Screening of single-donor apheresis platelets for bacterial contamination: the PASSPORT study results.

BACKGROUND: The PASSPORT study was an FDA-mandated surveillance of outdated 7-day apheresis platelets (APs) to assess the bacterial culture release test (RT) performance and the chance of transfusing APs containing viable bacteria compared to untested 5-day APs. STUDY DESIGN AND METHODS: Aerobic and anaerobic culture bottles were inoculated with 4 to 5 mL from APs 24 to 36 hours postcollection. APs were released after 24 hours if no growth was observed. Released APs were recalled for RT positives, and clinical services were notified. Day 8 APs were recultured (surveillance test [ST]). Initially positive RTs and STs were confirmed by AP reculture. RESULTS: A total of 388,903 RTs were accrued September 2005 through January 2008 from 52 regional blood centers: RT-positive APs interdicted before transfusion, 76 true positive (TP; 195/million; 95% confidence interval [CI], 154-244/million) and 57 indeterminate (IN); and RT-positive APs transfused, 14 TP and 242 IN. There were 14 reported septic transfusion reactions (STRs) from 13 AP collections (23 units) transfused on Days 3 through 7; three STRs were from Day 6 or 7 APs. There were two false-negative RTs causing STRs in three patients. No deaths were reported. STs had four TPs of 6039 tested (662/million; 95% CI, 180-1695/million). CONCLUSIONS: RT culturing prevents issuance of some bacterially contaminated APs. ST culture data and clinical reports suggest that this screening fails to detect all contaminated units. No fatalities were reported related to AP transfusion. Additional actions or testing may be required to further reduce the residual STR risk of RT APs, even with a 5-day storage limitation.

Purification of monocytes from cryopreserved mobilized apheresis products by elutriation with the Elutra device.

The Elutra biomedical device allows semi-automatic enrichment of monocytes by elutriation, using a single-use, closed and cGMP compliant tubing set, in a cost effective way. The procedure has been validated using fresh apheresis products from nonmobilized donors. We here evaluated the possibility of using Elutra to enrich monocytes from frozen/thawed apheresis products collected from mobilized healthy donors. Frozen apheresis products from 6 G CSF mobilized donors were thawed and used in 16 elutriation procedures. We compared the recovery and purity of enriched monocytes using different buffer compositions and elutriation profiles. Elutriated monocytes were cultured to generate mature dendritic cells (DCs). Depending in part of the initial granulocyte contamination in the apheresis product, the use of Desoxyribo Nuclease (DNAse) to avoid aggregation, was needed through only the initial steps or throughout the elutriation process. The average monocyte recovery was 85+/-31%. The average purity was 73+/-9%. The recovery of mature DC at d8 of culture was 20+/-6% of the input monocyte numbers. We conclude that Elutra allows the purification of monocytes from thawed mobilized apheresis. It requires no pre-processing of the cell product before elutriation, and allows the generation of phenotypically mature DC in quantities that are compatible with a clinical use.

Microsurgical penile replantation facilitated by postoperative HBO treatment.

Successful microsurgical replantation of a penis amputated at the level of the pubis is a rare occurrence worldwide. Moreover, the use of hyperbaric oxygen (HBO) for a postoperative replant Pseudomonas wound infection has not been reported. There is also disagreement regarding the importance of microsurgical repair of only the dorsal arteries or only the profundi arteries of the penis. A case is reported of penile replantation with a postoperative Pseudomonas wound infection treated with HBO to prevent potential replant loss, with a worldwide literature review. At 1-year postoperative follow-up, the patient has normal urinary flow and reports spontaneous erection, with the ability for intromission and a sensate glans. HBO facilitated the success of a penile replantation complicated by postoperative Pseudomonas wound infection. In addition, a literature review supports the microsurgical repair of at least a single isolated dorsal penile artery, but not a single or multiple profundi arteries.

Monitoring and isolation of blood dendritic cells from apheresis products in healthy individuals: a platform for cancer immunotherapy.

The fundamental role of dendritic cells (DC) in initiating and directing the primary immune response is well established. Furthermore, it is now accepted that DC may be useful in new vaccination strategies for preventing certain malignant and infectious diseases. As blood DC (BDC) physiology differs from that of the DC homologues generated in vitro from monocyte precursors, it is becoming more relevant to consider BDC for therapeutic interventions. Until recently, protocols for the isolation of BDC were laborious and inefficient; therefore, their use for investigative cancer immunotherapy is not widespread. In this study, we carefully documented BDC counts, yields and subsets during apheresis (Cobe Spectra), the initial and essential procedure in creating a BDC isolation platform for cancer immunotherapy. We established that an automated software package (Version 6.0 AutoPBPC) provides an operator-independent reliable source of mononuclear cells (MNC) for BDC preparation. Further, we observed that BDC might be recovered in high yields, often greater than 100% relative to the number of circulating BDC predicted by blood volume. An average of 66 million (range, 17-179) BDC per 10-l procedure were obtained, largely satisfying the needs for immunization. Higher yields were possible on total processed blood volumes of 15 l. BDC were not activated by the isolation procedure and, more importantly, both BDC subsets (CD11c(+)CD123(low) and CD11c(-)CD123(high)) were equally represented. Finally, we established that the apheresis product could be used for antibody-based BDC immunoselection and demonstrated that fully functional BDC can be obtained by this procedure.