Raman spectroscopic differentiation of activated versus non-activated T lymphocytes: An in vitro study of an acute allograft rejection model
| Autor: | Olena Palyvoda, Sandra L. Nehlsen-Cannarella, Scott A. Gruber, Omar R. Fagoaga, Jagdish S. Thakur, Gregory W. Auner, Kristian L. Brown |
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| Rok vydání: | 2009 |
| Předmět: |
Graft Rejection
T-Lymphocytes Receptor expression Immunology Lymphocyte Activation Spectrum Analysis Raman Article symbols.namesake chemistry.chemical_compound Humans Immunology and Allergy Medicine Creatinine business.industry T-cell receptor Mitomycin C Discriminant Analysis T lymphocyte Kidney Transplantation Molecular biology In vitro Transplantation chemistry symbols Raman spectroscopy business |
| Zdroj: | Journal of Immunological Methods. 340:48-54 |
| ISSN: | 0022-1759 |
| DOI: | 10.1016/j.jim.2008.10.001 |
| Popis: | Acute rejection (AR) remains a significant complication in renal transplant patients. Using serum creatinine for AR screening has proven problematic, and thus a noninvasive, highly sensitive and specific test is needed. T cells from human peripheral blood were analyzed using Raman spectroscopy. Fifty-one Mixed Lymphocyte Culture (MLC) activated T cells (ATC), 28 Mitomycin C inactivated T cells (ITC), and 35 resting T cells (RTC), were studied utilizing 785 and 514.5 nm wavelengths. Statistical analysis following subtraction of fluorescence used Student's t test to quantify peak ratio differences and discriminant function analysis (DFA), with three distinct sectors assigned for grouping purposes: Sector I, ITC; Sector II, ATC; Sector III, RTC. Differences between ATC and non-activated T cells (ITC and RTC) were found at 1182 and 1195 cm-1 peak positions for both wavelengths. Significant differences in peak ratios for 785 and 514.5 nm wavelengths existed between ATC and RTC ( p = 0.001 and p = 0.006, respectively) and ATC and ITC ( p = 0.001 and p = 0.001, respectively), with a trend in differences observed between ITC and RTC ( p = 0.07 and p = 0.08, respectively). Analysis of the DFA-derived sector distribution for the 785 and 514.5 nm wavelengths revealed a sensitivity of 95.7% and 89.3%, respectively, and a specificity of 100% and 93.8%, respectively. This data suggests that Raman spectroscopy can detect significant differences between activated and nonactivated T cells based upon cell-surface receptor expression, thereby establishing unique signatures that can aid in the development of a noninvasive AR screening tool with high sensitivity and specificity. |
| Databáze: | OpenAIRE |
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