Expression of Gangliosides GM3(NeuAc) and GM3(NeuGc) in Myelomas and Hybridomas of Mouse, Rat, and Human Origin1

Müthing, Johannes; Steuer, Heiko; Peter‐Katalinić, Jasna; Marx, Uwe; Bethke, Ulf; Neumann, Ulrich; Lehmann, Jürgen

doi:10.1093/oxfordjournals.jbchem.a124504

Cited by 37 publications

(21 citation statements)

References 75 publications

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“…P3-X63-Ag8.653 (X63) murine myeloma cell line (ATCC NCRL 1580), expressing NeuGcGM3 in their membrane (30), and H82 human lung carcinoma cell line (ATCC HTB-175), negative for the expression of the ganglioside, were grown in DMEM (Invitrogen) supplemented with 10% heat-inactivated FCS (HyClone), 2 mM L-glutamine, 25 mM HEPES, 100 U/ml penicillin, 100 g/ml streptomycin, and maintained at 37°C with 5% CO2.…”

Section: Gangliosides and Cellsmentioning

confidence: 99%

Characterization of the Antibody Response against NeuGcGM3 Ganglioside Elicited in Non-Small Cell Lung Cancer Patients Immunized with an Anti-Idiotype Antibody

Hernández¹,

Toledo²,

Martínez³

et al. 2008

The Journal of Immunology

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1E10 mAb is an anti-Id murine mAb (Ab2 mAb) specific for an Ab1 mAb that reacts with NeuGc-containing gangliosides, sulfatides, and Ags expressed in some human tumors. In preclinical studies, this Ab2 Ab was able to mimic NeuGc-containing gangliosides only in animals lacking expression of these Ags in normal tissues. In this study, we report on the immune responses elicited in 20 non-small cell lung cancer patients treated with 1 mg of aluminum hydroxide-precipitated 1E10 mAb. In the hyperimmune sera from 16 of 20 patients, a strong specific Ab response of both IgM and IgG isotypes against NeuGcGM3 ganglioside was observed. Patient immune sera were able to induce complement-independent cell death of NeuGcGM3-expressing X63 murine myeloma target cells. Significant immunoreactivity to NeuGcGM3 was still detected after the complete abrogation of the reactivity against 1E10 mAb by the adsorption of patient sera with this Ab. We hypothesize that Id−Ag+ Abs could reflect the activation of an autologous idiotypic cascade into the patients. Both Id+Ag+ and Id−Ag+ fractions were separated by affinity chromatography and characterized. Although IgG isotype Abs were found in both fractions, IgM isotype Abs were found only in the Id−Ag+ fraction. Both Id+Ag+ and Id−Ag+ Abs were able to specifically recognize and induce cell death in NeuGcGM3-expressing X63 myeloma target cells. Patients that developed IgG and/or IgM Abs against NeuGcGM3 showed longer median survival times.

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Section: Gangliosides and Cellsmentioning

confidence: 99%

Characterization of the Antibody Response against NeuGcGM3 Ganglioside Elicited in Non-Small Cell Lung Cancer Patients Immunized with an Anti-Idiotype Antibody

Hernández¹,

Toledo²,

Martínez³

et al. 2008

The Journal of Immunology

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“…Human granulocyte gangliosides (Fig. 1b, lane S 2 ) did not bind this antibody, indicating the high specificity of the anti-G M3 (Neu5Gc) antibody, which does not react with G M3 (Neu5Ac) or any ganglio-or neolacto-series gangliosides [24]. Staining with anti-G M3 (Neu5Ac) antibody showed a similar expression pattern, i.e.…”

Section: Expression Of G M3mentioning

confidence: 89%

“…Polyclonal chicken anti-G M3 (Neu5Ac) and anti-G M3 (Neu5Gc) antibodies have been characterized in our previous reports [24,27]. Rabbit anti-GgOse 4 Cer antiserum used for the detection of G M1b and G D1a has also been described in earlier publications [28,29].…”

Section: Antibodies and Cholera Toxin B Subunit (Choleragenoid)mentioning

confidence: 99%

“…A ganglioside mixture composed of G M3 (Neu5Ac), IV 3 Neu5Ac-nLcOse 4 Cer (IV 3 nLc4), IV 6 Neu5Ac-nLcOse 4 Cer (IV 6 nLc4) and VI 3 Neu5Ac-nLcOse 6 Cer (VI 3 nLc6) was isolated from human granulocytes as previously described [23]. G M3 (Neu5Gc) was purified from a mouse-mouse hybridoma [24]. Gangliosides from a murine T lymphoma YAC-1 and a lymphoreticular tumour cell line MDAY-D2 were isolated according to established procedures [25,26] (see Table 1).…”

Section: Thin-layer Chromatography and Reference Gangliosidesmentioning

confidence: 99%

“…The separation of individual murine gangliosides on HPTLC plates as double or triple bands is a common feature, due to the variation in the ceramide portion (C 16 -or C 24 -fatty acids) and oligosaccharide moiety (Neu5Ac and Neu5Gc) [24]. The upper band in our samples was composed of G M3 (Neu5Ac) with a C 24 -fatty acid, and the lower band was G M3 (Neu5Ac) with a C 16 -fatty acid and G M3 (Neu5Gc) with a C 24 -fatty acid.…”

Section: Expression Of G M3mentioning

confidence: 99%

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Ganglioside expression in tissues of mice lackingβ2-microglobulin

Markotić

Marušić

Tomac

et al. 2002

Clinical and Experimental Immunology

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SUMMARYThis study presents a comparative analysis of gangliosides from lymphoid (spleen and thymus) and other (brain, liver, lungs and muscle) tissues of C57BL/6 mice lacking the gene for b 2 -microglobulin (b 2 M), a constitutive component of the MHC class I molecule. Ganglioside fractions in the tissues of mice homozygous (b 2 M-/-) and heterozygous (b 2 M-/+) for the gene deletion were determined by high performance thin-layer chromatography (HPTLC), followed by immunostaining with specific polyclonal antibodies. Ubiquitous gangliosides G M3 (Neu5Ac) and G M3 (Neu5Gc) were the dominant gangliosides in the lungs of the control b 2 M-/+ mice, whereas the homozygous knockout mice had substantially decreased expression of these structures. The lungs of the b 2 M-/-mice also had reduced expression of T-lymphocyte-specific G M1b -type gangliosides (G M1b and GalNAc-G M1b ). b 2 M-deficient mice also had more G M1a and G D1a gangliosides in the liver, and several neolacto-series gangliosides were increased in the brain and lungs. This study provides in vivo evidence that the b 2 M molecule can influence the acquisition of a distinct ganglioside assembly in different mouse organs, implicating its non-immunological functions.

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