The Subclass Nature and Clinical Significance of the IgG Antibody Response in Patients Undergoing Allergen-Specific Immunotherapy

The purpose of this paper is to discuss the methodological difficulties in quantitation of human IgG subclass antibodies to allergens, to describe the subclass nature of the IgG antibody response in patients undergoing allergen-specific immunotherapy, and to discuss the possible immunological functions and clinical significance of allergen-specific IgG antibodies of different subclasses. Based on results obtained by use of assays with documented specificity it is concluded that the IgG antibody response during allergen-specific immunotherapy is IgG1 and IgG4 restricted, although low levels of IgG2 and IgG3 antibodies to some allergens may occur. In most patients the early IgG antibody response is IgG1 dominated and the late IgG4 dominated. A too early or too pronounced IgG4 dominated antibody response seems to indicate a poor clinical outcome of immunotherapy with inhalant allergens, whereas a pronounced early IgG1 antibody production has been found to be associated with a decrease in synthesis of IgE antibodies to an insect venom. It is therefore proposed that an early IgG1 dominated response is necessary to induce suppression of the ongoing IgE antibody production, which in its turn may be a prerequisite for long-lasting clinical effect. The possibility of induction of an early IgG1 dominated response in every patient by use of alternative immunotherapy procedures is discussed.     

Rapid detection of convallatoxin using five digoxin immunoassays

Context. Cardiac glycosides of plant origin are implicated in toxic ingestions that may result in hospitalization and are potentially lethal. The utility of commonly available digoxin serum assays for detecting foxglove and oleander ingestion has been demonstrated, but no studies have evaluated the structurally similar convallatoxin found in Convallaria majalis (lily of the valley) for rapid laboratory screening, nor has digoxin immune Fab been tested as an antidote for this ingestion. Objective. We aimed to (1) evaluate multiple digoxin assays for cross-reactivity to convallatoxin, (2) identify whether convallatoxin could be detected in vivo at clinically significant doses, and (3) determine whether digoxin immune Fab could be an effective antidote to convallatoxin. Materials and methods. Cross-reactivities of purified convallatoxin and oleandrin with five common digoxin immunoassays were determined. Serum from mice challenged with convallatoxin was tested for apparent digoxin levels. Binding of convallatoxin to digoxin immune Fab was determined in vitro. Results. Both convallatoxin and oleandrin were detectable by a panel of commonly used digoxin immunoassays, but cross-reactivity was variable between individual assays. We observed measurable apparent digoxin levels in serum of convallatoxin intoxicated mice at sublethal doses. Convallatoxin demonstrated no binding by digoxin immune Fab. Conclusion. Multiple digoxin immunoassays detect botanical cardiac glycosides including convallatoxin and thus may be useful for rapid determination of severe exposures, but neutralization of convallatoxin by digoxin immune Fab is unlikely to provide therapeutic benefit.     

血清抗体检测在一起本地传播新冠肺炎聚集性疫情溯源调查中的应用

目的 在深圳市一起本地传播COVID - 19聚集性疫情溯源调查中，对重点涉疫人群开展抗体检测，帮助明确疫情扩散范围，追踪可能的源头。方法 采用流行病学调查、大数据手段锁定重点涉疫人群，除对其开展SARS - COV - 2核酸检测外，采用化学发光法检测抗体，对阳性者进一步开展流行病学调查和中和抗体检测，判断其在疫情传播中的作用。结果 该疫情的9名个案在病程中均检测到IgM抗体、IgG抗体阳性。5 606名涉疫人员检测了抗体，26人阳性，初筛阳性率为0.47%，其中11人IgG抗体阳性，15人IgM抗体阳性；但中和抗体实验和核酸检测均阴性。3人曾经出现过呼吸道症状，均否认与深港货车司机、国外入境人员、冷冻进口货物和发热呼吸道病例接触史。某口岸附近小区阳性率高于其他场所，差异有统计学意义（χ2 = 8.29，P = 0.004）。结论 通过核酸和抗体检测结果可判断，本起疫情传播范围较为局限；化学发光抗体检测法有一定假阳性率，可以作为新冠肺炎疫情溯源调查的辅助方法，对有流行病学史的涉疫人员进行筛查，但不宜对一般人群进行大规模筛查。     

Demystifying Analytical Approaches for Urine Drug Testing to Evaluate Medication Adherence in Chronic Pain Management

ABSTRACT

This comprehensive review of analytical methods used for urine drug testing for the support of pain management describes the methods, their strengths and limitations, and types of analyses used in clinical laboratories today. Specific applications to analysis of opioid levels are addressed. Qualitative versus quantitative testing, immunoassays, chromatographic methods, and spectrometry are discussed. The importance of proper urine sample collection and processing is addressed. Analytical explanations for unexpected results are described. This article describes the scientific basis for urine drug testing providing information which will allow clinicians to differentiate between valid and questionable claims for urine drug testing to monitor medication adherence among chronic pain patients.     

Polymorphism in the C-reactive protein (CRP) gene affects CRP levels in plasma and one early marker of atherosclerosis in men: The Health 2000 Survey

The gastrointestinal hormone gastrin is measured in plasma in physiological, pathophysiological and diagnostic investigations. In the diagnosis of hypergastrinaemic diseases such as gastrinomas and gastric achlorhydria, measurement of gastrin concentrations in circulation is crucial. Gastrin circulates, however, not as a single peptide but as a mixture of peptides of different lengths and amino acid derivatizations. Moreover, in hypergastrinaemia the peptide pattern changes. Consequently, diagnostic gastrin measurements require immunoassays that recognize the pathological plasma patterns, which are characterized by a predominance of the large peptides (gastrin‐34 and gastrin‐71) and less, if any, of the shorter main form of gastrin in normal tissue, gastrin‐17. Alternatively, and in specific cases, “processing‐independent assays” (PIA) for progastrin may be considered, since hypersecreting gastrin cells also release substantial amounts of biosynthetic precursors and processing intermediates. Recently, gastrin kits that do not take the pathological plasma patterns into account have been marketed and may miss the diagnosis. Therefore, proper diagnosis of gastrinomas and other hypergastrinaemic diseases requires insight into cellular gastrin synthesis and peripheral metabolism, and also into the design of useful immunoassays. This review discusses the art of measuring gastrin in plasma with adequate diagnostic specificity.     

