Antigenicity of a Spontaneous Murine Mammary Adenocarcinoma during in Vitro Cultivation

1977 ◽  
Vol 63 (5) ◽  
pp. 415-427 ◽  
Author(s):  
Anwar A. Hakim
Keyword(s):  
Extracellular Protease ◽  
Mammary Adenocarcinoma ◽  
In Vitro Cultivation ◽  
Membrane Glycoproteins ◽  
Spleen Cells ◽  
Alpha 1 Antitrypsin ◽  
Immunogenic Properties ◽  
Cells Cultured ◽  
Immune Spleen Cells

The present study reports on the transplantability and immunogenic properties of a murine spontaneous mammary adenocarcinoma (AdCa) cultured under various conditions. When cultured under standard conditions for 300 days, AdCa cells became nononcogenic, and 74 % survived the effects of immune spleen cells. Cell cultures grown in presence of the « extracellular protease » retained their transplantability, and only 25 % survived the effects of immune spleen cells. In cotton-stoppered bottles without HEPES, the cells retained their transplantability and survived after incubation with immune spleen cells, whereas AdCa cells cultured in presence of human alpha-1-antitrypsin became less oncogenic and remained sensitive to the killing effects of immune spleen cells. When treated with serum of mice or rabbits immunized with AdCa cells, the cells cultured under standard conditions or grown in presence of the human antitrypsin became non-transplantable, whereas cells grown in cotton-stoppered glass bottles or in presence of the « extracellular protease » retained their oncogenicity. When cultured AdCa cells were incubated with cAMP and then used to immunize syngeneic hosts, tumors arising from a challenging dose of untreated AdCa cells grew faster in mice immunized with cAMP-treated cells than in mice immunized with untreated cells. On the other hand, immunization with AdCa cells treated with EDTA, iodoacetate, or heparin reduced the transplantability of the challenging AdCa cells. Treatment with vibrio cholera neuraminidase (VCN) at pH 5.6 increased the immunogenicity of AdCa cells: immunization with VCN-treated AdCa cells protected syngeneic hosts against challenge with untreated AdCa cells, whereas formalin-or glutaraldehydeflxed untreated AdCa cells gave a transient protection. Rabbit and mouse antisera raised against VCN-treated AdCa cells and against membrane glycoproteins of untreated AdCa cells showed definite though variable cytotoxic activity against cultured untreated AdCa cells.

scholarly journals Antigen-specific suppressor T-cell activity in genetically restricted immune spleen cells.

1978 ◽  
Vol 148 (5) ◽  
pp. 1271-1281 ◽  
Author(s):  
C W Pierce ◽  
J A Kapp
Keyword(s):  
T Cells ◽  
T Cell ◽  
Cell Activity ◽  
Antibody Responses ◽  
Spleen Cells ◽  
Suppressor T Cells ◽  
Specific Suppressor T Cells ◽  
Immune Spleen Cells

Virgin spleen cells develop comparable primary antibody responses in vitro to syngeneic or allogeneic macrophages (Mphi) bearing the terpolymer L-glutamic acid60-L-alanine30-L-tyrosine10 (GAT), whereas immune spleen cells primed with syngeneic or allogeneic GAT-Mphi develop secondary responses preferentially when stimulated with GAT-Mphi syngeneic to the GAT-Mphi used for priming in vivo. These restrictions are mediated by products of the I-A subregion of the H-2 complex and are operative at the level of the GAT-Mphi-immune helper T-cell interactions. To investigate why these immune spleen cells fail to develop a significant antibody response to GAT-Mphi other than those used for in vivo immunization and determine the mechanism by which the restriction is maintained, spleen cells from virgin and syngeneic or allogeneic GAT-Mphi-primed mice were co-cultured in the presence of GAT-Mphi of various haplotypes. Antibody responses to GAT developed only in the presence of GAT-Mphi syngeneic to the Mphi used for in vivo priming; responses in cultures with GAT-Mphi allogeneic to the priming Mphi, whether these Mphi were syngeneic or allogeneic with respect to the responding spleen cells, were suppressed. The suppression was mediated by GAT-specific radiosensitive T cells. Thus, development of GAT-specific suppressor T cells appears to be a natural consequence of the immune response to GAT in responder as well as nonresponder mice. The implications of stimulation of genetically restricted immune helper T cells, and antigen-specific, but unrestricted, suppressor T cells after immunization with GAT-Mphi in vivo are discussed in the context of regulatory mechanisms in antibody responses.

scholarly journals Therapeutic Effect of Clarithromycin on a Transplanted Tumor in Rats

