Publication Date:
2012-11-16
Description:
Abstract 4125 Introduction and Aims. CIK cells are CD3+/CD56+ T lymphocytes known for their antitumour effect against several haematological malignancies and solid tumours. CIK cells are obtained ex-vivo by stimulating peripheral blood mononucleated cells (MNC) with IFN-gamma (day 0), IL-2, anti-CD3 monoclonal-antibody (day 1) and IL2 every 3 days from day 1 to the 21st when maximum expansion of CD3+/CD56+ is expected as firstly described by Negrin. The percentage of CIK cells at the end of expansion represents a criteria for batch release: if CIK cells are less than 40% of the bulk population at the end of the culture, the batch should be considered suboptimal for transplantation. We have analyzed cell expansion dynamics of 30 samples evaluating the composition of cells constituting the bulk. In 11 samples (37%) CIK percentage reached plateau on day 17 instead of day 21, and then started to decrease rapidly. We believe that it is fundamental for the operator to predict in advance the harvest day in which CIK cells reach the maximal concentration in the bulk. Thus, the aim of this study was to introduce a new approach to control and optimize the expansion process based on multivariate statistical data analysis in order to improve its quality. Methods. Multivariate Batch Statistical Process Control (BSPC) and regression models based on Bidirectional-Orthogonal Projections to Latent Structures (O2PLS) were applied for monitoring the expansion process. Phenotypical analysis of cell populations was performed by flow cytometry by measuring the following different cellular subsets (11 variables): total T lymphocytes (CD3+), T-Helper lymphocytes (CD3+4+), T-cytotoxic lymphocytes (CD3+8+), CIK cells (CD3+56+), NK cells (CD3–56+), T lymphocytes (CD3+56-), monocytes (CD14+), B lymphocytes (CD19+), granulocytes (CD33+) and the undifferentiated subset CD3–56-. BSPC allowed us to produce suitable control charts while to estimate the level of CIK cells on days 17 and 21 we built different O2PLS regression models using as predictors the descriptions of the cellular population of the previous days. The chained use of the obtained regression models enabled us to predict in advance unsatisfactory expansions. Results. The expansions having a percentage of CIK cells ≥40% were used to build different types of control charts. In particular, the charts based on DModX and on T2 resulted predictive in the detection of unsatisfactory expansions. Indeed the expansions having CIK
Print ISSN:
0006-4971
Electronic ISSN:
1528-0020
Topics:
Biology
,
Medicine
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