Download PDF
Homeopathy 2003; 92(01): 11-18
DOI: 10.1054/homp.2002.0082
Original Paper
Copyright © The Faculty of Homeopathy 2003

Influence of the diluent on the effect of highly diluted histamine on basophil activation

I Lorenz
1   Forschungsinstitut KWALIS gGmbH, Dipperz, Germany
,
EM Schneider
2   Section Experimental Anaesthesiology, University Clinic of Ulm, Germany
,
P Stolz
1   Forschungsinstitut KWALIS gGmbH, Dipperz, Germany
,
A Brack
1   Forschungsinstitut KWALIS gGmbH, Dipperz, Germany
,
J Strube
1   Forschungsinstitut KWALIS gGmbH, Dipperz, Germany
› Author Affiliations
Subject Editor:
Further Information

Publication History

Received20 May 2002
revised18 September 2002

accepted18 November 2002

Publication Date:
27 December 2017 (online)

    Permissions and Reprints

Background: In modern pharmaceutical practice, it is common to use purified ethanol and purified water for the preparation of homeopathic dilutions. Hahnemann in 1827 recommended good brandy as a diluent. Brandy contains a lot of accompanying substances in addition to ethanol.

Purpose of the study: The research question was whether different diluents influence the effectiveness of high dilutions, especially above Avogadro's number. We compared two dilution media to investigate the diluent's influence. Within the limitations of the test-system, the dilution media were as similar to good brandy as possible and like purified ethanol. Dilutions of histamine were prepared with both media. As test-system, we used modified basophil activation in an in vitro cell system. Basophils are activated by anti-immunoglobulin E (anti-IgE). The activation of basophils is inhibited by prior incubation with histamine. The reduction in activation was measured with different dilutions of histamine. The test system used a 3-colour flow cytometric method. The interleukin-3 (IL-3) receptor CD123 was used to identify basophils in the leukocyte mixture. The CD63 surface marker was used for quantification of activated basophils.

Results: With higher concentrations of histamine, we observed inhibition on optimally anti-IgE-stimulated basophil activation with a clear concentration dependency. With low concentrations of histamine (up to 10-31), we also observed inhibition of IgE-mediated basophil activation. Differences were observed between the dilution media.

Conclusion: The preliminary results support the hypothesis that the dilution medium may influence the effects of high dilutions. This could be of importance for homeopathic pharmaceutical practice as well as for ultra-high dilution experiments. The refined basophil test system proved to be highly sensitive and reliable. Further studies are needed.

Keywords

basophil activation - anti-IgE - highly diluted histamine - diluent - ultra-high dilution effects
 

