Follicular Dendritic Cell-secreted Protein Research Articles

FDC-SP as a diagnostic and prognostic biomarker and modulates immune infiltrates in renal cell carcinoma

BackgroundRenal cell carcinoma (RCC), one of the top 10 causes of cancer death, is responsible for more than 90% of all cases of primary renal cancer worldwide. Follicular dendritic cell-secreted protein (FDC-SP) specifically binds to activated B cells and regulates the generation of antibodies. It is also thought to promote cancer cell invasion and migration, which could help with tumor metastases. This study aimed to assess the efficacy of FDC-SP in the diagnosis and prognosis of RCC and to investigate the relationship between immune infiltration in RCC and these outcomes.ResultsRCC tissues had significantly higher levels of FDC-SP protein and mRNA than normal tissues. The high level of FDC-SP expression was linked to the T stage, histological grade, pathological stage, N stage, M stage, and OS event. Functional enrichment analysis identified the major pathways that were enriched as immune response regulation, complement, and coagulation. Immunological checkpoints and immune cell infiltration were observed to substantially correlate with the levels of FDC-SP expression. FDC-SP expression levels showed the ability to precisely distinguish high-grade or high-stage renal cancer (area under the curve (AUC) = 0.830, 0.722), and RCC patients with higher FDC-SP expression levels had worse prognoses. The AUC values for one-, two-, and five-year survival rates were all greater than 0.600. Moreover, the FDC-SP expression is an independent predictive biomarker of OS in RCC patients.ConclusionFDC-SP may be a prospective therapeutic target in RCC as well as a possible diagnostic and prognostic biomarker associated with immune infiltration.

Complement Regulatory Protein CD46 Manifests a Unique Role in Promoting the Migration of Bladder Cancer Cells.

CD46 is a membrane-bound complement regulatory protein (mCRP) possessing a regulatory role with the complement system. CD46 protects the host cells from damage by complement. Expression of CD46 is also highly maintained in many cancers, including bladder cancers, and thus functions as a receptor for many cancer therapeutic viruses. In this study we report a unique role of CD46 as a progression factor of cancer cells in bladder cancers. Resulting data from a DNA microarray using CD46-altered HT1376 bladder cancers demonstrated a pool of target genes, including complement C3 α chain (C3α), matrix Gla protein (MGP), AFAP-AS1, follicular dendritic cell secreted protein (FDCSP), MAM domain containing 2 (MAMDC2), gamma-aminobutyric acid A receptor pi (GABRP), transforming growth factor, beta-induced (TGFBI), a family of cytochrome P450 (CYP24A1), sialic acid binding Ig-like lectin 6 (SIGLEC6), metallothionein 1E (MT1E), and several members of cytokeratins. Subsequent studies using quantitative RT-PCR and Western blot analyses confirmed CD46-mediated regulation of C3α, MGP, and keratin 13 (KRT13). MGP and KRT13 are known to be involved in cell migration and cancer cell metastasis. A cell migration assay demonstrated that CD46 enhanced migratory potential of bladder cancer cells. Taken all together, this report demonstrated that CD46 is generally overexpressed in bladder cancers and plays a unique role in the promotion of cancer cell migration. Further detailed studies are needed to be performed to clarify the action mechanism of CD46 and its application to cancer therapeutics.

FDCSP Is an Immune-Associated Prognostic Biomarker in HPV-Positive Head and Neck Squamous Carcinoma.

Background: Head and neck squamous carcinoma (HNSC) poses a major threat to human life. The role of human papillomavirus (HPV) infection in the initiation and progression of HNSC is becoming more widely accepted. HPV-positive (HPV+) HNSC has shown unique responses to cancer therapies, which may be due to differences in immune cell infiltration. It is critical to determine how the immune responses to HPV in HNSC are regulated. Methods: Transcriptome data of HNSC from The Cancer Genome Atlas (TCGA) and the Gene Expression Omnibus (GEO) database were analyzed. Then, the CIBERSORT algorithm was used to calculate immune cell infiltration in HNSC. FDCSP expression level was detected by qPCR in the HNSC tissues collected from the Nanfang Hospital. Results: Follicular dendritic cell secreted protein (FDCSP) was highly expressed in HPV+ HNSC, and higher expression of FDSCP was associated with a favorable prognosis. In HPV+ HNSC samples, FDCSP significantly increased the proportion of T follicular helper cells (TFHs). FDCSP expression was also found to be associated with TP53 mutation status in HPV+ HNSC. The function of FDCSP was intimately connected to chemokine pathways, particularly with the C-X-C motif chemokine ligand 13 (CXCL13). We verified that the high expression of FDCSP in HPV+ HNSC and higher FDCSP is closely related to prognosis in HNSC samples we collected by qPCR. Conclusions: Collectively, these findings may provide fresh evidence that FDCSP is a potential chemokine-associated prognostic biomarker in HPV+ HNSC.

