Low-molecular-weight heparin and unfractionated heparin decrease Th-1, 2, and 17 expressions

PLoS One. 2014 Nov 3;9(11):e109996. doi: 10.1371/journal.pone.0109996. eCollection 2014.

Abstract

Background: We evaluated the effects of T helper cell differentiation in a mite-allergic animal model treated with inhaled heparins of different molecular weight.

Method: BALB/c mice were divided into four groups: 1. Control, 2. Mite intratracheal (mIT), 3. Inhaled heparin (hIN), 4. Inhaled low-molecular-weight heparin (lmwhIN). Groups 2, 3, and 4 were sensitized twice with Der p allergen subcutaneously on day 1 and day 8. Der p allergen was administered intratracheally on day 15. Groups 3 and 4 were treated with heparin or low-molecular-weight (lmw) heparin intranasally from day 1 to 22. Splenocytes from sacrificed mice stimulated with 16 µg/ml of Der p were cultured for 72 hours. Supernatants of splenocyte were collected to analyze the effect of Interleukin (IL)17-A/F, Interferon(IFN)-γ, IL-4, IL-13, and IL-10. Serum was also collected for Der P-specific IgE level on day 23. Total RNA was extracted from spleen tissue for mRNA expression. Gene expression of Foxp3, IL-10 IFN-γ, GATA3, IL-5, and RORγt were analyzed.

Results: Both hIN and lmwhIN groups had lower serum IgE level than that of the mIT group (both p<0.0001). Both hIN and lmwhIN groups showed significantly decreased transcripts of GATA-3, IFN-γ, IL-5, and RORγt mRNA in their spleen. Regarding the supernatant of splenocyte culture stimulated with Der p, compared with the mIT group, there were significant decreases in IL-17A/F, IFN-γ, IL-4, IL-13, and IL-10 secretion in inhaled hIN and lmwhIN groups.

Conclusions: From this balb/c mice study, the analyses of mRNA and cytokines revealed that both intranasal heparin and lmw heparin treatment decreased the expression of Th1, Th2, and Th17 in spleen. The underlying mechanism(s) warrant further studies.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibody Specificity / immunology
  • Antigens, Dermatophagoides / immunology
  • Cytokines / biosynthesis
  • Gene Expression Regulation / drug effects
  • Heparin / administration & dosage
  • Heparin / pharmacology*
  • Heparin, Low-Molecular-Weight / administration & dosage
  • Heparin, Low-Molecular-Weight / pharmacology*
  • Immunoglobulin E / blood
  • Immunoglobulin E / immunology
  • Interleukin-17 / biosynthesis*
  • Interleukin-17 / genetics
  • Male
  • Mice
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism
  • Spleen / immunology
  • Th1 Cells / drug effects*
  • Th1 Cells / immunology
  • Th1 Cells / metabolism*
  • Th2 Cells / drug effects*
  • Th2 Cells / immunology
  • Th2 Cells / metabolism*

Substances

  • Antigens, Dermatophagoides
  • Cytokines
  • Heparin, Low-Molecular-Weight
  • Interleukin-17
  • RNA, Messenger
  • Immunoglobulin E
  • Heparin

Grants and funding

This work was supported by the Veterans General Hospitals, University System of Taiwan, Joint Research Program (grant numbers 101DFA0400013, 101DFA0400014, VGHUST100-G6-2-2, and VGHUST101-G6-2-2), as well as National Science Council (grant numbers 100-2622-B-007-001-CC1 and 99-3112-B-007-002). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.