The invention provides methods of making immune effector cells (e.g., T cells, NK cells) that can be engineered to express a chimeric antigen receptor (CAR), and compositions and reaction mixtures comprising the same.

Microfluidic filter devices and methods of fabricating such devices. A microfluidic filter device for capturing an object (e.g., a red blood cell) can include a filter structure having a plurality of through holes extending from a first side of the filter structure to a second side of the filter structure and arranged in a repeating pattern, the through holes sized to capture the object. The device further includes a substrate including a plurality of vanes that supports at least a portion of the filter structure, a plurality of electrodes comprising a set of electrodes associated with each through hole, each set of electrodes including at least a pair of electrodes associated with each through hole and aligned with its associated through hole to apply electrical forces to an object captured in the through hole, and electrical connections to each of the plurality of electrodes.

A method of treating a subject in need of a non-syngeneic cell or tissue graft is disclosed. The method comprising: (a) transplanting into a subject a dose of T cell depleted immature hematopoietic cells, wherein the T cell depleted immature hematopoietic cells comprise less than 510.sup.5 CD3.sup.+ T cells per kilogram body weight of the subject, and wherein the dose comprises at least about 510.sup.6 CD34+ cells per kilogram body weight of the subject; and subsequently (b) administering to the subject a therapeutically effective amount of cyclophosphamide, wherein the therapeutically effective amount comprises 25-200 mg per kilogram body weight, thereby treating the subject.

The present invention relates to the use of a combination of: (i) a macromolecular erythrocyte sedimentation enhancer, and (ii) dimethyl sulphoxide (DMSO), dimethylglycine (DMG) and/or valine;
to recover non-erythrocyte blood cells from a blood cell-containing sample and/or to prime non-erythrocyte blood cells to protect their integrity in subsequent cryopreservation step(s).

Described herein are methods for selecting lymphocytes for adoptive cell therapy based on P-glycoprotein expression and compositions comprising same.

The present invention provides a method of preparing a population of genetically modified cells which comprise a chimeric antigen receptor (CAR) or a transgenic T-cell receptor (TCR) comprising: providing a starting population of cells; depleting said starting population of cells which express a target antigen; and introducing into a cell in the depleted starting population a nucleic acid sequence which encodes a CAR or transgenic TCR against the target antigen. The present invention also provides genetically modified cells, pharmaceutical compositions and pharmaceutical compositions for use in the treatment and/or prevention of disease.

A method for determining immunogenicity of a protein agent. The method includes constructing a library of peripheral blood mononuclear cells having various HLA-DRB1 genotypes; culturing peripheral blood mononuclear cell CD14+ monocyte-derived immature dendritic cells for each genotype in a medium containing a protein to be measured, GM-CSF, IL-4, TNF-, IL-1, IL-6 and PGF.sub.2 to prepare mature dendritic cells; removing CD8+ T cells from the peripheral blood mononuclear cells for each genotype to prepare CD8+ T cell-free peripheral blood mononuclear cells; co-culturing the mature dendritic cells and the CD8+ T cell-free peripheral blood mononuclear cells at a cell count ratio of approximately 1:5 to 1:20; and quantifying the CD4+ T cells proliferated by co-cultivation per genotype.

The invention provides compositions and methods useful for the depletion of cells, such as CD45+, CD135+, CD34+, CD90+, and/or CD110+ cells, and for the treatment of various hematopoietic diseases, metabolic disorders, cancers, and autoimmune diseases, among others. Described herein are antibodies, antigen-binding fragments, ligands, and conjugates thereof that can be applied to effect the treatment of these conditions, for instance, by depleting a population of CD45+, CD135+, CD34+, CD90+, or CD110+ cells in a patient, such as a human. The compositions and methods described herein can be used to treat a disorder directly, for instance, by depleting a population of CD45+, CD135+, CD34+, CD90+, or CD110+ cancer cells or autoimmune cells. The compositions and methods described herein can also be used to prepare a patient for hematopoietic stem cell transplant therapy and to improve the engraftment of hematopoietic stem cell transplants by selectively depleting endogenous hematopoietic stem cells prior to the transplant procedure.

The object is to remove thrombocytes from apheresis blood. Mononuclear cells are prepared so as not to contain thrombocytes. The present invention provides a method for preparing mononuclear cells, which comprises (1) the step of mixing a non-cytotoxic and nonionic density-adjusting agent with apheresis blood to adjust density of plasma to be 1.066 to 1.078 g/ml, and (2) the step of centrifuging stratified layers including the following layers a and b to separate mononuclear cells:

a. a layer of a density gradient centrifugation medium for separation of mononuclear cells,

b. a layer of the density-adjusted apheresis blood layered under the layer a.

The present disclosure provides methods of hematopoietic stem cell transplantation (HSCT). In particular, the present disclosure provides a method of HSCT using a combination of an in-vivo T-cell depletion method, with an ex-vivo method of ?? T cell expansion and ?? T cell depletion. The in-vivo T-cell depletion method depletes (in-vivo) the alloreactive T cells that would otherwise increase the risk of GvHD.