concanavalin-a and Leukemia-P388

An in vitro cytotoxic screening of extracts of Rhaphidophora korthalsii indicated cytotoxicity in the ether fraction. ED50 values of the extract against P388, Molt 4, KB and SW 620 were 12, 14, 8 and 13 micrograms/ml, respectively. The extract was relatively more toxic on P388 and Molt 4 cell lines at concentrations of 50 micrograms/ml and 100 micrograms/ml. Screening with mouse splenocytes showed that the hot water extract had splenocytes stimulating activity.

Topics: Adenocarcinoma; Adjuvants, Immunologic; Animals; Antineoplastic Agents, Phytogenic; Carcinoma, Squamous Cell; Cell Division; Colonic Neoplasms; Coloring Agents; Concanavalin A; Dose-Response Relationship, Drug; Female; Humans; Leukemia; Leukemia P388; Lymphocyte Activation; Mice; Mice, Inbred BALB C; Nasopharyngeal Neoplasms; Plant Extracts; Plant Lectins; Plants, Medicinal; Spleen; Tetrazolium Salts; Thiazoles; Tumor Cells, Cultured; Water

The requirements for primary in vitro plaque-forming cell (PFC) development in cultures of purified rat splenic B cells have been examined. Rat B cells were directly responsive to the type 1 antigen trinitrophenyl-Brucella abortus (TNP-BA), but both T cells and adherent accessory cells were required for B cell responses to the type 2 antigen TNP-Ficoll and the T cell-dependent (TD) antigen sheep erythrocytes (SRBC). However, the cellfree supernatants from concanavalin A-induced spleen cells of rat or mouse origin replaced the requirement for T cells and macrophages, and resulted in PFC development in response to TNP-Ficoll and SRBC and augmented PFC numbers in response to TNP-BA. Culture supernatants from induced murine T cell and macrophage cell lines were used to partially deduce the molecular requirements for the support of PFC development by rat B cells to these three antigens. Supernatants from the EL-4 (EL-4 sup) and B151 K12 (B15 sup) T cell lines augmented TNP-BA responses, suggesting that B cell growth factor II (BCGF-II) mediated this effect. An admixture of purified interleukin 2 (IL 2) and B15 sup supported PFC development to SRBC; indicating that IL 2, BCGF-II, and the T cell-replacing factor in B15 sup (B15-TRF) were sufficient to support this response. In addition, the IL 2 plus B15 sup-supported anti-SRBC PFC response was increased by the addition of an interleukin 1-containing fraction from the supernatant of the macrophage line P388D1. PFC development in response to TNP-Ficoll had the most stringent requirements and only occurred in the presence of EL-4 sup and B15 sup (IL 2, BCGF-I, BCGF-II, EL-TRF, B15-TRF). These data indicate that different cellular and molecular requirements exist for PFC development in response to types 1, 2, and TD antigens by rat B cells.

Topics: Animals; Antigen-Presenting Cells; Antigens, T-Independent; B-Lymphocytes; Brucella abortus; Concanavalin A; Female; Ficoll; Hemolytic Plaque Technique; Interleukin-1; Interleukin-2; Interleukin-5; Leukemia P388; Lymphocyte Activation; Lymphokines; Male; Mice; Mice, Inbred C3H; Rats; Rats, Inbred F344; T-Lymphocytes; Trinitrobenzenes

Cell surface alteration was studied in a subline of murine lymphocytic leukaemia resistant to the broad-spectrum anticancer agent adriamycin (P388/ADR) employing concanavalin A(Con A)-induced agglutination and rearrangement of lectin receptors. Con A induced more agglutination of P388/ADR when compared to the drug-sensitive parental cell line (P388/S). Studies on the redistribution of Con A and Ricinus communis agglutinin-I revealed a high percentage of P388/ADR showing internalized fluorescence, while a majority of P388/S displayed a uniform distribution of fluorescence on the cell surface.

