Noncytolytic extraction of cell surface antigens using butanol

LeGrue, Stephen J.

doi:10.1007/bf00048096

Cited by 19 publications

(12 citation statements)

References 54 publications

(53 reference statements)

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“…Alternatively, the TATA may reside both in the cytoplasm and plasma membrane or the TATA may be very loosely bound to the cell membrane and shed during the membrane preparation. This last explanation is not supported by the low immunogenicity of the butanol extract, as 2.5% butanol extraction of tumour cells yields loosely bound peripheral components of the cell membrane [20]. Extraction of cells with 3 M KC1 also yields peripheral membrane components, but these extracts contain large amounts of intracellular proteins as well.…”

Section: Discussionmentioning

confidence: 94%

“…The extraction of viable tumour cells with a low concentration of butanol [20] yields only peripheral membrane components. Hydrophobic TAA that are more readily bound to the cell membrane, such as embryonic antigens from Simian-virus-40-and adenovirus-7-induced hamster sarcomas [6], and altered class I major histocompatibility TATA of an ultraviolet-light-induced tumour [35] can not be isolated by butanol extraction.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Reconstituted membranes of tumour cells (proteoliposomes) induce specific protection to murine lymphoma cells

Bergers

Otter

Groot

et al. 1992

Cancer Immunol Immunother

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Antigens presented on cell membranes or on liposomes are usually more immunogenic than antigens in soluble form, this being one of the reasons for the weak immunogenicity of extracted tumour-associated transplantation antigens (TATA). The main objective of this study is to solubilize TATA from tumour cells and to present them on a membrane-like structure to the immune system. Crude tumour cell membranes of SL2 lymphosarcoma cells (a spontaneously arising, weakly immunogenic tumour) were solubilized with octylglucoside or sodium deoxycholate, and reconstituted membranes (proteoliposomes) were prepared by detergent removal. Mice immunized s.c. with reconstituted membranes were protected against an i.p. challenge with tumour cells. Although octylglucoside solubilized only 41% of the membrane proteins, the reconstituted membranes were as immunoprotective as crude membranes. (Glyco)proteins were probably the major membrane components in the reconstituted membranes that induce immunoprotection, as mice immunized with preparations constituted of (glyco)lipids from SL2 cells could not reject SL2 cells. If Freund's complete adjuvant was used with the first immunization injection, no potentiation of the elicited immune responses was observed. Besides the membrane TATA, SL2 cells contained an apparently non-membrane-bound TATA, which was found in the cytoplasm. It is concluded that detergent solubilization of membranes and subsequent preparation of reconstituted membranes can be used to obtain membrane tumour-associated antigens that retain activity for induction of protective tumour immunity. The major advantage of this method is that membrane proteins are solubilized and are subsequently presented on a membrane-like structure that resembles the tumour cell membrane. On theoretical and practical grounds it provides a promising alternative for whole-cell vaccines.

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Section: Discussionmentioning

confidence: 94%

Section: Discussionmentioning

confidence: 99%

Reconstituted membranes of tumour cells (proteoliposomes) induce specific protection to murine lymphoma cells

Bergers

Otter

Groot

et al. 1992

Cancer Immunol Immunother

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“…We do not know how the 22S complex is anchored to the cell surface. According to LeGrue (17), butanol removes external but not integral membrane proteins from live cells. In this respect our complex resembles other external cell surface proteins, and it is probable that, like fibronectin, it is attached through a specific site to an integral membrane protein receptor (18,19).…”

Section: Figmentioning

confidence: 99%

Characterization of toposomes from sea urchin blastula cells: a cell organelle mediating cell adhesion and expressing positional information.

Noll¹,

Matranga²,

Cervello³

et al. 1985

Proc. Natl. Acad. Sci. U.S.A.

