Method of Measuring Adaptive Immunity

摘要

A method of measuring immunocompetence is described. This method provides a means for assessing the effects of diseases or conditions that compromise the immune system and of therapies aimed to reconstitute it. This method is based on quantifying T-cell diversity by calculating the number of diverse T-cell receptor (TCR) beta chain variable regions from blood cells.

主权项

1. A method, comprising:
generating a plurality of amplicons comprising rearranged TCR or IG CDR3 nucleic acid sequences from a first sample comprising nucleic acid molecules obtained from lymphocytes from a mammalian subject, using a plurality of V-segment primers and a plurality of J-segment primers in a single multiplex PCR, wherein each V-segment primer is complementary to one or more functional TCR or IG V-region gene segments, wherein each J-segment primer is complementary to one or more functional TCR or IG J-region gene segments, wherein said plurality of V-segment primers and said plurality of J-segment primers are capable of amplifying in said single multiplex PCR at least 104 distinct amplicons representing a diversity of rearranged TCR or IG CDR3 nucleic acid sequences present in said first sample; sequencing said plurality of amplicons; and determining a total diversity of rearranged TCR or IG CDR3 nucleic acid sequences from said plurality of sequenced amplicons of said first sample.