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Biomedical subjects

D J Phillips

Publications and source records attributed to D J Phillips.

At least 109 records · Page 6Linked to original sources

Isotype-specific enzyme immunoassay for influenza antibody with monoclonal antibodies to human immunoglobulins.

Mouse monoclonal antibodies specific for human immunoglobulin isotypes were investigated for use in an isotype-specific enzyme immunoassay for detection of antibody to influenza type A hemagglutinin (H1 and H3). The monoclonal antibody reagents were compared with isotype-specific, hyperimmune rabbit antisera from the National Institutes of Health. Endpoint titers for immunoglobulin G (IgG) obtained with the two reagents were within fourfold of each other 84% of the time (79 of 84) and within eightfold of each other 95% of the time (89 of 94). Regression analysis of the data gave a multiple correlation coefficient (r2) of 0.77 and a Spearman rank value of 0.83 (P less than 0.001). For IgA reagents, endpoint titers agreed within fourfold 77% of the time (88 of 114) and within eightfold 92% of the time (105 of 114). The r2 was 0.73, and Spearman rank was 0.83 (P less than 0.001). IgM antibody was detected in only 17 of 114 sera by either monoclonal or polyclonal reagents. Of these sera, 14 (82%) gave titers with the two reagents that were within fourfold of each other. A similar number of fourfold titer rises were detected with each reagent in paired sera showing hemagglutination inhibition titer rises. Monoclonal antibody reagents detected 27 IgA, 29 IgG, and 6 IgM rises, while polyclonal antisera detected 26 IgA, 31 IgG, and 7 IgM rises. These results show that monoclonal antibodies specific for human immunoglobulin isotypes are suitable as reagents for diagnostic assays. The advantages of monoclonal antibodies are their high degree of specificity and the ability to be standardized and produced in unlimited quantities. Moreover, the availability of immunoglobulin subclass- and allotype-specific monoclonal antibodies will enable a more detailed analysis of the antibody response to influenza as well as other infectious agents.

Animals↗

Evaluation of monoclonal antibodies having specificity for human IgG sub-classes: results of an IUIS/WHO collaborative study.

Seventy-four monoclonal antibodies (McAb) of putative specificity for human IgG (11), the IgG sub-classes (59) or Gm allotypes (4) have been evaluated for reactivity and specificity in eight laboratories employing different assay techniques or protocols. For the IgG, IgG3, IgG4, G1m(f) and G3m(u) specificities McAb have been produced that can be satisfactorily applied in most methodologies employed and have potential as reference reagents. The IgG1 and particularly IgG2 specificities proved problematical with all McAb evaluated demonstrating apparent assay restriction and whilst performing well in some assays proved to be poor or inactive reagents in others. However, the study identifies McAb individually suited to application within most commonly employed methodologies. Epitope display is the probable variability rather than capricious behaviour by the McAb. IgG1 and IgG2 were the least immunogenic of the sub-class proteins and there is evidence that epitope display is influenced by the physical and chemical procedures used to immobilize or fix antigen - a common requirement in the assay systems studied.

Antibodies, Monoclonal↗

Carotid endarterectomy. Relationship of outcome to early restenosis.

The results following carotid endarterectomy were prospectively evaluated in 134 patients (145 sides) by repeat ultrasonic duplex scanning and clinical evaluation extending for a period of 4 years. There were 107 men and 27 women in the study group. The perioperative stroke rate was 1.3% and the mortality rate, 0.7%. There were 9 late deaths, of which two were stroke related (1.4%). Focal symptoms occurred in 12 patients on the ipsilateral side, six of which were strokes (one lacunar). The remaining symptoms developed in the presence of moderate degrees of carotid stenosis (less than 50%). There were seven patients who had transient ischemic attacks (TIAs) referable to the operated side, but only two of these were associated with a recurrent high-grade stenosis. During follow-up 32 (22%) patients had recurrent high-grade stenosis. Restenosis regressed in seven, giving a persistent rate of 17.1%. The incidence of restenosis was significantly higher in women (p less than 0.01). By life-table analysis, restenosis occurred early, the majority within 24 months. There was no consistent association between the development of symptoms and the occurrence of restenosis. Therefore, it is concluded that there is no justification for reoperation based on the degree of narrowing observed to prevent subsequent TIAs and strokes.

Aged↗

IgG subclass antibodies to herpes simplex virus.

Several mouse monoclonal antibodies specific for human IgG1, IgG2, IgG3, and IgG4 were evaluated by enzyme-linked immunosorbent assay to detect human IgG subclass antibodies to herpes simplex virus (HSV) antigens. The variable results with different monoclonal antibodies point to the need for well-characterized reagents in the study of antibody responses to infectious agents. The 204 sera tested were obtained from 157 patients with various forms of clinically manifest HSV infections and from several controls. IgG1 antibodies were demonstrated in almost all HSV-infected subjects and were the first antibodies to appear in primary genital infections. IgG2, IgG3, and IgG4 antibodies were detected in acute-phase sera, most often in patients with recurrent genital herpes but in none of those with primary infections. IgG4 antibodies occurred significantly more frequently in sera from men than in those from women with recurrent genital infections.