Follicular dendritic cell-secreted protein gene expression is upregulated and spread in nifedipine-induced gingival overgrowth.

Follicular dendritic cell-secreted protein (FDC-SP) is secreted protein expressed in follicular dendritic cells, periodontal ligament and junctional epithelium (JE). Its expression could be controlled during inflammatory process of gingiva; however, responsible mechanism for gingival overgrowth and involvement of FDC-SP in clinical condition is still unclear. We hypothesized that JE-specific genes are associated with the initiation of drug-induced gingival enlargement (DIGE) called gingival overgrowth, and investigated the changes of JE-specific gene's expression and their localization in overgrown gingiva from the patients. Immunohistochemical analysis revealed that the FDC-SP localization was spread in overgrown gingival tissues. FDC-SP mRNA levels in GE1 and Ca9-22 cells were increased by time-dependent nifedipine treatments, similar to other JE-specific genes, such as Amelotin (Amtn) and Lamininβ3 subunit (Lamβ3), whereas type 4 collagen (Col4) mRNA levels were decreased. Immunocytochemical analysis showed that FDC-SP, AMTN, and Lamβ3 protein levels were increased in GE1 and Ca9-22 cells. Transient transfection analyses were performed using luciferase constructs including various lengths of human FDC-SP gene promoter, nifedipine increased luciferase activities of -345 and -948FDC-SP constructs. These results raise the possibility that the nifedipine-induced FDC-SP may be related to the mechanism responsible for gingival overgrowth does not occur at edentulous jaw ridges.

Triptolide inhibits tonsillar IgA production by upregulating FDC-SP in IgA nephropathy.

IgA nephropathy (IgAN) is primarily resulted of qualitative abnormality of IgA. The occurrence of IgAN is associated with affected tonsils which enhances the IgA production via IgA class switching and immuno-activation. Follicular dendritic cell-secreted protein (FDC-SP) was found to be a negative effect for IgA production in tonsil. The previous studies suggested that Triptolide might reduce IgA production by its immunosuppression role. Given this background, this study investigated the mechanisms underlying the role of Triptolide and FDC-SP in the generation of IgA and IgA class switching in tonsil of IgAN patients. Immunohistochemistry and reverse transcription-polymerase chain reaction revealed that the expression of FDC-SP was increased in the tonsils of IgAN patients with Triptolide treatment compared with those without treatment. Meanwhile, the expression of FDC-SP was negatively correlated with IgA inducing cytokines in the tonsils of IgAN patients treated with Triptolide, due to the significant decreased IgA-bearing cells. The expression of FDC-SP in tonsillar tissue was confirmed by double immunofluorescence. Importantly, Triptolide promoted FDC-SP secretion, and correlated negatively with decreased IgA production in isolated FDC-associated clusters, which had been isolated from patients without TW treatment previously. Our study demonstrated that Triptolide might have an impact on FDC-SP production and downregulation of IgA synthesis in the tonsils of IgAN patients, which could be a promising strategy for therapeutic intervention in IgAN patients.

