Induction of antigen-specific unresponsiveness by glioblastoma culture supernatants (GCS)

A method of selectively inhibiting an immune response to one or more selected autoantigenic protein antigens of multiple sclerosis, comprising:

• exposing purified or isolated antigen presenting cells (APCs), comprising macrophages, monocytes, dendritic cells, and/or B cells, to an immunosuppressive composition comprising one or more factors secreted by one or more glioblastoma cells,
• wherein the APCs present an autoantigenic protein antigen of multiple sclerosis against which selective inhibition of an immune response is desired;
• and wherein incubation of the APCs with the one or more factors results in effects comprising:
• (a) decreasing expression of MHC class II antigens and CD 80/86 on the surface of the monocytes and the dendritic cells,
• (b) increasing expression of IL-10 in the monocytes and dendritic cells, and
• (c) decreasing the expression of IL-12 in the monocytes and dendritic cells;
• and wherein the one or more factors secreted by the glioblastoma cells have the following characteristics:
• (1) induce APCs to induce T cells to undergo apoptosis;
• (2) molecular weight greater than about 40 kDa;
• (3) ability to bind to anion, but not cation, exchange columns;
• (4) maintain an ability to induce APCs to induce T cells to undergo apoptosis (i) within the pH range of 2 to 11, (ii) following heat exposure up to 56° C., and (iii) following immunoprecipitation of TGF-β1, TGF-β2, TGF-β3, IL-6, calcitonin gene related peptide (CGRP), and M-CSF from the composition;
• and
• (5) lose the ability to induce APCs to induce T cells to undergo apoptosis (i) following heat exposure above 56° C., or (ii) after exposure to trypsin;
• and
• introducing a therapeutically effective amount of the APCs exposed to the immunosuppressive composition into a subject in whom a selectively inhibited immune response to the antigen is desired, wherein introduction of the APCs inhibits the immune response of the subject to the antigen.

A method of selectively inhibiting an immune response to one or more selected autoantigenic protein antigens of multiple sclerosis comprising:

• exposing purified or isolated antigen presenting cells (APCs), which present an autoantigenic protein antigen of multiple sclerosis against which selective inhibition of an immune response is desired, to an immunosuppressive composition comprising one or more factors secreted by a glioblastoma cell, wherein the one or more factors secreted by the glioblastoma cell have the following characteristics:
• (a) induce APCs to induce T cells to undergo apoptosis;
• (b) molecular weight greater than about 40 kDa;
• (c) ability to bind to anion, but not cation, exchange columns;
• (d) maintain an ability to induce APCs to induce T cells to undergo apoptosis (i) within the pH range of2 to 11, (ii) following heat exposure up to 56° C., and (iii) following immunoprecipitation of TGF-β1, TGF-β2, TGF-β3, IL-6, calcitonin gene related peptide (CGRP), and M-CSF from the composition;
• and
• (e) lose the ability to induce APCs to induce T cells to undergo apoptosis (i) followings heat exposure above 56° C., or (ii) after exposure to trypsin;
• and
• introducing a therapeutically effective amount of the APCs exposed to the immunosuppressive composition into a subject in whom a selectively inhibited immune response to the antigen is desired, wherein introduction of the APCs inhibits the immune response of the subject to the antigen;
• wherein the glioblastoma cell is selected from the group consisting of SNB 19 (DSMZ no. 325), A172 (ATCC no. CRL-1620), U87 MG (ATCC no. HTB-14), U138 MG (ATCC no. HTB-16), U373 MG (ECACC no. 89081403), T98G (ATCC no. CRL-1690), DBTRG-05MG (ATCC no. CRL-2020), M059K (ATCC no. CRL-2365), M059J (ATCC no. CRL-2366), and U118 MG (ATCC no. HTB-15).