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Biomedical subjects

S Junqua

Publications and source records attributed to S Junqua.

13 recordsLinked to original sources

Decreased accessibility of globotriaosylceramide associated with decreased tumorigenicity in Burkitt's lymphoma variants induced by immunoselection.

To investigate a role for globotriaosylceramide (Gb3) as a tumor-associated antigen, variant cells resistant to treatment with complement and monoclonal antibody 38-13, which recognizes Gb3, were selected from a Burkitt's lymphoma cell line, Ramos. Variant cells displayed a clear decrease of antibody-binding capacity whereas the amount of Gb3 at their plasma membrane was not significantly different from that of Ramos parental cells. This demonstrated a reduced accessibility of Gb3 at the surface of variant cells. In parallel, no changes in other surface antigens were recorded as compared to those in Ramos cells. No changes of proliferative properties in suspension culture or of c-myc expression were observed although variant cells showed a decreased colony-forming capacity in agar. Variant cells showed a significant reduction in tumorigenic potential when injected s.c. into nude mice. The decreased tumorigenicity appeared related to the low antibody-binding capacity because both tumorigenicity and Gb3 antigenicity were recovered in parallel in revertant cells growing in suspension culture. In vivo, after two transplantations of variant cells into mice, cells isolated from the few induced tumors still retained the low antibody-binding capacity.

Animals

Kinetic analysis of choriocarcinoma cell intoxication induced by ricin and ricin A chain immunotoxin.

The kinetics of protein synthesis inhibition was studied in the choriocarcinoma-derived BeWo cell line treated with ricin and an immunotoxin (IT) constructed by linking a specific antibody to the A chain of ricin. The IT was specifically cytotoxic to BeWo and other choriocarcinoma cells. The multistep process underlying this kinetics was explored using two mathematical models where the protein synthesis-inactivation step is preceded by one or two processing rate-limiting steps. Theoretical curves were computed for different concentrations of toxic reagents. Two processing steps were found necessary to predict the duration of the observed latent period before initiation of protein synthesis inhibition. With this model, a satisfactory fit was obtained. A mathematical modeling of the intoxication induced by ricin or ricin A chain IT thus requires two processing steps to account for the data observed. In addition, the data suggest that the cytoplasmic internalization of ricin is a slow process compensated by an extremely fast enzymatic inactivation of ribosomal activity. This implies that in this and similar systems, endocytosis might not be involved in the cytoplasmic internalization of the few molecules of ricin A chain responsible for cell intoxication.

Antibodies, Monoclonal

Comparison of inhibitory effect of galactose analogs on the binding and cytotoxicity of an anti-globotriaosylceramide monoclonal antibody coupled or not coupled to pokeweed antiviral protein.

The results described here provided an example of a human IgM monoclonal antibody against a tumor-associated glycolipid and of the unusual properties of its corresponding immunotoxin (IT). The monoclonal antibody referred to as 38-13 has been previously described and reacted with the globotriaosylceramide [Gb3:Gal(alpha 1----4)-Gal(beta 1----4)-Glc(beta 1----1)ceramide] specifically expressed on surface membrane of Burkitt's lymphoma (BL) cells. An immunotoxin (38-13 IT) combined with the pokeweed antiviral protein (PAP) toxin via S-S bridges showed paradoxically a lower cytotoxic effect in BL Ramos cells than in non-BL cells such as leukemic mouse L1210 cells, while these cells appeared not to be involved by flow cytometric analysis and complement-dependent cytotoxicity. Consequently, the inhibitory effect of selective galactose analogs on the binding and the cytotoxicity of 38-13 antibody conjugated or not to PAP toxin was compared on BL and non-BL cells. Only the galactose blocked in alpha configuration provided a fine inhibition of 38-13 binding on BL Ramos cells and both alpha and beta-galactose allowed us to establish a clear distinction between the pathway entry of 38-13 IT in BL and non-BL cells; in close correlation with the 38-13 binding specificity the 38-13 IT cytotoxic effect in Ramos BL cells could also be prevented by alpha-Gal only, suggesting that this toxic action is probably mediated through the IT binding to Gb3 antigenic sites. In contrast, on apparently irrelevant L1210 cells, 38-13 IT showed a cytotoxic effect which was inhibited preferentially by lactose (Gal in beta configuration). It was discussed that IT binding alone to either antigenic sites which are inhibited by the hapten alpha-Gal, or nonspecific sites which can compete with the hapten beta-Gal is unable to induce efficient killing of cells. But cooperation of both bindings might give an attractive explanation of IT cytotoxic effect. It was concluded that the unexpected activity of 38-13 IT in non-BL cells probably could be mediated through an active macromolecular transport process which could implicate a beta-galactoside-binding protein (lectin).

Animals

Antitumor activity against L1210 mouse leukemia of immunotoxins containing a monoclonal antibody to the glycolipid globotriaosylceramide.

Since immunotoxin (IT) containing the antiglobotriaosylceramide monoclonal antibody was found to be cytotoxic in murine L1210 leukemia cells, its potential antitumor activity could be evaluated in animals using the L1210 model. In vitro, L1210 cells incubated IT before grafting in DBA/2 mice failed to induce leukemia. All tumor cells were neutralized by IT. In animals, a significant but limited therapeutic effect on leukemic mice was obtained when IT was injected i.p. shortly after L1210 cell grafting. In contrast, no toxic effect of IT was observed in non-leukemic mice at doses far above those used in our therapeutic treatment. The potentiation effect of chloroquine on IT was moderated when a cloning efficiency assay was used, but 70% of the mice grafted with in vitro chloroquine-treated L1210 cells were cured with IT treatment.

Animals

Determination of antibody-complement mediated cytotoxicity using ATP release induced by a monoclonal antibody against the Burkitt lymphoma associated globotriaosylceramide antigen.

The kinetics of ATP release from Burkitt lymphoma target cells following complement activation by rat monoclonal IgM antibody have been used to develop a standard assay for antibody-complement mediated cytotoxicity. This assay permitted the study of the reactivity of the monoclonal IgM with Burkitt and non-Burkitt cell lines and to perform IgM-complement mediated cytotoxicity inhibition experiments with oligosaccharides. This led to a precise definition of the antigenic determinant recognized by the monoclonal IgM on globotriaosylceramide.

Adenosine Triphosphate

[Regulation of the biosynthesis of macromolecules of the intercellular corneal matrix].

The influence of a 1M CaCl2 extract and of a collagenase digest of corneal stroma of the biosynthesis of the macromolecules of corneal stroma was investigated. Calf corneas were incubated "in vitro" with radioactive tracers (14C-L-proline; 3H-D-glucosamine or 35SO4) in the presence or absence of the above extracts. After incubation the corneas are submitted to a fractional extraction in order to separate the major macromolecular fractions of the stroma. An increase in incorporation of all tracers is observed in the 1M CaCl2 (CTC), TCA and urea-extracts (containing resp. the diffusible macromolecules, proteoglycans, polymeric collagen and structural glycoproteins) in the presence of the macromolecular extracts and also with the collagenase-hydrolysate of cornea. These results show the existence of a stimulation of the biosynthesis of intracellular matrix macromolecules of the cornea by corneal extracts, probably through positive "feedback" type of mechanism.

Animals