Interleukin-21 enhances rituximab activity in a cynomolgus monkey model of B cell depletion and in mouse B cell lymphoma models.

Rituximab, a monoclonal antibody targeting CD20 on B cells, is currently used to treat many subtypes of B cell lymphomas. However, treatment is not curative and response rates are variable. Recombinant interleukin-21 (rIL-21) is a cytokine that enhances immune effector function and affects both primary and transformed B cell differentiation. We hypothesized that the combination of rIL-21 plus rituximab would be a more efficacious treatment for B cell malignancies than rituximab alone. We cultured human and cynomolgus monkey NK cells with rIL-21 and found that their activity was increased and proteins associated with antibody dependent cytotoxicity were up-regulated. Studies in cynomolgus monkeys modeled the effects of rIL-21 on rituximab activity against CD20 B cells. In these studies, rIL-21 activated innate immune effectors, increased ADCC and mobilized B cells into peripheral blood. When rIL-21 was combined with rituximab, deeper and more durable B cell depletion was observed. In another series of experiments, IL-21 was shown to have direct antiproliferative activity against a subset of human lymphoma cell lines, and combination of murine IL-21 with rituximab yielded significant survival benefits over either agent alone in xenogeneic mouse tumor models of disseminated lymphoma. Therefore, our results do suggest that the therapeutic efficacy of rituximab may be improved when used in combination with rIL-21.

Function allocation in complex systems: reframing an old problem.

In this article, we offer a new, macroergonomics perspective on the long-debated issue of function allocation. We believe thinking in this domain needs to be realigned, moving away from the traditional microergonomics conceptualisation, concerned predominantly with task-based decisions, and towards a macroergonomics approach, viewing function allocation choices as central to effective systems design. We frame our arguments within a systems perspective, advocating that function allocation issues need to be on the agenda of all individuals with a wider interest in the human and organisational aspects of complex work systems, including people who commission, sponsor, design, implement and use such systems. We also argue that allocation decisions should form a transparent, explicit stage early in the systems design and development process, involve multiple stakeholders (including end-users), be evidence-based, framed within the language of risk and utilise iterative methods (e.g. scenarios planning techniques). PRACTITIONER SUMMARY: This article presents a macroergonomics approach to function allocation, advocating its importance in effective systems design. Adopting a systems mindset, we argue function allocation should form an explicit stage early in the design process, involve multiple stakeholders, be evidence-based, framed within the language of risk and utilise iterative methods.

Interlaboratory comparison of extraction efficiency of pesticides from surface and laboratory water using solid-phase extraction disks.

A continuation of an earlier interlaboratory comparison was conducted (1) to assess solid-phase extraction (SPE) using Empore disks to extract atrazine, bromacil, metolachlor, and chlorpyrifos from various water sources accompanied by different sample shipping and quantitative techniques and (2) to compare quantitative results of individual laboratories with results of one common laboratory. Three replicates of a composite surface water (SW) sample were fortified with the analytes along with three replicates of deionized water (DW). A nonfortified DW sample and a nonfortified SW sample were also extracted. All samples were extracted using Empore C(18) disks. After extraction, part of the samples were eluted and analyzed in-house. Duplicate samples were evaporated in a 2-mL vial, shipped dry to a central laboratory (SDC), redissolved, and analyzed. Overall, samples analyzed in-house had higher recoveries than SDC samples. Laboratory x analysis type and laboratory x water source interactions were significant for all four compounds. Seven laboratories participated in this interlaboratory comparison program. No differences in atrazine recoveries were observed from in-house samples analyzed by laboratories A, B, D, and G compared with the recovery of SDC samples. In-house atrazine recoveries from laboratories C and F were higher when compared with recovery from SDC samples. However, laboratory E had lower recoveries from in-house samples compared with SDC samples. For each laboratory, lower recoveries were observed for chlorpyrifos from the SDC samples compared with samples analyzed in-house. Bromacil recovery was <65% at two of the seven laboratories in the study. Bromacil recoveries for the remaining laboratories were >75%. Three laboratories showed no differences in metolachlor recovery; two laboratories had higher recoveries for samples analyzed in-house, and two other laboratories showed higher metolachlor recovery for SDC samples. Laboratory G had a higher recovery in SW for all four compounds compared with DW. Other laboratories that had significant differences in pesticide recovery between the two water sources showed higher recovery in DW than in the SW regardless of the compound. In comparison to earlier work, recovery of these compounds using SPE disks as a temporary storage matrix may be more effective than shipping dried samples in a vial. Problems with analytes such as chlorpyrifos are unavoidable, and it should not be assumed that an extraction procedure using SPE disks will be adequate for all compounds and transferrable across all chromatographic conditions.