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Cell adhesion in the sea urchin blastula is mediated by a 22S genus-specific glycoprotein complex consisting initially of six 160-kDa subunits that are processed proteolytically as development proceeds. Noncytolytic removal of the 22S particle from the surface with either 2.5% butanol or trypsin renders dissociated cells reaggregation incompetent, and addition restores reaggregation and development. Polyclonal antibodies against the 22S complex prevent reaggregation in a genus-specific manner while monoclonal antibodies stain cell surface structures in a pattern consistent with a code that specifies the position of a cell in the embryo by a unique combination of subunits in its cell adhesion particles. The existence of similar particles in Drosophila and amphibian embryos suggests that these glycoprotein complexes are a general class of organelles, the toposomes, that in the embryo mediate cell adhesion and express positional information.Sea urchin embryos are uniquely suited for the study of cell-cell interaction in embryogenesis because removal of Ca2+ and Mg2' causes them to dissociate into single cells that, upon restoration of these ions, reaggregate spontaneously to reform a developing embryo (1). Reaggregation is blocked in a strictly genus-specific manner by antibodies (or their Fab fragments) against purified membranes or against butanol extracts from purified membranes (2, 3). Equally inhibitory are antibodies raised against the components extracted noncytolytically from live cells with 2.5% (vol/vol) butanol (2). The proteins solubilized by butanol from purified membranes or from the cell surface neutralize the inhibiting antibodies, stimulate reaggregation, and reconstitute cells that had been rendered reaggregation incompetent by noncytolytic extraction with butanol (2).Attempts to purify the active component gave results suggesting that the activity was associated with a large glycoprotein complex. Thus, when the butanol extract was chromatographed on phenyl-Sepharose or DEAE-Sepharose, the reaggregation-promoting activity was eluted as a single peak that in NaDodSO4 gels was resolved into a defined set of glycoproteins ranging from about 70 to 180 kDa that was identical for the two purification methods (4). Here we report that all the cell adhesion activity in sea urchin blastula embryos detectable in our bioassays is indeed the property of an oligomeric glycoprotein particle. Similar particles have been isolated previously from a number of other sea urchin genera (cited in ref. 5). While the biological function of this particle remained obscure, it was found to originate in yolk granules and to undergo developmentally regulated processing into a number of fragments held together by S-S bridges and noncovalent bonds (5).After the studies reported here were completed, strikingly similar glycoprotein complexes were discovered on the surface of imaginal disc cells of Drosophila by screening for monoclonal antibodies with positional specificity (6). The similarity of the results in sea urchins...

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“…For example, highly malignant lymphoma cells (RAW117) have been shown to inhibit both T-cell and B-cell mitogen-induced proliferation of normal murine spleen cells. Interestingly, a similar inhibition was observed when RAW117 cells were extracted with butanol 28, and this extract was used instead of whole RAW117 cells 29. In another study where cell surface molecules from lymphoma cells were extracted with butanol, an inhibitory effect of this extract on natural killer and lymphokine activated killer cell mediated cytotoxicity was also observed 30.…”

Section: Cell Surface Molecules and Cancer Immunitymentioning

confidence: 58%

Cellular Cancer Vaccines: an Update on the Development of Vaccines Generated from Cell Surface Antigens

Lokhov¹,

Balashova²

2010

J. Cancer

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A recent advance in anti-cancer therapies has been the use of cancer cells to develop vaccines. However, immunization with cancer cell-based vaccines has not resulted in significant long-term therapeutic benefits. A possible reason for this is that while cancer cells provide surface antigens that are targets for a desired immune response, they also contain a high abundance of housekeeping proteins, carbohydrates, nucleic acids, lipids, and other intracellular contents that are ubiquitous in all mammalian cells. These ubiquitous molecules are not the intended targets of this therapy approach, and thus, the immune response generated is not sufficient to eliminate the cancer cells present. In this review, a discussion of the cell surface of cancer cells is presented in relation to the goals of improving antigen composition of cancer cell-based vaccines. Strategies to enrich vaccines for cancer-specific antigens are also discussed.

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Noncytolytic extraction of cell surface antigens using butanol

Cited by 19 publications

References 54 publications

Reconstituted membranes of tumour cells (proteoliposomes) induce specific protection to murine lymphoma cells

Reconstituted membranes of tumour cells (proteoliposomes) induce specific protection to murine lymphoma cells

Characterization of toposomes from sea urchin blastula cells: a cell organelle mediating cell adhesion and expressing positional information.

Cellular Cancer Vaccines: an Update on the Development of Vaccines Generated from Cell Surface Antigens

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