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G Berke

Publications and source records attributed to G Berke.

At least 109 records · Page 6Linked to original sources

T lymphocyte-mediated cytolysis: dissociation of the binding and lytic mechanisms of the effector cell.

To determine functional relationships between the cytotoxic T lymphocyte (CTL) receptor for target binding and the lytic mechanism, we have studied the reaction between two immunized CTL populations (AalphaB and BalphaA), both at the population and the single-cell level. When studied at the population level, the reaction of AalphaB with BalphaA (bidirectional system) resulted in formation of AalphaB/BalphaA conjugates and bidirectional cytolysis. However, when the viability of cells in individual AalphaB/BalphaA conjugates was analyzed, unidirectional instead of bidirectional lysis occurred. These results indicate that under conditions that are conducive to lysis, binding of a potentially lytic cell to its target does not necessarily result in target lysis. Short heat treatment of CTL (44 degrees C, 10 min) totally abolished their lytic activity, without affecting their capacity to bind specifically, thus dissociating the binding from the lytic activity of the CTL. The cytolytic activity is probably associated with, or triggered by the CTL-binding unit. The binding unit, on the other hand, appears to be a functional receptor of the CTL, which is involved in but not sufficient to bring about lysis.

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Activity of tumor-associated lymphoid cells at short intervals after administration of irradiated syngeneic and allogeneic tumor cells.

After a single intraperitoneal injection of irradiated tumor cells, host cells capable of responding against syngeneic tumors were detected in peritoneal exudates of mice. Although irradiation of the injected tumor prevented its overgrowth, it did not significantly alter the antigenicity of the tumor. Immunologic activities of tumor-associated host cells in the peritoneal cavity were continuously monitored, starting 48 hr after tumor administration. In vitro cell-mediated lysis of syngeneic tumors appeared as early as 3 days after irradiated tumor administration. In addition, peritoneal exudate cells from inoculated mice were capable of adoptively transferring immunity. Purification of these peritoneal exudate cells on nylon wool columns yielded a nonadherent Ig-negative lymphocyte fraction whose cytolysis was tumor-specific and T cell-associated. The macrophage-free lymphocyte fraction exhibited a higher in vitro activity against tumors than unpurified peritoneal exudates. This tumor-host system allowed the study of cells which directly interact with the tumor cells in vivo, starting shortly after tumor administration. The results reported in this paper show that tumor-associated lymphoid cells capable of mounting anti-tumor response in vivo and in vitro can be demonstrated as early as 3 days after tumor inoculation.

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Possible role of nucleus-membrane interaction in capping of surface membrane receptors.

Interaction of multivalent ligands and cell surface receptors can induce redistribution of these receptors to form patches and caps. In this study, we have investigated the role of nucleus-membrane interaction in the capping of membrane components. Mouse L cells and leukemia EL4 cells were enucleated with the aid of cytochalasin B, yielding cytoplasts and karyoplasts. Capping of surface receptors was induced by allo- and hetero-immune sera followed by fluorescein-conjugated antiglobulin serum, or by the plant lectin concanavalin A. Capping could easily be induced in intact cells, but virtually no capping was detected in the nucleus-free cytoplasts. Interestingly, karyoplasts, which posses cell-membrane components but very little cytoplasm, could be easily induced to cap their surface antigens. Hence, cap formation of membrane components seems not to be an autonomous membrane process. The data suggest that interaction of surface membranes and inner cell components associated with the nucleus is involved in the movement of surface membrane receptors.

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Energy requirements of the binding and lytic steps of T lymphocyte-mediated cytolysis of leukemic cells in vitro.

The process of T lymphocyte-mediated destruction of target cells in vitro is temperature dependent and is suppressed by inhibitors of energy metabolism, but the energy requirements for the conjugation and lytic steps of the process are different. The conjugation step of lymphocytes and target cells is prevented by metabolic inhibitors and is dependent on temperature, increasing in the range from 2 degrees C to 22 degrees C. However, after lymphocyte-target cell conjugates are formed, they are stable at low temperatures (0 degree C) and in the presence of metabolic inhibitors. Thus, the formation but not the maintenance of lymphocyte-target cell conjugates is an energy-dependent process. In contrast to the conjugation step, which is influenced both by metabolic inhibitors and by temperature, the lytic step, which occurs only between 22 degrees C and 37 degrees C, is not affected by inhibitors of energy metabolism. The sigmoidal behavior of lysis as a function of temperature, showing a sharp inflection around 20 degrees C, can be explained in terms of the alteration of the viscosity of the membrane matrix of the target cells rather than as a general metabolic effect on the killer and target cells.

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Nature of lymphocyte-tumor interaction. A general method for cellular immunoabsorption.

The binding of sensitized lymphocytes to tumor cells that leads to tumor cell lysis in vitro has been investigated using poly-L-lysine-fixed tumor cell monolayers and lymphocytes obtained from the anatomical site of tumor allograft rejection. The results show that magnesium is an important prerequisite for this interaction and that the extent of lymphocyte-tumor cell binding depends upon temperature as well as pH. Binding can occur in the absence of serum, although serum factors are necessary for the completion of the cytolytic process. The poly-L-lysine technique is applicable to the formation of confluent monolayers with virtually any normal or neoplastic cell type, including those that are otherwise nonadherent to surfaces. Cells immobilized by this technique can be used for the specific immunoabsorption and subsequent recovery of effector lymphocytes sensitized against transplantation or tumor cell antigens.

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