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Immunoglobulin G Expression in Human Sperm and Possible Functional Significance.

ABSTRACT: Immunoglobulin G (IgG), the major molecule of the immune system, which was traditionally thought to be produced by differentiated B-lymphocytes, had recently been found in non-immune cells including spermatozoa of rabbit testis. To study if human sperms could produce IgG that might play a role in fertilization, we employed immunofluorescent staining, Western blot, in situ hybridization, RT-PCR (reverse transcription polymerase chain reaction) and immunoelectron microscope and found that human sperms were capable of synthesizing IgG. IgG protein and mRNA were detected in the cytoplasm, mainly the neck region of the sperm and IgG immunoreactivity was found to cover the entire sperm cell. The essential enzymes necessary for IgG synthesis and class switching, RAG1 (recombination activating gene 1), RAG2 (recombination activating gene 2) and AID (activation-induced cytidine deaminase), were also detected in the sperm cells. Furthermore, we found that anti-IgG antibody could inhibit sperm from penetrating Zona-free hamster egg with statistical significance. These discoveries suggested that immunoglobulin G could be produced by human sperms and it might play a role during fertilization.

SUBMITTER: Yan M

PROVIDER: S-EPMC4735602 | biostudies-other | 2016 Feb

REPOSITORIES: biostudies-other

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Immunoglobulin G Expression in Human Sperm and Possible Functional Significance.

Yan Meiling M Zhang Xiaoyu X Pu Qinxue Q Huang Tao T Xie Qingdong Q Wang Yan Y Li Jing J Wang Yun Y Gu Huan H Huang Tianhua T Li Zhiling Z Gu Jiang J

Scientific reports 20160201

Immunoglobulin G (IgG), the major molecule of the immune system, which was traditionally thought to be produced by differentiated B-lymphocytes, had recently been found in non-immune cells including spermatozoa of rabbit testis. To study if human sperms could produce IgG that might play a role in fertilization, we employed immunofluorescent staining, Western blot, in situ hybridization, RT-PCR (reverse transcription polymerase chain reaction) and immunoelectron microscope and found that human sp ...[more]

PMID: 26833114

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Project description:BACKGROUND:High expression of the polymeric immunoglobulin receptor (PIGR) has previously been associated with a favourable prognosis in a few cancer forms, but its expression and relationship with clinical outcome in epithelial ovarian cancer (EOC) has not yet been reported. The aim of this study was therefore to examine the clinicopathological correlates and prognostic significance of PIGR expression in EOC. METHODS:After an initial screening in the Human Protein Atlas portal, a validated antibody was selected for extended analysis of immunohistochemical PIGR expression in tissue microarrays with tumours from 154 incident cases of EOC from two pooled prospective population-based cohorts. Subsets of corresponding benign-appearing fallopian tubes (n?=?38) and omental metastases (n?=?33) were also analysed. Kaplan-Meier analysis and Cox regression analysis were applied to examine the impact of PIGR expression on overall survival (OS) and ovarian cancer-specific survival (OCSS). RESULTS:PIGR expression was significantly higher in fallopian tubes compared to primary tumours and metastases (p?<?0.001) and lower in carcinoma of the serous subtype compared to other carcinomas (p?<?0.001). PIGR expression was significantly associated with lower grade (p?=?0.001), mucinous histological subtype (p?=?0.002), positive progesterone receptor expression (p?=?0.009) and negative or low Ki-67 expression (p?=?0.003). Kaplan-Meier analysis revealed a significantly improved OS (p?=?0.013) and OCSS (p?=?0.009) for patients with tumours displaying high expression of PIGR. These associations were confirmed in unadjusted Cox regression analysis (HR?=?0.48; 95% CI 0.26-0.87; p?=?0.015 for OS and HR?=?0.43, 95% CI 0.22-0.82; p?=?0.011 for OCSS) but did not remain significant after adjustment for age, grade and clinical stage. CONCLUSIONS:This study provides a first demonstration of PIGR expression in human fallopian tubes, primary EOC tumours and metastases. High tumour-specific expression of PIGR was found to be associated with a favourable prognosis in unadjusted, but not in adjusted, analysis. These findings are novel and merit further investigation.

