_:b36340856 "Patients meeting the revised American\u2013European consensus criteria for SjS (n = 4) [>>7<<] as well as age and sex-matched healthy volunteers (n = 4) were recruited for the study from The Ohio State University Wexner Medical Center (OSUWMC) clinics, the Research Match program at OSUWMC, and the American Red Cross." . _:b566904888 "2"^^ . _:b566904894 . _:b566904891 "2"^^ . _:b566904889 . _:b566904896 . _:b566904890 "2"^^ . _:b566904888 . _:b36340874 "In concordance, previous work has demonstrated that CD4+ T-cells predominate the salivary gland infiltrate of SjS patients [>>1<<] and that these cells are largely responsible for mediating glandular destruction [3]." . _:b566904891 . _:b566904901 "2"^^ . _:b36340871 . . _:b566904900 "2"^^ . _:b566904890 . "PMC0" . . _:b36340864 "glands from SjS patients is comprised mostly of CD4+ T-cells with CD8+ T-cells and B-cells detected to a lesser extent, which suggests that T-cells are the primary effector cells of target organ inflammation and glandular dysfunction [>>11<<,12]. To characterize the salivary and lacrimal gland infiltrates of our chimeric mice, immunohistochemistry was performed to detect T-cells (CD4 and CD8), B-cells (CD20), NK cells (CD56), and macro phages (CD68)." . _:b566904901 . _:b566904903 "2"^^ . . _:b36340852 . _:b566904902 "2"^^ . _:b566904900 . _:b36340874 . . . _:b566904897 "2"^^ . _:b566904903 . _:b36340872 . _:b566904896 "2"^^ . _:b566904902 . "10.1016%2Fj.clim.2014.10.004" . . _:b566904897 . _:b566904899 "2"^^ . . _:b36340852 "This mouse has been shown to achieve successful human engraftment using 10-fold fewer human peripheral blood mononuclear cells (PBMCs) than the preceding humanized mouse strains [>>4<<]. Moreover, NSG chimeras display no symptoms of graft versus host disease with transfers of up to 20 \u00D7 106 human PBMCs for at least 30 days, allowing a 4\u20135 week window for investigation [5]. In addition, the NOD mouse has been shown to" . _:b36340854 . _:b566904896 . _:b566904898 "2"^^ . . _:b566904899 . _:b36340849 . _:b36340862 _:b36340864 . _:b36340862 _:b36340865 . _:b36340853 "Moreover, NSG chimeras display no symptoms of graft versus host disease with transfers of up to 20 \u00D7 106 human PBMCs for at least 30 days, allowing a 4\u20135 week window for investigation [>>5<<]. In addition, the NOD mouse has been shown to spontaneously develop sialoadenitis with infiltrates consisting primarily of CD4+ T-cells and adoptive transfer of these cells into NOD-scid mice recapitulated this autoimmune phenotype [6]," . . _:b36340857 "PBMCs were isolated under Ficoll gradient centrifugation as previously described [>>8<<]." . _:b566904898 . _:b36340876 "with previously published studies demonstrating possible roles for other immune cells in SjS, B-cells were present at significantly greater levels in lacrimal and salivary gland infiltrates when compared to transfers from healthy donors [>>2<<]." . . . . . _:b566904905 "2"^^ . . _:b566904904 "2"^^ . . _:b36340866 _:b36340876 . _:b36340852 . _:b36340861 . . _:b566904905 . _:b36340866 _:b36340872 . _:b36340866 _:b36340873 . _:b36340853 . . _:b36340866 _:b36340874 . _:b566904898 . _:b566904906 "2"^^ . _:b566904904 . _:b36340866 _:b36340875 . _:b36340866 _:b36340868 . _:b36340866 _:b36340869 . _:b36340854 . _:b36340866 _:b36340870 . _:b36340866 _:b36340871 . . _:b36340855 . _:b36340866 _:b36340867 . _:b566904906 . _:b36340859 "XT eSlide capture device (Aperio, Vista, CA), and analyzed by Aperio ImageScope digital analysis software (v9.1) as detailed formerly to quantitate inflammation by H&E and to determine lymphocyte localization by immunohistochemistry [>>9<<]." . . _:b36340848 . _:b566904886 . _:b36340873 . _:b36340849 . _:b36340850 . _:b36340861 "Salivary gland function was tested following pilocarpine stimulation (1 \u03BCg/g body weight) according to previously described methods [>>10<<]. Briefly, mice were restrained, injected with pilocarpine intraperitoneally, and observed until the appearance of saliva, which was collected for 5 min and pooled