Letter to the Editor
Basophil activation test in oral desensitization to cow’s milk allergy
United European Gastroenterology Journal 2016, Vol. 4(5) 714–715 ! Author(s) 2016 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav DOI: 10.1177/2050640615614793 ueg.sagepub.com
Salvatore Chirumbolo
Abstract The recent paper by Nucera et al., showed that the basophil activation test (BAT) in flow cytometry is able to monitor an acquired tolerance induced by a desensitization treatment in food allergy. The paper by Nucera et al. reported two standpoints in the CD63 response to food allergy and OAT and their large difference in CD63 response before and after suggests for the optimal performance of a CD123/HLADR/CD63 BAT in oral food allergy immunotherapy.
Keywords Cow’s milk, food allergy, basophil activation test, CD63, immunotherapy, food allergy immunotherapy Received: 9 July 2015; accepted: 13 September 2015
Dear Sir, The recent paper by Nucera et al.,1 in the latest issue of this journal, showed that the basophil activation test (BAT) in flow cytometry is able to monitor an acquired tolerance induced by a desensitization treatment in food allergy. BAT is a cellular test able to shed a light on the basophil response to molecules able to elicit an allergy-related mechanism, even in allergen immunotherapy.2 In the paper by Nucera et al.,1 the authors aimed to show whether the analysis of cell activation markers usually used in BAT, such as CD63, would result in a reliable approach for monitoring the acquisition of clinical tolerance by specific oral desensitization in food allergy, such as egg or cow’s milk. Yet, Nucera et al. did not show any BAT result, as related to any of the three patients enrolled in the study. Although they employed a well suited test,3,4 they were limited to presenting an exemplificative figure showing a BAT with CCR3 (CD193) gating strategy, despite the CD123-PE/HLA-DR-PerCP protocol indicated in Methods. They certainly stated about the finding of a significant reduction in CD63 expression level at the end of the cow’s milk desensitization protocol, but to what extent? Aside from data expressed in Table 2, without any statistical variance report, it would be very interesting to know the extent of this reduction and if the same is related to the
CD63% parameter and its cutoff value, for which critical issues were recently reported.5 In this circumstance, though restricted to a very narrow range of cases, BAT might give important insights about the ‘‘effectiveness’’ of an oral allergen therapy (OAT), particularly if BAT is able to follow up therapy as related to the allergen dosage and exposure times.1,6 The authors showed a great difference in CD63 expression before and after OAT, using defined allergens such as a-lactoalbumin, b-lactoglobulin, and casein. In this circumstance, a more detailed dose–response and time– response evaluation of OAT in children should elucidate how the reported CD63 decrease in basophil membrane expression was due to OAT rather than other mechanisms involved in the spontaneous and physiological membrane recycling of CD63, particularly for allergens such as casein.7,8 The rate of CD63 membrane upregulation, which should affect the CD63% evaluation at a BAT, depends strictly on cell endowment
Department of Medicine, University Physiopathology of Obesity, Verona, Italy
of
Verona
Laboratory
of
Corresponding author: Salvatore Chirumbolo, Department of Medicine, University of Verona Laboratory of Physiopathology of Obesity, LURM est Policlinico GB Rossi, Piazzale AL Scuro 10, 37134 Verona, Italy. Email: [email protected]
Chirumbolo of CD63-intracellular pool and the ability to respond to defined allergens and different type of allergens;8 therefore, during the OAT follow up, the CD63% effect of immunotherapy should be described in a dose- and time-dependent fashion. Furthermore, basophils are at a late time involved in the mechanism and particularly through the Th2-cell mediated immune response. A first response should be retrieved at the mucosal-IgA immunity. CD63 most probably is not easily activated by immune reactions in an IgE-mediated mechanism in mucosal tissues,9 contrary to past reports showing an IgA-mediated basophil activation.10,11 Yet, a close association between secretory-IgAs and pediatric food allergy was recently reported,12 and that serum IgE, as well as IgG1 or IgG4, might not correlate with food allergy, as well as IgA, depending on the type of food allergen.13 This would mean that the reactivity of basophils to food allergens, measured by CD63 membrane expression, needs to be further evaluated on the basis of the different allergen, allergy individual systemic and mucosal response and OAT protocol, a perspective which compels the investigator to highlight OAT successful outcome by tracing CD63 expression throughout the whole OAT treatment and later follow up, as the membrane recycling and updisplacement of CD63 depends on this panoply of immune and humoral mechanisms. The paper by Nucera et al. reported two standpoints in the CD63 response to food allergy and OAT and their large difference in CD63 response before and after suggests for the optimal performance of a CD123/HLADR/CD63 BAT in oral food allergy immunotherapy. Funding This research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors.
Conflict of interest The author states he has no conflict of interest.
References 1. Nucera E, Pecora V, Buonomo A, et al. Utility of basophil activation test for monitoring the acquisition of clinical tolerance after oral desensitization to cow’s milk: pilot study. United Eur Gastroenterol J 2015; 3: 272–276.