Serological reactivity of baculovirus-expressed Ebola virus VP35 and nucleoproteins.

Ebola virus (EBOV) is a member of the family Filoviridae and is classified as a biosafety level 4 virus. This classification makes the preparation of antigen and performance of diagnostic assays time-consuming and complicated. The objective of this study was to evaluate the value of EBOV immunoassays based on recombinant nucleoprotein (r-NP) and recombinant VP35 (r-VP35) using large serum panels of African origin and from primates. Furthermore, we investigated whether the results obtained with EBOV r-VP35 enzyme-linked immunosorbent assay (ELISA) could improve on the findings obtained with the EBOV r-NP ELISA. The full-length EBOV NP and VP35 of the EBOV subtype Zaire were expressed as histidine-tagged recombinant proteins in the baculovirus expression system. The antigenic reactivity and specificity of these recombinant proteins were determined by Western blotting and ELISA using EBOV specific monoclonal antibodies. The results obtained with the r-NP and r-VP35 ELISAs were compared with the results obtained in an indirect immunofluorescence assay based on native EBOV subtype Zaire. EBOV specific monoclonal antibodies reacted specifically with the respective proteins in both Western blot and ELISA. Five hundred and twenty six samples from humans and primates were tested with r-NP and r-VP35 ELISAs. Monkey serum samples positive for EBOV subtype Reston and Zaire were both positive in the EBOV r-NP ELISA, whereas only the EBOV Zaire infected monkeys were positive in the r-VP35 ELISA. The sensitivity and specificity values of the EBOV recombinants' ELISAs compared to those of the immunofluorescence assay were 92% and 99% for r-NP and 44% and 100% for r-VP35. r-NP ELISA proved to be a sensitive and specific assay for EBOV diagnosis and for epidemiological studies for both EBOV subtypes Reston and Zaire. The use of r-VP35 in an ELISA format has no additional value for EBOV serodiagnosis.

Interlaboratory comparison of pesticide recovery from water using solid-phase extraction disks and gas chromatography.

An interlaboratory study was conducted to assess the suitability of C18 solid-phase extraction disks to retain and ship different pesticides from water samples. Surface and deionized water samples were fortified with various pesticides and extracted using C18 disks. Pesticides were eluted from disks and analyzed in-house, or disks were sent to another laboratory where they were eluted and analyzed. Along with the disks, a standard pesticide solution in methanol was also shipped to be used for fortification, extraction, and analysis. The highest recovery from deionized or surface water using shipped disks was obtained for cyanazine (>97%), followed by metalaxyl (>96%), and atrazine (>92%). Although <40% of the bifenthrin, chlorpyrifos, and chlorothalonil fortified in surface water was recovered from shipped disks, recoveries from deionized water were >70%. From in-house eluted disks, bifenthrin and chlorpyrifos were recovered at 118 and 105%, whereas chlorothalonil showed 71% recovery, indicating that poor recovery from surface water was due to loss during shipping rather than low retention by the C18 disks. There was no consistent relationship between recovery from C18 disk and physicochemical properties for the pesticides included in this study. For most of the 13 pesticides tested, there were no differences in recovery between in-house extracted disks and shipped disks, indicating the suitability of disks to concentrate and transport pesticides extracted from water samples.