The present disclosure relates generally to the manufacture of regulatory T cells (Tregs) for use in immunotherapy. In particular, the present disclosure relates to robust approaches for the expansion of alloantigen-reactive Tregs ex vivo. Alloantigen-reactive Tregs produced in this way are suitable for the induction and/or maintenance of immunologic tolerance in recipients of allogeneic transplants.

Provided herein are, inter alia, methods and compositions for treating, preventing, or reducing the risk of dysbiosis, inflammation, inflammatory diseases, childhood obesity, and premature birth. Included are methods and compositions for increasing or promoting healthy or normal immune system maturation. In aspects, provided herein are methods and compositions for detecting and isolating bacterial strains. Isolated bacterial strains and culture methods are also provided.

Several embodiments disclosed herein relate to methods and processes for the co-expansion of multiple types of immune cells, in order to generate a mixed cell population. Some embodiments relate to the use of various stimuli specific to the various subpopulations to achieve expansion of those subpopulations at a particular time in a culturing process in order to generate an expanded population of immune cells having a desired ratio of the various subpopulations. In several embodiments, such mixed cell populations exhibit desirable characteristics, such as cytotoxic effects against tumor cells that enhance the efficacy of cancer immunotherapy.

Provided herein are methods for determining the presence of cancer (malignant versus benign), monitoring the progression of cancer, monitoring cancer relapse, monitoring the response to cancer therapy, or cancer staging in a subject, by evaluating CD33.sup.+/HLA-DR.sup.low, CD14.sup.+/HLA-DR.sup.low, CD66b.sup.+/HLA-DR.sup.low or, CD11b.sup.+/HLA-DR.sup.low MDSC for activation of a transcription factor. Transcription factors include, but are not limited to, STAT3, pSTAT3, HIF1α, or C/EBPβ. The MDSC phenotype can be CD33.sup.+HLA-DR.sup.lowHIF1α.sup.+/STAT3.sup.+, CD14.sup.+HLA-DR.sup.lowHIF1α.sup.+/STAT3.sup.+/pSTAT3.sup.+/C/EBPb.sup.+, CD66b.sup.+HLA-DR.sup.lowHIF1α.sup.+/STAT3.sup.+/pSTAT3.sup.+/C/EBPb.sup.+, CD33.sup.+HLA-DR.sup.lowHIF1α.sup.+/STAT3.sup.+/pSTAT3.sup.+/C/EBPb.sup.+, CD11b.sup.+HLA-DR.sup.lowHIF1α.sup.+/STAT3.sup.+/pSTAT3.sup.+/C/EBPb.sup.+, or CD11b.sup.+HLA-DR.sup.lowC/EBPβ.sup.+. Also provided herein are methods for inducing human MDSC from healthy donor peripheral blood mononuclear cells (PBMC) by co-culturing PBMC with human solid tumor cell lines and subsequently measuring their suppressive ability.

Provided are antigen-density sensing molecular circuits and methods of using the same. Aspects of such circuits will generally include an antigen-triggered switch component and a therapeutic component specific for the same antigen as the antigen-triggered switch component. The circuits will generally be configured such that expression of the therapeutic component is induced by the antigen-triggered switch component when the switch is activated by binding the antigen. Nucleic acids, expression constructs, vectors and the like encoding such circuits, and cells genetically modified to include an antigen-density sensing molecular circuit are also provided. Also provided are methods of making antigen-density sensing molecular circuits, methods of inducing expression of high affinity therapeutics specific to an antigen expressed by a target cell, methods of activating an immune response to a target cell, methods of treating a subject for a cancer expressing an antigen, and the like, where such methods involve antigen-density sensing molecular circuits.

The present invention relates to therapeutic agents, particularly to therapeutic polypeptides and nucleic acids having the capacity for selective expression under conditions of hypoxia, cells incorporating the nucleic acids and their use in therapy, in particular in methods requiring selective expression under conditions of hypoxia, such as typically found in solid cancers. The nucleic acids encode novel hypoxia-responsive chimeric antigen receptors (CARs). The invention also relates to hypoxia-responsive regulatory nucleic acids.

To provide a technique for selecting a target cell producing a target substance that specifically binds to a desired cell membrane protein more rapidly and efficiently. A substrate 1 having a plurality of microwells 2 is provided. A first cell 3 expressing a target cell membrane protein on its surface is allowed to adhere to each of the microwells 2. One or two second cells 5 as a candidate of a target cell are introduced into each microwell 2, and are allowed to coexist with the first cell 3 in the microwell 2, and target substance 6 secreted by the second cell 5 is brought into contact with the first cell 3. A microwell 2 including the first cell 3 to which the target substance 6 binds is identified. The second cell 5 as the target cell is recovered from the identified microwell 2. One example of the target substance 6 is an antibody. Visualization may be performed by adding a label substance 7.

The present invention relates to peptides, proteins, nucleic acids and cells for use in immunotherapeutic methods. In particular, the present invention relates to the immunotherapy of cancer. The present invention furthermore relates to tumor-associated T-cell peptide epitopes, alone or in combination with other tumor-associated peptides that can for example serve as active pharmaceutical ingredients of vaccine compositions that stimulate anti-tumor immune responses, or to stimulate T cells ex vivo and transfer into patients. Peptides bound to molecules of the major histocompatibility complex (MHC), or peptides as such, can also be targets of antibodies, soluble T-cell receptors, and other binding molecules.

Provided herein are methods for obtaining tumor-targeting T cells, as well as for identifying tumor-targeting TCRs.

Provided herein are compositions and methods for adoptive cell therapy comprising engineered immune cells that express a tumor antigen-targeted chimeric antigen receptor and a SIRPα polypeptide.