Triptolide Inhibits Tonsillar IgA Class Switching by Upregulating FDC-SP in IgA Nephropathy

IgA nephropathy (IgAN) is characterized by qualitative abnormality of IgA in affected tonsils, where present enhancement of IgA production by IgA class switching and immuno-activation. Follicular dendritic cell-secreted protein (FDC-SP) was found to be effective in tonsillar IgA production of IgAN. Triptolide, a core medicinal component of Tripterygium wilfordii, has been suggested to control IgA production by its' immunosuppression effect. Given this background, this study investigated the mechanisms underlying the role of Triptolide and FDC-SP in the generation of IgA and IgA class switching in tonsil of IgAN patients. Immunohistochemistry and reverse transcription-polymerase chain reaction revealed that the expression of FDC-SP were increased and was correlated negatively with decreased IgA inducing cytokines in the tonsils of IgAN patients with Triptolide treatment compared with those without treatment, followed by significantly decreased of IgA-bearing cells. The location of FDC-SP in tonsillar tissue was confirmed by double immunofluorescence. Importantly, Triptolide promoted FDC-SP secretion, and correlated negatively with decreased IgA production in isolated FDC-associated clusters. Our study demonstrated that Triptolide may be involved in IgA production in the tonsils of IgAN patients, inhibiting IgA class switching in IgAN patients through the cooperative roles of FDC-SP, highlighting a promising strategy for therapeutic intervention in IgAN. Funding Statement: This work was supported by the National Nature Science Foundation of China (81600539), Natural Science Foundation of Heilongjiang Province of China (QC2012C041, LC2016038), Nn10 program of Harbin Medical University Cancer Hospital (Nn10 2017-03), Postdoctoral Science Start-up Foundation of Heilongjiang Province (Hongxue Meng), Harbin Special fund project for Science and technology innovation(2016RAQXJ203), Youth elite training Foundation of Harbin Medical University Cancer Hospital (JY2016-06) and Outstanding Youth Foundation of Harbin Medical University Cancer Hospital (JCQN-2018-05)，Clinical Scientific Research Foundation of China Medical and Health Development Foundation (Hongxue Meng), Harbin Medical University Postgraduate Research and Practice Innovation Project (Huining Li). Declaration of Interests: All authors have read the journal's policy on disclosure of potential conflicts of interest and have none to declare. Ethics Approval Statement: This research was carried out according to the principles of the Declaration of Helsinki. Written informed consent was obtained from all participants in this study. Approval for this study was obtained from the Medical Ethics Committees of First Affiliated Hospital of Hei Longjiang University of Chinese Medicine (HZYLLBA201714).

Molecular Assessment of Human Peri-implant Mucosal Healing at Laser-Modified and Machined Titanium Abutments.

To compare, by gene profiling analysis, the molecular events underscoring peri-implant mucosa formation at machined vs laser-microgrooved implant healing abutments. Forty endosseous implants were placed by a one-stage approach in 20 healthy subjects in nonadjacent sites for single-tooth restorations. In a split-mouth design, machined smooth and laser-microgrooved healing abutments were randomly assigned in each subject. Peri-implant mucosa adjacent to healing abutments was harvested by tissue punch biopsy at either 1, 2, 4, or 8 weeks following abutment placement. Total RNA was isolated from the peri-implant transmucosal soft tissues. A whole genome microarray using the Affymetrix Human Gene 2.1 ST Array was performed to describe gene expression profiles in relation to abutment topography and healing time duration. Data analysis was completed using GeneSpring software v.12.6. Differential gene expression was revealed at all time points and among surfaces. Five hundred one genes were differentially expressed (fold change ≥ 2.0) at machined versus laser-modified abutments, and 459 of these were statistically significant (P ≤ .05). At 1 week, unique expression of IL-24 and MMP1 was observed in tissues from laser-treated surfaces. At 2, 4, and 8 weeks, mRNAs encoding keratins and protective proteins of cornified epithelium were upregulated in tissues from laser-modified abutments. At 4 weeks, upregulation (> 2-fold) of mRNAs encoding proteins associated with collagen fibril formation and function was observed in tissue from laser-modified abutments. In both tissues of machined and laser-modified abutments, mRNAs encoding junctional epithelium-specific proteins, ostogenic ameloblast associated protein (ODAM) and follicular dendritic cell secreted protein (FDCSP) were highly upregulated throughout weeks 2 to 8. Peri-implant abutment mucosal wound healing involves selective differentiation of epithelium and induction of the junctional epithelium. Laser-mediated alterations in abutment topography enhance collagen fibril-associated gene expression and alter epithelium/junctional epithelial gene expression. Clinically, shallower probing depths are measured at laser-mediated versus machined implant abutments.

Tumor necrosis factor-α regulates human follicular dendritic cell-secreted protein gene transcription in gingival epithelial cells.