715 2. Chirumbolo S. Immunotherapy in allergy and cellular tests: state of art. Hum Vaccin Immunother 2014; 10: 1595–1610. 3. Chirumbolo S, Ortolani R and Vella A. CCR3 as a single selection marker compared to CD123/HLADR to isolate basophils in flow cytometry: some comments. Cytometry A 2011; 79: 102–106. 4. Chirumbolo S, Vella A, Ortolani R, et al. Differential response of human basophil activation markers: a multi-parameter flow cytometry approach. Clin Mol Allergy 2008; 6: 12. 5. Chirumbolo S. Major pitfalls in BAT performance may be caused by gating protocols and CD63% cut off evaluation. Cytometry A 2014; 85: 382–385. 6. Chirumbolo S, Zanoni G, Ortolani R, et al. In vitro biphasic effect of honey bee venom on basophils from screened healthy blood donors. Allergy Asthma Immunol Res 2011; 3: 58–61. 7. Ford LS, Bloom KA, Nowak-Wegrzyn AH, et al. Basophil reactivity, wheal size, and immunoglobulin levels distinguish degrees of cow’s milk tolerance. J Allergy Clin Immunol 2013; 131: 180–186. 8. Chirumbolo S. Basophil activation test in allergy: time for an update? Int Arch Allergy Immunol 2012; 158: 99–114. 9. Bucci C, Zingone F, Russo I, et al. Gliadin does not induce mucosal inflammation or basophil activation in patients with non-celiac gluten sensitivity. Clin Gastroenterol Hepatol 2013; 11: 1294–1299. 10. Iikura M, Yamaguchi M, Miyamasu M, et al. Secretory IgA-mediated basophil activation. II. Roles of GTPbinding regulatory proteins and phosphatidylinositol 3kinase. Biochem Biophys Res Commun 1999; 264: 575–579. 11. Iikura M, Yamaguchi M, Fujisawa T, et al. Secretory IgA induces degranulation of IL-3-primed basophils. J Immunol 1998; 161: 1510–1515. 12. Hogendorf A, Stan´czyk-Przyluska A, SieniwiczLuzen´czyk K, et al. Is there any association between secretory IgA and lactoferrin concentration in mature human milk and food allergy in breastfed children? Med Wieku Rozwoj 2013; 17: 47–52. 13. Guhsl EE, Hofstetter G, Lengger N, et al. IgE, IgG4 and IgA specific to Bet v 1-related food allergens do not predict oral allergy syndrome. Allergy.2015 70: 59–66.
Basophil activation test in oral desensitization to cow’s milk allergy
United European Gastroenterology Journal 2016, Vol. 4(5) 714–715 ! Author(s) 2016 Reprints and permissions: sagepub.co.uk/journalsPermissions.nav DOI: 10.1177/2050640615614793 ueg.sagepub.com
Salvatore Chirumbolo
Abstract The recent paper by Nucera et al., showed that the basophil activation test (BAT) in flow cytometry is able to monitor an acquired tolerance induced by a desensitization treatment in food allergy. The paper by Nucera et al. reported two standpoints in the CD63 response to food allergy and OAT and their large difference in CD63 response before and after suggests for the optimal performance of a CD123/HLADR/CD63 BAT in oral food allergy immunotherapy.
Keywords Cow’s milk, food allergy, basophil activation test, CD63, immunotherapy, food allergy immunotherapy Received: 9 July 2015; accepted: 13 September 2015
Dear Sir, The recent paper by Nucera et al.,1 in the latest issue of this journal, showed that the basophil activation test (BAT) in flow cytometry is able to monitor an acquired tolerance induced by a desensitization treatment in food allergy. BAT is a cellular test able to shed a light on the basophil response to molecules able to elicit an allergy-related mechanism, even in allergen immunotherapy.2 In the paper by Nucera et al.,1 the authors aimed to show whether the analysis of cell activation markers usually used in BAT, such as CD63, would result in a reliable approach for monitoring the acquisition of clinical tolerance by specific oral desensitization in food allergy, such as egg or cow’s milk. Yet, Nucera et al. did not show any BAT result, as related to any of the three patients enrolled in the study. Although they employed a well suited test,3,4 they were limited to presenting an exemplificative figure showing a BAT with CCR3 (CD193) gating strategy, despite the CD123-PE/HLA-DR-PerCP protocol indicated in Methods. They certainly stated about the finding of a significant reduction in CD63 expression level at the end of the cow’s milk desensitization protocol, but to what extent? Aside from data expressed in Table 2, without any statistical variance report, it would be very interesting to know the extent of this reduction and if the same is related to the
CD63% parameter and its cutoff value, for which critical issues were recently reported.5 In this circumstance, though restricted to a very narrow range of cases, BAT might give important insights about the ‘‘effectiveness’’ of an oral allergen therapy (OAT), particularly if BAT is able to follow up therapy as related to the allergen dosage and exposure times.1,6 The authors showed a great difference in CD63 expression before and after OAT, using defined allergens such as a-lactoalbumin, b-lactoglobulin, and casein. In this circumstance, a more detailed dose–response and time– response evaluation of OAT in children should elucidate how the reported CD63 decrease in basophil membrane expression was due to OAT rather than other mechanisms involved in the spontaneous and physiological membrane recycling of CD63, particularly for allergens such as casein.7,8 The rate of CD63 membrane upregulation, which should affect the CD63% evaluation at a BAT, depends strictly on cell endowment