  • References

  • 1 Davenas E, Beauvais F, Amara J. Human basophil degranulation triggered by very dilute antiserum against IgE. Nature 1988; 333: 816-818.
    CrossrefPubMedGoogle Scholar
  • 2 Gilbert HS, Ornstein L. Basophil counting with a new staining method using alcian blue. Blood 1975; 42: 279-286.
    PubMedGoogle Scholar
  • 3 Sainte-Laudy J, Vallon C, Guerin JC. [Analysis of membrane expression of the CD63 human basophil activation marker. Applications to allergologic diagnosis]. Allerg Immunol (Paris) 1994; 26: 211-214.
    PubMedGoogle Scholar
  • 4 Knol EF, Mul FP, Jansen H, Calafat J, Roos D. Monitoring human basophil activation via CD63 monoclonal antibody 435. J Allergy Clin Immunol 1991; 88: 328-338.
    CrossrefPubMedGoogle Scholar
  • 5 Sainte-Laudy J, Belon P. Inhibition of human basophil activation by high dilutions of histamine. Agents Actions 1993; 38: C245-C247.
    CrossrefPubMedGoogle Scholar
  • 6 Belon P, Cumps J, Ennis M. Inhibition of human basophil degranulation by successive histamine dilutions: results of a European multi-centre trial. Inflamm Res 1999; 48: S17-S18.
    CrossrefPubMedGoogle Scholar
  • 7 Poitevin B, Davenas E, Benveniste J. In vitro immunological degranulation of human basophils is modulated by lung histamine and Apis mellifica . Br J Clin Pharmacol 1988; 25: 439-444.
    CrossrefPubMedGoogle Scholar
  • 8 Sainte-Laudy J, Belon P. Analysis of immunosuppressive activity of serial dilutions of histamine on human basophil activation by flow cytometry. Inflamm Res 1996; 45: S33-S34.
    CrossrefPubMedGoogle Scholar
  • 9 Brown V, Ennis M. Flow-cytometric analysis of basophil activation: inhibition by histamine at conventional and homeopathic concentrations. Inflamm Res 2001; 50: S47-S48.
    PubMedGoogle Scholar
  • 10 Lorenz I, Schneider EM, Stolz P, Brack A, Strube J. Sensitive flow cytometric method to test activation influenced by homeopathic histamine dilutions. Homeopathy 2002
    PubMedGoogle Scholar
  • 11 Valent P. Immunophenotypic characterization of human basophils and mast cells. Chem Immunol 1995; 61: 34-48.
    PubMedGoogle Scholar
  • 12 Agis H, Fureder W, Bankl HC. Compara-tive immunophenotypic analysis of human mast cells, blood basophils and monocytes. Immunology 1996; 87: 535-543.
    CrossrefPubMedGoogle Scholar
  • 13 Donnenberg VS, O'Connell PJ, Logar AJ. Rare-event analysis of circulating human dendritic cell subsets and their presumptive mouse counterparts. Transplantation 2001; 72: 1946-1951.
    CrossrefPubMedGoogle Scholar
  • 14 Hahnemann S. Reine Arzneimittellehre von Samuel Hahnemann. Dresden: Heidelberg Haug; 1991. p. 333.
    Google Scholar
  • 15 Bohme, H, Hartke, K, Deutsches Arzneibuch/DAB7, 7, 1968, Wissenschafliche Verlagsgesellschaft Stuttgart, Gori Verlag Frankfurt, 1977.
    PubMed
  • 16 Hildebrand A. Deutsches Arzneibuch: amtliche Ausgabe/DAB 10. Stuttgart: Dt. Apotheker Verlag; 1991.
    Google Scholar
  • 17 Nakagawa T, Stadler BM, de Weck AL. Flow-cytometric analysis of human basophil degranulation. I. Quantification of human basophils and their degranulation by flow-cytometry. Allergy 1981; 36: 39-47.
    CrossrefPubMedGoogle Scholar
  • 18 MacGlashan Jr DW. Graded changes in the response of individual human basophils to stimulation: distributional behavior of events temporally coincident with degranulation. J Leukoc Biol 1995; 58: 177-188.
    CrossrefPubMedGoogle Scholar
  • 19 Leon JA, Mesa-Tejada R, Gutierrez MC. Increased surface expression and shedding of tumor associated antigens by human breast carcinoma cells treated with recombinant human interferons or phorbol ester tumor promoters. Anticancer Res 1989; 9: 1639-1647.
    PubMedGoogle Scholar
  • 20 Borowski A, van Valen F, Ulbrecht M. Monomorphic HLA class I-(non-A, non-B) expression on Ewing's tumor cell lines, modulation by TNF-alpha and IFN-gamma. Immunobiology 1999; 200: 1-20.
    CrossrefPubMedGoogle Scholar
  • 21 Falcone FH, Haas H, Gibbs BF. The human basophil: a new appreciation of its role in immune responses. Blood 2000; 96: 4028-4038.
    CrossrefPubMedGoogle Scholar
  • 22 Kepley CL, Wilson BS, Oliver JM. Identification of the Fc epsilonRI-activated tyrosine kinases Lyn, Syk, and Zap-70 in human basophils. J Allergy Clin Immunol 1998; 102: 304-315.
    CrossrefPubMedGoogle Scholar
  • 23 Amano T, Furuno T, Hirashima N, Ohyama N, Nakanishi M. Dynamics of intracellular granules with CD63-GFP in rat basophilic leukemia cells. J Biochem (Tokyo) 2001; 129: 739-744.
    CrossrefPubMedGoogle Scholar
  • 24 Knol EF, Mul FP, Kuijpers TW, Verhoeven AJ, Roos D. Intracellular events in anti-IgE nonreleasing human basophils. J Allergy Clin Immunol 1992; 90: 92-103.
    CrossrefPubMedGoogle Scholar
  • 25 Kepley CL, Youssef L, Andrews RP, Wilson BS, Oliver JM. Multiple defects in Fc epsilon RI signaling in Syk-deficient nonreleaser basophils and IL-3-induced recovery of Syk expression and secretion. J Immunol 2000; 165: 5913-5920.
