Abstract: Construct-complexes of a hemoglobin, a hepatocyte modifying substance bound to the hemoglobin, and a haptoglobin bound to the hemoglobin, are provided, for administration to mammalian patients. The construct-complex may be formed ex vivo, or a hemoglobin-hepatocyte modifying substance combination may be administered to the patient so that haptoglobin in the mammalian body bonds thereto to form the construct-complex in vivo. Disorders of the liver may be diagnosed and treated using construct-complexes described herein.

Abstract: Methods for the expansion of CD4, CD8, and DP T-cells from HIV infected patients are disclosed which allow the maintenance of low levels of HIV. The invention further discloses methods for the inhibition of HIV gene expression. Also disclosed are methods for the rapid and efficient screening of cells derived from HIV-infected patients to assess the suitability of various antiviral treatments. The invention further provides a means for the generation of cell banks for use in immune reconstitution and means of treating or modifying expanded cell populations prior to infusion to enhance or modulate therapeutic effectiveness.

Abstract: Hemoglobin conjugates useful as a hemoglobin-based oxygen carriers are prepared by reacting hemoglobin with oxidatively ring-opened polysaccharides such as hydroxyethyl starch or dextran, and storing the resultant conjugate under conditions which allow it to transform to a lower molecular weight product, after conjugation. The conjugate is then reductively stabilized to form secondary amino bonds between the hemoglobin and the polysaccharide, and formulated as an HBOC.

Abstract: The method for obtaining and expanding TcR&ggr;&dgr;+ T cells in culture is described. The method involves: 1) culturing cells from a sample containing TcR&ggr;&dgr;+ T cells or precursors thereof in a first culture medium comprising a T cell mitogen and at least two cytokines and 2) culturing the cells obtained in step 1) in a second culture medium comprising at least two cytolines. Preferably, the method comprises 1) culturing the cells in a first culture medium comprising (a) a T cell mitogen, (b) interleukin-2 and (c) interleukin-4, and 2) culturing the cells obtained in step 1) in a second culture medium comprising (i) interleukin-2 and (ii) interleukin-4 to obtain TcR&ggr;&dgr;+ T cells. The TcR&ggr;&dgr;+ T cells obtained by the method can be used in a variety of experimental, therapeutic and commercial applications.

Abstract: Construct-complexes of a hemoglobin, a hepatocyte modifying substance bound to the hemoglobin, and a haptoglobin bound to the hemoglobin, are provided, for administration to mammalian patients. The construct-complex may be formed ex vivo, or a hemoglobin-hepatocyte modifying substance combination may be administered to the patient so that haptoglobin in the mammalian body bonds thereto to form the construct-complex in vivo. Disorders of the liver may be diagnosed and treated using construct-complexes described herein.

Abstract: Hemoglobin conjugates useful as a hemoglobin-based oxygen carriers are prepared by reacting hemoglobin with oxidatively ring-opened polysaccharides such as hydroxyethyl starch or dextran, and storing the resultant conjugate under conditions which allow it to transform to a lower molecular weight product, after conjugation. The conjugate is then reductively stabilized to form secondary amino bonds between the hemoglobin and the polysaccharide, and formulated as an BOC.

Abstract: The present invention describes a serum-free medium that promotes the growth and differentiation of erythroid cells, cells that are highly transducible by a non-viral method and genes which increase the growth of erythroid cells and decrease their dependency on Epo. This invention can be used in the expansion of hematopoietic stem cells to produce cultures of erythroid cells as a source of erythroid-specific proteins such as hemoglobin. Hematopoietic stem cells are cultured ex vivo in a serum-free culture medium with the addition of IL-3, SCF and EPO. Cells transfected with the gene described in the present invention can be cultured in the serum-free culture medium with decreased dependency on Epo and other cytokines, thereby reducing the cost of the production of hemoglobin.

Abstract: The invention is directed to methods for the production of selected populations of lymphocytes. Lymphocytes produced can be isolated and purified using well known and established procedures to provide a consistent lymphocyte source which one of ordinary skill in the art can modify to provide an appropriate type or an optimal level of a desired lymphocyte. The availability of such cell populations allows for not only for the complete reconstitution of the depleted, defective or missing lymphocyte population in a patient, but also provides the flexibility of having sufficient cells to permit multiple or cyclic treatments. These methods for expanding target cell populations are broadly applicable to the selective expansion of several types of lymphocytes and are demonstrated to maintain phenotype as well as antigen specificity.

Abstract: A chemically modified, crosslinked hemoglobin product suitable for use as a hemoglobin based oxygen carrier comprises a mixture of hemoglobin species and consists essentially of about 40% tetrameric hemoglobin units of molecular weight about 64,000 daltons, up to 5% dimeric hemoglobin units of molecular weight about 32,000 daltons, and the balance oligomeric hemoglobin units of molecular weight up to about 60,000 daltons, the mixed product containing no polymeric hemoglobin species of molecular weight greater than 600,000 daltons. The product can be made directly by a crosslinking reaction under controlled conditions, without the need for separating therefrom high molecular weight species.

Abstract: A preselected hemoglobin species is separated from contaminants having a different acidity from that of the preselected hemoglobin species, by an overload displacement chromatography process. To remove more acidic contaminants, the process is conducted under anion exchange conditions. To remove more basic contaminants, the process is conducted under cation exchange conditions. In either case, the exchange column is overloaded to displace the hemoglobin species therefrom with contaminants having greater affinity for the column, and using the impure hemoglobin solution as the displacer.

Abstract: Hemoglobin for forming the basis of blood substitute is modified by crosslinking it by reaction with a polyaldehyde produced by ring opening oxidation of a di/tri-saccharide such as raffinose, wherein the o-saccharide solutions are maintained and the reaction is conducted at a pH from about 5.0-7.0, and at a stoichiometry from about 1:1-4:1 based upon the molar ratio of di/tri-saccharide to hemoglobin tetramers, followed by appropriate reduction of the Schiff base linkages so formed to stabilize them. Careful control of the pH of ring opening of the di/tri-saccharide ensures consistent production of the polyaldehyde without unwanted side products and reaction of this product with the hemoglobin at the specified stoichiometry leads to high yields of stabilized, tetrameric hemoglobin and oligomers thereof of predetermined composition and beneficial properties.

Abstract: Hemoglobin is purified from a crude solution thereof, to obtain an aqueous solution containing at least 99% of a preselected hemoglobin species, by a two stage displacement chromatography process. One of the stages is conducted under anionic exchange conditions, and the other under cationic exchange conditions. In both stages, the exchange column is overloaded to displace the hemoglobin species therefrom with contaminants having greater affinity for the column, and using the impure hemoglobin solution as the displacer. Normally, anionic exchange is conducted first, with contaminants more acidic than the hemoglobin displacing the hemoglobin from the column and themselves remaining attached to the column for separation. The cationic exchange process is conducted second, on the eluent from the first column, and in this stage, the more basic contaminants displace the hemoglobin from the column under overload conditions, to yield a substantially pure hemoglobin solution.