scholarly journals Expression of hybrid Ia molecules on the cell surface of reticulum cell sarcomas that are undetectable on host SJL/J lymphocytes

1981 ◽  
Vol 153 (3) ◽  
pp. 501-513 ◽  
Author(s):  
SM Wilbur ◽  
B Bonavida
Keyword(s):  
Cell Surface ◽  
Reticulum Cell ◽  
Cell Binding ◽  
Spleen Cells ◽  
Normal Spleen ◽  
Tumor Recognition ◽  
Α Chain ◽  
Chain Synthesis ◽  
Β Chain

SJL/J (H-2 (8)) lymphocytes, primed in vitro against primary, cultured, and transplantable syngeneic reticulum cell sarcomas (RCS) were found to recognize and bind to the tumor without subsequent cytolysis. Additional data showed that the recognition was also directed against Ia molecules of the H-2(d), but not H-2(k), haplotype. Normal spleen cells of DBA/2, B 10.D2, and B 10.OL mice were bound, whereas those of CBA, B 10.BR, B 10.A, B 10.GD, and D2.GD were not. Furthermore, the Ia molecules were in the form of a hybrid, because spleen cells from F(1) progeny of a B10.A and a B10.GD parent were recognized and bound as effectively as the RCS. Recognition was not restricted solely to the H-2(d) haplotype. Spleen cells from B10.S(9R) mice were also significantly bound. This result suggested that the RCS expresses a hybrid Ia molecule containing a β-chain of the H-2(8) haplotype. Recognition of this hybrid Ia molecule by the host resulted in a cross- reactive recognition of H-2(d) specificities. Further analysis revealed that the RCS express on their cell surface an α-chain of the hybrid Ia molecule which is involved in host anti-tumor recognition. Preincubation of the RCS with monoclonal antibody directed against the Ia.7 specificity on the α-chain could block lymphocyte-to-tumor cell binding. The blocking activity could be removed by preabsorption of the antibody on the RCS, as well as normal Ia.7-bearing lymphocytes, but not on lymphocytes that do not express Ia.7, such as SJL/J. The data suggest that the hybrid Ia molecules expressed on the RCS, and recognized by tumor-primed syngeneic lymphocytes, are composed of both a syngeneic and an alien chain. The component alien to the SJL/J host is the Ia.7-bearing α-chain. Normal SJL/J cells synthesize but do not express the β-chain. In the RCS, however, alien α-chain synthesis permits expression of the syngeneic β-chain in the form of a hybrid Ia molecule.

scholarly journals Specific elimination of cytotoxic effector cells. I. Adsorptive behavior of effectors and their precursors and spleen cell monolayers.

1976 ◽  
Vol 144 (4) ◽  
pp. 996-1008 ◽  
Author(s):  
J R Neefe ◽  
D H Sachs
Keyword(s):  
Mouse Spleen ◽  
Spleen Cells ◽  
Effector Cells ◽  
Cell Monolayers ◽  
Specific Suppression ◽  
Normal Spleen ◽  
Adsorptive Behavior ◽  
Cytotoxic Effector ◽  
Primed Mouse

Monolayers formed of normal mouse spleen cells attached to polystyrene coated with poly-L-lysine were tested for their ability to bind specifically antigen-reactive cells in normal or primed mouse spleen. 88 to greater than 98% of the activity of cytotoxic populations was removed by a single adsorption. However, normal spleen cells or spleen cells previously primed in vitro could not be depleted of their capacity to be sensitized, even when adsorption effectively removed all residual cytotoxic activity from the same previously primed population. In fact, exposure to an immunoadsorbent augmented the ultimate cytotoxicity generated in a nonspecific fashion. This augmentation was especially dramatic in the case of a previously primed population and may have reflected the removal of a nonspecific suppressor. If antigen-reactive precursors cannot be removed efficiently by adsorption, other approaches to the generation of tolerant lymphoid populations, such as specific suppression of precursor differentiation must be sought.

scholarly journals HETEROGENEITY OF THE EFFECTOR CELLS IN THE CYTOTOXIC REACTION AGAINST ALLOGENEIC LYMPHOMA CELLS

1973 ◽  
Vol 137 (2) ◽  
pp. 369-386 ◽  
Author(s):  
James Forman ◽  
Sven Britton
Keyword(s):  
Immune Cells ◽  
Host Response ◽  
Marked Reduction ◽  
Cytotoxic Effect ◽  
Immune Serum ◽  
Spleen Cells ◽  
Effector Cells ◽  
Normal Spleen ◽  
Peak Phase

