Molecular differences between the divergent responses of ovalbumin-specific CD4 T cells to alum-precipitated ovalbumin compared to ovalbumin expressed by Salmonella

Mol Immunol. 2008 Aug;45(13):3558-66. doi: 10.1016/j.molimm.2008.05.010. Epub 2008 Jun 25.


CD4 T helper (Th) cell differentiation defined by in vitro cytokine-directed culture systems leaves major gaps in our knowledge of the mechanisms driving divergent Th differentiation. This is evident from our analysis of the response of mouse ovalbumin-specific CD4 T cells to different forms of ovalbumin that induce markedly distinct responses in vivo. We show that live attenuated ovalbumin-expressing Salmonella (SalOVA) induce Th1-associated T-bet and IFN-gamma. Conversely, alum-precipitated ovalbumin (alumOVA) induces the Th2-associated GATA-3 and IL-4. The early diversity occurring within these CD4 T cells isolated 3 days after immunization was assessed using real-time RT-PCR microfluidic cards designed with 384 selected genes. The technique was validated both at the population and single cell levels at different stages of the responses, showing beta2-microglobulin to be a more stably expressed reference mRNA than either beta-actin or 18S RNA. SalOVA was then shown selectively to induce the OVA-specific CD4 T cells to produce many chemokines and pro-inflammatory cytokines, contrasting with alumOVA-induced cells that only produced a few Th2-associated cytokines. Several cytokines and features associated with follicular helper functions were induced in the OVA-specific CD4 T cells by both antigens. Finally, IL-17RB is strongly associated with OVA-specific CD4 T cells responding to alumOVA, suggesting that alum may promote Th2 immune response through a role for the IL-25/IL-17RB pathway.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't
  • Validation Study

MeSH terms

  • Alum Compounds / pharmacology*
  • Animals
  • CD4-Positive T-Lymphocytes / drug effects
  • CD4-Positive T-Lymphocytes / immunology
  • CD4-Positive T-Lymphocytes / metabolism*
  • Cell Differentiation / drug effects
  • Cell Differentiation / genetics
  • Cell Differentiation / immunology
  • Gene Expression Profiling
  • Gene Expression Regulation / immunology
  • Immunization
  • Immunoprecipitation
  • Interferon-gamma / metabolism
  • Interleukin-4 / metabolism
  • Mice
  • Mice, Inbred C57BL
  • Mice, Transgenic
  • Oligonucleotide Array Sequence Analysis
  • Ovalbumin / drug effects
  • Ovalbumin / immunology*
  • Ovalbumin / metabolism*
  • Salmonella / immunology*
  • Salmonella / metabolism
  • Salmonella Infections / genetics
  • Salmonella Infections / immunology
  • Salmonella Infections / metabolism
  • Salmonella Infections / therapy
  • Th1 Cells / immunology
  • Th1 Cells / metabolism
  • Th1 Cells / physiology
  • Th2 Cells / immunology
  • Th2 Cells / metabolism
  • Th2 Cells / physiology


  • Alum Compounds
  • Interleukin-4
  • aluminum sulfate
  • Interferon-gamma
  • Ovalbumin