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Updated: Aug 8, 2026
Isolation and Characterization of Dendritic Cells and Macrophages from the Mouse Intestine
Published on: May 21, 2012
Dendritic cell heterogeneity in vivo: two functionally different dendritic cell populations in rat intestinal lymph
1Sir William Dunn School of Pathology, University of Oxford, United Kingdom.
Insights
Two distinct dendritic cell (DC) populations migrate from the intestine. These L-DC subsets differ in phenotype and function, suggesting separate lineages or microenvironmental influences.
Area of Science:
- Immunology
- Cell Biology
- Dendritic Cell Biology
Background:
- Intestinal dendritic cells (L-DC) exhibit varied surface marker expression.
- Subpopulations of L-DC display distinct morphological and biochemical characteristics.
Purpose of the Study:
- To characterize the phenotypic and functional differences between L-DC subsets.
- To investigate the implications of these differences for immune responses.
Main Methods:
- Flow cytometry analysis of L-DC surface markers (CD4, OX41, B7, MHC class II, ICAM-1).
- Assessment of nonspecific esterase activity and morphology.
- In vitro mixed lymphocyte reactions (MLRs) and in vivo T cell priming assays.
- Antigen processing and presentation assays.
Main Results:
- Two L-DC populations identified: CD4+/OX41+ and CD4-/OX41-.
- CD4+/OX41+ L-DC are potent antigen-presenting cells (APCs) for both naive and sensitized T cells.
- CD4-/OX41- L-DC are less effective at native antigen presentation but strongly stimulate MLRs.
- Differential expression of B7 and invariant chain observed between subsets.
- Stable turnover rates suggest distinct lineages rather than precursor-progeny relationships.
Conclusions:
- Two functionally and phenotypically distinct dendritic cell populations migrate from the intestine.
- These L-DC subsets may represent separate lineages or be modulated by distinct microenvironmental cues.
- The findings have implications for understanding intestinal immunity and immune tolerance.
Abstract:
DC derived from rat pseudo-afferent lymph (L-DC) vary in CD4, CD11b/c, Thy1, and OX41 expression. CD4 and OX41 are expressed by the same subpopulation (50-60%) of L-DC. CD4+/OX41+ L-DC express short fine processes and low nonspecific esterase, whereas CD4- DC/OX41- express long pseudopodia, high nonspecific esterase, and many cytoplasmic inclusions. These differences are stable in culture. Both populations express similar amounts of MHC class II, ICAM-1, CD11b/c and OX62. Most CD4-/OX41- L-DC are strongly positive for B7, but CD4+ L-DC express less B7, and some may be negative. Both populations express invariant chain, but both the absolute numbers and levels of expression were higher for CD4- DC. Surprisingly, CD4+ L-DC are more potent APC than CD4- cells in MLRs, for sensitized T cells in vitro and for naive T cells in vivo. Cultured CD4+/OX41+ DC can still process and present native Ag. Cultured CD4-/OX41- cells cannot present native Ag but can stimulate strong MLRs. CD4- DC invariant chain expression decreases in culture, whereas expression by CD4+ DC is stable for 48 h. CD4+ and CD4- L-DC have similar turnover times in vivo, suggesting that one population is not the precursor of the other. Thus, two separate DC populations that differ functionally and phenotypically migrate from intestine to mesenteric nodes. This may reflect distinct DC lineages or differentiation modulated by different microenvironmental stimuli.