Follicular dendritic cell-secreted protein (FDC-SP) is a secreted protein expressed in follicular dendritic cells, periodontal ligament and junctional epithelium. To elucidate the transcriptional regulation of the human FDC-SP gene by tumor necrosis factor-α (TNF-α), we conducted real-time PCR, Western blotting, transient transfection analyses with chimeric constructs of the FDC-SP gene promoter linked to a luciferase reporter gene, gel mobility shift and chromatin immunoprecipitation assays using Ca9-22 gingival epithelial cells. TNF-α (10ng/ml) induced FDC-SP mRNA and protein levels at 3hr and reached maximum at 12hr. In transient transfection assays, TNF-α (12hr) increased the LUC activities of constructs between -116FDCSP and -948FDCSP including the human FDC-SP gene promoter. Transcriptional stimulations by TNF-α were partially inhibited in the -345FDCSP constructs that included 3-bp mutations in the YY1, GATA, CCAAT enhancer-binding protein 2 (C/EBP2) and C/EBP3. Transcriptional activities induced by TNF-α were inhibited by tyrosine kinase, MEK1/2 and phosphoinositide 3-kinase inhibitors. The results of ChIP assays showed that YY1, GATA and C/EBPβ transcription factors interacted with the YY1, GATA, C/EBP2 and C/EBP3 elements that were increased by TNF-α. These studies show that TNF-α stimulates human FDC-SP gene transcription by targeting YY1, GATA, C/EBP2 and C/EBP3 in the FDC-SP gene promoter.

Effects of interleukin-1β on human follicular dendritic cell-secreted protein gene expression in periodontal ligament cells.

Follicular dendritic cell-secreted protein (FDC-SP) is expressed in FDCs, human periodontal ligament (HPL) cells, and junctional epithelium. To evaluate the effects of interleukin-1 beta (IL-1β) on FDC-SP gene expression in immortalized HPL cells, FDC-SP mRNA and protein levels in HPL cells following stimulation by IL-1β were measured by real-time polymerase chain reaction and Western blotting. Luciferase (LUC), gel mobility shift, and chromatin immunoprecipitation (ChIP) analyses were performed to study the interaction between transcription factors and promoter regions in the human FDC-SP gene. IL-1β (1 ng/mL) induced the expression of FDC-SP mRNA and protein levels at 3 h, and reached maximum levels at 12 h. IL-1β increased LUC activities of constructs (-116FDCSP - -948FDCSP) including the FDC-SP gene promoter. Transcriptional inductions by IL-1β were partially inhibited by 3-base-pair (3-bp) mutations in the Yin Yang 1 (YY1), GATA, CCAAT-enhancer-binding protein2 (C/EBP2), or C/EBP3 in the -345FDCSP. IL-1β-induced -345FDCSP activities were inhibited by protein kinase A, tyrosine-kinase, mitogen-activated protein kinase (MEK)1/2, and PI3-kinase inhibitors. The results of gel shift and ChIP assays revealed that YY1, GATA, and C/EBP-β interacted with the YY1, GATA, C/EBP2, and C/EBP3 elements that were increased by IL-1β. These studies demonstrate that IL-1β increases FDC-SP gene transcription in HPL cells by targeting YY1, GATA, C/EBP2, and C/EBP3 in the human FDC-SP gene promoter.

Effect of follicular dendritic cell secreted protein on gene expression of human periodontal ligament cells

ObjectiveThe objective of this study was to investigate the specific roles of follicular dendritic cell secreted protein (FDC-SP), a protein exists in saliva, in the inhibition of calcium precipitation during periodontal regeneration, as well as affect phenotype expression of human periodontal ligament cells (hPDLCs) during the differentiation process. DesignTo investigate this, we applied microarray technology to identify gene expression changes in hPDLCs transfected with FDC-SP and then clustered them according to their biological functions. ResultsOne hundred seventy-one genes were found differentially expressed by at least two-fold between FDC-SP -transfected and empty vector-transfected cells. Besides, genes encoding cell-cycle proteins, blood-related and cell differentiation-related proteins tended to be up-regulated after FDC-SP transfection, whereas cytokine/growth factors, signal transduction and metabolism-related genes tended to be down-regulated in hPDLCs overexpression FDC-SP. ConclusionsThe present study investigated FDC-SP’s roles in hPDLCs’ phenotype expression, via comparing the gene expression profiles between FDC-SP -transfected hPDLCs and empty vector-transfected cells upon microarray analysis. hPDLCs overexpression FDC-SP appear to display different gene expression patterns. In all, these observations showed a potential of FDC-SP in the maintenance of PDL homeostasis and its ultimate contribution to periodontal would-healing processes.