Serologically indicated pneumococcal pneumonia in children: a population-based study in primary care settings

The aim of the study was to assess age-specific incidences of community-acquired pneumonia (CAP) caused by Streptococcus pneumoniae and diagnosed serologically in a child population. The study was population-based, and prospective, and performed in primary health care settings. During a surveillance period of 12 months from 1981-1982, all pneumonia cases in a defined child population (57% urban residents) were registered prospectively. In total, 201 CAP cases were diagnosed (mean age 5.6 years; 57% boys; 58% urban residents). S. pneumoniae etiology was studied by antibody and immune complex (IC) assays to C-polysacchride (C-PS), type-specific capsular polysaccharides (CPS), and to pneumolysin (Ply), in acute and convalescent sera. Serologic evidence of S. pneumoniae etiology was indicated in 57(28%) cases, 35(61%) being mixed infections with other microbes. The distribution of pneumococcal cases was 44%, 30% and 26% in the three 5-year age groups, respectively. There were 33 (58%) males and 34 (60%) urban residents. In total, 26 cases were identified by antibody assays and 35 cases by IC assays, 26/35 being positive in acute sera. Responses to C-PS, CPS and Ply, when antibody and IC results are combined, were found equally often in 23-25 cases. The total annual incidence of pediatric S. pneumoniae CAP was 6.4/1000/year. S. pneumoniae etiology was found in 28% of the children and was similar at all ages. The incidence of pneumococcal CAP was assessed for the first time, being high (19/1000/year) in 0- to 4-year-old urban boys and rather stable (5-9/1000/year) in all other groups by age, sex and residence.     

Phenytoin immunoassay measurements in serum samples from patients with renal insufficiency: comparison with high-performance liquid chromatography

The debate continues regarding the possible interference of phenytoin metabolites in phenytoin immunoassays, and its clinical importance for patients with renal failure. The aim of this study was to compare the results obtained using the Abbott fluorescence polarization immunoassay (FPIA), Dade enzyme-multiplied immunoassay technique (EMIT), and high-performance liquid chromatography (HPLC) to establish the significance of the differences in conditions of renal failure. Thirty-six adult patients who had been treated with phenytoin and whose renal function ranged from normal to severely impaired were chosen for this study. In accordance with previously established validation criteria for analytical methods for the determination of drugs, a 15% bias from the HPLC phenytoin values was considered an acceptable limit. The mean (+/-SEM) glomerular filtration rate (GFR) of the patients was 37.5+/-4.6 mL/min (range = 10-102 mL/min).The mean values found using FPIA (10.8+/-1.2 microg/mL) and EMIT (10.8+/-1.3 microg/mL) presented acceptable deviations with respect to HPLC (10.5+/-1.2 microg/mL), and a high correlation was found among the results (N = 36) of the different methods (r > or = 0.987, P < 0.001). An FPIA deviation above the 15% bias limit with respect to HPLC was found only in two cases with very low serum phenytoin concentrations and low GFR values (< 20 mL/min), although it does not appear to be important in terms of adjusting drug dosage. According to our data, FPIA and EMIT gave accurate results for total phenytoin in serum samples from patients with renal failure.     

International study to compare antigen-specific methods used for the measurement of antiplatelet autoantibodies

Platelet-associated and plasma autoantibodies against platelet glycoproteins (GP) have been demonstrated in patients with autoimmune thrombocytopenia (AITP) using various methods. Eight laboratories in seven countries participated in this international study to evaluate the interlaboratory agreement using glycoprotein-specific immunoassays for these autoantibodies. The participating laboratories received blind samples of frozen washed platelets and plasma from 22 normal donors and 22 AITP patients. Platelet-associated and plasma autoantibodies against GPIIb–IIIa and GPIb–IX were measured by MAIPA, immunobead assay or modified antigen capture assay. Of the control samples, 96.0% and 97.2% of all results for platelet-associated and plasma autoantibodies to GPIIb–IIIa/GPIb–IX, respectively, were negative. The mean variation coefficient of the control samples of platelet-associated and plasma autoantibodies was 89.5% (range 11.1–272.9%) and 46.5% (range 21.0–78.0%), respectively. In 20/22 patient samples, platelet-associated autoantibodies to either glycoprotein were noted by at least two laboratories. The mean degree of agreement in these samples was 74.0%. There was a significant correlation in the individual antibody measurements between all laboratories (Kendall coefficient of concordance 0.60 and 0.38, P < 0.001; Spearman rank order test, range of correlation coefficient 52.3–94.0% and 42.2–85.0%, P < 0.05, for anti-GPIIb–IIIa and anti-GPIb–IX, respectively). In contrast, plasma autoantibodies to either glycoprotein were noted by at least two laboratories in only 13/22 patient samples. Moreover, the degree of agreement was poor (50.1%) and a significant correlation was noted between only six pairs of laboratories. We conclude that methods used in this study yield good interlaboratory agreement in measuring platelet-associated autoantibodies against GPIIb–IIIa and GPIb–IX. In contrast, poor agreement was found in detecting plasma autoantibodies to the same glycoproteins.