Adult↗

Cryptosporidiosis in hospital personnel. Evidence for person-to-person transmission.

An intern responsible for the care of a patient with chronic cryptosporidiosis developed acute diarrhea and serologic evidence of cryptosporidium infection. Sera from 26 hospital personnel exposed to the patient and 18 personnel with no exposure were examined with an indirect immunofluorescent antibody procedure for the presence of antibodies to Cryptosporidium. Eight (31%) exposed personnel--5 nurses, 2 house officers, and 1 student--had positive antibody titers (1:10 or more). The frequency of positivity in the nurse-housestaff-student group (8 of 18, 45%) was significantly greater (p less than 0.05) than that in the attending physicians and respiratory therapists (0 of 8). The former group had significantly more exposure to the patient's feces than did the latter group (p less than 0.01). Three of eighteen control personnel (17%) had positive cryptosporidium antibody titers. These findings suggest that Cryptosporidium may be transmitted from person to person in the hospital environment and that serologic evidence of infection is common among hospital personnel.

Adult↗

Immunoassay for IgG rheumatoid factor with a murine monoclonal anti-Fd antibody.

Conventional radioimmunoassays for IgG rheumatoid factors (RF) detect the binding of IgG RF (or their F(ab')2 fragments) to solid-phase human IgG Fc fragments or rabbit IgG. Binding is detected with a radiolabeled antibody that is IgG-specific but is nonreactive with human Fc (i.e., an alpha-Fd reagent) or with rabbit IgG. Anti-Fd reagents are quite laborious to produce. We developed a murine monoclonal alpha-Fd antibody, confirmed its specificity, and compared it with conventional rabbit alpha-Fd in the IgG RF radioimmunoassay. We also adapted the assay to an enzyme-linked assay with both human Fc and rabbit IgG as antigen. We compared the four assay methods with each other and examined their relationship to certain clinical and laboratory features of rheumatoid arthritis.

Animals↗

Phase I study of intravenous gamma globulin in multiple myeloma.

Seventeen patients with multiple myeloma were given intravenous immunoglobulin at doses ranging from 150 mg/kg to 500 mg/kg in a phase I study. The intravenous immunoglobulin was well tolerated with only three transient episodes of mild clinical toxicity during 27 infusions. In no instance was hepatic or renal toxicity seen. Marked biologic variability over the one month study period in total IgG levels in patients with non-IgG myeloma and IgG subclasses in many of the patients was observed, making intravenous immunoglobulin half-life determinations based on IgG or IgG subclass levels problematical. The decay of functional antibody to hepatitis B surface antigen was determined. Analysis of the hepatitis antibody data suggested that intravenous immunoglobulin half-life was in the range of seven to 20 days for the entire study group and was not related to the isotype of the myeloma paraprotein or to the baseline levels of IgG. No infections were observed in the study group during the study period, but the potential for infection prophylaxis by intravenous immunoglobulin in myeloma patients must be evaluated in a randomized, prospective, controlled phase III study.

Adult↗

Application of a solid-phase immunofluorometric assay to the selection of monoclonal antibody specific for the adenovirus group-reactive hexon antigen.

An immunofluorometric assay (IFMA) has been evaluated as a screening assay to detect monoclonal antibodies to the group-specific antigen of the adenovirus hexon component. The antibodies were produced as mouse ascitic fluids from hybridoma cells generated from Balb/C mice immunized with purified adenovirus type 2 hexon component and crude adenovirus type 3 culture supernatants. The purified IgG fractions from all monoclonal ascitic fluids tested were identified as the IgG1 K mouse isotype. Antibody titers ranged from 102,400 to 204,800 by the IFMA, from 200 to 12,800 by indirect FA, and were generally nonreactive in counterelectrophoresis, complement fixation, hemagglutination-inhibition, serum neutralization, and immune electron microscopy titrations. The IFMA is a reliable method for quantitating low levels of specific antibody in large numbers of test samples, and is therefore ideal as a screening assay for monoclonal antibody in tissue culture fluids and in mouse ascitic fluids.

Adenoviruses, Human↗

Evaluation of thirty-one mouse monoclonal antibodies to human IgG epitopes.

Stable clones of 31 mouse hybridomas that produce monoclonal antibodies (MAbs) against human IgG antigenic determinants were obtained. The number of hybridomas of different specificity described are: 2 anti-IgG1 Fc, 1 anti-IgG2 Fc, 1 anti-IgG2 Fd, 2 anti-IgG3 Fc, 2 anti-IgG3 hinge, 3 anti-IgG4 Fc, 3 anti-IgG4 Fd, 2 anti-nG4m(b), 4 anti-IgGFc, 2 anti-IgGFd, 1 anti-kappa, 1 anti-lambda, 1 anti-non IgG1, 2 anti-non IgG2, 2 anti-non-IgG3, 2 anti-non-IgG4. Evidence is presented validating their specificity. Some MAbs demonstrated to be avid, potent, and specific for well defined IgG-subclass epitopes may be partially or completely inactive in other assay systems, presumably because of different presentations of antigen epitopes. In general, this problem requires careful writing of protocols describing the use of MAbs.