as a measurement of salivary gland function." . . _:b36340851 . . _:b566904884 . _:b36340860 . . _:b36340855 "materials and methods" . _:b36340855 . _:b36340848 . _:b36340861 . _:b566904903 . _:b36340863 . _:b36340862 . _:b36340862 . _:b566904885 . _:b36340863 . . _:b36340856 . _:b36340859 . _:b36340857 . _:b36340858 . _:b566904899 . _:b36340858 . _:b36340875 "In concordance, previous work has demonstrated that CD4+ T-cells predominate the salivary gland infiltrate of SjS patients [1] and that these cells are largely responsible for mediating glandular destruction [>>3<<]. Consistent with previously published studies demonstrating possible roles for other immune cells in SjS, B-cells were present at significantly greater levels in lacrimal and salivary gland infiltrates when compared to transfers from" . _:b36340859 . _:b36340870 "IFN-\u03B3 and TNF-\u03B1 are major cytokines involved in the promotion of Th1 pro-inflammatory responses and have been shown to be elevated in the saliva of SjS patients [>>15<<]. Moreover, mouse models of SjS have shown that IFN-\u03B3 deficiency prevents clinical onset and tissue-specific autoimmune responses [16]. Despite the traditional role as a Th2 anti-inflammatory cytokine, IL-10 levels are elevated in the" . _:b36340860 . _:b36340868 . _:b36340856 . _:b36340869 . _:b36340870 . _:b566904887 . _:b36340871 . . _:b566904890 . _:b566904902 . _:b36340864 . _:b36340876 . . _:b36340867 . _:b36340865 . . _:b566904906 . . _:b36340850 "Despite the detection of B-cells in advanced lesions [>>2<<], T-cells predominate the infiltrate and appear to mediate the cytotoxic pathology and inflammatory cytokine biology associated with SjS (reviewed in [3])." . . _:b36340866 . _:b566904900 . _:b36340870 . _:b36340854 "In addition, the NOD mouse has been shown to spontaneously develop sialoadenitis with infiltrates consisting primarily of CD4+ T-cells and adoptive transfer of these cells into NOD-scid mice recapitulated this autoimmune phenotype [>>6<<], which suggests that there may already be an environment conducive to SjS-like disease in NSG mice." . . _:b36340869 . _:b36340853 . _:b36340867 . _:b36340876 . _:b36340868 . . . _:b566904900 . _:b566904901 . _:b36340848 "introduction" . _:b566904902 . _:b566904903 . _:b566904896 . _:b566904897 . _:b566904898 . _:b566904899 . . _:b566904901 . _:b566904904 . _:b36340872 . _:b566904905 . . _:b566904906 . . _:b566904884 . _:b36340873 . . _:b566904885 . _:b566904886 . _:b566904887 . _:b36340868 "This may be attributable to the propensity of NOD mice to develop sialoadenitis and that this autoimmune phenotype is transferrable to NOD-scid mice [>>6<<], which indicates that an environment permissive to an inflammatory response is presumably present to some extent already in the NSG mice." . _:b36340874 . _:b36340869 "Accordingly, both IFN-\u03B3 and IL-10 were found to be elevated in peripheral blood T-cells of SjS patients [>>14<<]. IFN-\u03B3 and TNF-\u03B1 are major cytokines involved in the promotion of Th1 pro-inflammatory responses and have been shown to be elevated in the saliva of SjS patients [15]. Moreover, mouse models of SjS have shown that IFN-\u03B3 deficiency" . . _:b36340873 "the traditional role as a Th2 anti-inflammatory cytokine, IL-10 levels are elevated in the serum [17] of SjS patients and correlate with disease activity, thus suggesting a potentially pathogenic role for IL-10 in the presence of IFN-\u03B3 [>>18<<]. In addition, IL-6 and IL-17 are cytokines involved in the Th17 pathway and have been implicated in multiple autoimmune diseases, including SjS." . _:b36340851 "Despite the detection of B-cells in advanced lesions [2], T-cells predominate the infiltrate and appear to mediate the cytotoxic pathology and inflammatory cytokine biology associated with SjS (reviewed in [>>3<<]). While the trigger of the cellular infiltration remains unknown, the pathogenesis of SjS is currently thought to involve genetic, neuro-hormonal and environmental factors, and to require participation of the innate and the