Expression and shedding of major histocompatibility complex product and blood group antigens by cells in monolayer cultures

Plesser, Y M; Weiss, David; Markson, Y.; Doljanski, F.

doi:10.1016/0008-8749(80)90273-7

Cited by 9 publications

(2 citation statements)

References 15 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…A more sophisticated analytical approach using photoreactive probes [33] may be required to confirm this contention. In view of the relatively firm membrane association of gp87, and its expression on the membrane of melanoma cells [8, lo], gp87 may be released by the process of membrane shedding such as that described for histocompatibility antigens [34]. A process similar to the antigen shedding from cultured melanoma cells may well operate in vivo.…”

Section: Discussionmentioning

confidence: 99%

Immunopurification, characterization, and nature of membrane association of human melanoma‐associated oncofetal antigen gp87 defined by monoclonal antibody 140.240

Liao

Kwong

Khosravi

1985

J of Cellular Biochemistry

View full text Add to dashboard Cite

A melanoma-associated oncofetal antigen, gp87 (a p97-like molecule), defined by the monoclonal antibody (MoAb) 140.240 has been purified to homogeneity from the spent medium of cultured melanoma cells by a two-step immunoadsorbent procedure. The first immunoadsorbent step using glutaraldehyde-insolubilized MoAb 140.240 (ascites fluid) resulted in a 13-fold enrichment with 93% recovery in the bound material. In the second immunoadsorbent step constructed by the purified IgG2a of MoAb 140.240 (culture fluid) coupled to CNBr-activated Sepharose 4B the bound material from the first step was further purified resulting in a 330-fold purification with 90% recovery. SDS-polyacrylamide gel electrophoretic analysis of the final purified material revealed a single band migrating as a polypeptide with an approximate molecular weight of 87 Kd, consistent with the size of the molecule immunoprecipitated by MoAb 140.240 from lysates of radiolabelled melanoma cells. Preliminary amino acid analysis indicates a particularly high proportion of phenylalanine in gp87. We have also compared gp87 with two well defined antigens, HLA-A,B,C (integral membrane protein) and "94K" melanoma/carcinoma-associated antigen (peripheral membrane protein) with respect to antigen extractability from melanoma cells using phosphate-buffered saline, 0.1 M urea, 3 M NaCl, or nonionic detergent (NP-40). The results showed that whereas 94K antigen was extractable by each of the four different solutions, gp87, similar to HLA-A,B,C antigens, could only be extracted with NP-40, strongly suggesting that gp87 is an integral melanoma cell component.

show abstract

Section: Discussionmentioning

confidence: 99%

Immunopurification, characterization, and nature of membrane association of human melanoma‐associated oncofetal antigen gp87 defined by monoclonal antibody 140.240

Liao

Kwong

Khosravi

1985

J of Cellular Biochemistry

View full text Add to dashboard Cite

show abstract

“…Proteins such as membrane-associated immunoglobulins (15), mouse alloantigens encoded by the major histocompatibility complex (16), human histocompati-* Corresponding author. bility antigens (40), and blood group substances (40) have been shown to be shed from the surface of normal cells. Cell surface tumor-associated antigens are shed by murine (8) and human (9) melanoma cells and by methylcholanthreneinduced rat sarcoma cells (1, 12).…”

mentioning

confidence: 99%

Dynamic nature of the association of large tumor antigen and p53 cellular protein with the surfaces of simian virus 40-transformed cells

Santos¹,

Butel²

1984

J Virol

View full text Add to dashboard Cite

A molecular complex of simian virus 40 large tumor antigen (T-Ag) and p53 cellular protein is present on the surface of simian virus 40-transformed mouse cells. The stability of the association of the two proteins with the cell surface was characterized. Cells were either surface iodinated by the lactoperoxidase technique or metabolically labeled with [35S]methionine, and surface antigens were detected by differential immunoprecipitation with specific antibodies immediately after labeling or after incubation at 37°C. A rapid, concomitant disappearance of TAg and p53 from the cell surface was observed. The half-life of iodinated surface TAg was less than 30 min, whereas that of [35S]methionine-labeled surface TAg was 1 to 2 h. Although TAg and p53 were rapidly lost, both were also rapidly replaced on the cell surface, since newly exposed molecules could be detected when cells were reiodinated after a 2-h chase period. Control experiments established that the loss of the surface molecules was not induced by the iodination reaction. The appearance of surface TAg was prevented when cellular protein synthesis was inhibited with cycloheximide. The disappearance and replacement of TAg and p53 appeared to be energy-independent processes, as neither was inhibited by sodium azide or 2,4-dinitrophenol. Incubation of iodinated cells at 4°C did block the loss of TAg and p53. These observations suggest that TAg and p53 are coordinately turned over in the plasma membrane. The nature of the association of the TAg -p53 complex with the cell surface can best be described as highly dynamic.

show abstract

Presence of normal human cell surface antigens in plasma of athymic mice bearing a human colon carcinoma and in normal human plasma

Markson

Weiss²,

Weiss³

et al. 1985

Cancer Immunol Immunother

View full text Add to dashboard Cite

The mixed haemadsorption (MHA) method was employed for detection of several normal antigenic components on the surface of human colon carcinoma cells (HT-29). The antigens were expressed by cells in monolayer cultures and in suspensions prepared by monolayer trypsinization, and by cells of tumours growing progressively in athymic mice. The plasma of such animals bearing medium sized and large, non-necrotic tumours contained all the antigens, as determined by the radial diffusion immune haemolysis method (RDIH); the plasma of animals with small or large heavily necrotic tumours did not contain detectable amounts of any of the determinants. The half-life of the determinants in the circulation as extracellular entities was ca. 20 h. The same antigens, and fibronectin, were found to be ubiquitously represented in normal human plasma. It is proposed that the presence of membrane antigens in plasma is the result of physiological shedding of cell surface constituents by living cells.

show abstract

Expression and shedding of major histocompatibility complex product and blood group antigens by cells in monolayer cultures

Cited by 9 publications

References 15 publications

Immunopurification, characterization, and nature of membrane association of human melanoma‐associated oncofetal antigen gp87 defined by monoclonal antibody 140.240

Immunopurification, characterization, and nature of membrane association of human melanoma‐associated oncofetal antigen gp87 defined by monoclonal antibody 140.240

Dynamic nature of the association of large tumor antigen and p53 cellular protein with the surfaces of simian virus 40-transformed cells

Presence of normal human cell surface antigens in plasma of athymic mice bearing a human colon carcinoma and in normal human plasma

Contact Info

Product

Resources

About