The Scripps Research Institute

Ortho-mcneil Pharmaceutical Corp.

Janssen Pharmaceuticals, Inc.

Michael R Jackson

Per A Peterson

Zeling Cai

Juli Degraw

Wei-Xing Shi

Yan Kong

Anders Brunmark

Jonathan Sprent

Ann M Moriarty

Lars Karlsson

Didier J Leturcq

Pierre Langlade-Demoyen

Susan R Webb

Ola Winqvist

Alain Luxembourg

Marja Heiskala

Jon M Richards

Alain T Luxembourg

Ola Wingvist

Weixing Shi

T cell responses are often diminished in humans with a compromised immune system. We have developed a method to isolate, stimulate and expand na&#xef;ve cytotoxic T lymphocyte precursors (CTLp) to antigen-specific effectors, capable of lysing tumor cells in vivo. This ex vivo protocol produces fully functional effectors. Artificial antigen presenting cells (AAPCs;) transfected with human HLA class I and defined accessory molecules, are used to stimulate CD8T cells from both normal donors and cancer patients. The class I molecules expressed to a high density on the surface of thecells are empty, allowing for efficient loading of multiple peptides that results in the generation of polyclonal responses recognizing tumor cells endogenously expressing the specific peptides. The responses generated are robust, antigen-specific and reproducible if the peptide epitope is a defined immunogen. This artificial antigen expression system can be adapted to treat most cancers in a significant majority of the population.

Cell therapy method for the treatment of tumors

T cell responses are often diminished in humans with a compromised immune system. We have developed a method to isolate, stimulate and expand na&#xef;ve cytotoxic T lymphocyte precursors (CTLp) to antigen-specific effectors, capable of lysing tumor cells in vivo. This ex vivo protocol produces fully functional effectors. Artificial antigen presenting cells (AAPCs;) transfected with human HLA class I and defined accessory molecules, are used to stimulate CDT cells from both normal donors and cancer patients. The class I molecules expressed to a high density on the surface of thecells are empty, allowing for efficient loading of multiple peptides that results in the generation of polyclonal responses recognizing tumor cells endogenously expressing the specific peptides. The responses generated are robust, antigen-specific and reproducible if the peptide epitope is a defined immunogen. This artificial antigen expression system can be adapted to treat most cancers in a significant majority of the population.

Cytotoxic T lymphocytes (CTLs) specific for antigenic peptides derived from IgE molecule can be generated in vitro by stimulating resting naive CD8 T cells with IgE peptides presented by artificial antigen presenting cells. The IgE specific CTLs lyse the target cells loaded with IgE peptides in vitro and inhibit antigen specific IgE response in vivo. In addition, adoptive transfer of the IgE specific CTL to an asthmatic mouse model can inhibit the development of lung inflammation and airway hypersensitivity. IgE specific CTL provides a treatment for allergic asthma and other IgE-mediated allergic diseases. Antigenic peptides identified from non-tumor self-antigens induce specific cytotoxic T lymphocyte (CTL) in vitro. The CTL induced by peptides identified from CD40L can kill activated CD4 T cells. In vitro generated CTL specific for CD40L inhibit CD4-dependent antibody responses of all isotypes in vivo. In contrast, CTL induced by antigenic peptides derived from IgE specifically inhibit IgE responses, and adoptive transfer of CD40L-specific CTL to NOD mice at early age delay the development of diabetes in NOD mice. In vitro generated CTL specific for non-tumor self-antigens expressed on activated CD4 T cells regulate immune responses in vivo.

Ex-vivo priming for generating cytotoxic T lymphocytes specific for non-tumor antigens to treat autoimmune and allergic disease

IgE antigenic peptides

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The present invention relates to synthetic antigen-presenting matrices, their methods of making and their methods of use. One such matrix is cells that have been transfected to produce MHC antigen-presenting molecules with one or more accessory molecules. The matrices are used to activate naive CD4T cells as well as shift the ongoing activation state into a preferred differentiated population of either Th1 or Th2 cells.

A new method to capture, purify and expand antigen-specific T lymphocytes has been developed using magnetic beads coated with recombinant MHC class I molecules. This method was optimized using homogenous populations of naive T cells purified from mice transgenic for the 2C T cell receptor (TCR). These T cells were captured on beads coated with MHC class I molecules and the relevant antigenic peptides. MHC and peptide specificity was confirmed by the usage of irrelevant MHC peptide combinations. An enrichment of 800 to 1600 fold was measured, using 2C T cells mixed with irrelevant T cells, starting from a 2C T cell frequency of 1/3000. The same approach was used to purify antigen-specific CD8T cells from total CD8T cells from naive mice. The recovered cells could be expanded and specifically kill target cells in vitro; they had a significant effect in vivo as well. We expect this procedure to be suitable to purify and expand in vitro tumor- and virus-specific killer T cells for use in cell therapy.

Purification of antigen-specific T cells

MHC class II antigen-presenting systems and methods for activatind CD4+T cells

The present invention relates to synthetic antigen-presenting matrices, their methods of making and their methods of use. One such matrix is cells that have been transfected to produce MHC antigen-presenting molecules and assisting molecules such as co-stimulatory molecules. The matrices can be used to activate CD8 T-cells to produce cytokines and become cytotoxic.

Antigen presenting system and methods for activation of T-cells

Growing community of inventors

Del Mar, CA, United States of America

Michael R Jackson