Diagnostic relevance of the lymphocyte transformation test for sensitization to beryllium and other metals (IUPAC Technical Report)

The lymphocyte transformation test (LTT) has been proven useful especially in the diagnosis of drug-induced allergic disorders. It is an in vitro test which is based on the fact that lymphocytes, which have been sensitized by a certain antigen, transform into blasts and proliferate when they are again exposed to this antigen. This proliferation is determined by measurement of the incorporation of [3H ]-thymidine or bromodeoxyuridine into replicating DNA. The test has the advantage over skin tests of avoiding re-exposure of individuals, and it was, therefore, hoped that it may also help to diagnose metal allergies and especially sensitization toward beryllium. However, the LTT measures only the sensitization of lymphocytes, but not the effector reaction, i.e., there may be positive results in exposed individuals even in the absence of clinical symptoms. There are several reports evaluating the LTT toward gold salts (Au), amalgam (Hg), nickel (Ni), beryllium (Be), and several other metals. With metals other than Be, the LTT appears to be of little use. In contrast, the LTT with Be may, indeed, define patients at risk of developing chronic beryllium disease (CBD), which affects mainly the respiratory tract and may even cause death. Beryllium sensitization progresses to CBD at a rate of 7-11 % per year. Since the Be-LTT can detect sensitization in workers who have not yet developed a disease it is an important diagnostic tool to detect individuals at risk. In conclusion, the LTT can detect a cell-mediated immunological response of an individual to metals. However, for most metals its usefulness is questionable, with the exception of Be; a positive Be-LTT can identify not only patients with CBD, but also persons at risk of developing CBD in later years.

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Drug-Induced Allergic Cytopenia: In Vitro Confirmation by the Lymphocyte Transformation Test

Archives of Internal Medicine ◽

10.1001/archinte.160.14.2218-a ◽

2000 ◽

Vol 160 (14) ◽

pp. 2218-a-2219

Author(s):

B. Sachs

Keyword(s):

Lymphocyte Transformation ◽

Lymphocyte Transformation Test ◽

Drug Induced

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Diagnostic methods for confirming drug-allergy – the lymphocyte transformation test in dermatology

Orvosi Hetilap ◽

10.1556/oh.2008.28215 ◽

2008 ◽

Vol 149 (24) ◽

pp. 1107-1114

Author(s):

Sarolta Makó ◽

Réka Lepesi-Benkő ◽

Márta Marschalkó ◽

Gyöngyvér Soós ◽

Sarolta Kárpáti

Keyword(s):

Drug Allergy ◽

Lymphocyte Transformation ◽

Lymphocyte Transformation Test ◽

Diagnostic Methods

A gyógyszermellékhatások felismerése és a tüneteket kiváltó gyógyszer oki szerepének bizonyítása komoly felkészültséget igényel. E közlemény célja a gyógyszerallergiás reakciók diagnosztikai lehetőségeinek rövid áttekintése és a lymphocytatranszformációs teszt gyógyszer-hiperszenzitivitási reakciókban való bizonyító szerepének bőrgyógyászati szempontok szerinti értékelése. A lymphocytatranszformációs teszt azon a megfigyelésen alapul, hogy a gyógyszerrel való első találkozáskor kialakult antigénspecifikus T-sejtek osztódni kezdenek az antigénnel való in vitro megismételt találkozás után. A szenzibilizációt az osztódó T-sejtekbe történő 3 H-timidin-beépülés mértéke jelzi. A hatóanyag-specifikus T-sejtek szinte mindig részt vesznek a gyógyszerallergiás reakciókban, ezért a vizsgálat előnye, hogy sok gyógyszernél és különböző immunreakciók eseteiben egyaránt jól alkalmazható. Hátránya a munkaigényesség, valamint az, hogy specificitásának és szenzitivitásának bizonyításához hiányoznak a széles körű, nagy beteganyagon elvégzett tanulmányok. Emiatt a teszt nem egyértelműen elfogadott a gyógyszerallergia igazolására. Hiányosságai ellenére azonban, jobb prediktív értékű egyéb vizsgálatok hiányában, a lymphocytatranszformációs tesztnek fontos szerepe van a gyógyszerallergiák diagnosztizálása terén.

