Abstract
Impairment of intestinal epithelial barriers contributes to the progression of HIV/SIV infection and leads to generalized HIV-induced immune-cell activation during chronic infection. Rhesus macaques are the major animal model for studying HIV pathogenesis. However, detailed characterization of isolated rhesus epithelial cells (ECs) from intestinal tissues is not well defined. It is also not well documented whether isolated ECs had any other cell contaminants from intestinal tissues during the time of processing that might hamper interpretation of EC preparations or cultures. In this study, we identify and characterize ECs based on flow cytometry and immunohistochemistry methods using various enzymatic and mechanical isolation techniques to enrich ECs from intestinal tissues. This study shows that normal healthy ECs differentially express HLA-DR, CD23, CD27, CD90, CD95 and IL-10R markers. Early apoptosis and upregulation of ICAM-1 and HLA-DR in intestinal ECs are thought to be the key features in SIV mediated enteropathy. The data suggest that intestinal ECs might be playing an important role in mucosal immune responses by regulating the expression of different important regulatory and adhesion molecules and their function.
Highlights
The intestinal mucosal immune response in healthy individuals is characterized by a balance between immunity, which protects mucosal surfaces from harmful microbes, and tolerance, which permits the intestinal mucosa to interact in a nonpathogenic way with the commensal bacteria and dietary antigens to which it is constantly exposed [1,2,3]
This study shows that epithelial cells (ECs) are positive for HLA-DR, CD23, CD27, CD90, CD95 and IL-10R phenotypes
Similar percentages of single positive ECs were evident in EDTA treated upper (70.2%65.6%) and lower (70.7%610.6%) layer cells isolated from percoll gradient where no prior DTT treatment was followed (Figs. 2, 5 & 6)
Summary
The intestinal mucosal immune response in healthy individuals is characterized by a balance between immunity, which protects mucosal surfaces from harmful microbes, and tolerance, which permits the intestinal mucosa to interact in a nonpathogenic way with the commensal bacteria and dietary antigens to which it is constantly exposed [1,2,3]. Detailed isolation and characterization of rhesus ECs from intestinal tissues are poorly documented, whereas the rhesus macaque (RM) model is well recognized for understanding HIV/SIV pathogenesis, disease progression and HIV vaccine development [14] It is not well documented whether isolated ECs have other cell contaminants from intestinal tissues during the time of processing or whether isolation methods could be improved or optimized to reduce contamination that might hamper the study design using EC cultures. These ECs in normal uninfected and SIV infected RMs were not characterized with respect to memory and/or effector status, adhesion, antigen presentation, or regulatory receptor expression compared to intestinal CD45+ leukocytes
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