T Cell Sciences, Inc.

Ortho Diagnostic Systems Inc.

T Cell Diagnostics, Inc.

Michael C Brown

Stephen H Ip

Patrick C Kung

Stefan J Kirchanski

A R Azad

Linda A McKeen

Linda A MacKeen

Linda A Mackeen

Cell-free or released T cell antigen receptors, immunoassays which allow the detection of the released T cell antigen receptor in cell culture supernatants, cell lysates, and biological fluids, as well as diagnostic and therapeutic compositions and methods for monitoring and treating certain diseases or disorders which elicit or involve a T cell response are described. The released T cell antigen receptors described herein differ from the cell membrane bound antigen receptors and appear to exist in a variety of forms, some of which may be complexed with other T cell determinants such as the T3 antigen. Despite the size heterogeneity of the released T cell antigen receptors or receptor/complexes, these may be reliably identified using anti-receptor antibodies, including, but not limited to, anti-major framework, anti-minor frarmework and anti-clonotypic antibodies which define particular epitopes of the released T cell receptor or receptor/complex. The immunoassays described herein may be used to test for the presence of the released receptor or receptor/complex in biological samples and may be used to diagnose or monitor patients with disorders or diseases that elicit or involve T cell response who exhibit elevated serum level of the receptor or receptor/complex. In addition, the cell free T cell antigen receptors themselves may be used diagnostically and/or therapeutically for diseases or conditions that are not easily addressed using immunoglobulins.

Cell free T cell antigen receptor beta chain and its clinical utilities

The present invention is directed to the measurement of soluble T cell growth factor receptors, soluble T cell differentiation antigens, or related soluble molecules or fragments thereof, and the use of such measurements in the diagnosis, staging, and therapy of diseases and disorders. Specific embodiments involve the diagnosis and monitoring of therapy using absolute values of such soluble molecules. Further embodiments involve detecting a change in the levels of such soluble molecules, in the diagnosis and therapy of diseases and disorders. In specific embodiments, measurements of interleukin-2 receptor levels can be made to detect lung cancer, or to stage squamous cell lung carcinoma. In other embodiments, detection of increases in both soluble IL2R and creatinine in the body fluid of a transplant patient can be used to differentially diagnose renal allograft rejection from infection. The invention is also directed to methods for measurement of soluble CD4 antigens, which measurements can be used, in a specific embodiment, to diagnose a state of immune activation, to diagnose rheumatoid arthritis, to monitor therapeutic efficacy (e.g. of AIDS treatments), or to stage adult T cell leukemia in a patient. In another aspect, the invention relates to the detection, staging, and monitoring of therapy of diseases and disorders by measuring a plurality of soluble T cell markers.

Therapeutic and diagnostic methods using soluble T cell surface molecules

The present invention is directed to the measurement of soluble T cell growth factor receptors, soluble T cell differentiation antigens, or related soluble molecules or fragments thereof, and the use of such measurements in the diagnosis and therapy of diseases and disorders. The measurement of such molecules can be valuable in monitoring the effect of a therapeutic treatment on a subject, detecting and/or staging a disease in a subject, and in differential diagnosis of a physiological condition in a subject. These measurements can also aid in predicting therapeutic outcome and in evaluating and monitoring the immune status of patients. In specific embodiments, measurements of serum or plasma interleukin-2 receptor levels can be made, to detect or stage leukemia or lymphoma. In other embodiments, IL2R levels, or CD8 levels, can be used to differentially diagnose renal allograft rejection, as distinguished from Cyclosporin A nephrotoxicity. In another embodiment, CD8 levels can be measured to differentially diagnose rheumatoid arthritis, as distinct from other joint diseases. In particular embodiments, measurements of the soluble T cell surface molecules can be accomplished by sandwich enzyme immunuoassays.

Methods are provided for detecting and determining the amount in a sample of an antigenic determinant from an antigen receptor derived and released from a T cell or NK cell. Methods are also provided for detecting and determining the amount in a sample of an antigenic determinant from a complex of at least a portion of an antigen receptor derived from and released from a T cell or NK cell and a protein complex. These methods form the bases for methods of diagnosing and monitoring in a subject a disease characterized by the presence or an amount different from a normal subject of one of these antigenic determinants in a body fluid. A soluble antigen receptor or complex thereof derived from a T cell or a NK cell but free of such T cell or NK cell is also provided.

Assay systems for detecting cell-free T cell antigen receptor related

Lysing reagents for use in flow cytometry instruments. Lysing reagents suitable for lysing red blood cells are provided thereby permitting 3-part differential counting of white blood cells. The reagents comprise a leukoprotective agent for preserving the lymphocyte cellular integrity during analysis, and buffers adjusted to provide the correct pH environment for optimal lysis and activity. Preferred embodiments will additionally incorporate non-ionic preservatives in very low concentrations.

Flow cytometry lysing reagent with leukoprotective agent for producing a

An immunological substance detecting reagent is provided which, in the preferred embodiment, employs an immunological homolog specific for the immunological substance to be detected coupled to a water-soluble polymer having a net charge not greater than zero. The water-soluble polymer further has associated a plurality of marker substances such as fluorophore molecules thereby providing a reagent of increased sensitivity for the detection of an antigen-antibody immunological reaction.

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Wayland, MA, United States of America

Michael C Brown