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TO010A NEW IMMUN-TOXICOLOGICAL TEST TO DETECT POLYSULFONE HYPERSENSITIVITY IN HEMODIALYSIS PATIENTS

Nephrology Dialysis Transplantation ◽

10.1093/ndt/gfaa141.to010 ◽

2020 ◽

Vol 35 (Supplement_3) ◽

Author(s):

Joachim Beige ◽

Ralph Wendt ◽

Despina Rüssmann ◽

Karl-Peter Ringel

Keyword(s):

False Negative ◽

False Negative Rate ◽

Lymphocyte Transformation ◽

Clinical Problem ◽

Lymphocyte Transformation Test ◽

Laboratory Research ◽

Humoral Immune ◽

Before And After ◽

Positive Results

Abstract Background and Aims Incompatibility of dialysis procedure due to hypersensitivity against dialyzer material which currently is mainly based on polysulfone and derivatives can not be assessed by routine laboratory tests. Although the frequency of such symptoms is suspected to be low (below 2%) such resembles an important clinical problem because dialysis procedures are frequently accompanied by symptoms of non-tolerability with reasons not being entirely clear while circulatory reasons are suspected to play a major role. Method To enlighten the role of polysulfone hypersensitivity, we adapted known standardized material immune-toxicological tests (lymphocyte transformation test, basophil degranulation test) to the specific conditions of dialysis and polysulfone material sensitivity. We developed a method of polysulfone micronisation and measured humoral immune response of isolated patient´s lymphocytes when incubated with polysulfone dispersion. Results 39 samples from 103 patients with suspected polysulfone hypersensitivity showed positive results for type 1 (n=19), type 4 (n=18) or both type (n=2) reactions. There were no significant differences in the level of stimulation measured for DI, SI and lymphogenesis before and after dialysis (average delta -0.4; -0.28; - 1.74, p = 0.71; 0.34; 0.37) and with different dialyzer materials (Tab. 1). Patients with pos. type 4 results (LTT and lymphogenesis) showed highly correlated results in either LTT or lymphogenesis test (Fig. 1, R=0.87, p<0.0001). 8 out of 8 samples from patients with repeated test on different PS showed positive results on either PS. One patient tested positive on PS showed no hypersensitivity with another non-PS (PMMA) material. Conclusion This is the first methodological report showing plausible in-vitro results of patients samples concerning polysulfone intolerance. On the first superficial view, a “false-negative” rate of 60% looks rather disappointing, because all samples derived from patients with suspicion of PS hypersensitivity. However, due to the clinical variability of intolerance symptoms and the high prevalence of any problems after HD initiation, mainly of circulatory origin after initiating extracorporeal circuit, this rate may obviously express the true frequency of isolated PS material hypersensitivity in suspected patients. Alternative pathophysiological pathways of material sensitivity like complement activation, remain to be elucidated and incorporated into a comprehensive future testing panel. Further clinical and laboratory research is needed to define true polysulfone hypersensitivity and to enlighten the field of hypothetic subclinical material incompatibility in patients with impaired dialysis tolerability.

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Drug-Induced Thrombocytopenia

Archives of Pathology & Laboratory Medicine ◽

10.5858/133.2.309 ◽

2009 ◽

Vol 133 (2) ◽

pp. 309-314

Author(s):

Barton Kenney ◽

Gary Stack

Keyword(s):