Animals↗

Natural history of carotid artery disease on the side contralateral to endarterectomy.

The natural history of the nonoperated carotid artery opposite an endarterectomy was examined in 134 patients by means of ultrasonic duplex scanning over a period extending to 48 months. None of the nine deaths that occurred during follow-up was stroke related. A total of 22 arteries showed progression of disease over this period. By life-table analysis the mean annual rate of progression for all categories of disease was 12.6% and 7.4% for progression to a diameter reduction greater than 50%. Disease progression was more rapid in patients under 65 years of age. Symptoms occurred in 13 patients for an overall incidence of 10% and a mean annual rate estimated at 5%. All symptoms indicated transient ischemic attacks; there were no strokes. There was a strong relationship between the development of symptoms and stenoses greater than 80% either at the initial examination or secondary to progression. No correlation was found between the presence of bruits or their change over time and the progression or appearance of symptoms. Conservative management of nonoperated vessels opposite an endarterectomy appears appropriate until symptoms develop or a lesion greater than 80% is detected.

Adult↗

Diversification strategies for hospital pharmacies.

Several ways used by the pharmacy department of a large university hospital to generate revenue through diversification are described. The department offers its facilities and staff as a resource in training medical service representatives for several pharmaceutical manufacturers, which is projected to provide $85,000 in net income for fiscal year (FY) 1983-84. The pharmacy department also conducts a six-month program for training pharmacy technicians, which yields a small net profit. The pharmacy department actively participates in educational programs such as college courses and clerkships earning extra income. An apothecary-style outpatient pharmacy was set up under a for-profit corporation. Services have been expanded to include the preparation of i.v. solutions that support home care. A durable medical equipment (DME) business is planned. The ambulatory and home-care programs are expected to generate approximately $165,000 in net profit next year. Contract pharmaceutical services are provided to another hospital. The net income generated through diversification in this pharmacy department will exceed $250,000 in FY 1983-84.

Allied Health Personnel↗

Six-month hospital-pharmacy-based technician training program.

A six-month training program for hospital pharmacy technicians is described. The development of this program was stimulated by the need to use departmental resources more efficiently and the desire of local hospital pharmacy directors to have trained technicians. Financial support was sought and received from the local Private Industry Council. The department selects 10 students and two alternates for each class on the basis of test performance and an interview. The program consists of 1014 scheduled hours of instruction. The first week is spent orienting the students to the program and general hospital pharmacy concepts. The next six weeks are spent in a laboratory where the students acquire their basic skills. The rest of the program, approximately 4.5 months, is spent in on-the-job training, additional didactic lectures, and field trips. The students are evaluated routinely throughout the program. At the end of the program, each participant is granted a certificate. Attrition has been low; three classes have completed the program, and 28 technicians have graduated. Twenty-five of these technicians are now employed full-time. Positive feedback has been received from the technicians' employers. This program meets the need of area hospital pharmacies to recruit trained technicians.

Curriculum↗

The impact of Duplex scanning on the evaluation of patients with asymptomatic bruits in the region of the carotid arteries.

Between January 1980 and January 1981, 95 patients with 'asymptomatic' bruits in the region of the carotid arteries were evaluated at the University of Washington with an ultrasonic Duplex scanner combined with spectral analysis. Based upon the interpretation of a hardcopy of the spectral analysis, the artery of interest was classified into one of five categories: A normal; B 5-15% diameter reduction; C 16-49% diameter reduction; D 50-99% diameter reduction, and E total occlusion. Forty-eight patients had unilateral and 47 patients had bilateral bruits; 142 bruits were evaluated. The distribution of disease in the internal carotid artery was: A: 2% (3/142); B: 30% (42/142); C: 37% (53/142); D: 29% (41/142), and E: 2% (3/142). If the presence of a high grade stenosis of the internal carotid artery (50-99% diameter reduction) is an indication for invasive intervention, then 71% of these patients would not require contrast arteriography. The role of a direct ultrasonic study, such as Duplex scanning, in patients with asymptomatic bruits is discussed.

Auscultation↗

Hospital-based training for pharmaceutical manufacturers' representatives.

A hospital-based training program for pharmaceutical manufacturers' representatives is described. The pharmacy department of a large teaching institution established a training program for new sales representatives of a major pharmaceutical company. Goals were outlined by the sales training manager and the pharmacy department. The sales training personnel, department of pharmacy, and the cooperating departments of medicine, surgery, and nursing worked together to formulate objectives for the sessions, and teaching responsibilities were delegated to members of all these departments. The program length varied from one to five days. A formal contract was developed specifying content, program dates, and reimbursement. The institution is reimbursed for the use of the facility, materials, and administrative overhead. The program's success has led to the development of similar programs with several other companies. The extra income has enabled the pharmacy to create a new division within the department. Evaluations from more than 500 sales representatives who have participated in the programs have been consistently positive. The pharmacy department in a teaching institution has the resources to provide a training program for sales representatives that can be an additional source of income.

Commerce↗