Identification of novel follicular dendritic cell sarcoma markers, FDCSP and SRGN, by whole transcriptome sequencing.

Follicular dendritic cell (FDC)-sarcoma is a rare neoplasm with morphologic and phenotypic features of FDCs. It shows an extremely heterogeneous morphology, therefore, its diagnosis relys on the phenotype of tumor cells. Aim of the present study was the identification of new specific markers for FDC-sarcoma by whole transcriptome sequencing (WTS). Candidate markers were selected based on gene expression level and biological function. Immunohistochemistry was performed on reactive tonsils, on 22 cases of FDC-sarcomas and 214 control cases including 114 carcinomas, 87 soft tissue tumors, 5 melanomas, 5 thymomas and 3 interdigitating dendritic cell sarcomas. FDC secreted protein (FDCSP) and Serglycin (SRGN) proved to be specific markers of FDC and related tumor. They showed better specificity and sensitivity values than some well known markers used in FDC sarcoma diagnosis (specificity: 98.6%, and 100%, respectively; sensitivity: 72.73% and 68.18%, respectively). In our cohorts CXCL13, CD21, CD35, FDCSP and SRGN were the best markers for FDC-sarcoma diagnosis and could discriminate 21/22 FDC sarcomas from other mesenchymal tumors by linear discriminant analysis. In summary, by WTS we identified two novel FDC markers and by the analysis of a wide cohort of cases and controls we propose an efficient marker panel for the diagnosis of this rare and enigmatic tumor.

Localization and expression pattern of amelotin, odontogenic ameloblast-associated protein and follicular dendritic cell-secreted protein in the junctional epithelium of inflamed gingiva.

The purpose of this study is to elucidate the localization of amelotin (AMTN), odontogenic ameloblast-associated protein (ODAM) and follicular dendritic cell-secreted protein (FDC-SP) at the junctional epithelium (JE) in Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans infected mice and inflamed and non-inflamed human gingiva. We performed immunostaining to determine the localization and expression pattern of AMTN, ODAM and FDC-SP. AMTN, ODAM and FDC-SP in A. actinomycetemcomitans infected mice did not change dramatically compared with non-infected mice. AMTN and FDC-SP expressions were observed stronger in P. gingivalis infected mice at early stage. However, at the following stage, the coronal part of the AMTN expression disappeared from the JE, and FDC-SP expression decreased due to severe inflammation by P. gingivalis. ODAM expressed internal and external basal lamina, and the expression increased not only at early stage but also at the following stage in the inflammatory JE induced by P. gingivalis. In the human gingival tissues, AMTN was detected at the surface of the sulcular epithelium and JE in the non-inflamed and inflamed gingiva, and the localization did not change the process of inflammation. ODAM and FDC-SP were more widely detected at the sulcular epithelium and JE in the non-inflamed gingiva. In the inflamed gingiva, localization of ODAM and FDC-SP was spread into the gingival epithelium, compared to AMTN. These studies demonstrated that the expression pattern of AMTN, ODAM and FDC-SP at the JE were changed during inflammation process and these three proteins might play an important role in the resistance to inflammation.