The cytotoxic effect of spleen cells from H-2 allogeneic mice was tested in vitro against an A strain leukemia (YAC) labeled with [125I]iododeoxyuridine. After the mice were primed with tumor cells, significant and specific H-2 immunity was detected on day 3 and peak cytotoxicity was observed between 7 and 14 days after priming. Two effector cells appear to be involved in the host response, because spleens taken from mice soon after priming were not sensitive to antitheta sera and complement while those taken during the peak stages of the response showed a marked reduction in cytotoxicity after treatment. Macrophages were not involved, since removal of these cells by the carbonyl iron method did not result in any reduction in cytotoxicity. Immune serum that was capable of inducing cell-mediated cytotoxicity in normal spleen cell populations also augmented cytotoxicity of spleen cells taken from mice primed 3 days previously. However, when spleen cells were taken from mice during the peak phase of the immune response, the same serum at the same dilutions inhibited the preexisting cytotoxicity. A difference was also detected in the killing efficiencies between early and late immune cells.

scholarly journals SYNERGY BETWEEN SUBPOPULATIONS OF MOUSE SPLEEN CELLS IN THE IN VITRO GENERATION OF CELL-MEDIATED CYTOTOXICITY

1974 ◽  
Vol 140 (6) ◽  
pp. 1646-1659 ◽  
Author(s):  
Richard J. Hodes ◽  
Barry S. Handwerger ◽  
William D. Terry
Keyword(s):  
Mixed Lymphocyte Culture ◽  
Lymphocyte Culture ◽  
Specific Cell ◽  
Spleen Cells ◽  
Normal Spleen ◽  
In Vitro Induction ◽  
Cytotoxic Effector ◽  
Nylon Wool

Two subpopulations separated from normal spleen have been shown to synergize as responding cells in the in vitro induction of specific cell-mediated cytotoxicity during the mixed lymphocyte culture (MLC). The synergizing populations are a nylon wool column-adherent and a nylon wool column-nonadherent fraction, enriched for B lymphocytes and T lymphocytes, respectively. When a mixture of these fractions is used as the responding cell population in MLC, greater cytotoxicity is generated than would be expected from the sum of activities generated in the two subpopulations sensitized separately. The synergy appears to occur at the sensitization rather than the effector phase. The synergizing cell which is contained in the nylon-adherent subpopulation is distinct from the cytotoxic effector T lymphocyte, is resistant to lysis by rabbit antimouse brain serum, and is unresponsive to phytohemagglutinin; its synergizing function could not be replaced by either plastic-adherent spleen cells or peritoneal exudate cells. These results suggest a role of a non-T-cell nonmacrophage population in the generation of cytotoxic activity.

scholarly journals In vitro generation of tumor-specific cytotoxic lymphocytes. Secondary allogeneic mixed tumor lymphocyte culture of normal murine spleen cells

1977 ◽  
Vol 146 (2) ◽  
pp. 468-482 ◽  
Author(s):  
S Gillis ◽  
KA Smith
Keyword(s):  
Cell Surface ◽  
Lymphocyte Culture ◽  
Leukemia Cells ◽  
Cytotoxic Lymphocytes ◽  
Mixed Tumor ◽  
Spleen Cells ◽  
Syngeneic Tumor ◽  
Murine Spleen ◽  
Murine Leukemia

In vivo or in vitro immunity to murine leukemia virus (MuLV)-induced leukemia cells which do not effectively produce virus, has been difficult to demonstrate. Because immunizations with allogeneic murine leukemia cells have been used to confer syngeneic tumor immunity to virus- producing cells, we attempted to generate lymphocytes, cytotoxic to syngeneic nonproducer leukemia cells, by stimulating normal murine spleen cells with allogeneic nonproducer leukemia cells in mixed tumor lymphocyte culture (MTLC) reactions in vitro. Secondary allogeneic MTLC of normal C57BL/6 or DBA/2 spleen cells effectively produced syngeneic tumor-specific cytotoxic lymphocytes. Target cells lysed in lymphocyte- mediated cytolysis (LMC) assays, included both Friend and Rauscher virus- induced syngeneic murine leukemia cells and chemically-induced hematopoietic tumor cells. Syngeneic tumor cells were lysed regardless of whether they produced infectious MuLV or expressed viral antigens gp-71, p-30, or p-12 at the cell surface. Syngeneic normal cells (thymus, lymph node, or Concanavalin A-stimulated spleen cells) used as targets in LMC assays were uneffected by lymphocytes harvested from secondary allogeneic MTLC. Several other in vitro culture treatments including secondary syngeneic MTLC and repetitive mixed lymphocyte culture stimulations were incapable of generating tumor-specific cytotoxic lymphocytes. Based upon these results, we propose that secondary MTLC stimulation of normal spleen cells with allogeneic nonproducer leukemia cells selects for the proliferation of two subpopulations of antigen-specific cytotoxic lymphocytes. The population capable of effecting syngeneic tumor cell lysis is directed against tumor-associated cell surface antigens which may be distinct from viral structural proteins or glycoproteins. The growth of these tumor-specific cytotoxic lymphocytes may be enhanced by a soluble allogeneic effect factor produced by the proliferation of the second subpopulation of lymphocytes generated in repetitive allogeneic MTLC, namely those lymphocytes with specificities directed against differing histocompatibility antigens.