Topics: Animals; Cell Aggregation; Cell Line; Cell Membrane; Cell Membrane Permeability; Concanavalin A; Doxorubicin; Drug Resistance; Leukemia P388; Leukemia, Experimental; Mice; Mutation; Receptors, Concanavalin A

The constitutive culture supernatant (SN) of the macrophage tumor line P388D1 (P388 SN) and the concanavalin A (Con A)-induced culture supernatant of the T cell hybridoma FS6-14.13 (FS6 Con A SN) were shown to contain nonspecific factors capable of inducing increased Ia expression by normal resting B cells in a dose-dependent manner. In six consecutive experiments the relative increase in Ia expression induced by P388 SN was 4.9 +/- 0.9, with FS6 Con A SN 10.7 +/- 1.5, and with a combination of both preparations 13.0 +/- 1.7. This increase in Ia expression was observed to occur in virtually all the B cells, reaching maximum levels within 24 h of culture. The interleukin-induced increase in B cell Ia expression occurred in the absence of ancillary signals provided by ligand-receptor Ig cross-linking and despite the fact that virtually all the control B cells, cultured in the absence of factors, remained in G0. These results suggest that functional receptors for at least some interleukins are expressed on normal resting B cells and their effects can be manifest in the absence of additional activating signals. The increased Ia expression induced by the nonspecific factor preparations was shown to be correlated with enhanced antigen-presenting capacity by the B cells to T cell hybridomas. The nature of the interleukins responsible for these effects remains to be definitively determined, however, the activity of FS6 Con A SN was shown to correlate with B cell growth factor activity and increased B cell Ia expression was not observed using interleukin 2 (IL-2) or interferon-gamma, prepared by recombinant DNA technology.

Topics: Animals; Antigens; B-Lymphocytes; Cell Cycle; Concanavalin A; Histocompatibility Antigens Class II; Interferon-gamma; Interleukin-1; Interleukin-2; Kinetics; Leukemia P388; Lymphocyte Cooperation; Lymphokines; Mice; Mice, Inbred Strains; Ovalbumin

While the combination of L1210 murine leukemia cell vaccine (L1210 vaccine) with 6-mercaptopurine (6-MP) or 6-thioguanine produces a therapeutic response greater than that induced by either of these agents alone, its combination with cyclophosphamide, N4-behenoyl-1-beta-D-arabinofuranosylcytosine, or 5-fluorouracil does not produce such a response. The administration of cyclophosphamide, N4-behenoyl-1-beta-D-arabinofuranosylcytosine, or 5-fluorouracil alone resulted in a response as great as, or greater than, that induced by 6-MP alone. This and the finding that the 6-MP-induced response was more pronounced upon its delayed rather than its early administration indicate that 6-MP-induced reduction of the tumor burden does not explain this augmentation. The combination of 6-MP and L1210 vaccine was not effective in mice bearing 6-MP-resistant L1210 leukemia; however, an augmented response occurred when the tumor burden was reduced by N4-behenoyl-1-beta-D-arabinofuranosylcytosine, indicating that reduction of the tumor burden by 6-MP was only partially associated with augmentation of the therapeutic response. Augmentation was associated with vaccine-induced antitumor immunity because it was induced by the combination of 6-MP and concanavalin A-bound, but not concanavalin A-free L1210 vaccine. This augmentation was dependent on the timing of the L1210 vaccine administration. The combination was not effective in mice bearing P388 leukemia, indicating the tumor specificity of the augmentation. These results show that 6-MP not only reduced the tumor burden but also potentiated the vaccine-dependent antitumor immunity, resulting in the induction of an augmented therapeutic response.

Topics: Animals; Concanavalin A; Dose-Response Relationship, Drug; Drug Administration Schedule; Drug Synergism; Immunotherapy; Leukemia; Leukemia L1210; Leukemia P388; Leukemia, Experimental; Male; Mercaptopurine; Mice; Thioguanine

Antigen-induced T cell proliferation requires T cell interaction with antigen in the context of MHC I region-compatible accessory cells. The resulting activation and proliferation of T cells involves the production and utilization of several lymphokines or interleukins. This report describes experiments wherein these events could be separated into two phases, T cell activation and T cell proliferation. The first phase was achieved by stimulating antigen-specific T cell lines with antigen-pulsed ultraviolet light-irradiated accessory cells. T cell proliferation (second phase) could then be initiated by the addition of a soluble lymphokine with the characteristics of interleukin 1 (IL 1). These effects were only observed with homologous antigen and accessory cells syngeneic to the T cells at the I-A and E/C subregion of the MHC. This report has two applications in the study of lymphocyte-lymphokine interactions. First, T cell recognition of antigen and antigen-induced T cell proliferation can be examined as physically separate events. Secondly, this system may be used as a specific and sensitive means of measuring the effects of IL 1 on T cells.

