Multimeric humanized varicella-zoster virus antibody fragments to gH neutralize virus while monomeric fragments do not.

Murine monoclonal antibody 206 (MAb mu206) binds to gH, the varicella-zoster virus (VZV) fusogen, neutralizing the virus in vitro in the absence of complement and inhibiting cell-to-cell spread and egress of VZV in cultured cells. We have humanized this antibody to generate MAb hu206 by complementarity determining region grafting. MAb hu206 retained binding and in vitro neutralizing activity, as well as cross-reactivity with ten different VZV strains. Single-chain antibody fragments (scAb) derived from MAb hu206 were produced in Escherichia coli. These scAb retained the binding properties of the whole antibody. However, monomeric scAb exhibited markedly reduced neutralizing activity compared to the bivalent parental MAb hu206. Shortening the peptide linker joining the V(H) to the V(kappa) domain from 14 to 5 or even 0 residues encouraged multimerization and increased neutralizing efficacy. The fact that Fab fragments enzymatically generated from whole MAb hu206 lost their neutralizing potency lent support to the proposal that valency is important for VZV neutralization at this epitope.

V(D)J germline gene repertoire analysis of monoclonal D antibodies and the implications for D epitope specificity.

BACKGROUND: The D antigen is a highly immunogenic human RBC antigen. Alloimmunization against the D antigen produces high-affinity antibodies that cause hemolytic transfusion reactions and HDN. STUDY DESIGN AND METHODS: Cloning and subsequent sequence analysis of 11 new samples of monoclonal anti-D was performed in an attempt to identify V(D)J germline gene usage. Sequences were compared and analyzed with 37 previously published samples of anti-D for identification of V(H) and V(L) pairings, canonical structures, and conformation of restricted germline gene usage. RESULTS: The V(H) and V(L) pairings used by the new D MoAbs resulted in seven canonical combinations, three of which had not been described previously. Preferential usage of gene segments from the VH3 and VH4 families and of D3, D6, JH6, and DPK9 germline gene segments was also determined. Three samples of anti-D from different donors were found to use similar V(H) and V(kappa) germline genes, despite the fact that two of the antibodies recognized epD6/7 and the third recognized epD1. From the cumulative analysis of the anti-D IgG, 24 V(H) and V(L) gene pairings were identified, resulting in only 10 canonical structures. CONCLUSIONS: Despite the potential for diversity, only a minority of V(H) and V(L) germline genes are used by anti-D. Consequently, V(H) and V(L) pairings and the resulting canonical structures are similarly restricted.

Comparison between hybridoma and Fab/phage anti-RhD: their V gene usage and pairings.

Our 11 anti-RhD's in conjunction with 37 previously published RhD antibodies, produced by hybridoma technology were analysed for germline gene usage and restriction in VH and VL pairings. The 17 VH germline genes used by the hybridoma anti-RhD IgG were derived from 4 VH families (VH1, VH2, VH3 and VH4). Eighteen kappa chains were restricted to only 5 germline genes from only 2 V kappa families (V kappa 1 and V kappa 3). However, the 13 lambda chains were not as restricted, using 10 V lambda germline genes from 4 families (V lambda 1, V lambda 2, V lambda 3 and V lambda 8). Fifty six unique Fab/phage anti-RhD were also analysed. In all cases the Fab/phage VH germline genes were derived from the VH3 family (41/41). The 29 kappa chains were restricted to 4 germline genes primarily from V kappa 1 (97%) and the 24 lambda chains used 10 V lambda germline genes from 5 families (V lambda 1, V lambda 2, V lambda 3, V lambda 4 and V lambda 7). The VH germline genes of the Fab/phage were restricted to 4 of the 17 used by the hybridoma anti-RhD IgG (DP46, DP49, DP50 and DP77). Ninety percent of the Fab/phage were restricted to 1 of the 5 V kappa germline genes used by the IgG (DPK9). However, the repertoire of the V lambda germline genes used in these two systems is different, with analysis showing greater diversity in V lambda gene usage with 8 unique germline genes used by 76% Fab/phage compared to 4 unique genes used by 46% of the IgG hybridoma anti-RhD.

Management forecast: optimizing the use of organizational and individual knowledge.

Knowledge management provides a means for sharing data, lessons learned, and accumulated knowledge throughout an organization or within an entire industry. Gone are the days of hoarding knowledge to ensure job security; today's workers and managers must work together to find new and innovative ways to use what they know and optimize how that knowledge is accessed. Knowledge management's new approach to shared intellectual resources has implications for workers and managers in all fields and promises to redraw the management landscape. But are healthcare organizations setting the stage for reengineering themselves all over again?

Policy issues associated with analyzing outcomes of care.

Policy questions associated with the conduct of outcomes studies are addressed and outcomes management and outcomes research are differentiated. Multiple outcomes initiatives are being undertaken in the United States, from the institutional to national and international levels. Nurses have had limited involvement, thus far-although the array of outcomes being examined is considerable. Members of an American Academy of Nursing expert panel on quality health care discuss the environmental context and multiple focuses of outcomes analysis. They provide an overview of current activities, present the challenges confronting nursing in its pursuit of a quality assessment agenda, and make recommendations for increasing nurses' participation in quality initiatives.

Financial perspectives for the perioperative nurse.

This article describes the preparation of the operating room budget and identifies six components that contribute to cost per case. The successful budget can be achieved by following a six-step budgetary process. The budget process is shown by sample tables.

Principles, values, and ethics set the stage for managed care nursing.

Principles, values, and ethics constitute an ethical system that provides nurses a context in which to make ethical decisions. As the managed care environment generates new ethical issues and intensifies already existing ones, ethical education and decision-making skills become even more critical to nursing professionals, as well as physicians and hospital administrators.

A perioperative acuity measurement system.

An integrative acuity measurement system has been developed to increase efficiency and effectiveness of care delivery. Patients are classified into groups on the basis of nursing care required, thus facilitating both determination of actual nursing cost and accurate projection of staffing needs. It provides a universal language among perioperative departments for measuring similar outcomes.

Service integration in the reform era.

Fifty years of health care reform have centered around "fixing up" existing operational processes by examining the tasks attached to those processes. Today, restructuring seeks to identify the core processes of hospitals, such as patient care, and reorganize around those processes. Reorganization is painful, but also exhilarating. If hospitals are to use this exhilaration as a source of growth in a new economic climate, management must redefine itself and its relationship to staff.