1999 ◽  
Vol 43 (1) ◽  
pp. 67-72 ◽  
Author(s):  
Kazuhiko Sassa ◽  
Yutaka Mizushima ◽  
Takashi Fujishita ◽  
Rokuo Oosaki ◽  
Masashi Kobayashi
Keyword(s):  
Body Weight ◽  
Tumor Cells ◽  
Death Rate ◽  
Antitumor Effect ◽  
Transforming Growth Factor ◽  
Mammary Adenocarcinoma ◽  
Spleen Cells ◽  
Therapeutic Antitumor Effect ◽  
Allogeneic Tumor

ABSTRACT The therapeutic antitumor effect of clarithromycin (CAM) was examined with the 13762NF mammary adenocarcinoma and F-344 rat system. When CAM treatment at a dosage of 2 mg/kg of body weight orally for 21 days was commenced after inoculation of the tumor, no significant decrease in death rate was observed, although the loss in body weight was less than that in the untreated group. When tumor-bearing (TB) rats were treated with CAM in combination with carboplatin or cyclophosphamide, a significant decrease in the death rate was obtained, although neither treatment alone proved to be effective. A beneficial effect was also observed when CAM treatment was combined with surgical treatment. CAM showed no direct cytotoxicity to this tumor in vitro according to the MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay. Spleen cells obtained from TB rats receiving CAM treatment showed a stronger tumor-neutralizing activity than those from rats which had not received CAM treatment (Winn assay). Enhanced induction of cytotoxic cells to allogeneic tumor was also observed in rats immunized with allogeneic tumor cells together with CAM treatment (51Cr release assay). The 13762NF tumor produces transforming growth factor-β (TGF-β), tumor necrosis factor alpha, and matrix metalloproteinase-9, and treatment of tumor cells with CAM in vitro for 24 h significantly inhibited the expression of the genes coding for these proteins (reverse transcription-PCR). Levels of expression of the TGF-β and interleukin-6 genes of spleen cells obtained from CAM-treated TB rats were both significantly lower than those of spleen cells from CAM-untreated TB rats. This study suggests that CAM has biological response modifier activities resulting in a beneficial therapeutic antitumor effect and might be useful for the treatment of human cancers.

scholarly journals MECHANISMS OF RECOVERY FROM A GENERALIZED VIRAL INFECTION: MOUSEPOX

1971 ◽  
Vol 133 (5) ◽  
pp. 1074-1089 ◽  
Author(s):  
R. V. Blanden
Keyword(s):  
Antiviral Activity ◽  
Immune Serum ◽  
Mononuclear Phagocytes ◽  
Host Responses ◽  
Spleen Cells ◽  
Hyperimmune Serum ◽  
Recovery Mechanisms ◽  
Interferon Responses ◽  
Immune Spleen Cells

The following passive transfer experiments evaluated the contributions of the various host responses in recovery from mousepox. (a) Immune spleen cells transferred highly efficient antiviral activity, but preinfected recipients of these cells made no detectable splenic interferon or antibody in the 24 hr interval after cell transfer. (b) Passively administered interferon was ineffective. (c) Recipients of hyperimmune serum had much more antibody than recipients of immune spleen cells but significantly less antiviral activity. (d) Immune spleen cell populations with antiviral activity contained mediators of CMI to virus antigens. (e) The antiviral activity of immune spleen cells was specific; it was inhibited by in vitro treatment with ATS, anti-light chain serum, and anti-theta ascitic fluid, but not by removal of mononuclear phagocytes from the immune population. These results are interpreted to mean that recovery mechanisms conferred by immune spleen cells were triggered by specifically sensitized, thymus-derived lymphocytes, and that antibody and interferon responses were of less importance. A radiosensitive recipient component was necessary for the full expression of the antiviral activity of both immune cells and immune serum. It seemed likely that this component was the blood monocyte.

scholarly journals Cell interactions in the suppression of in vitro antibody responses.

1976 ◽  
Vol 143 (6) ◽  
pp. 1421-1428 ◽  
Author(s):  
C E Calkins ◽  
S Orbach-Arbouys ◽  
O Stutman ◽  
R K Gershon
Keyword(s):  
B Lymphocytes ◽  
Immune Cells ◽  
Suppressor Cells ◽  
Antibody Responses ◽  
Secondary Antibody ◽  
Spleen Cells ◽  
Homeostatic Mechanism ◽  
Normal Spleen ◽  
Cells Cultured

