[Omeo33] Art 1021 - Inflamm Res, 2009, doi: 10.1007/s00011-009-0044-4

[Gino Santini]

Inhibition of CD203c membrane up-regulation in human basophils by high  
dilutions of histamine: a controlled replication study
Salvatore Chirumbolo, Maurizio Brizzi, Riccardo Ortolani, Antonio  
Vella and Paolo Bellavite

Objective: Previous research suggests that human basophil activation  
may be inhibited by histamine even at extremely low doses (high  
dilutions). However, uncertainties about the nature of the phenomenon  
and its reproducibility mean that further, rigorously controlled  
studies are necessary.
Methods: Serial 1:100 (v:v) histamine dilutions (centesimal dilutions,  
C) and water controls were tested on human basophil responsiveness to  
anti-IgE antibodies, using flow cytometry. Each dilution step was  
followed by vertical mechanical shaking (also designed as succussion)  
at 20 strokes/s. Basophil-enriched buffy coats from healthy blood  
donors were incubated with 10−4 mol/l histamine (2C) and with  
serially diluted preparations from 10−20 mol/l (10C) to 10−32 mol/l  
(16C), then incubated for 30 min with 1 μg/ml goat monoclonal anti- 
human IgE and basophils stained for immunophenotyping.
Results: Membrane up-regulation of CD203c, which in these experimental  
conditions proved to be a more consistent activation marker than CD63,  
was significantly inhibited in samples treated with histamine at the  
dilutions of 2C (P = 0.001), 12C (P = 0.047), 14C (P = 0.003), 15C (P  
= 0.036) and 16C (P = 0.009). Control water dilutions/succussions did  
not show any significant effect.
Conclusion: Using a strictly standardized flow cytometry protocol and  
a new dilution/succussion procedure, we have shown that low and high  
dilutions of histamine inhibit CD203c up-regulation in anti-IgE  
stimulated basophils.

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