Establishment of a basophil activation test in BN rats

Yanru Guo; Rong Sun; Wei Li; Zhaohua Liu; Yanru Guo; Rong Sun; Wei Li; Zhaohua Liu

doi:10.3934/Allergy.2020003

AIMS Allergy and Immunology

2020, Volume 4, Issue 2: 20-31. doi: 10.3934/Allergy.2020003

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Research article

Establishment of a basophil activation test in BN rats

1.
Center for New Drug Evaluation, School of Pharmaceutical Sciences, Cheeloo college of medicine, Shandong University, Jinan 250012, China
2.
Shandong Academy of Chinese Medicine, Jinan 250014, China

Received: 27 February 2020 Accepted: 11 May 2020 Published: 14 May 2020

Allergies induced by low molecular weight compounds (LMWCs) have become a major problem in clinical medication. Effective preclinical prediction of sensitization can solve this problem; however, there are currently no reliable methods to predict sensitization, and it is necessary to establish a more effective model. The basophil activation test (BAT) has been widely used clinically to diagnose and research allergic diseases with high sensitivity. The purpose of this study was to detect the expression of CD63 in basophils in OVA-sensitized brown Norway (BN) rats, establishing a model for the BAT to assess the sensitization of the low molecular weight compound PG. The number of basophils in rats was detected by flow cytometry and blood smear analysis. The peripheral blood of OVA-sensitized rats was incubated with OVA, and the increase in CD63 on basophils was greater than that with PBS in vitro. After penicillin G-conjugated OVA (PG/OVA) was incubated with the peripheral blood of penicillin G-conjugated BSA (PG/BSA)-sensitized rats, the expression of CD63 on basophils was significantly higher than that with PBS. The basophils in the sensitized rats were again exposed to the same antigen in vitro, and flow cytometry detected a significant increase in CD63, demonstrating that the rat BAT model was successfully built, and it can be used to evaluate the potential sensitization of low molecular weight compounds.
- basophil activation test,
- CD63,
- low molecular weight compounds,
- immediate hypersensitivity reactions,
- safety evaluation in preclinical
Citation: Yanru Guo, Rong Sun, Wei Li, Zhaohua Liu. Establishment of a basophil activation test in BN rats[J]. AIMS Allergy and Immunology, 2020, 4(2): 20-31. doi: 10.3934/Allergy.2020003

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Abstract

Allergies induced by low molecular weight compounds (LMWCs) have become a major problem in clinical medication. Effective preclinical prediction of sensitization can solve this problem; however, there are currently no reliable methods to predict sensitization, and it is necessary to establish a more effective model. The basophil activation test (BAT) has been widely used clinically to diagnose and research allergic diseases with high sensitivity. The purpose of this study was to detect the expression of CD63 in basophils in OVA-sensitized brown Norway (BN) rats, establishing a model for the BAT to assess the sensitization of the low molecular weight compound PG. The number of basophils in rats was detected by flow cytometry and blood smear analysis. The peripheral blood of OVA-sensitized rats was incubated with OVA, and the increase in CD63 on basophils was greater than that with PBS in vitro. After penicillin G-conjugated OVA (PG/OVA) was incubated with the peripheral blood of penicillin G-conjugated BSA (PG/BSA)-sensitized rats, the expression of CD63 on basophils was significantly higher than that with PBS. The basophils in the sensitized rats were again exposed to the same antigen in vitro, and flow cytometry detected a significant increase in CD63, demonstrating that the rat BAT model was successfully built, and it can be used to evaluate the potential sensitization of low molecular weight compounds.

Acknowledgments

We would like to thank the centre for new drug safety evaluation for establishing a platform to exchange ideas and protocols. Furthermore, we wish to thank all colleagues. This work forms part of the master thesis. The project was supported by the Shandong Province Independent Innovation and Transformation of Scientific & Technological Achievements Project, China (2014ZZCX02104) and the Shandong Provincial Natural Science Foundation, China (ZR2012HM056). The funder had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Conflict of interests

All authors declare no conflicts of interest in this paper.

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