adaptive" . _:b566904882 . _:b566904883 . _:b566904892 . _:b36340875 . _:b566904893 . _:b36340872 "Despite the traditional role as a Th2 anti-inflammatory cytokine, IL-10 levels are elevated in the serum [>>17<<] of SjS patients and correlate with disease activity, thus suggesting a potentially pathogenic role for IL-10 in the presence of IFN-\u03B3 [18]." . _:b566904894 . _:b566904895 . _:b566904888 . _:b566904889 . _:b566904890 . _:b566904891 . _:b566904892 . . _:b566904883 . _:b36340857 . _:b36340866 . _:b566904904 . _:b36340871 "Moreover, mouse models of SjS have shown that IFN-\u03B3 deficiency prevents clinical onset and tissue-specific autoimmune responses [>>16<<]. Despite the traditional role as a Th2 anti-inflammatory cytokine, IL-10 levels are elevated in the serum [17] of SjS patients and correlate with disease activity, thus suggesting a potentially pathogenic role for IL-10 in the presence" . _:b36340875 . _:b566904882 . . . _:b36340858 "Serial paraffin sections were used for immunohistochemistry and hematoxylin and eosin (H&E) staining as previously described [>>9<<]. Briefly, all slides were stained in Richard Allan Scientific Hematoxylin (Thermo Scientific, Waltham, MA) and Eosin-Y (Thermo Scientific) with the Leica Autostainer (Leica Biosystems, Buffalo Grove, IL). Immunohistochemistry was" . . _:b566904893 . . _:b566904885 "2"^^ . _:b36340862 "results" . _:b566904894 . _:b36340867 "Since graft versus host inflammation is not observed in the NSG mouse strain for at least 30 days following adoptive transfer [>>5<<], we exploited this 4-week timeframe to explore human SjS autoimmune responses in our mouse chimeras." . _:b36340855 _:b36340856 . _:b36340860 "H&E-stained paraffin sections of lacrimal and salivary glands were subjected to blinded histopathological analysis by a board-certified veterinary pathologist (BB) as described previously [>>9<<]. Inflammation and acinar epithelial necrosis were scored 0\u20134:" . _:b566904884 "2"^^ . _:b36340855 _:b36340857 . _:b566904895 . _:b36340855 _:b36340858 . _:b36340865 . _:b36340855 _:b36340859 . _:b36340855 _:b36340860 . _:b566904887 "2"^^ . . _:b566904888 . _:b566904885 . _:b36340855 _:b36340861 . _:b36340851 . _:b566904884 . _:b566904886 "2"^^ . _:b36340866 "discussion" . _:b566904887 . _:b36340848 _:b36340852 . _:b566904886 . _:b36340848 _:b36340853 . . _:b36340848 _:b36340854 . _:b566904891 . _:b566904883 "2"^^ . _:b36340848 _:b36340849 . _:b36340848 _:b36340850 . _:b36340848 _:b36340851 . _:b36340864 . _:b566904882 "2"^^ . . _:b566904905 . _:b566904893 "2"^^ . _:b566904883 . . _:b566904889 . _:b566904897 . _:b566904892 "2"^^ . . _:b566904882 . . . _:b36340850 . _:b36340862 _:b36340863 . _:b566904895 "2"^^ . _:b566904893 . _:b36340863 "To establish a novel chimeric mouse model of SjS, immuno-deficient NSG mice were adoptively transferred with 5 \u00D7 106 PBMCs isolated from healthy volunteers and patients diagnosed with primary SjS [>>7<<]. At the time of transfer, all human PBMC populations expressed similar and detectable levels of markers for T-cells (CD3, CD4, and CD8), B-cells (CD20), Natural killer cells (NK; CD56), and macrophages (CD68), as determined by flow" . . _:b566904892 . _:b566904894 "2"^^ . _:b36340865 "from SjS patients is comprised mostly of CD4+ T-cells with CD8+ T-cells and B-cells detected to a lesser extent, which suggests that T-cells are the primary effector cells of target organ inflammation and glandular dysfunction [11,>>12<<]. To characterize the salivary and lacrimal gland infiltrates of our chimeric mice, immunohistochemistry was performed to detect T-cells (CD4 and CD8), B-cells (CD20), NK cells (CD56), and macro phages (CD68)." . _:b36340849 "Glandular histopathology is primarily characterized by a CD4+ T-cell infiltrate [>>1<<]. Despite the detection of B-cells in advanced lesions [2], T-cells predominate the infiltrate and appear to mediate the cytotoxic pathology and inflammatory cytokine biology associated with SjS (reviewed in [3]). While the trigger of the" . _:b566904889 "2"^^ . _:b566904895 .