is referred to as serotype Y. A murine immunoglobulin G3 (IgG3) monoclonal antibody, SYA/J6, specific for the PS O antigen of the S. flexneri Y LPS, was developed by Bundle (7) and Carlin and coworkers (8). The O-antigen Y PS is a linear heteropolymer with a tetrasaccha- -(13] and is recognized by the monoclonal antibody SYA/J6 (7, 9). The oligosaccharides of S. flexneri serotype Y have been well studied by nuclear magnetic resonance techniques, which have provided a three-dimensional model of the determinant in solution (4,30,31,32) and the identity of the biological repeating unit (9).Glycoconjugate vaccines to prevent shigellosis, focused on oligosaccharide analogues related to Shigella flexneri 2a and other serotypes, have been synthesized and evaluated (3,10,29,38). An interesting approach to vaccine design is the use of molecules that mimic the immunogenic element of interest (11,20,28). Carbohydrate-mimetic peptides have potential as surrogate ligands for traditional carbohydrate vaccines, providing more discriminating immune responses (19,20,28). However, there are few examples of immunological responses with peptide-based PS mimics (18,20,28). Therefore, the requirements for cross-reactivity are not fully understood and are certainly not predictable, because of the limited data set available.In order to further exploit this principle, a carbohydratemimetic peptide of the S. flexneri Y O-antigen PS, MDWNM HAA, cross-reactive with the anti-S. flexneri Y O-PS monoclonal antibody, SYA/J6, was identified by phage library screening (15). The structures of complexes of the antibody SYA/J6 Fab fragment with synthetic deoxytrisaccharide and pentasaccharide ligands, related to the S. flexneri Y O antigen, and with the carbohydrate-mimetic peptide have been determined by X-ray crystallography (17,33,34,35). The structure of the Fab complex with MDWNMHAA revealed differences, and few similarities, with respect to the oligosaccharide complexes (34), providing the first evidence that the modes of binding of the pentasaccharide and octapeptide differ considerably and that few aspects of structural mimicry exist (34).

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Section: Cross-reactivity: Anti-ps (Ps-tt) Antibodies Bind To Mdw Nmhmentioning

confidence: 70%

“…Antibody titers to MDWNMHAA and LPS in sera from mice were determined before vaccination and 2 weeks after each vaccination by ELISA as described previously (39). The titer was defined as the highest dilution yielding an absorbance of Ն0.1 after twice the average background reading was subtracted.…”

Section: Psmentioning

confidence: 99%

Immunological Evidence for Functional Rather than Structural Mimicry by aShigella flexneriY Polysaccharide-Mimetic Peptide

Borrelli

Hossany

Pinto

2008

Clin Vaccine Immunol

Self Cite

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“…These studies examined metabolic engineering with a maximum substrate concentration of 40 μM, and it is possible that higher levels of ManNPr, ManNBu or ManNiBn could produce detectable levels of modified GM3. Other in vitro studies have used precursor concentrations of up to 3-20 mM for metabolic engineering, and these conditions result in incorporation of ManNPr and ManNBu by tumor cells (34,53,55). However, such high concentrations may be difficult to achieve in vivo.…”

Section: Metabolic Engineering Of Cancer Cells Was Tested With Severamentioning

confidence: 99%

Efficient Metabolic Engineering of GM3 on Tumor Cells by N-Phenylacetyl-d-mannosamine

et al. 2006

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Abnormal carbohydrates expressed on tumor cells, which are referred to as tumor-associated carbohydrate antigens (TACAs), are potential targets for development of cancer vaccines. However, immune tolerance to TACAs has severely hindered progress in this area. To overcome this problem, we have developed a novel immunotherapeutic strategy based on synthetic cancer vaccines and metabolic engineering of TACAs on tumor cells. One critical step of this new strategy is metabolic engineering of cancer, namely to induce expression of an artificial form of a TACA by supplying tumors with an artificial monosaccharide precursor. To identify the proper precursor for this application, N-propionyl, N-butanoyl, N-iso-butanoyl and N-phenylacetyl derivatives of Dmannosamine were synthesized, and their efficiency as biosynthetic precursors to modify sialic acid and induce expression of modified forms of GM3 antigen on tumor cells was investigated. For this purpose, tumor cells were incubated with different N-acyl-D-mannosamines, and modified forms of GM3 expressed on tumor cells were analyzed by flow cytometry using antigen-specific antisera. Nphenylacetyl-D-mannosamine was efficiently incorporated in a time and dose dependent manner to bioengineer GM3 expression by several tumor cell lines including K562, SKMEL-28 and B16-F0. Moreover, these tumor cell lines also showed ManPAc-dependent sensitivity to cytotoxicity medicated by anti-PAcGM3 immune serum and complement. These results provide an important validation for this novel therapeutic strategy. Because N-phenylacetyl GM3-protein conjugates are particularly immunogenic, the combination of an N-phenylacetyl GM3 conjugate vaccine with systemic N-phenylacetyl-D-mannosamine treatment is a promising immunotherapy for future development and application to melanoma and other GM3-bearing tumors. KeywordsMetabolic engineering; cancer vaccine; GM3; D-mannosamine; sialic acid Sialic acids are unique nine-carbon acidic monosaccharides ubiquitously distributed in high animals and human beings (1). The most common form of sialic acids is N-acetyl-D-neuraminic acid (Neu5Ac, also knows as N-acetyl sialic acid). Neu5Ac is a fascinating sugar in that it usually appears at the nonreducing end of cell surface glycans and thus plays forefront roles in various biological functions such as cell proliferation, immune recognition and pathogenic invasion (2-8).

