Methods and compositions for diagnostic assays for measuring cell mediated immune response

摘要

Described are compositions and methods for detecting or monitoring the ability of an individual to mount a cell mediated immune response to a target antigen. Methods rely in part upon the physical association, e.g., by fusion, of a Lethal Factor (LF) polypeptide with a target antigen. The LF polypeptide moiety, including, for example, an LFn polypeptide moiety, serves as a transport factor to deliver target antigens, including full length target polypeptides, to the cytosol of an intact, living immune cell from an individual. Measurement of a cytokine response by the immune cell from the individual provides a read out of a cell mediated immune response. The methods and compositions described provide diagnostic as well as prognostic information and can guide the direction of therapy.

主权项

1. A kit for measuring a cell mediated immune (CMI) response to a TB antigen polypeptide of a biological sample from a subject, the kit comprising:
a. a panel of at least two fusion polypeptides wherein each fusion polypeptide comprises a portion of a LF polypeptide lacking LF enzymatic activity but sufficient to promote transmembrane delivery of said fusion polypeptide to a cell, fused to a TB-specific antigen polypeptide, wherein said portion of a LF polypeptide comprises at least the 60 carboxy-terminal amino acids of SEQ ID NO: 3, or a conservative substitution variant thereof that promotes transmembrane delivery, wherein the panel of at least two fusion polypeptides comprises at least two different TB-specific antigen polypeptides, wherein one of the TB-specific antigen polypeptide is TB1 (CFP) polypeptide of SEQ ID NO: 7and wherein one of the TB-specific antigen polypeptide is TB2 (ESAT) polypeptide of SEQ ID NO: 6; and b. antibody-based detection means to detect INF-γreleased from the biological sample.