Follicular dendritic cell-secreted protein may enhance osteoclastogenesis in periodontal disease

ABSTRACTPurpose of the study: Follicular dendritic cell-secreted protein (FDC-SP) has been found to be expressed in periodontal ligament (PDL), a layer of soft connective tissue between tooth root and alveolar bone, and involved in immunoreaction. This study was performed to explore the potential role of FDC-SP in periodontal disease. Materials and Methods: The human periodontal ligament cells (hPDLCs) were stimulated with Porphyromonas gingivalis (P. gingivalis) lipopolysaccharide (LPS) and FDC-SP expression was examined by real-time PCR and western blot. Then this molecule was overexpressed or silenced in hPDLCs by transfection of FDC-SP expression plasmids or its small-interfering (si) RNA, respectively, and the effects of FDC-SP on expression of osteogenesis- and osteoclastogenesis-related genes in hPDLCs were analyzed by real-time PCR and western blot. Results: Our results showed that P. gingivalis LPS upregulated FDC-SP expression in hPDLCs. Overexpression of FDC-SP could decrease the expression of osteogenesis-related genes, increase the expression of osteoclastogenesis-related genes and RANKL/OPG ratio in hPDLCs. Meanwhile, silence of FDC-SP expression in hPDLCs remarkably inversed the above results. Conclusions: LPS-induced upregulation of FDC-SP expression in hPDLCs may enhance osteoclastogenesis in periodontal disease.

Decreased expression of follicular dendritic cell–secreted protein correlates with increased immunoglobulin A production in the tonsils of individuals with immunoglobulin A nephropathy

Immunoglobulin A nephropathy (IgAN) is characterized by a qualitative abnormality of IgA in the circulation and IgA deposition in the renal mesangium. Recent research has indicated that pathogenic IgA may originate from affected tonsils. Follicular dendritic cell-secreted protein (FDC-SP), a small novel secretory protein that may regulate the induction of B-cell responses, has been suggested to control IgA production. Given this background, this study investigated the expression of FDC-SP and its correlation with IgA production in the tonsils of IgAN patients. Immunohistochemistry and reverse transcription-polymerase chain reaction were used to compare the expression of FDC-SP in the tonsils of IgAN patients with tonsillitis and of non-IgAN patients with chronic tonsillitis. The location of FDC-SP in tonsillar tissue was confirmed by double immunofluorescence. We found that FDC-SP expression significantly decreased and was correlated negatively with enhanced IgA production in the tonsils of IgAN patients. FDC-SP secreted by follicular dendritic cells may act on germinal center B cells and participate in the modulation of IgA generation in the tonsils. Our study demonstrated that FDC-SP may be involved in IgA production in the tonsils of IgAN patients, making this protein an attractive candidate immunomodulator, and highlighting a promising strategy for therapeutic intervention in IgAN.

Transfection With Follicular Dendritic Cell Secreted Protein to Affect Phenotype Expression of Human Periodontal Ligament Cells

Follicular dendritic cell secreted protein (FDC-SP), has been found to inhibit osteogenic differentiation of human periodontal ligament cells (hPDLCs) in recent studies. Based on these findings, we further investigate its effect on phenotype expression of hPDLCs in the present study, aiming to contribute to a better understanding of the biological functions governing FDC-SP-induced hPDLC differentiation. hPDLCs were firstly identified with immunocytochemical staining, followed by transfection with FDC-SP lentiviral vector. Western blot analysis was used to confirm the expression of FDC-SP. Then the influence of FDC-SP transfection on hPDLC proliferation, osteogenic and fibrogenic phenotype expression was evaluated at the mRNA and protein level. Procollagen type I c-peptide production was measured and alizarin red staining was then conducted to demonstrate effect of FDC-SP on functional differentiation. We found that hPDLCs could be successfully transfected with FDC-SP. Cell proliferation and cell cycle tests indicated that transfection with FDC-SP did not affect hPDLC proliferation. Moreover, according to real-time PCR and Western blot results, expression levels of type 1 collagen alpha 1, type 1 collagen alpha 2 and type 3 collagen were upregulated while that of osteocalcin, osteopontin, and bone sialoprotein were downregulated in FDC-SP transfected cells. In addition, hPDLCs overexpressing FDC-SP exhibited higher PIP production than the controls. Our findings demonstrate that transfection with FDC-SP has negligible adverse effect on proliferation of hPDLCs and imply the biological function of FDC-SP as a fibroblastic phenotype stabilizer by inhibiting hPDLCs differentiation into mineralized tissue-forming cells, thus regulating regeneration in periodontal tissue engineering.

Follicular dendritic cell secreted protein FDC-SP controls IgA production.