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“…The mAb used in this study was purified by protein G affinity chromatography (Pierce, Rockford, USA). The potency of the antibody in ascites was 5×10 4 .…”

Section: Monoclonal Antibody Productionmentioning

confidence: 99%

“…Polysialic acid (PSA), a linear homopolymer composed of α-(2-8)-linked N-acetyl-neuraminic acid (NeuAc) residues, is a unique biological form of sialic acid that is an important cancer-associated antigen [2] . Several studies have taken advantage of the permissiveness of the sialic acid and PSA biosynthetic pathways to remodel the cell-surface landscape of tumors both in vitro and in vivo [3][4][5] . This remodeling can be performed by replacing N-acetyl-mannosamine (ManAc), the physiological precursor of sialic acid, with an exogenous source of unnatural N-acyl mannosamines, which results in the introduction of these unnatural sialosides into surface glycoconjugates [6] .…”

Section: Introductionmentioning

confidence: 99%

N-Propionyl polysialic acid precursor enhances the susceptibility of multiple myeloma to antitumor effect of anti-NprPSA monoclonal antibody

Xiong

Liang

et al. 2012

Acta Pharmacol Sin

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Aim: To study the antitumor effect of anti-NprPSA monoclonal antibody (mAb) in combination with ManNPr, a precursor of N-propionyl PSA, in multiple myeloma (MM), and to explore the mechanisms of the action. Methods: Human multiple myeloma cell line RPMI-8226 was tested. The cells were pre-treated with ManNPr (1, 2, and 4 mg/mL), and then incubated with anti-NprPSA mAb (1 mg/mL). Cell apoptosis in vitro was detected using MTT assay and flow cytometry. BALB/c nude mice were inoculated sc with RPC5.4 cells. On 5 d after the injection, the mice were administered sc with anti-NprPSA mAb (200 μg/d) and ManNPr (5 mg/d) for 8 d. The tumor size and body weight were monitored twice per week. TUNEL assay was used for detecting apoptosis in vivo. The apoptotic pathway involved was examined using Western blot analysis and caspase inhibitor. Results: Treatment of RPMI-8226 cells with anti-NprPSA mAb alone failed to inhibit cell growth in vitro. In RPMI-8226 cells pretreated with ManNPr, however, the mAb significantly inhibited the cell proliferation, decreased the viability, and induced apoptosis, which was associated with cleavage of caspase-3, caspase-8, caspase-9, and poly(ADP-ribose) polymerase. In the mouse xenograft model, treatment with the mAb in combination with ManNPr significantly inhibited the tumor growth, and induced significant apoptosis as compared to treatment with the mAb alone. Moreover, apoptosis induced by the mAb in vivo resulted from the activation of the caspases and poly(ADP-ribose) polymerase. Conclusion: The anti-NprPSA mAb in combination with ManNPr is an effective treatment for in vitro and in vivo induction of apoptosis in multiple myeloma.

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Bioengineering of Surface GD3 Ganglioside for Immunotargeting Human Melanoma Cells

Cited by 51 publications

References 47 publications

Immunological Evidence for Functional Rather than Structural Mimicry by aShigella flexneriY Polysaccharide-Mimetic Peptide

Immunological Evidence for Functional Rather than Structural Mimicry by aShigella flexneriY Polysaccharide-Mimetic Peptide

Efficient Metabolic Engineering of GM3 on Tumor Cells by N-Phenylacetyl-d-mannosamine

N-Propionyl polysialic acid precursor enhances the susceptibility of multiple myeloma to antitumor effect of anti-NprPSA monoclonal antibody

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