Passive Cutaneous Anaphylaxis

2011 ◽  
Keyword(s):  
Passive Cutaneous Anaphylaxis

The Development and Interlaboratory Validation of a Quantitative Antibody Capture ELISA for the Measurement of Specific Guinea-pig IgG1: An Alternative to the Passive Cutaneous Anaphylaxis Assay

1996 ◽  
Vol 24 (1) ◽  
pp. 73-79
Author(s):  
Laura S. Babcock ◽  
Thomas T. Kawabata ◽  
Victor S. Moore ◽  
Catherine M. Condo ◽  
Annette Prentø
Keyword(s):  
Guinea Pig ◽  
Passive Cutaneous Anaphylaxis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protein Antigens ◽  
Capture Elisa ◽  
Elisa Format ◽  
Interlaboratory Validation ◽  
Antibody Capture ◽  
Development And Validation

Specific guinea-pig IgG1 has traditionally been measured by using an in vivo guinea-pig passive cutaneous anaphylaxis (PCA) assay. This paper describes the development and validation of a quantitative enzyme-linked immunosorbent assay (ELISA) for specific IgG1 against two protein antigens (Alcalase and Enzyme B) as an alternative to the PCA assay. The ELISA format involved a rabbit antibody bound to microtitre plates to capture the antigen. The test sera is added to this, followed sequentially by goat anti-guinea-pig IgG1 and rabbit anti-goat-IgG-alkaline phosphatase conjugate. Aliquots of each serum sample from immunised guinea-pigs were analysed with the ELISA for specific IgG1 titres in three laboratories and were compared to titres determined by using the PCA assay. The findings demonstrate that there is a good correlation between the ELISA and the PCA assay and that the ELISA shows good interlaboratory reproducibility. Thus, the antibody capture ELISA described in this report is a valid and robust replacement for the guinea-pig PCA assay.

scholarly journals MiR-154-5p-MCP1 Axis Regulates Allergic Inflammation by Mediating Cellular Interactions

2021 ◽  
Vol 12 ◽  
Author(s):  
Misun Kim ◽  
Hyein Jo ◽  
Yoojung Kwon ◽  
Myeong Seon Jeong ◽  
Hyun Suk Jung ◽  
...  
Keyword(s):  
Allergic Inflammation ◽  
Passive Cutaneous Anaphylaxis ◽  
Allergic Reactions ◽  
Dependent Manner ◽  
Cellular Interactions ◽  
Array Analysis ◽  
Molecular Features ◽  
Rat Basophilic Leukemia Cells

In a previous study, we have demonstrated that p62, a selective receptor of autophagy, can regulate allergic inflammation. In the present study, microRNA array analysis showed that miR-154-5p was increased by antigen (DNP-HSA) in a p62-dependent manner in rat basophilic leukemia cells (RBL2H3). NF-kB directly increased the expression of miR-154-5p. miR-154-5p mediated in vivo allergic reactions, including passive cutaneous anaphylaxis and passive systemic anaphylaxis. Cytokine array analysis showed that antigen stimulation increased the expression of MCP1 in RBL2H3 cells in an miR-154-5p-dependent manner. Reactive oxygen species (ROS)-ERK-NF-kB signaling increased the expression of MCP1 in antigen-stimulated RBL2H3 cells. Recombinant MCP1 protein induced molecular features of allergic reactions both in vitro and in vivo. Anaphylaxis-promoted tumorigenic potential has been known to be accompanied by cellular interactions involving mast cells, and macrophages, and cancer cells. Our experiments employing culture medium, co-cultures, and recombinant MCP1 protein showed that miR-154 and MCP1 mediated these cellular interactions. MiR-154-5p and MCP1 were found to be present in exosomes of RBL2H3 cells. Exosomes from PSA-activated BALB/C mouse induced molecular features of passive cutaneous anaphylaxis in an miR-154-5p-dependent manner. Exosomes from antigen-stimulated RBL2H3 cells enhanced both tumorigenic and metastatic potentials of B16F1 melanoma cells in an miR-154-5p-dependent manner. Exosomes regulated both ROS level and ROS mediated cellular interactions during allergic inflammation. Our results indicate that the miR-154-5p-MCP1 axis might serve as a valuable target for the development of anti-allergy therapeutics.

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