At Risk ◽

19Th Century ◽

Clinical Management ◽

Drug Induced ◽

Medication Withdrawal ◽

Platelet Antibodies ◽

Patients At Risk ◽

Drug Dependent ◽

The 19Th Century ◽

Prompt Diagnosis

Abstract Drug-induced thrombocytopenia was first described in the 19th century, yet our understanding of its pathogenesis continues to evolve. The list of drugs implicated in drug-induced thrombocytopenia is extensive and growing. Many, if not most, of these medications induce thrombocytopenia by immune mechanisms. Because the degree of thrombocytopenia can put patients at risk for serious bleeding, a prompt diagnosis is key to clinical management. The laboratory approach to diagnosing drug-induced thrombocytopenia is 2-pronged. First, nondrug causes of thrombocytopenia must be ruled out. Second, testing for drug-dependent platelet antibodies, available at specialized reference laboratories, often can identify the offending medication, although usually not in time for initial clinical management. Once a medication is suspected of causing thrombocytopenia, it must be discontinued promptly, and the patient should be monitored closely. Thrombocytopenia generally resolves quickly after offending medication withdrawal, and the prognosis of drug-induced thrombocytopenia is then excellent.

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Recombinant in vitro test T-SPOT.TB as a screening method for early diagnosis of tuberculosis infection

Tuberculosis and lung diseases ◽

10.21292/2075-1230-2020-98-4-48-52 ◽

2020 ◽

Vol 98 (4) ◽

pp. 48-52

Author(s):

E. P. Eremenko ◽

E. A. Borodulina ◽

I. A. Sergeeva ◽

D. A. Kudlay ◽

B. E. Borodulin

Keyword(s):

Screening Method ◽

Latent Tuberculosis ◽

Tuberculosis Infection ◽

Mantoux Test ◽

Skin Tests ◽

In Vitro Tests ◽

In Vitro Test ◽

Healthy Children ◽

Vitro Test

In addition to standard skin tests (Mantoux test with 2 TU PPD-L and diaskintest) for the diagnosis of tuberculosis infection, in vitro tests are used. One of these tests is T-SPOT.TB being more widely used in recent years.The objective: to evaluate the effectiveness of T-SPOT.TB test for early detection of tuberculosis infection in children and adolescents in Samara Region.Subjects and methods. From 2016 to 2019, results of T-SPOT.TB tests performed in 596 children aged 2 to 17 years inclusive were analyzed; those children had no immunodiagnosis of tuberculosis infection using skin tests since their parents refused to have it.Results. It was found out that the major reason for refusing skin tests was the “fear” of visiting a TB dispensary if the result had been positive — 38.43% (n = 229). The latent tuberculosis infection according to the results of T-SPOT.TB among children with concomitant pathology made 2.6%, among healthy children – 0.7%.Conclusion. T-SPOT.TB test may be used as an alternative method for diagnosis of tuberculosis infection, should the parent refuse to have skin tests. In children with concomitant pathology, T-SPOT.TB test can serve as a leading method for immunodiagnosis of tuberculosis.The authors state that they have no conflict of interests.

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Development of a modified lymphocyte transformation test for diagnosing drug-induced liver injury associated with an adaptive immune response

Journal of Immunotoxicology ◽

10.1080/1547691x.2016.1254305 ◽

2017 ◽

Vol 14 (1) ◽

pp. 31-38 ◽

Cited By ~ 13

Author(s):

Jessica Whritenour ◽

Mira Ko ◽

Qing Zong ◽

Jianying Wang ◽

Karrie Tartaro ◽

...

Keyword(s):

Immune Response ◽

Liver Injury ◽

Adaptive Immune Response ◽

Lymphocyte Transformation ◽

Lymphocyte Transformation Test ◽

Drug Induced ◽

Drug Induced Liver Injury ◽

Adaptive Immune

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Eosinophilic Peritonitis and Drug-Induced Lymphocyte Transformation in Vitro

Peritoneal Dialysis International ◽

10.1177/089686089501500117 ◽

1995 ◽

Vol 15 (1) ◽

pp. 76-77 ◽

Cited By ~ 4

Author(s):

Takashi Sakano ◽

Hiroko Takahashi ◽

Mikio Mori ◽

Takashi Hamasaki

Keyword(s):

Lymphocyte Transformation ◽

Drug Induced

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Quinolone Allergy

Pharmacy ◽

10.3390/pharmacy7030097 ◽

2019 ◽

Vol 7 (3) ◽

pp. 97 ◽

Cited By ~ 9

Author(s):

Edoabasi U. McGee ◽

Essie Samuel ◽

Bernadett Boronea ◽

Nakoasha Dillard ◽

Madison N. Milby ◽

...