    CrossrefPubMedGoogle Scholar
  • 26 Hamamoto K, Ohga S, Nomura S, Yasunaga K. Cellular distribution of CD63 antigen in platelets and in three megakaryocytic cell lines. Histochem J 1994; 26: 367-375.
    CrossrefPubMedGoogle Scholar
  • 27 Beinert T, Munzing S, Possinger K, Krombach F. Increased expression of the tetraspanins CD53 and CD63 on apoptotic human neutrophils. J Leukoc Biol 2000; 67: 369-373.
    CrossrefPubMedGoogle Scholar
  • 28 Fureder W, Agis H, Semper H. Differential response of human basophils and mast cells to recombinant chemokines. Ann Hematol 1995; 70: 251-258.
    CrossrefPubMedGoogle Scholar
  • 29 Sainte-Laudy J, Sabbah A, Drouet M, Lauret MG, Loiry M. Diagnosis of venom allergy by flow cytometry. Correlation with clinical history, skin tests, specific IgE, histamine and leukotriene C4 release. Clin Exp Allergy 2000; 30: 1166-1171.
    CrossrefPubMedGoogle Scholar
  • 30 Abuaf N, Rajoely B, Ghazouani E. Validation of a flow cytometric assay detecting in vitro basophil activation for the diagnosis of muscle relaxant allergy. J Allergy Clin Immunol 1999; 104: 411-418.
    CrossrefPubMedGoogle Scholar
  • 31 Monneret G, Gutowski MC, Bienvenu J. Detection of allergen-induced basophil activation by expression of CD63 antigen using a tricolour flow cytometric method. Clin Exp Immunol 1999; 115: 393-396.
    CrossrefPubMedGoogle Scholar
  • 32 Sanz ML, Sanchez G, Gamboa PM. Allergen-induced basophil activation: CD63 cell expression detected by flow cytometry in patients allergic toDermatophagoides pteronyssinus and Lolium perenne . Clin Exp Allergy 2001; 31: 1007-1013.
    CrossrefPubMedGoogle Scholar
  • 33 Baumgartner S, Heusser P, Thurneysen S. Methodo-logical standards and problems in preclinical homoeo-pathic potency research. Forsch Komplementarmed 1998; 5: 27-32.
    PubMedGoogle Scholar
  • 34 Pearce FL. Biological effects of histamine: an overview. Agents Actions 1991; 33: 4-7.
    CrossrefPubMedGoogle Scholar
  • 35 Clough GF, Church MK. Histamine synthesis, action and inactivation: is the balance disturbed in allergy?. Clin Exp Allergy 1999; 29: 9-11.
    CrossrefPubMedGoogle Scholar
  • 36 Lichtenstein LM, Gillespie E. Inhibition of histamine release by histamine controlled by H2 receptor. Nature 1973; 244: 287-288.
    CrossrefPubMedGoogle Scholar
  • 37 Lichtenstein LM, Gillespie E. The effects of the H1 and H2 antihistamines on ‘allergic’ histamine release and its inhibition by histamine. J Pharmacol Exp Ther 1975; 192: 441-450.
    PubMedGoogle Scholar
  • 38 Tedeschi A, Lorini M, Arquati M, Miadonna A. Regulation of histamine release from human basophil leucocytes: role of H1, H2 and H3 receptors. Allergy 1991; 46: 626-631.
    CrossrefPubMedGoogle Scholar
  • 39 Knol EF, Verhoeven AJ, Roos D. Stimulus secretion coupling in human basophilic granulocytes. Clin Exp Allergy 1993; 23: 471-480.
    CrossrefPubMedGoogle Scholar
  • 40 Parsons ME. Histamine receptors: an overview. Scand J Gastroenterol Suppl 1991; 180: 46-52.
    PubMedGoogle Scholar
  • 41 Del Valle J, Gantz I. Novel insights into histamine H2 receptor biology. Am J Physiol 1997; 273: G987-G996.
    PubMedGoogle Scholar
  • 42 Sachs B, Hertl M, Merk HF. Histamine receptors on lymphocytes: distribution and functional significance. Skin Pharmacol Appl Skin Physiol 2000; 13: 313-323.
    CrossrefPubMedGoogle Scholar
  • 43 Sainte-Laudy J. Modulation of allergen and anti-igE induced human basophil activation by serial histamine dilutions. Inflamm Res 2000; 49: S5-S6.
    CrossrefPubMedGoogle Scholar
  • 44 Samal S, Geckeler KE. Unexpected solute aggregation in water on dilution. Chem Commun (Camb) 2001; 21: 2224-2225.
    PubMedGoogle Scholar
  • 45 Kurimoto Y, De Weck AL, Dahinden CA. The effect of interleukin 3 upon IgE-dependent and IgE-independent basophil degranulation and leukotriene generation. Eur J Immunol 1991; 21: 361-368.
    CrossrefPubMedGoogle Scholar
  • 46 Hook WA, Siraganian RP. Influence of anions, cations and osmolarity on IgE-mediated histamine release from human basophils. Immunology 1981; 43: 723-731.
    PubMedGoogle Scholar
  • 47 Zimatkin SM, Anichtchik OV. Alcohol-histamine interactions. Alcohol Alcohol 1999; 34: 141-147.
    CrossrefPubMedGoogle Scholar
  • 48 Dyck LE. Isoenzymes of aldehyde dehydrogenase in human lymphocytes. Alcohol Clin Exp Res 1990; 14: 534-538.
    CrossrefPubMedGoogle Scholar
  • 49 Grisham MB, Granger DN. Neutrophil-mediated mucosal injury. Role of reactive oxygen metabolites. Dig Dis Sci 1988; 33: 6S-15S.
    CrossrefPubMedGoogle Scholar
  • 50 Sainte-Laudy J, Belon P. Application of flow cytometry to the analysis of the immunosuppressive effect of histamine dilutions on human basophil activation: effect of cimetidine. Inflamm Res 1997; 46: S27-S28.
    CrossrefPubMedGoogle Scholar