US 6,586,243 B2Granted

Directed maturation of CD34 negative stem cells to programmable antigen presenting cells

OwnerCem Cezayirli, Mel Silversinventors

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Abstract

A method is described whereby dendritic cells derived from the CD34+ and CD 34− hematopoietic cell lineages are directed to become programmable antigen presenting cells. The programmed cells may be pulsed with tumor cell RNA or tumor cell RNA expression products. The protocol provides for directing the maturation of dendritic cells to become antigen presenting cells. The protocol further provides for isolating tumor cell RNA from biopsy material that has been prepared in paraffin block storage. The directed dendritic cell is provided with a plurality of tumor markers by using tumor RNA in toto, the poly A+RNA fraction or the expression product of such RNA. Once activated the dendritic cells are incubated with T4 and T8 lymphocytes to stimulate and sensitize the T lymphocytes which upon introduction either into a donor host or a nondonor recipient will provide immune response protection.

Claims14

Dependent claims

A method for producing a disease-specific immunogenic cell composition capable of stimulating an immune response to said disease, the method comprising the steps of:

a. isolating CD34 negative hematopoietic stem cells from a donor;
b. incubating the stem cells in a cell culture medium comprising GM-CSF, SCF and IL-4, whereby the stem cells are directed to develop into programmable antigen presenting cells (pAPCs);
c. mixing the pAPCs with cellular material derived from cells positive for said disease, the cellular material selected from the group consisting of cell lysate, cell RNA, cell poly A+ RNA, cDNA derived from cell RNA, and cDNA derived from cell RNA incorporated into an expression vector;
d. incubating the pAPCs with the cellular material, whereby loaded pAPCs are produced;
and
e. incubating the loaded pAPCs with TNF-α, whereby disease-specific programmed super antigen presenting cells (pSAPCs) capable of stimulating an immune response to said disease are produced.

3 more elements

Dependencies are inferred from the claim text.

Facts

Application: 09/747,153Filed 20 Dec 2000 · continuation
Priority: 7 Feb 1997
Published: US 2001/0005506 A128 Jun 2001
Granted: 1 Jul 2003
Term ends: Not estimatedThe source records don’t support an estimate.

Classification12 codes

A61K 38/20
Cytokines; Lymphokines; Interferons: interleukins [IL]
A61K 38/193
Cytokines; Lymphokines; Interferons: colony stimulating factors [CSF]
A61K 40/19
Characterised by the cell type used: dendritic cells
A61K 40/24
Characterised by the effect or the function of the cells: antigen-presenting cells [APC]

Family

Application60/037,435Us provisional application · filed 7 Feb 1997
Application60/038,736Us provisional application · filed 14 Feb 1997
Application60/042,110Us provisional application · filed 26 Mar 1997
Application09/017,842Parent of this continuation · filed 3 Feb 1998
US 2001/0005506 A1Pre-grant publication · published 28 Jun 2001
US 6,251,665 B1Parent of this continuation
US 6,586,243 B2This patent

From the US filing history. Foreign family members aren’t listed.