Pentamers and streptamers for the monitoring of CMV-specific CD8+ T-cell immune-reconstitution after allogeneic stem cell transplantation: a comparative analysis
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1
Fundación Miguel Servet-Navarrabiomed, Spain
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2
Hospital de Navarra, Spain
Reconstitution of CMV-specific CD8+ T-cells (CTLs) is crucial for patients after allogeneic Stem Cell Transplantation (allo-SCT). The aim of this study is to evaluate the correlation between staining with two different multimers (Pentamers (PM) and Streptamers (ST)) in the monitoring of immune reconstitution.
A total of 94 PBMCs samples were isolated from 14 CMV-seropositive HLA-A*02:01 patients. To form the ST complex, 0.75 ug PE-labelled Strep-Tactin and 0.5 ug Streptamer HLA I were mixed. In addition, 0.2ug HLA-A*02:01/STpp65495-503 or 5 ul HLA-A*02:01/PMpp65495-503 were incubated with 1x106 human AB serum-blocked PBMCs for 45 minutes at 4ºC or 10 minutes at room temperature, respectively. After multimeric staining, anti-human CD3/CD8/CD45/CD4 were added and 500,000 cells were acquired in a FACSCanto II. CMV-specific CTLs response was monitored monthly during a median of 11.4 months after SCT with a median of 8 samples analysed per patient. Correlation between both techniques was determined by Rho Spearman test.
The median frequencies of CMV-specific CTLs detected by PM and ST were 31.58x105/L (IQR=4.04-267.24) and 42.85x105/L (IQR=3.52-288.31), respectively. Both techniques correlate each other (rSpearman = 0.982; p<0.01). Nevertheless, ST showed higher mean values than PM looking at the median value of the difference PM-ST (-1.42 [-23.5, 1.35]. The correlation between the difference PM-ST and the PM/ST mean showed that the higher CMV-specific CTLs quantity the larger differences PM-ST (rSpearman=-0.331; p<0.01).
We concluded that both techniques may be equally useful to examine immune reconstitution, although detection levels are higher with ST staining possibly due to a larger sensitivity of this technique.
Keywords:
CMV,
Stem Cell Transplantation,
pentamer,
streptamer,
specific T cells
Conference:
15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013.
Presentation Type:
Abstract
Topic:
Translational immunology and immune intervention
Citation:
CIAURRIZ
M,
Ramirez
N,
Beloki
L,
Mansilla
C,
Zabalza
A,
Bandres
E,
Rodríguez
M and
Olavarria
E
(2013). Pentamers and streptamers for the monitoring of CMV-specific CD8+ T-cell immune-reconstitution after allogeneic stem cell transplantation: a comparative analysis.
Front. Immunol.
Conference Abstract:
15th International Congress of Immunology (ICI).
doi: 10.3389/conf.fimmu.2013.02.00924
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Received:
27 Jun 2013;
Published Online:
22 Aug 2013.
*
Correspondence:
Mr. Eduardo Olavarria, Fundación Miguel Servet-Navarrabiomed, Pamplona, Spain, eduardo.olavarria.lopez@cfnavarra.es