Follicular dendritic cell secreted protein (FDC-SP) is a secreted peptide predominantly expressed in mucosal tissues. We previously reported that FDC-SP transgenic mice have altered B-cell responses to systemic immunization; however, the role of FDC-SP in mucosal immunity is unknown. Here, we report that FDC-SP functions in regulating immunoglobulin A production. FDC-SP transgenic mice show decreased IgA levels in serum, saliva, and bronchoalveolar lavage fluid. Reciprocally, FDC-SP-deficient mice show significantly increased IgA levels in serum and intestinal lavage, associated with accumulation of IgA+ cells in blood, bone marrow, Peyer's patches, and lymph nodes. FDC-SP-deficient mice generated higher titers of antigen-specific IgA but normal IgG1 responses upon immunization. Purified FDC-SP transgenic B cells generated decreased IgA responses to transforming growth factor β (TGFβ)+interleukin 5 (IL5) stimulation. Consistent with a direct effect of FDC-SP on B cells, recombinant FDC-SP suppressed B-cell IgA production in vitro. Six- to 14-month-old FDC-SP-deficient mice show IgA deposition in kidney glomeruli, which was associated with proteinuria and pathology consistent with mild IgA nephropathy (IgAN). Our results demonstrate a novel biological activity of FDC-SP in controlling B-cell IgA production and identify FDC-SP-deficient mice as a novel mouse model of IgAN.

Osteogenic differentiation of human periodontal ligament cells after transfection with recombinant lentiviral vector containing follicular dendritic cell secreted protein

Follicular dendritic cell secreted protein (FDC-SP), has been identified in human periodontal ligament (PDL) in a recent study. It is suggested that the expression of FDC-SP might be associated with the osteogenic differentiation and mineralization of human periodontal ligament cells (hPDLCs). However, the intrinsic mechanism regarding this is still unclear. The aim of this study was to establish hPDLCs with safe and efficient overexpression of FDC-SP and to elucidate the influence of FDC-SP transfection on hPDLC osteogenesis in periodontal regeneration. We first applied a recombinant lentiviral vector containing FDC-SP to transfect hPDLCs via different multiplicity of infection (MOI) levels (1, 10, 20, 50 and 100). Western blot was performed to confirm the expression of FDC-SP. MTT assay was employed to evaluate the proliferation status of transfected cells. Then, the extent of osteogenic differentiation was investigated by simultaneous monitoring of alkaline phosphatase (ALP) activity assessment, immunofluorescent staining, the expression patterns of osteoblastic markers and mineralization staining. We found that hPDLCs transfected via MOI 20, 50 and 100 exhibited expression of FDC-SP protein compared with MOI 1 and 10. There was no significant effect of FDC-SP transfection (at different MOI levels of 1, 10 and 20) on the proliferation of hPDLCs, whereas higher MOI levels (50 and 100) inhibited cell proliferation ability. In addition, ALP activity decreased significantly in FDC-SP-transfected hPDLCs at day 7. When stained with alizarin red, cells overexpressing FDC-SP formed less mineralized nodules at 21d post-induction of differentiation, compared with the control cultures. Osteogenic inhibition was also confirmed by ALP immunostaining. Moreover, mRNA expression levels of osteoblastic markers decreased after FDC-SP transfection, which were in accordance with western blot results. Our data suggest that MOI 20 is optimal to transfect hPDLCs, which achieves safe and efficient overexpression of FDC-SP in transfected cells. Moreover, FDC-SP overexpression inhibits osteogenic differentiation of hPDLCs. The present study contributes to a better understanding of the biological functions governing FDC-SP-induced hPDLC differentiation.

Follicular dendritic cell‐secreted protein is decreased in experimental periodontitis concurrently with the increase of interleukin‐17 expression and the Rankl/Opg mRNA ratio

