scholarly journals Immunogenic variants obtained by mutagenesis of mouse mastocytoma P815. II. T lymphocyte-mediated cytolysis.

1980 ◽  
Vol 152 (5) ◽  
pp. 1184-1193 ◽  
Author(s):  
T Boon ◽  
J Van Snick ◽  
A Van Pel ◽  
C Uyttenhove ◽  
M Marchand
Keyword(s):  
Tumor Cell ◽  
T Lymphocyte ◽  
Peritoneal Exudate ◽  
Antigenic Specificity ◽  
Spleen Cells ◽  
Peritoneal Exudate Cells ◽  
Specific Antigens

Tumor cell variants that were rejected by syngeneic mice (tum-) were obtained from mastocytoma P815 by mutagenesis (as described in the accompanying report (13). A considerable T lymphocyte-mediated lysis was observed upon incubation of these tum- variants with peritoneal exudate cells collected a few days after an intraperitoneal challenge of immune animals. Spleen cells from these animals were cytolytic after stimulation in vitro with the immunizing variant. New antigens, absent from the original P815 tum+ cells, were detected on 15 of the 21 tum- variants that were tested. All these antigens appeared to be different. No new antigen was detected on any of 10 mutagenized P815 clones that had retained their ability to form tumors. We compared the evidence obtained in vivo and in vitro for the presence of specific antigens on five tum- variants. Three variants were shown both in vivo and in vitro to carry an individual antigen. One showed no specificity either in vivo or in vitro. However, for one variant, no specificity was observed in vivo, although cytolysis tests demonstrated the existence of a singular antigenic specificity.

scholarly journals Regulatory mechanisms in cell-mediated immune responses. II. A genetically restricted suppressor of mixed lymphocyte reactions released by alloantigen-activated spleen cells.

1975 ◽  
Vol 142 (6) ◽  
pp. 1391-1402 ◽  
Author(s):  
S S Rich ◽  
R R Rich
Keyword(s):  
Immune Responses ◽  
Mixed Lymphocyte Reaction ◽  
Antigenic Specificity ◽  
Spleen Cells ◽  
Suppressor Factor ◽  
Homology Relationship ◽  
Cell Suppression ◽  
Mixed Lymphocyte Reactions

The mechanism of alloantigen-activated spleen cell suppression of mixed lymphocyte reaction (MLR) is explored in this report. Activated murine suppressor spleen cells elaborated a soluble noncytotoxic factor which suppressed MLR responses by 55-95%. Generation of suppressor factor required both in vivo alloantigen sensitization and specific in vitro restimulation. Suppressor factor was not produced by activated spleen cells which had been treated with anti-Thy-1.2 serum and complement. Antigenic specificity toward alloantigens of the stimulator cells was not demonstrable. In contrast, suppressor factor effectively inhibited MLR response only of responder cells of those strains that shared the D-end and the I-C subregion of the H-2 complex with the cells producing suppressor factor. Therefore, active suppression appears to require an MHC-directed homology relationship between regulating and responder cells in MLR.

A Technique for Sequential Examination of in Vitro Macrophage-Tumor Interaction using Lm, Sem and Tem

1976 ◽  
Vol 34 ◽  
pp. 350-351
Author(s):  
C. Bucana ◽  
B. Hobbs ◽  
L. C. Hoyer ◽  
M. G. Hanna
Keyword(s):  
Guinea Pig ◽  
Tumor Cell ◽  
Tumor Immunology ◽  
Time Lapse ◽  
Cell Interaction ◽  
Peritoneal Exudate Cells ◽  
Cytotoxicity Assays ◽  
Sem And Tem

A sequential examination of macrophage-tumor cell interaction in vitro, using time-lapse cinematography followed by SEM and TEM of the same cells, is a valuable asset in the interpretation of in vivo and in vitro cell-mediated cytotoxicity assays that are widely used in tumor immunology. Previous reports on sequential examination of biological specimens involved large tissue blocks. This report describes a technique that allows sequential examination of cell - cell interaction using three modes of microscopy.Guinea pig hepatoma (L-10) cells were plated sparsely on 25mm glass coverslips 24hr before adding guinea pig peritoneal exudate cells (PEC). PEC were allowed to settle for 1 l/2hr; the coverslips were then transferred to a Dvorak-Stotler chamber and examined with a Zeiss photomicroscope III.

scholarly journals STUDIES ON ANTIBODY FORMATION BY PERITONEAL EXUDATE CELLS IN VITRO

1960 ◽  
Vol 111 (4) ◽  
pp. 573-600 ◽  
Author(s):  
John M. McKenna ◽  
Kingsley M. Stevens
Keyword(s):  
Tissue Culture ◽  
Cell Types ◽  
Peritoneal Exudate ◽  
Rabbit Serum ◽  
Normal Rabbit Serum ◽  
Peritoneal Exudate Cells ◽  
Carbon Particles ◽  
Secondary Systems