scholarly journals Absolute Rates of Globin Chain Synthesis in Thalassemia

Blood ◽  
1968 ◽  
Vol 31 (2) ◽  
pp. 226-233 ◽  
Author(s):  
ARTHUR BANK ◽  
ALBERT S. BRAVERMAN ◽  
JOYCE V. O’DONNELL ◽  
PAUL A. MARKS
Keyword(s):  
Peripheral Blood ◽  
Erythroid Cell ◽  
Absolute Rate ◽  
Globin Chain ◽  
Globin Chains ◽  
The Absolute ◽  
Α Chain ◽  
Chain Synthesis ◽  
Globin Chain Synthesis ◽  
Β Chain

Abstract The absolute rate of α chain synthesis per erythroid cell in the peripheral blood of patients with β thalassemia has been shown to be normal while that of β chains is markedly decreased or absent. The results indicate that α chains do not require the presence of β chains for their normal synthesis and release. In addition, γ chain synthesis does not compensate for the decreased β chain synthesis. A marked heterogeneity in the amount of β globin chains produced by different patients with β thalassemia is also prominent.

scholarly journals INDUCTION OF IMMUNITY AND TOLERANCE IN VITRO BY HAPTEN PROTEIN CONJUGATES

1972 ◽  
Vol 135 (4) ◽  
pp. 735-753 ◽  
Author(s):  
Marc Feldmann
Keyword(s):  
B Cells ◽  
Inhibitory Effect ◽  
Spleen Cells ◽  
Protein Conjugates ◽  
Normal Spleen ◽  
High Concentrations ◽  
Induced Tolerance ◽  
Thymus Cells ◽  
Relationship Of

Of many dinitrophenylated (DNP) protein conjugates tested, only DNP conjugated to polymerized flagellin (DNP-POL) (or the structurally related bacterial flagella) elicited a primary anti-DNP response in vitro. Other DNP proteins, such as DNP-monomeric flagellin (DNP-MON), were capable of inducing secondary responses in vitro. The capacity of DNP-POL to immunize spleen cell suspensions devoid of thymus-derived cells was the reason for the greater immunogenicity of DNP-POL, since even large numbers of flagellin-reactive activated thymus cells did not increase the anti-DNP response of normal spleen cells immunized with DNP-POL, whereas the thymus-dependent response to DNP-MON was markedly increased. The capacity of various batches of DNP-POL to immunize normal spleen cells in vitro varied markedly, depending on the number of DNP groups conjugated. DNP-POL with few DNP groups conjugated was immunogenic, but even at very high concentrations did not induce tolerance. In contrast, highly conjugated DNP-POL did not immunize, but readily induced tolerance. DNP-POL with intermediate degrees of conjugation were, like unconjugated polymeric flagellin, capable of inducing both immunity and tolerance. Since DNP-POL immunizes bone marrow-derived lymphocytes (B cells) directly the reduced response was not due to a masking of carrier determinants, necessary for cell collaboration. By using mixed DNP-5-(dimethylamino)-1-naphthalyl (dansyl)-POL conjugates, it was found that the inhibitory effect of a high degree of hapten conjugated was hapten specific. Depolymerization of DNP-POL to DNP-MON, which does not induce primary anti-DNP responses, was excluded by centrifugation analysis and electron microscopy. The relationship of the degree of hapten conjugation on DNP-POL to the capacity to induce tolerance and immunity in B cells has clarified the mechanism of immunological triggering of these cells. A model of the mechanism of "signal" discrimination between immunity and tolerance in B cells, based on these findings, is proposed.

scholarly journals LYMPHOCYTE MEMBRANE DYNAMICS

1971 ◽  
Vol 134 (6) ◽  
pp. 1373-1384 ◽  
Author(s):  
Robert E. Cone ◽  
John J. Marchalonis ◽  
Ronald T. Rolley
Keyword(s):  
Cell Surface ◽  
Disc Electrophoresis ◽  
Membrane Dynamics ◽  
Surface Proteins ◽  
Dynamic State ◽  
Cellular Respiration ◽  
Spleen Cells ◽  
Cell Surface Proteins ◽  
Before And After