Normal T and immune B lymphocytes interact in a fashion that leads to suppression of the immune response. Normal spleen cells added to cultures of primed spleen cells specifically suppressed both the IgM and IgG secondary antibody response of the primed cells to less than 30% of the response of the immune cells cultured alone. Cell crowding as a possible in vitro artifact was ruled out. The suppression was specific for the priming antigen, even when the specific and nonspecific antigens were included in the same cultures. Suppression required both normal T and immune B cells to be present in culture. We suggest that the immune population produces a signal that can induce normal T cells to become specific suppressor cells. This form of interaction may represent an important regulatory (homeostatic) mechanism in the immune system.

scholarly journals CELLS MEDIATING SPECIFIC IN VITRO CYTOTOXICITY

1972 ◽  
Vol 135 (4) ◽  
pp. 890-906 ◽  
Author(s):  
Pierre Golstein ◽  
Hans Wigzell ◽  
Henric Blomgren ◽  
Erik A. J. Svedmyr
Keyword(s):  
T Cells ◽  
In Vitro Cytotoxicity ◽  
Cell Populations ◽  
Target Cells ◽  
Present System ◽  
Spleen Cells ◽  
A Cell ◽  
Thymus Cells ◽  
Immune Spleen Cells

In order to investigate whether only T cells are involved in a cell-mediated cytotoxic system in vitro, we tested the cytotoxicity of immune killing cell populations as deprived as possible of B cells. Educated thymus cells, immune spleen cells purified by filtration through a column of beads coated with antimouse Ig antiserum, and finally educated thymus cells further purified by filtration through such a column fully retained their specific cytotoxic activity. This very strongly suggests that only T cells are involved in the killing of target cells by allogeneic immune cells in vitro, in this system. Receptor-bearing cells involved in killing in the present system are thus very probably T cells. This point was further strengthened by the demonstration of specific adsorption, on the relevant monolayers, of each of the three above mentioned killing cell populations.

scholarly journals Changes in suppressor mechanisms during postnatal development in mice.

1978 ◽  
Vol 147 (1) ◽  
pp. 87-97 ◽  
Author(s):  
C E Calkins ◽  
O Stutman
Keyword(s):  
Antibody Response ◽  
Postnatal Development ◽  
Suppressor Cells ◽  
Glass Beads ◽  
Spleen Cells ◽  
Immune Spleen Cells ◽  
Nylon Wool

The activity of suppressor cells from spleens of mice of varying ages was assessed by their addition to cultures of normal or SRBC immune spleen cells together with a challenge of SRBC. 1-wk and adult spleen cells were highly suppressive of the secondary in vitro antibody response to SRBC. 3-wk spleen cells were less active in suppressing this response. The nature of the suppression and the character of the suppressor cells changed in this period. Whereas adult spleen cells demonstrated specificity, 1-wk cells nonspecifically suppressed all responses tested. Further, unlike adult suppressor cells (which are Thy.1.2 positive), 1-wk suppressor cells are insensitive to anti-Thy.1.2 treatment in this system. Both cells are nonadherent to glass beads and nylon wool and are undetectable in the normal thymus.

scholarly journals Monoclonal antibody to murine gamma interferon inhibits lymphokine-induced antiviral and macrophage tumoricidal activities.

1984 ◽  
Vol 159 (5) ◽  
pp. 1560-1565 ◽  
Author(s):  
G L Spitalny ◽  
E A Havell
Keyword(s):  
Monoclonal Antibody ◽  
Antiviral Activity ◽  
Recombinant Dna ◽  
Gamma Interferon ◽  
Spleen Cells ◽  
Tumoricidal Activity ◽  
Antiviral Activities ◽  
Immune Spleen Cells ◽  
Mouse Myeloma

Fusion of rat immune spleen cells with mouse myeloma cells resulted in the formation of a stable hybridoma that secretes monoclonal antibody (MAb) directed against murine gamma interferon ( MuIFN -gamma). This MAb specifically neutralized the antiviral activity of a variety of MuIFN -gamma preparations, including a sample produced by recombinant DNA technologies. In contrast, the antiviral activities of a mixture of MuIFN -alpha plus MuIFN -beta, as well as those of rat or human IFN-gamma, were not neutralized by this antibody. The ability of the MAb to inhibit lymphokine-induced macrophage activation was also tested. It was found that in relation to the quantity of antibody needed to completely neutralize antiviral activity, much higher concentrations of MAb were required to abolish the capacity of lymphokine preparations to induce macrophage tumoricidal activity in vitro. The MAb was also coupled to cyanogen bromide-activated Sepharose beads and used as an immunoadsorbent. By reacting lymphokines with MAb coupled to an insoluble matrix, it was possible to show that this immobilized antibody completely and specifically removed from the lymphokine preparations the ability both to invoke macrophage tumoricidal activity and to mediate antiviral activity.

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