T-helper type 17 (Th17) cells produce interleukin-17 (IL-17) and help to protect against inflammation and infection in periodontal disease. Furthermore, while follicular dendritic cell-secreted protein (FDC-SP) may be involved in the inflammation of periodontal tissue, the biological role of FDP-SP in periodontal disease is still unknown. The purpose of the present study was to clarify the expression of IL-17 and FDC-SP in experimental periodontitis in rats. Seven-week-old male Wistar rats were divided into baseline control, sham and test groups. Experimental periodontitis was induced by placing a ligature in the mesiopalatal area, and untreated rats served as a baseline control group. Morphological changes in alveolar bone were investigated 7, 14 and 28 d after treatment. Expression of the Rankl, osteoprotegerin (Opg) and Il17 genes was analyzed 5 and 7 d after the induction of experimental periodontitis. Alveolar bone resorption progressed in the test group for 7 d, but not thereafter. At 5 d after the induction of periodontitis, the Rankl/Opg mRNA ratio and the expression of IL-17 in the test group were significantly increased compared with the respective values in the baseline control group; however, there were no significant differences between the test and control groups at 7 d. The expression of FDC-SP was significantly decreased in the test group compared with the baseline control group at 5 and 7 d after the induction of periodontitis, and this value had returned to normal levels at 14 and 28 d. These results suggest that both IL-17 and FDC-SP could be involved in the inflammatory response, and FDC-SP in the junctional epithelium might play an important role in the Th17 cell-related immune response.

Lipopolysaccharide induces rapid loss of follicular dendritic cell‐secreted protein in the junctional epithelium

Oshiro A, Iseki S, Miyauchi M, Terashima T, Kawaguchi Y, Ikeda Y, Shinomura T. Lipopolysaccharide induces rapid loss of follicular dendritic cell-secreted protein in the junctional epithelium. J Periodont Res 2012; 47: 689-694. © 2012 John Wiley & Sons A/S Background and Objective: We have previously reported that mRNA encoding follicular dendritic cell-secreted protein (FDC-SP) is expressed specifically in the junctional epithelium at the gingival crevice. Other tissues, such as tonsil, prostate gland and trachea, also express high levels of FDC-SP. These tissues participate in a range of functions closely related to innate immunity. Therefore, it is hypothesized that FDC-SP plays a crucial role in close association with the host defense system within the gingival crevice. Accordingly, the main aim of this study was to investigate the expression and localization of FDC-SP in and around the junctional epithelium and to observe the dynamic changes of FDC-SP in experimental inflammation. We examined, immunohistochemically, the expression of FDC-SP in the junctional epithelium using a specific antibody raised in rabbit after immunization with a synthetic peptide derived from the hydrophilic region of FDC-SP. Experimental inflammation was induced in the upper molars of Wistar rats by applying bacterial lipopolysaccharide (LPS; 5 mg/mL in sterile saline) for 1 h. We confirmed that FDC-SP is present in the junctional epithelium in a pattern that is consistent with the expression of FDC-SP mRNA. Of special interest is that no FDC-SP was detectable in the junctional epithelium 3 h after transient topical treatment with LPS. The presence of FDC-SP in the junctional epithelium and its loss after LPS treatment strongly support our hypothesis of FDC-SP playing a crucial role in close association with the host defense system within the gingival crevice.

Effect of FDC-SP on the phenotype expression of cultured periodontal ligament cells

IntroductionRecently, a novel protein, follicular dendritic cell secreted protein (FDC-SP), has been identified in human periodontal ligament (PDL) tissue and a biomolecular study suggested that the expression of FDC-SP might be associated with the expression of the PDL phenotype. The purpose of this study was to test the effect of FDC-SP on the proliferation and phenotype of PDL cells.Material and methodsPeriodontal ligament cells obtained following the 3rd passage were exposed to various concentrations of FDC-SP. The cell proliferation was monitored by 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide(MTT) assay. Then, as a measure of osteogenic activity, the alkaline phosphatase (ALP) activity was recorded after 4, 7, and 14 days using p-nitrophenylphosphate as a substrate. Finally, total RNA was extracted and RT-PCR was performed for gene analysis.ResultsThe results indicated that PDL cells exposed to 50 ng/ml FDC-SP could proliferate more rapidly. RT-PCR results showed that the mRNA expression of epidermal growth factor receptor (EGFR) was obviously upregulated and the mRNA expression of osteocalcin (OCN) and bone sialoprotein (BSP) were downregulated in PDL cells exposed to FDC-SP. Moreover, two groups of PDL cells exposed to FDC-SP showed a significant decrease of ALP activity during all the culture days.ConclusionsIn sum, the findings observed in this study suggest that FDC-SP in PDL cells could positively affect the proliferation and act as a fibroblastic phenotype stabilizer by inhibiting their differentiation into mineralized tissue-forming cells.