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Allergic sensitization increases the amount of extracellular ATP hydrolyzed by guinea pig leukocytes

  • Jaime ChávezEmail author
  • Mario H. Vargas
  • Jesús Martínez-Zúñiga
  • Ramcés Falfán-Valencia
  • Enrique Ambrocio-Ortiz
  • Verónica Carbajal
  • Rosa Sandoval-Roldán
Original Article

Abstract

Increased levels of ATP have been found in the bronchoalveolar lavage of patients with asthma, and subjects with this disease, but not healthy subjects, develop bronchospasm after nebulization with ATP. Because the main mechanism for controlling the noxious effects of extracellular ATP is its enzymatic hydrolysis, we hypothesized that allergic sensitization is accompanied by a decreased functioning of such hydrolysis. In the present study, peripheral blood leukocytes from sensitized and non-sensitized guinea pigs were used for determining the extracellular metabolism (as assessed by inorganic phosphate production) of ATP, ADP, AMP, or adenosine, and for detecting possible changes in the expression (qPCR and Western blot) of major ectonucleotidases (NTPDase1, NTPDase3, and NPP1) and purinoceptors (P2X1, P2X7, P2Y4, and P2Y6). Contrary to our hypothesis, we found that leukocytes from allergic animals produced higher amounts of inorganic phosphate after stimulation with ATP and ADP, as compared with leukocytes from non-sensitized animals. Although at first glance, this result suggested that sensitization caused higher efficiency of ectonucleotidases, their mRNA and protein expressions were unaffected. On the other hand, after sensitization, we found a significant increase in the protein expression of P2X7 and P2Y4, two purinoceptors known to be responsible for ATP release after activation. We concluded that allergic sensitization increased the amount of ATP hydrolyzed by ectonucleotidases, the latter probably not due to the enhanced efficiency of its enzymatic breakdown, but rather due to an increased release of endogenous ATP or other nucleotides, partly mediated by enhanced expression or P2X7 and P2Y4 receptors.

Keywords

Extracellular ATP ATP hydrolysis Ectonucleotidase Purinoceptors Allergic sensitization Guinea pig asthma model 

Notes

Compliance with ethical standards

Conflict of interest

Jaime Chávez declares that he has no conflict of interest.

Mario H. Vargas declares that he has no conflict of interest.

Jesús Martínez-Zúñiga declares that he has no conflict of interest.

Ramcés Falfán-Valencia declares that he has no conflict of interest.

Enrique Ambrocio-Ortiz declares that he has no conflict of interest.

Verónica Carbajal declares that she has no conflict of interest.

Rosa Sandoval-Roldán declares that she has no conflict of interest.

Ethical approval

Animal management was done according with the 2011 Guide for the Care and Use of Laboratory Animals. All procedures performed in guinea pigs were in accordance with the ethical standards of the institution, and the study protocol was approved by our institutional scientific and bioethics committees, with the approval number B23-12.

Supplementary material

11302_2019_9644_MOESM1_ESM.doc (34 kb)
ESM 1 (DOC 33 kb)
11302_2019_9644_MOESM2_ESM.pptt (53 kb)
ESM 2 (PPTT 52 kb)

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  • Jaime Chávez
    • 1
    Email author return OK on get
  • Mario H. Vargas
    • 1
  • Jesús Martínez-Zúñiga
    • 1
  • Ramcés Falfán-Valencia
    • 2
  • Enrique Ambrocio-Ortiz
    • 2
  • Verónica Carbajal
    • 1
  • Rosa Sandoval-Roldán
    • 1
  1. 1.Departamento de Investigación en Hiperreactividad BronquialInstituto Nacional de Enfermedades Respiratorias Ismael Cosío VillegasMexico CityMexico
  2. 2.Laboratorio HLAInstituto Nacional de Enfermedades Respiratorias Ismael Cosío VillegasMexico CityMexico

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