Regulatory glycoprotein for immune response and the use there of in the production of T-cell growth factor

Claim 2

A process for producing a serum-free and mitogen-free T-cell growth factor (Il-2) comprising incubating a serum-free and mitogen-free liquid tissue culture medium containing therein lymphocyte cells, a biologically active blood serum glycoprotein having a molecular weight of about 90,000 and being able to cause T-cell growth factor production in the presence of Interleukin-1 and T-lymphocytes; said glycoprotein being further characterized by at least one sialic acid moiety as a biologically active site; by retaining more than one-half of its biological activity after being heated to 80° C. for 30 minutes; by its specific binding affinity to wheat germ agglutinin and a triazine die, color index 61211, of chemical formula 1-amino-4-[[4-[[4-chloro-6-[[3(or 4)-sulfophenyl]amino]-1,3,5-triazin-2-yl]amino]-3-sulfophenyl]amino]-9,10-dihydro-9,10-dioxo-2-anthracene sulfonic acid; and by its lack of specific binding affinity to lysine, and a serum-free and mitogen-free Interleukin-1 preparation and allowing the T-cell growth factor synthesized by the T-lymphocyte cells to collect in the liquid phase of the tissue culture medium.

Claim 3

A process for producing a serum-free and mitogen-free T-cell growth factor (Il-2) comprising adding a biologically active blood serum glycoprotein having a molecular weight of about 90,000 and being able to cause T-cell growth factor production in the presence of Interleukin-1 and T-lymphocytes; said glycoprotein being further characterized by at least one sialic acid moiety as a biologically active site; by retaining more than one-half of its biological activity after being heated to 80° C. for 30 minutes; by its specific binding affinity to wheat germ agglutinin and a triazine die, color index 61211, of chemical formula 1-amino-4-[[4-[[4-chloro-6-[[3(or 4)-sulfophenyl]amino]-1,3,5-triazin-2-yl]amino]-3-sulfophenyl]amino]-9,10-dihydro-9,10-dioxo-2-anthracene sulfonic acid; and by its lack of specific binding affinity to lysine to a serum-free and mitogen-free liquid tissue culture medium; preparing a serum-free and mitogen-free Interleukin-1 preparation by incubating pro-macrophage cells which have been activated with a mitogen in a serum-free and mitogen-free liquid tissue culture medium to synthesize Interleukin-1 by the activated pro-macrophage cells and collect the Interleukin-1 in the liquid phase of the tissue culture medium; mixing the liquid tissue culture medium containing said blood serum glycoprotein and the serum-free and mitogen-free Interleukin-1 preparation; incubating said mixture in the presence of T-lymphocytes to synthesize T-cell growth factor from said T-cell lymphocytes and collect the T-cell growth factor in the liquid phase of the tissue culture medium; separating the cells from liquid phase and recovering the serum-free and mitogen-free liquid phase containing the T-cell growth factor.

Claim 4

A process for producing a serum-free and mitogen-free T-cell growth factor (Il-2) comprising adding a biologically active blood serum glycoprotein having a molecular weight of about 90,000 and being able to cause T-cell growth factor production in the presence of Interleukin-1 and T-lymphocytes; said glycoprotein being further characterized by at least one sialic acid moiety as a biologically active site; by retaining more than one-half of its biological activity after being heated to 80° C. for 30 minutes; by its specific binding affinity to wheat germ agglutinin and a triazine die, color index 61211, of chemical formula 1-amino-4-[[4-[[4-chloro-6-[[3(or 4)-sulfophenyl]amino]-1,3,5-triazin-2-yl]amino]-3-sulfophenyl]amino]-9,10-dihydro-9,10-dioxo-2-anthracene sulfonic acid; and by its lack of specific binding affinity to lysine to a serum-free and mitogen-free liquid tissue culture medium; preparing a serum-free and mitogen-free Interleukin-1 preparation by incubating pro-macrophage cells which have been activated with a mitogen in a serum-free and mitogen-free liquid tissue culture medium to synthesize Interleukin-1 by the activated pro-macrophage cells and collect the Interleukin-1 in the liquid phase of the tissue culture medium; mixing the liquid tissue culture medium containing said blood serum glycoprotein and the serum-free and mitogen-free Interleukin-1 preparation; incubating said mixture in the presence of T-lymphocytes to synthesize T-cell growth factor from said T-cell lymphocytes and collect the T-cell growth factor in the liquid phase of the tissue culture medium; separating the cells from liquid phase and recovering the serum-free and mitogen-free liquid phase containing the T-cell growth factor.

Claim 5

The process of claim 1 wherein the liquid tissue culture medium containing the blood serum glycoprotein is mixed with the serum-free and mitogen-free Interleukin-1 preparation at a volume mixing ratio of about 1:1.