Alteration of lymphocyte surface properties by insertion of foreign functional components of plasma membrane.

Abstract

The lymphocyte plasma membrane plays a key role in the functions of the immune system. Much effort is being made to understand the correlation between the composition and structure of the lymphocyte plasma membrane and the associated cell functions. Current approaches to this problem are based primarily on studies of isolated membrane constituents and characterization of cell surface structures in intact cells, mainly done on isolated lymphocyte subpopulations.A promising approach involves the transfer of membrane components from one cell type (the donor) to another (the acceptor) in order to study the effect of the new membranal composition on the ability of the acceptor cell to respond to extracellular signals. Present techniques for the transfer of membrane proteins and lipids into plasma membranes of eukaryotic cells involve cell-cell and membrane-cell fusion with fusogenic agents, in particular Sendai virus and polyethylene glycol (1, 2). The transfer of receptors for hormones (3, 4) and asialoglycoproteins (5) to tissue culture cells by these methods has been demonstrated. The methods have also been applied to the transfer of isolated membrane vesicles obtained from highly metastatic cells to the plasma membrane of poorly metastatic sublines (6). However, application of these procedures to the transfer of membrane components to primary cells such as lymphocytes has never been reported, presumably because of the inability of the latter to undergo massive cell-cellThe publication costs of this article were defrayed in part by page charge payment. This article must therefore be hereby marked "advertisement" in accordance with 18 U. S. C. §1734 solely to indicate this fact. 7247 fusion by Sendai virus (1) and their poor viability after treatment with polyethylene glycol (2).In this study we report the successful coreconstitution of lymphocyte membrane components with Sendai virus envelopes (7,8) (5 Ci/mmol) were from the Radiochemical Center (Amersham, England). Soybean agglutinin was prepared (9) and converted to its 125I-labeled derivative as described (10). Fluorescein isothiocyanate-conjugated limulin was a gift from Michel Monsigny

Implantation of mouse histocompatibility antigens into membranes of cultured tumor cells

“…This leads to the incorporation of vesicle components into plasma membranes of target cells [4][5][6].…”

“…Purified membranes (200-500 mg) or crude extracts were mixed with Triton X-100 extract of SV [4][5][6] containing 1 mg of envelope proteins. The mixture was dialyzed at 4 "C in Spectrofor-2 dialysis tubing (Spectrum Medical Industries, Los Angeles, CA) until the complete removal of Triton X-100 [5] under the conditions previously described [ a ] .…”

“…This induced a specific C1-exchange activity in FELC, without affecting the viability of the recipient cells [4]. Using a similar technique, receptors for Epstein-Barr virus (EBV) [5] and receptors for lectins [6] have been implanted into membranes of receptor-negative cells [5, 61. This allowed infection by EBV of the previously nonsusceptible cells like mouse lymphocytes [5].…”

Human B lymphocytes form rosettes after insertion of T lymphocyte membrane constituents

“…We have recently developed a method for the transfer of plasma membrane components between primary murine lymphocytes using fusogenic membrane vesicles composed of donor plasma membranes and envelope glycoproteins of Sendai virus [5]. Using this method, we have demonstrated that inserted H-2 gene membrane products mediate the immune response phenotype of antigen-presenting cells [6].…”

Direct transfer of antigen-specific cytolytic activity to noncytolytic cells upon fusion with liposomes derived from cytolytic T cell clones.

“…Liposomes were prepared by a modification of previously published procedures (13)(14)(15). Briefly, donor cells were extensively washed in phosphatebuffered saline (PBS) to remove medium and serum contaminants, followed by a 30 min incubation at 4°C in 0.5% NP-40-Tris-HCl-saline buffer, pH 7.4.…”