Cells from peritoneal exudates of rabbits sacrificed 3 days after an intraperitoneal injection of sterile mineral oil were grown in tissue cultures in medium 199 (75 per cent); normal rabbit serum (25 per cent). Antibody produced by the cells was assayed by an hemagglutination technique in which the antigens used were adsorbed to formalinized tanned sheep erythrocytes. These sensitized cells agglutinate in the presence of antibody specific to the adsorbed antigen. It has been demonstrated that: Peritoneal exudate cells produced hemagglutinating antibody to bovine gamma globulin (BGG) in a replicating tissue culture system for approximately 3 weeks when taken from animals given either primary or secondary injections of BGG. The mean hemagglutinating titer was 30 for the primary and 32 for the secondary systems. Since the other cell types did not persist, it is felt that monocytes were responsible for these results. Monocytes taken from normal rabbits and exposed to either BGG or egg albumen (EA) in vitro produced titers of 28 for about 2 weeks. Monocytes taken from rabbits given hyperimmunizing injections of BGG produced titers of 147 for about 1 week. Endotoxin from Salmonella typhosa caused the monocytes to form antibody as if they had been taken from hyperimmunized rabbits. This was true both when the antigen was given in vivo together with the endotoxin as well as when the cells were exposed to antigen in vitro. The titers were 223 and 97, respectively. Neither freshly harvested nor cultured monocytes were phagocytic for carbon particles or bacteria in vitro. Monocytes in tissue culture appeared to assume the morphology of fibroblasts, but did not stain with the characteristics of fibroblasts. The morphologic changes and staining characteristics of monocytes in tissue culture have been described. The implications of these findings have been discussed and an attempt made to integrate them into general biological theory.

scholarly journals IMMUNOCHEMICAL STUDIES ON THE SPECIFICITY OF CELLULAR HYPERSENSITIVITY

1968 ◽  
Vol 128 (6) ◽  
pp. 1451-1459 ◽  
Author(s):  
John R. David ◽  
Stuart F. Schlossman
Keyword(s):  
Cell Migration ◽  
Lymphoid Cell ◽  
Specific Binding ◽  
Peritoneal Exudate ◽  
Cellular Receptor ◽  
Anamnestic Response ◽  
Peritoneal Exudate Cells ◽  
Exquisite Specificity

The immunochemical specificity of antigen-induced inhibition of peritoneal exudate cell migration was studied in animals sensitized to chemically defined α,DNP(Lys)18 peptides. It was shown that sensitized peritoneal exudate cells could discriminate between various DNP-oligolysines. Only immunogenic members of the homologous series of α,DNP-L-lysines equal to or larger in size than the heptamer inhibited the migration of specifically sensitized peritoneal exudate cells. In contrast, nonimmunogenic α,DNP-L-lysines, a D-lysine containing stereoisomer of α,DNP L(Lys)9 (α,DNP-L4DL4) and (Lys)9ϵ, DNP were not inhibitory to the migration of peritoneal exudate cells derived from animals immunized to α,DNP(Lys)18. The exquisite specificity of the in vitro reaction of sensitized cells with antigen contrasts with the previously observed in vivo or in vitro specificity of anti-α,-DNP(Lys)n antibody, but parallels the specificity of the in vivo delayed or anamnestic response. These results suggest the presence of a still undefined but highly specific binding site, which functions as the cellular receptor for antigen on the sensitized lymphoid cell or on some "processing" cell.

A Molecular Approach to Immunoscintigraphy: A Study of the T-Antigen Conformation on the Surface of Tumors

1987 ◽  
Vol 26 (01) ◽  
pp. 1-6 ◽  
Author(s):  
S. Selvaraj ◽  
M. R. Suresh ◽  
G. McLean ◽  
D. Willans ◽  
C. Turner ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Tumor Cell ◽  
T Antigen ◽  
Human Carcinomas ◽  
Animal Tumor ◽  
Synthetic Antigens

The role of glycoconjugates in tumor cell differentiation has been well documented. We have examined the expression of the two anomers of the Thomsen-Friedenreich antigen on the surface of human, canine and murine tumor cell membranes both in vitro and in vivo. This has been accomplished through the synthesis of the disaccharide terminal residues in both a and ß configuration. Both entities were used to generate murine monoclonal antibodies which recognized the carbohydrate determinants. The determination of fine specificities of these antibodies was effected by means of cellular uptake, immunohistopathology and immunoscintigraphy. Examination of pathological specimens of human and canine tumor tissue indicated that the expressed antigen was in the β configuration. More than 89% of all human carcinomas tested expressed the antigen in the above anomeric form. The combination of synthetic antigens and monoclonal antibodies raised specifically against them provide us with invaluable tools for the study of tumor marker expression in humans and their respective animal tumor models.

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