Production of a tumor-specific xenoantiserum from partially purified immunoprotective tumor antigen

Tanaka, Tatsuhide; Yamagishi, Hisakazu; Pellis, Neal R.; Kahan, Barry D.

doi:10.1007/bf00200200

Cited by 3 publications

(9 citation statements)

References 18 publications

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“…The ability of butanol-extracted antigen to associate functionally with cell membranes and alter the antigenic phenotype was further demonstrated in vitro using a xenoantiserum which was functionally specific for MCA-F in an indirect immunofluorescence assay [29]. Membrane immunofluorescence was completely ablated from MCA-F cells by first extracting the targets in 2.5% 1-butanol ( Table 2).…”

Section: Adsorption Of Antigen Onto Cells and Vesiclesmentioning

confidence: 96%

Section: Indirect Immunofluorescence Assaymentioning

confidence: 99%

“…The assays were performed as described by Tanaka et al [29], using xenoantibodies raised in rabbits against the electrofocused, 3 M KCl-extracted MCA-F antigen. This antiserum binds to the homotypic tumor, MCA-F, but displays little or no binding to the antigenically distinct tumor, MCA-D [29]. Specific membrane immunofluorescence was calculated as the fluorescence index, defined as: % Unstained cells in normal rabbit serum -% Unstained cells in antiserum % Unstained cells in normal rabbit serum…”

Section: Indirect Immunofluorescence Assaymentioning

confidence: 99%

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Carrier and adjuvant properties of liposome-borne tumor-specific antigens

LeGrue¹

1984

Cancer Immunol Immunother

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The purpose of this study was to examine (1) the association of tumor extract proteins with phospholipid vesicles of varying physiochemical properties, and (2) the adjuvant and carrier properties of liposome-borne tumor antigens in the in vivo induction of an antitumor immune response. Cell surface antigens of the 3-methylcholanthrene-induced fibrosarcoma of C3H/HeJ mice, MCA-F, were extracted using 2.5% 1-butanol. Crude and electrofocused antigen preparations capable of eliciting a protective antitumor immune response were used to prepare liposome vaccines. The incorporation of extract proteins into liposomes formed by butanol dialysis (BVD) was three- to five-fold greater than the encapsulation of protein into the aqueous compartment of multilamellar vesicles (MLV). The electrochemical properties of the BDV had a significant effect on the induction of an antitumor response: Antigens borne on negatively charged, but not uncharged, liposomes were effective in protecting hosts against supralethal tumor challenge, and displayed a specific activity 20- to 50-fold greater than soluble antigen. Antigens carried by MLV were not effective in generating an immunoprotective response. The lipophilic characteristics of butanol-extracted antigens allowed (1) the passive adsorption of immunoprotective tumor antigen onto the surface of preformed vesicles, and (2) adsorption of MCA-F antigen onto the surface of an antigenically distinct tumor MCA-D. In the latter experiment, adsorption of MCA-F-specific antigen onto MCA-D cells resulted in a change in the membrane antigen phenotype as measured by indirect immunofluorescence. Although butanol released a lipophilic moiety from cells which spontaneously reassociated with phospholipid bilayers, no evidence for a lipoidal antigen was obtained when tumor-derived lipids were used as immunogens. This study demonstrates that butanol-extracted tumor antigen is lipophilic without being a lipid, and that negatively charged liposomes can be effective as carriers and adjuvants for tumor antigens in the induction of an antitumor immune response.

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Section: Adsorption Of Antigen Onto Cells and Vesiclesmentioning

confidence: 96%

Section: Indirect Immunofluorescence Assaymentioning

confidence: 99%

Section: Indirect Immunofluorescence Assaymentioning

confidence: 99%

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Carrier and adjuvant properties of liposome-borne tumor-specific antigens

LeGrue¹

1984

Cancer Immunol Immunother

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“…Rabbit antisera were prepared as described by Tanaka et al [34]. White New Zealand rabbits were immunized SC with TSTA preparations emulsified in incomplete Freund's adjuvant.…”

Section: Isotachophoresismentioning

confidence: 99%

“…Rabbit antisera were raised against the following antigens: 3 M KCl-extracted, pIEF-purifled TSTA; butanol-extracted, pIEF-purified TSTA; and crude butanol extracts. These antisera were functionally specific for MCA-F tumor cells as assessed by an indirect membrane immunofluorescence assay [34]. Antisera were produced in a xenogeneic host because of the difficulty in generating antibodies to TSTA in the syngeneic host [7,9,16].…”

Section: Isotachophoresismentioning

confidence: 99%

Purification of immunoprotective tumor antigens by preparative isotachophoresis

Saunders

Kahan

Pellis

1983

Cancer Immunol Immunother

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Tumor-specific transplantation antigens (TSTA) were purified from 3 M KCl and butanol extracts of C3H/HeJ 3-methylcholanthrene-induced fibrosarcomas by preparative isotachophoresis (pITP). Fractions from pITP which reacted with antisera to TSTA preparations in an enzyme-linked immunospecific assay were tested in vivo for induction of resistance to the growth of transplanted tumor cells. Isotachophoresis of crude 3 M KCl extracts yielded TSTA that was immunogenic at doses between 17 and 124 micrograms. Isotachophoresis of TSTA partially purified from crude 3 M KCl or butanol extracts by preparative isoelectric focusing (pIEF) of 3 M KCl and butanol extracts yielded TSTA that was immunogenic over a two-fold log dose range. As little as 10 ng purified TSTA reduced tumor growth by 50%. Tumor growth reduction was specific, and immunized animals survived longer than non-immunized controls. A purification of 10,000-fold over crude 3 M KCl extracts and 2,500-fold over crude butanol extracts was obtained. These results suggest that TSTA from murine tumor cells is preparatively purified by following extraction with pIEF and then pITP.

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Noncytolytic extraction of cell surface antigens using butanol

LeGrue

1985

Cancer Metast Rev

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Incubation of viable tumor cells in single-phase aqueous solutions of 1-butanol releases a subset of peripherally-associated membrane proteins. Among the extracted components are tumor-specific and tumor-associated antigens of several human and experimental neoplasms. The extraction technique is noncytolytic: extracted cells remain viable and proliferate both in vitro and in vivo. Examination of the denuded cells has elucidated several important differences in cell-surface phenotype that have proved useful in studying the biology of such diverse systems as cellular communications and the mechanisms of hematogenous metastasis. It is anticipated that noncytolytic butanol extraction will continue to prove a powerful approach for the isolation and characterization of a variety of cell-surface antigens.

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Production of a tumor-specific xenoantiserum from partially purified immunoprotective tumor antigen

Cited by 3 publications

References 18 publications

Carrier and adjuvant properties of liposome-borne tumor-specific antigens

Carrier and adjuvant properties of liposome-borne tumor-specific antigens

Purification of immunoprotective tumor antigens by preparative isotachophoresis

Noncytolytic extraction of cell surface antigens using butanol

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