{"search_session":{},"preferences":{"l":"en","queryLanguage":"en"},"patentId":"135-524-968-520-335","frontPageModel":{"patentViewModel":{"ref":{"entityRefType":"PATENT","entityRefId":"135-524-968-520-335"},"entityMetadata":{"linkedIds":{"empty":true},"tags":[],"collections":[{"id":8906,"type":"PATENT","title":"Univ Pennsylvania Patent Portfolio","description":"","access":"OPEN_ACCESS","displayAvatar":true,"attested":false,"itemCount":24238,"tags":[],"user":{"id":91044780,"username":"Cambialens","firstName":"","lastName":"","created":"2015-05-04T00:55:26.000Z","displayName":"Cambialens","preferences":"{\"usage\":\"public\",\"beta\":false}","accountType":"PERSONAL","isOauthOnly":false},"notes":[{"id":8221,"type":"COLLECTION","user":{"id":91044780,"username":"Cambialens","firstName":"","lastName":"","created":"2015-05-04T00:55:26.000Z","displayName":"Cambialens","preferences":"{\"usage\":\"public\",\"beta\":false}","accountType":"PERSONAL","isOauthOnly":false},"text":"
Search applicants and owners= \"Univ Pennsylvania\", \"Pennsylvania Univ\", \"Univ Penns*\", \"Penn* Univ\", \"University of Pennsylvania\", \"Pennsylvania University\".
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Search applicants and owners= \"Univ Pennsylvania\", \"Pennsylvania Univ\", \"Univ Penns*\", \"Penn* Univ\", \"University of Pennsylvania\", \"Pennsylvania University\".
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selecting a sample of human CD4+ T cells;\n
contacting said sample with an anti-CD25 antibody;\n
isolating cells that bind to said anti-CD25 antibody from said sample using a double column magnetic antibody cell sorting (MACS) purification procedure, to produce an isolated population of human CD4+CD25+ Treg cells;\n
culture-expanding said population of human CD4+CD25+ Treg cells comprising contacting said isolated population of human CD4+CD25+ Treg cells with immobilized anti-CD3 antibody and immobilized anti-CD28 antibody, further culture-expanding said isolated population of human CD4+CD25+ Treg cells in the presence of an irradiated CD4+ feeder cell or the irradiated CD4+ feeder cell conditioned medium, thereby producing culture-expanded therapeutic human Treg cells, wherein said culture-expanded Treg cells are CD62L+/CD27+ and are capable of inhibiting proliferation of CD4+CD25− responding T cells in a Mixed Lymphocyte Reaction (MLR) assay by at least 90%."],"number":1,"annotation":false,"title":false,"claim":true},{"lines":["The method of claim 1, wherein said isolating step comprises contacting the selected human CD4+ T cells with 2 μl of anti-CD25 magnetic microbeads per 107 total cells, and wherein the double column purification procedure comprises purifying by running the bead/cell composition over a magnetic column to separate bead-bound cells, washing, and re-eluting over a second magnetic column, and again washing until <1-2% of nonsuppressor cells remain in the isolated population of human CD4+CD25+ Treg cells."],"number":2,"annotation":false,"title":false,"claim":true},{"lines":["The method of claim 1, wherein said culture-expanding step produces an effective amount of suppressor cells to achieve therapeutic suppression of an immune or autoimmune response in a human subject."],"number":3,"annotation":false,"title":false,"claim":true},{"lines":["The method of claim 1, wherein said culture-expanding step further comprises contacting said isolated population of human CD4+CD25+ Treg cells with IL-2."],"number":4,"annotation":false,"title":false,"claim":true},{"lines":["The method of claim 1, wherein said isolated population of human CD4+CD25+ Treg cells are expanded at least 10-20 fold in 14 days of culture in said culture-expanding step."],"number":5,"annotation":false,"title":false,"claim":true},{"lines":["The method of claim 5, wherein said isolated population of human CD4+CD25+ Treg cells are expanded at least 100-fold by culturing for an additional 1-2 weeks."],"number":6,"annotation":false,"title":false,"claim":true},{"lines":["The method of claim 1, further comprising generating therapeutic human Treg cell lines that retain long term down-regulatory suppressor function."],"number":7,"annotation":false,"title":false,"claim":true},{"lines":["The method of claim 1, wherein the sample of human CD4+ T cells is selected from the group consisting of whole or partially purified blood or hematopoietic cells, wherein said hematopoietic cells are selected from the group consisting of peripheral blood mononuclear cells, peripheral blood lymphocytes, spleen cells, tumor-infiltrating lymphocytes and lymph node cells, and bone marrow cells."],"number":8,"annotation":false,"title":false,"claim":true},{"lines":["The method of claim 7, wherein said therapeutic human Treg cell lines retain long term down-regulatory suppressor function for at least three weeks."],"number":9,"annotation":false,"title":false,"claim":true},{"lines":["The method of claim 1, wherein said anti-CD25 antibody is directly conjugated to a magnetic microbead."],"number":10,"annotation":false,"title":false,"claim":true},{"lines":["The method of claim 1, wherein said MACS purification procedure is an indirect method, wherein said isolating step further comprises contacting said sample to magnetic microbeads conjugated to a secondary agent that binds to said anti-CD25 antibody."],"number":11,"annotation":false,"title":false,"claim":true},{"lines":["The method of claim 11, wherein said isolating step further comprises substantially enhancing CD4+CD25bright cells in said isolated population, while substantially depleting CD25dim cells in said isolated population."],"number":12,"annotation":false,"title":false,"claim":true},{"lines":["The method of claim 12, wherein said isolating step comprises contacting the selected, anti-CD25 antibody-contacted human CD4+ T cells with 2 μl of said magnetic microbeads per 107 total cells, and wherein the double column purification procedure comprises purifying by running the bead/cell composition over a magnetic column to separate bead-bound cells, washing, and re-eluting over a second magnetic column, and again washing until <1-2% of nonsuppressor cells remain in the purified isolate."],"number":13,"annotation":false,"title":false,"claim":true},{"lines":["The method of claim 11, wherein said anti-CD25 antibody is conjugated to FITC and said secondary agent is an anti-FITC antibody."],"number":14,"annotation":false,"title":false,"claim":true}]}},"filters":{"npl":[],"notNpl":[],"applicant":[],"notApplicant":[],"inventor":[],"notInventor":[],"owner":[],"notOwner":[],"tags":[],"dates":[],"types":[],"notTypes":[],"j":[],"notJ":[],"fj":[],"notFj":[],"classIpcr":[],"notClassIpcr":[],"classNat":[],"notClassNat":[],"classCpc":[],"notClassCpc":[],"so":[],"notSo":[],"sat":[]},"sequenceFilters":{"s":"SEQIDNO","d":"ASCENDING","p":0,"n":10,"sp":[],"si":[],"len":[],"t":[],"loc":[]}}