Topics: Animals; Concanavalin A; Epitopes; gamma-Globulins; Interleukin-1; Leukemia P388; Lipopolysaccharides; Lymphocyte Activation; Lymphokines; Macrophages; Mice; Mice, Inbred A; Monokines; Ovalbumin; Protein Biosynthesis; Proteins; Rats; Rats, Inbred Lew; T-Lymphocytes

We characterized an assay system to study the lymphokine-mediated induction of antigen-presenting ability in P388D1 cells. The ability of lymphokine-induced P388D1 macrophages to present antigen plus Id was measured by their ability to induce interleukin 2 production by antigen-specific, Id-restricted T cell hybridomas in the presence of the appropriate antigen. The production of IL 2 by the T cell hybridomas is known to be dependent on the expression of Ia antigens by the antigen-presenting cells. The results obtained suggest that a factor present in the supernatant of the T cell hybridoma FS7-20.6.18 is responsible for inducing the appearance of I-Ad and I-Ed on P388D1, measured by immunofluorescence, and the ability of the cell to present antigen in association with I-Ad or I-Ed. The factor mediating the induction of antigen-presenting ability is thought to be gamma-interferon, because the hybridoma FS7-20.6.18 is known to produce this lymphokine and the factor is sensitive to pH 2 incubation. gamma-Interferon produced by recombinant DNA technology was found to induce antigen-presenting ability in this assay; however, alpha- and beta-interferon were inactive. This observation suggests a unique immunoregulatory role for gamma-interferon. Using many T cell hybridomas in the assay, we were able to distinguish three groups: a) high avidity hybridomas that respond to antigen presented by uninduced P388D1 but show an enhanced response to antigen plus induced P388D1; b) medium avidity hybridomas that do not respond to antigen presented by uninduced P388D1; and c) low avidity hybridomas that show a limited response to antigen presented by induced P388D1, but the response of which increases if the P388D1 cells are induced for longer periods of time. These different patterns of response are believed to be dependent on the Ia antigen density expressed by the gamma-interferon-induced presenting cells, and suggest that the T cell receptors for Ag/Id display marked heterogeneity in their avidities for Ag/Id.

Topics: Animals; Cell Line; Concanavalin A; Dose-Response Relationship, Immunologic; Epitopes; Histocompatibility Antigens Class II; Hybridomas; Hydrogen-Ion Concentration; Interferon-gamma; Kinetics; Leukemia P388; Leukemia, Experimental; Lymphokines; Macrophages; Mice; Mice, Inbred BALB C; T-Lymphocytes

The antitumor effect of protein-bound polysaccharide, EA6, derived from fruit bodies of Flammulina velutipes (Curt. ex Fr.) Sing., when combined with a vaccine treatment was studied by the challenge test in BDF1 mice and L1210 murine leukemia system. Intensification of the antitumor effect of EA6 was dependent on doses, timing, and frequency of intraperitoneal administration of the material to the immunization by concanavalin A and/or glutaraldehyde treated L1210 vaccine. Administration of EA6 prior to the injection of the vaccine, or repeated injection of more than 4 times did not increase the life span of the animals. But when EA6 was given (40 mg/kg) after the injection of the vaccine, marked prolongation of the life span (ILS of 223%) was observed against challenging of 1 x 10(2) cells of L1210. Combined treatment of EA6 with vaccine exhibited prolonged ILS in the mice challenged with 1 x 10(3) cells of L1210. The specific immunity for L1210 induced by the vaccine was not affected by EA6.

Topics: Agaricales; Animals; Antigens, Fungal; Concanavalin A; Female; Immunity; Immunization; Leukemia L1210; Leukemia P388; Leukemia, Experimental; Mice; Polysaccharides; Time Factors; Vaccines

Topics: Adenosine; Adenosine Triphosphate; Animals; Biological Transport; Coformycin; Concanavalin A; Dipyridamole; Dogs; Hypoxanthines; Kinetics; Leukemia P388; Leukemia, Experimental; Leukocytes; Mice; T-Lymphocytes