Keyword(s):

Clinical Manifestations ◽

Provocation Test ◽

Cross Reactivity ◽

Skin Tests ◽

Diagnostic Tools ◽

In Vitro Tests ◽

Allergic Reactions ◽

Drug Induced ◽

Delayed Reaction

Quinolones are the second most common antibiotic class associated with drug-induced allergic reactions, but data on quinolone allergy are scarce. This review article discusses the available evidence on quinolone allergy, including prevalence, risk factors, diagnosis, clinical manifestations, cross-reactivity, and management of allergic reactions. Although the incidence of quinolone allergy is still lower than beta-lactams, it has been increasingly reported in recent decades, most likely from its expanded use and the introduction of moxifloxacin. Thorough patient history remains essential in the evaluation of quinolone allergy. Many diagnostic tools have been investigated, but skin tests can yield false-positive results and in vitro tests have not been validated. The drug provocation test is considered the test of choice to confirm a quinolone allergy but is not without risk. Evidence regarding cross-reactivity among the quinolones is limited and conflicting. Quinolone allergy can be manifested either as an immediate or delayed reaction, but is not uniform across the class, with moxifloxacin posing the highest risk of anaphylaxis. Quinolone should be discontinued when an allergic reaction occurs and avoided in future scenarios, but desensitization may be warranted if no alternatives are available.

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Development and Application of a Transcriptomic Signature of Bioactivation in an Advanced In Vitro Liver Model to Reduce Drug-induced Liver Injury Risk Early in the Pharmaceutical Pipeline

Toxicological Sciences ◽

10.1093/toxsci/kfaa094 ◽

2020 ◽

Vol 177 (1) ◽

pp. 121-139 ◽

Cited By ~ 4

Author(s):

Wen Kang ◽

Alexei A Podtelezhnikov ◽

Keith Q Tanis ◽

Stephen Pacchione ◽

Ming Su ◽

...

Keyword(s):

Liver Injury ◽

Human Model ◽

Reactive Metabolites ◽

In Vitro Test ◽

Drug Induced ◽

Vitro Assay ◽

Drug Induced Liver Injury ◽

Drug Candidates ◽

Transcriptomic Signature

Abstract Early risk assessment of drug-induced liver injury (DILI) potential for drug candidates remains a major challenge for pharmaceutical development. We have previously developed a set of rat liver transcriptional biomarkers in short-term toxicity studies to inform the potential of drug candidates to generate a high burden of chemically reactive metabolites that presents higher risk for human DILI. Here, we describe translation of those NRF1-/NRF2-mediated liver tissue biomarkers to an in vitro assay using an advanced micropatterned coculture system (HEPATOPAC) with primary hepatocytes from male Wistar Han rats. A 9-day, resource-sparing and higher throughput approach designed to identify new chemical entities with lower reactive metabolite-forming potential was qualified for internal decision making using 93 DILI-positive and -negative drugs. This assay provides 81% sensitivity and 90% specificity in detecting hepatotoxicants when a positive test outcome is defined as the bioactivation signature score of a test drug exceeding the threshold value at an in vitro test concentration that falls within 3-fold of the estimated maximum drug concentration at the human liver inlet following highest recommended clinical dose administrations. Using paired examples of compounds from distinct chemical series and close structural analogs, we demonstrate that this assay can differentiate drugs with lower DILI risk. The utility of this in vitro transcriptomic approach was also examined using human HEPATOPAC from a single donor, yielding 68% sensitivity and 86% specificity when the aforementioned criteria are applied to the same 93-drug test set. Routine use of the rat model has been adopted with deployment of the human model as warranted on a case-by-case basis. This in vitro transcriptomic signature-based strategy can be used early in drug discovery to derisk DILI potential from chemically reactive metabolites by guiding structure-activity relationship hypotheses and candidate selection.

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