Journal of Clinical Immunology

, Volume 31, Issue 2, pp 228–239

Functional Invariant NKT Cells in Pig Lungs Regulate the Airway Hyperreactivity: A Potential Animal Model

  • Gourapura J. Renukaradhya
  • Cordelia Manickam
  • Mahesh Khatri
  • Abdul Rauf
  • Xiangming Li
  • Moriya Tsuji
  • Gireesh Rajashekara
  • Varun Dwivedi
Article

DOI: 10.1007/s10875-010-9476-4

Cite this article as:
Renukaradhya, G.J., Manickam, C., Khatri, M. et al. J Clin Immunol (2011) 31: 228. doi:10.1007/s10875-010-9476-4

Abstract

Important roles played by invariant natural killer T (iNKT) cells in asthma pathogenesis have been demonstrated. We identified functional iNKT cells and CD1d molecules in pig lungs. Pig iNKT cells cultured in the presence of α-GalCer proliferated and secreted Th1 and Th2 cytokines. Like in other animal models, direct activation of pig lung iNKT cells using α-GalCer resulted in acute airway hyperreactivity (AHR). Clinically, acute AHR-induced pigs had increased respiratory rate, enhanced mucus secretion in the airways, fever, etc. In addition, we observed petechial hemorrhages, infiltration of CD4+ cells, and increased Th2 cytokines in AHR-induced pig lungs. Ex vivo proliferated iNKT cells of asthma induced pigs in the presence of C-glycoside analogs of α-GalCer had predominant Th2 phenotype and secreted more of Th2 cytokine, IL-4. Thus, baby pigs may serve as a useful animal model to study iNKT cell-mediated AHR caused by various environmental and microbial CD1d-specific glycolipid antigens.

Keywords

iNKT cellsairway hyperreactivitypigscytokines

Supplementary material

10875_2010_9476_MOESM1_ESM.pdf (53 kb)
Figure S1Acute AHR induced pig iNKT cells present in the PBMCs and lung-MNCs were preferentially proliferated in to CD4+ phenotype and secreted more of IL-4 than IFNg ex vivo. Pigs were intratracheally inoculated with a-GalCer or vehicle and euthanized at post-24 hr of inoculation. PBMCs (a, c, e) and lung-MNCs (b, d, f) isolated from pigs were cultured for eight days in the presence of vehicle, a-GalCer, GCK127, or GCK152 at indicated concentrations. Cells were immunostained using fluorochrome conjugated anti-pig CD3e, CD4a, and mouse empty or PBS-57-loaded CD1d tetramers, and subjected to flow cytometry. CD3+ gated cells were analyzed for mouse CD1d tetramer+ and CD4a+/- iNKT cells (a, b). The supernatants harvested from cultures were analyzed for IFNg (c and d) and IL-4 (e, f) by ELISA. Each bar represents the average percentage of iNKT cells or amounts of cytokine from three pigs +/- SEM in one independent experiment. Similar results were observed in another independent experiment. (PDF 52 kb)
10875_2010_9476_MOESM2_ESM.pdf (328 kb)
Figure S2.C-glycoside analogues of α–GalCer (GCK127 and GCK152) differentially induce pig iNKT cell proliferation ex vivo in acute AHR induced pigs. Pigs were intratracheally inoculated with a-GalCer or vehicle and euthanized at post-24 hr. PBMC and (b) lung-MNC isolated from pigs was cultured for eight days in the presence of vehicle, α-GalCer, GCK127, or GCK152 (1 μg/ml). Cells were immunostained using fluorochrome conjugated anti-pig CD3e, CD4α, and mouse empty or PBS-57-loaded CD1d tetramers, and the data was acquired by flow cytometry and analyzed using FlowJo software. CD3+ gated cells were analyzed for CD1d tetramer and CD4a. The data shown are from a representative pig (n= 3 pigs per group) in one independent experiment. Similar results were observed in another independent experiment. (PDF 327 kb)

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Gourapura J. Renukaradhya
    • 1
  • Cordelia Manickam
    • 1
  • Mahesh Khatri
    • 1
  • Abdul Rauf
    • 1
  • Xiangming Li
    • 2
  • Moriya Tsuji
    • 2
  • Gireesh Rajashekara
    • 1
  • Varun Dwivedi
    • 1
  1. 1.Food Animal Health Research Program, Ohio Agricultural Research and Development Center, Department of Veterinary Preventive MedicineThe Ohio State UniversityWoosterUSA
  2. 2.HIV and Malaria Vaccine Program, Aaron Diamond AIDS Research CenterThe Rockefeller UniversityNew YorkUSA