Cell culture modeling of specialized tissue: identification of genes expressed specifically by follicle-associated epithelium of Peyer's patch by expression profiling of Caco-2/Raji co-cultures.
Peyer's patch follicle-associated epithelium (FAE) regulates intestinal antigen access to the immune system in part through the action of microfold (M) cells which mediate transcytosis of antigens and microorganisms. Studies on M cells have been limited by the difficulties in isolating purified cells, so we applied TOGA mRNA expression profiling to identify genes associated with the in vitro induction of M cell-like features in Caco-2 cells and tested them against normal Peyer's patch tissue for their expression in FAE. Among the genes identified by this method, laminin beta3, a matrix metalloproteinase and a tetraspan family member, showed enriched expression in FAE of mouse Peyer's patches. Moreover, the C. perfringens enterotoxin receptor (CPE-R) appeared to be expressed more strongly by UEA-1(+) M cells relative to neighboring FAE. Expression of the tetraspan TM4SF3 gene and CPE-R was also confirmed in human Peyer's patch FAE. Our results suggest that while the Caco-2 differentiation model is associated with some functional features of M cells, the genes induced may instead reflect the acquisition of a more general FAE phenotype, sharing only select features with the M cell subset.
Lo
David
D
Digital Gene Technologies, Inc., La Jolla, CA 92037, USA. davidlo@dgt.com
Tynan
Wendy
W
Dickerson
Janet
J
Scharf
Melinda
M
Cooper
Judith
J
Byrne
Daragh
D
Brayden
David
D
Higgins
Lisa
L
Evans
Claire
C
O'Mahony
Daniel J
DJ
D000818:Animals; D018938:Caco-2 Cells; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000276:immunology; D017931:DNA Primers; D004847:Epithelial Cells / Q000502:physiology*; D020869:Gene Expression Profiling; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D010581:Peyer's Patches / Q000166:cytology* / Q000502:physiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013045:Species Specificity
D017931:DNA Primers