Stability of cryopreserved white blood cells (WBCs) prepared for donor WBC infusions.

BACKGROUND: White blood cells (WBCs) collected from hematopoietic stem cell transplant donors are often given to the recipient to speed immune recovery or treat disease relapse. The postthaw recovery and viability of cryopreserved donor WBCs, stored for as long as 7 years, were assessed. STUDY DESIGN AND METHODS: Total nucleated cell (TNC) cell recovery, CD3+ cell recovery, and TNC viability were measured in 311 clinical donor WBC products: 168 products were unmanipulated or minimally manipulated and 143 products were extensively manipulated. An additional 45 products were selected because they were stored for a longer duration; these were tested using both standard methods and global transcriptional analysis. All products were cryopreserved in 5% dimethyl sulfoxide (DMSO) plus 6% pentastarch and stored in liquid nitrogen. RESULTS: The mean duration of storage of the 311 products was 143 days. Their TNC recovery was 92 ± 17%, CD3+ cell recovery was 76 ± 19%, and the TNC viability was 84 ± 6%. Duration of storage had no effect on TNC recovery, CD3+ cell recovery, or TNC viability of the 311 products. The mean duration of storage of the long-term stored products was 5.2 years; their TNC recovery (93 ± 14%) and the TNC viability (78 ± 13%) did not differ from the 311 products, but their CD3 cell recovery was greater (86 ± 22%; p = 0.0042). Gene expression profiles of the long-term-stored products revealed no differences due to storage duration. CONCLUSIONS: Donor WBC products cryopreserved in 5% DMSO and 6% pentastarch can be stored in liquid nitrogen for at least 7 years.

Gene and microRNA analysis of neutrophils from patients with polycythemia vera and essential thrombocytosis: down-regulation of micro RNA-1 and -133a.

BACKGROUND: Since the V617F mutation in JAK2 may not be the initiating event in myeloprofilerative disorders (MPDs) we compared molecular changes in neutrophils from patients with polycythemia vera (PV) and essential thrombocythosis (ET), to neutrophils stimulated by G-CSF administration and to normal unstimulated neutrophils METHODS: A gene expression oligonucleotide microarray with more than 35,000 probes and a microRNA (miR) expression array with 827 probes were used to assess neutrophils from 6 MPD patients; 4 with PV and 2 with ET, 5 healthy subjects and 6 healthy subjects given G-CSF. In addition, neutrophil antigen expression was analyzed by flow cytometry and 64 serum protein levels were analyzed by ELISA. RESULTS: Gene expression profiles of neutrophils from the MPD patients were similar but distinct from those of healthy subjects, either unstimulated or G-CSF-mobilized. The differentially expressed genes in MPD neutrophils were more likely to be in pathways involved with inflammation while those of G-CSF-mobilized neutrophils were more likely to belong to metabolic pathways. In MPD neutrophils the expression of CCR1 was increased and that of several NF-kappaB pathway genes were decreased. MicroRNA miR-133a and miR-1 in MPD neutrophils were down-regulated the most. Levels of 11 serum proteins were increased in MPD patients including MMP-10, MMP-13, VCAM, P-selectin, PDGF-BB and a CCR1 ligand, MIP-1alpha. CONCLUSION: These studies showed differential expression of genes particularly involved in inflammatory pathways including the NF-kappaB pathway and down-regulation of miR-133a and miR-1. These two microRNAs have been previous associated with certain cancers as well as the regulation of hyperthrophy of cardiac and skeletal muscle cells. These changes may contribute to the clinical manifestations of the MPDs.