Department of Medicine, University Physiopathology of Obesity, Verona, Italy
of
Verona
Laboratory
of
Corresponding author: Salvatore Chirumbolo, Department of Medicine, University of Verona Laboratory of Physiopathology of Obesity, LURM est Policlinico GB Rossi, Piazzale AL Scuro 10, 37134 Verona, Italy. Email: [email protected]
Chirumbolo of CD63-intracellular pool and the ability to respond to defined allergens and different type of allergens;8 therefore, during the OAT follow up, the CD63% effect of immunotherapy should be described in a dose- and time-dependent fashion. Furthermore, basophils are at a late time involved in the mechanism and particularly through the Th2-cell mediated immune response. A first response should be retrieved at the mucosal-IgA immunity. CD63 most probably is not easily activated by immune reactions in an IgE-mediated mechanism in mucosal tissues,9 contrary to past reports showing an IgA-mediated basophil activation.10,11 Yet, a close association between secretory-IgAs and pediatric food allergy was recently reported,12 and that serum IgE, as well as IgG1 or IgG4, might not correlate with food allergy, as well as IgA, depending on the type of food allergen.13 This would mean that the reactivity of basophils to food allergens, measured by CD63 membrane expression, needs to be further evaluated on the basis of the different allergen, allergy individual systemic and mucosal response and OAT protocol, a perspective which compels the investigator to highlight OAT successful outcome by tracing CD63 expression throughout the whole OAT treatment and later follow up, as the membrane recycling and updisplacement of CD63 depends on this panoply of immune and humoral mechanisms. The paper by Nucera et al. reported two standpoints in the CD63 response to food allergy and OAT and their large difference in CD63 response before and after suggests for the optimal performance of a CD123/HLADR/CD63 BAT in oral food allergy immunotherapy. Funding This research received no specific grant from any funding agency in the public, commercial, or not-for-profit sectors.
Conflict of interest The author states he has no conflict of interest.
References 1. Nucera E, Pecora V, Buonomo A, et al. Utility of basophil activation test for monitoring the acquisition of clinical tolerance after oral desensitization to cow’s milk: pilot study. United Eur Gastroenterol J 2015; 3: 272–276.
715 2. Chirumbolo S. Immunotherapy in allergy and cellular tests: state of art. Hum Vaccin Immunother 2014; 10: 1595–1610. 3. Chirumbolo S, Ortolani R and Vella A. CCR3 as a single selection marker compared to CD123/HLADR to isolate basophils in flow cytometry: some comments. Cytometry A 2011; 79: 102–106. 4. Chirumbolo S, Vella A, Ortolani R, et al. Differential response of human basophil activation markers: a multi-parameter flow cytometry approach. Clin Mol Allergy 2008; 6: 12. 5. Chirumbolo S. Major pitfalls in BAT performance may be caused by gating protocols and CD63% cut off evaluation. Cytometry A 2014; 85: 382–385. 6. Chirumbolo S, Zanoni G, Ortolani R, et al. In vitro biphasic effect of honey bee venom on basophils from screened healthy blood donors. Allergy Asthma Immunol Res 2011; 3: 58–61. 7. Ford LS, Bloom KA, Nowak-Wegrzyn AH, et al. Basophil reactivity, wheal size, and immunoglobulin levels distinguish degrees of cow’s milk tolerance. J Allergy Clin Immunol 2013; 131: 180–186. 8. Chirumbolo S. Basophil activation test in allergy: time for an update? Int Arch Allergy Immunol 2012; 158: 99–114. 9. Bucci C, Zingone F, Russo I, et al. Gliadin does not induce mucosal inflammation or basophil activation in patients with non-celiac gluten sensitivity. Clin Gastroenterol Hepatol 2013; 11: 1294–1299. 10. Iikura M, Yamaguchi M, Miyamasu M, et al. Secretory IgA-mediated basophil activation. II. Roles of GTPbinding regulatory proteins and phosphatidylinositol 3kinase. Biochem Biophys Res Commun 1999; 264: 575–579. 11. Iikura M, Yamaguchi M, Fujisawa T, et al. Secretory IgA induces degranulation of IL-3-primed basophils. J Immunol 1998; 161: 1510–1515. 12. Hogendorf A, Stan´czyk-Przyluska A, SieniwiczLuzen´czyk K, et al. Is there any association between secretory IgA and lactoferrin concentration in mature human milk and food allergy in breastfed children? Med Wieku Rozwoj 2013; 17: 47–52. 13. Guhsl EE, Hofstetter G, Lengger N, et al. IgE, IgG4 and IgA specific to Bet v 1-related food allergens do not predict oral allergy syndrome. Allergy.2015 70: 59–66.
