title: Histamine releasibility and expression of Lyn and Syk kinases in Indian subjects and role of less potent IL-3 in non-releaser basophils.
abstract: BACKGROUND
  Allergen-mediated activation of the IgE signal pathway in basophils and mast cells leads to release of mediators in-vitro and in-vivo systems. However, basophils from 10% to 20% of the population do not release histamine and other mediators on activation of the IgE signal transduction pathway and this has been attributed to the absence of tyrosine kinases Lyn and Syk. Interestingly, when these non-releaser basophils are incubated with the IL-3, it leads to the recovery of the histamine releasibility.
  
  
  OBJECTIVE
  To investigate histamine releasibility in the Indian population and to evaluate the role of IL-3 with reference to non-releaser phenotypes.
  
  
  METHODS
  Peripheral blood basophils from healthy adults were purified by density gradient centrifugation and negative immuno-selection. Histamine release assay was performed fluorometrically. Assessment of Lyn and Syk expression were carried out by flow-cytometry. SNP analysis in the IL-3 gene was carried by sequencing analysis.
  
  
  RESULTS
  Histamine release after ConA challenge varied greatly from 0% to 100% in Indian subjects. Eighteen percent subjects showed less than 5% histamine release (non-releasers). Flow-cytometric analysis revealed a significantly reduced expression of Lyn and Syk kinases in basophils (p<0.05). Histamine release also significantly correlated with expression of Lyn and Syk kinase (p<0.05). Non-releasers showed the presence of SNP at +79 (T-C), which leads to the one amino acid change at 8th position in the mature IL-3 from serine to proline.
  
  
  CONCLUSIONS
  About 18% of the Indian subjects studied showed non-releaser phenotype and also had reduced Lyn and Syk kinase expression. Non-releasers have also shown the presence of less potent isoform of IL-3/P8, which is suspected to be responsible for the non-releaser phenotype. This needs to be extended to a larger sample size and could be a potential target for the development of therapeutics for allergic patients.
authors.affiliation: Allergy and Aerobiology Laboratory, Institute of Genomics and Integrative Biology, Delhi University Campus, Delhi 110007, India.
authors.forename: P
authors.initials: P
authors.lastname: Kumar
authors.forename: B
authors.initials: B
authors.lastname: Singh
authors.forename: R
authors.initials: R
authors.lastname: Lal
authors.forename: G W
authors.initials: GW
authors.lastname: Rembhotkar
authors.forename: A B
authors.initials: AB
authors.lastname: Singh
mesh_terms: D000328:Adult; D001491:Basophils / Q000502:physiology*; D005260:Female; D015870:Gene Expression; D006636:Histamine Release / Q000502:physiology*; D006801:Humans; D007194:India; D007377:Interleukin-3 / Q000235:genetics / Q000502:physiology*; D047908:Intracellular Signaling Peptides and Proteins; D008297:Male; D008875:Middle Aged; D020641:Polymorphism, Single Nucleotide; D011505:Protein-Tyrosine Kinases / Q000096:biosynthesis*; D000072377:Syk Kinase; D044465:White People / Q000235:genetics; D019061:src-Family Kinases / Q000096:biosynthesis*
chemical_list: C508596:IL3 protein, human; D007377:Interleukin-3; D047908:Intracellular Signaling Peptides and Proteins; D011505:Protein-Tyrosine Kinases; C000606896:SYK protein, human; D000072377:Syk Kinase; C060902:lyn protein-tyrosine kinase; D019061:src-Family Kinases