Cell surface proteins of normal and neoplastic lymphocytes were labeled with iodide-125I by lactoperoxidase-catalyzed iodination. Incubation of 125I-labeled iodide cells in vitro resulted in the release of iodinated surface proteins at a rapid rate which was dependent on cellular respiration and protein synthesis. Comparisons by disc electrophoresis showed a marked similarity between urea-soluble surface proteins extracted from iodinated cells and iodinated material released by the cells during in vitro incubation. The rate of release of cell surface proteins from thymus cells was three times faster than that of spleen cells or bone marrow-derived thoracic duct lymphocytes. In addition, different proteins were released at different rates as evidenced by the rate of release of 125I of rabbit anti-mouse immunoglobulin specifically bound to mouse spleen cells and comparisons by disc electrophoresis of urea-soluble iodinated surface proteins extracted from cells before and after incubation. The results suggest that a dynamic state exists at the cell surface. The possible role of the release of cell surface proteins in cell regulation and communication is discussed.

scholarly journals RAPID BREAKING OF TOLERANCE AGAINST ESCHERICHIA COLI LIPOPOLYSACCHARIDE IN VIVO AND IN VITRO

1972 ◽  
Vol 135 (4) ◽  
pp. 850-859 ◽  
Author(s):  
Olof Sjöberg
Keyword(s):  
Escherichia Coli ◽  
Immune Responsiveness ◽  
Spleen Cells ◽  
E Coli ◽  
Normal Spleen ◽  
Escherichia Coli Lipopolysaccharide ◽  
Tolerant State

The breaking of tolerance against the lipopolysaccharide from E. coli 055:B5 was studied. It was found that immune responsiveness recovered very slowly in vivo, tolerance still existing 3 wk after the last tolerizing injection. However, if spleen cells from tolerant mice were transferred into irradiated syngeneic recipients, the tolerant state was readily broken. Spleen cells transferred 3 days after the last tolerance-maintaining dose did not respond, whereas cells transferred on day 5 or 7 responded equally well as normal spleen cells. It was also possible to break tolerance by incubating tolerant spleen cells, which did not respond after transfer, for 20 hr in vitro before transfer into irradiated recipients. The results suggest that there exist reversibly inactivated cells in tolerant animals and that these cells can be reactivated upon removal of the cells to a neutral environment.

scholarly journals Suppressor T cells activated in a primary in vitro response to non-major histocompatibility alloantigens.

1982 ◽  
Vol 156 (5) ◽  
pp. 1398-1414 ◽  
Author(s):  
S Macphail ◽  
O Stutman
Keyword(s):  
Mixed Lymphocyte Reaction ◽  
Cytotoxic T Lymphocyte ◽  
Suppressor Cells ◽  
Suppressor Cell ◽  
Spleen Cells ◽  
Suppressor T Cells ◽  
Normal Spleen ◽  
In Vitro Response

Normal mouse spleen cells are not capable of mounting a primary cytotoxic T lymphocyte (Tc) response to non-H-2 alloantigens in vitro, although a good secondary H-2-restricted response is observable after in vivo immunization of the responder animals. Suppressor cells are generated in such a primary responses provided a Mls incompatibility exists between the responder and stimulator. These suppressors are not antigen specific, are Thy-1+, Lyt-1+, 2-, I-J-, and are highly radiosensitive. The suppressor cell precursors in normal spleen express the same phenotype. These suppressor cells are probably implicated in the lack of a primary Tc response in a primary mixed lymphocyte reaction across non-H-2 incompatibilities that include an Mls difference.

scholarly journals Globin Synthesis in Iron-deficiency Anemia

Blood ◽  
1974 ◽  
Vol 44 (4) ◽  
pp. 551-555 ◽  
Author(s):  
Isaac Ben-Bassat ◽  
Miriam Mozel ◽  
Bracha Ramot
Keyword(s):  
Iron Deficiency ◽  
Iron Deficiency Anemia ◽  
Deficiency Anemia ◽  
Globin Chains ◽  
Iron Deficient ◽  
Membrane Bound ◽  
Α Chain ◽  
Globin Synthesis ◽  
Chain Synthesis ◽  
Β Chain

Abstract The α to β globin chain ratio has been determined in the peripheral red cells of 11 patients with iron deficiency anemia. The mean ratio was found to be 0.74 ± 0.07, which is significantly lower than the ratio of 0.97 ± 0.07 obtained in normals. When the stroma-free hemolysates were purified prior to globin preparation the α to β ratio did not change. On the other hand, globin extracted from whole iron-deficient cells, including the stroma, had a higher α to β ratio, 0.88 ± 0.04, but still significantly lower than normal. These results suggest that in iron deficiency there is a decrease in α-chain synthesis relative to β-chain and that there are membrane-bound globin chains, but no excessive increase in the free α-chain pool. Similar findings have been reported previously in other states of heme deficiency like sideroblastic anemia and lead poisoning.

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