Differentiation of alloreactive versus CD3/CD28 stimulated T-lymphocytes using Raman spectroscopy: a greater specificity for noninvasive acute renal allograft rejection detection

Cytometry A. 2009 Nov;75(11):917-23. doi: 10.1002/cyto.a.20797.

Abstract

Acute rejection (AR) remains problematic in renal transplantation. As a marker, serum creatinine is limited, warranting a more effective screening tool. Raman spectroscopy (RS) can detect T-cell activation with high sensitivity. In this study we explore its specificity. Seventy-five inactivated, 40 alloantigen-activated, and 75 CD3/CD28-activated T cells were analyzed using RS. CD3/CD28-activated peak magnitudes (PM) were 4.3% to 23.9% lower than inactivated PM at positions: 903, 1031, 1069, 1093, 1155, 1326, and 1449 cm(-1), with a difference in peak ratio (PR) observed at the 1182:1195 cm(-1) position (0.91 +/- 0.06 vs. 1.2 +/- 0.01, respectively: P = 0.006). Differences in CD3/CD28- and alloantigen-activated PM were observed at: 903, 1031, 1093, 1155, 1326, and 1449 cm(-1), with no PR differences at the 1182:1195 cm(-1) position (0.91 +/- 0.06 vs. 0.86 +/- 0.09: P = 0.8). Spectral signature separation of CD3/CD28-and alloantigen-activated groups was 100% specific and sensitive. We conclude that RS can differentiate T cells activated by different stimuli with high sensitivity and specificity.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • CD28 Antigens / biosynthesis*
  • CD3 Complex / biosynthesis*
  • Cell Differentiation
  • Cell Membrane / metabolism
  • Cell Separation
  • Graft Rejection / diagnosis*
  • Graft Rejection / pathology
  • Humans
  • Kidney Transplantation / methods*
  • Leukocytes, Mononuclear / cytology
  • Lymphocyte Activation / immunology
  • Spectrum Analysis, Raman / methods*
  • T-Lymphocytes / immunology*
  • T-Lymphocytes / metabolism

Substances

  • CD28 Antigens
  • CD3 Complex