Project description:Hemoglobin (Hb) is a major protein involved in transport of oxygen (O2). It consists of Hb-? and Hb-? subunits, which are normally expressed by cells of erythroid lineage. However, till recently, it was not known whether non-erythroid cells like vaginal cells synthesize Hb and whether it has any functional significance. Therefore, we designed the following objectives: (1) to establish in-vitro culture system of human primary vaginal epithelial cells (hPVECs), (2) to determine whether Hb-? and Hb-? proteins are truly synthesized by hPVECs, (3) to evaluate the effect of LPS (lipopolysaccharide) on the expression of Hb-? and Hb-? proteins (4) to decipher the significance of the Hb-? and Hb-? expression in hPVECs and (5) to determine the molecular mechanism regulating the expression of Hb-? in hPVECs. To accomplish these studies, we applied a battery of assays such as RT-PCR, qRT-PCR, Flow cytometry, western blot, and immunofluorescence, Electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP). The results revealed the expression of Hb-? and Hb-? at both mRNA and protein level in hPVECs. The expression was significantly upregulated following LPS treatment (10?g/ml for 6 hrs) and these results are comparable with the expression induced by LPS in human vaginal epithelial cell line (VK2/E6E7). These cells constitutively produced low levels of pro-inflammatory (IL-6) and anti-inflammatory (IL-10) cytokines. Also, the response of phosphorylated (p65)-NF-?B to LPS was upregulated with increased expression of IL-6, Toll-like receptor-4 (TLR4) and human beta defensin-1 (hBD-1) in hPVECs and VK2/E6E7 cells. However, Bay 11-7082 treatment (5?M for 24 hrs) could neutralize the effect of LPS-induced p65-NF-?B activity and represses the production`of Hb-? and Hb-?. The results of EMSA revealed the presence of putative binding sites of NF-?B in the human Hb-? promoter region (nt-115 to -106). ChIP analysis confirmed the binding of NF-?B to Hb-? promoter. In conclusion, the present findings revealed for the first time that hPVECs synthesized Hb-? and Hb-? and the expression is comparable with the expression of VK2/E6E7 cells. The identification of NF-?B regulatory sequences in Hb-? promoter, whose activation is associated with immune response of hPVECs, indicating Hb-? and Hb-? may act as an endogenous antimicrobial defense protein against vaginal inflammation/infections.

Project description:INTRODUCTION: The polymeric immunoglobulin receptor (PIGR) has been proposed to be a candidate prognostic biomarker in a few cancer forms, and one previous study reported that reduced PIGR expression signifies more aggressive tumours of the distal esophagus and gastroesophageal junction (GEJ). In the present study, we examined the expression, clinicopathological correlates and prognostic significance of PIGR expression in an extended cohort of adenocarcinoma of the upper gastrointestinal tract. MATERIALS AND METHODS: Immunohistochemical PIGR expression was examined in a consecutive cohort of patients with surgically resected, radio-chemonaive adenocarcinoma of the esophagus, GE-junction and stomach (n?=?173), including paired samples of benign-appearing squamous epithelium (n?=?51), gastric mucosa (n?=?114), Barrett's esophagus (BE) or intestinal metaplasia (IM) (n?=?57) and lymph node metastases (n?=?75). Non-parametric tests were applied to explore associations between PIGR expression in primary tumours and clinicopathological characteristics. Classification and regression tree analysis was applied for selection of prognostic cut-off. The impact of PIGR expression on overall survival (OS) and recurrence-free survival (RFS) was assessed by Kaplan-Meier analysis and hazard ratios (HR) calculated by adjusted and unadjusted Cox proportional hazards modelling. RESULTS: PIGR expression was significantly higher in intestinal metaplasia (BE or gastric IM) compared to normal tissues and cancer (p?<?0.001). Reduced PIGR expression in primary tumours was significantly associated with more advanced tumour stage (p?=?0.002) and inversely associated with involved margins (p?=?0.034). PIGR expression did not differ between primary tumours and lymph node metastases. There was no significant difference in PIGR expression between tumours with and without a background of intestinal metaplasia. High PIGR expression was an independent predictor of a prolonged OS (HR?=?0.60, 95% CI 0.36-0.99) and RFS (HR?=?0.49, 95% CI 0.27-0.90) in patients with radically resected (R0) primary tumours and of an improved RFS (HR?=?0.32, 95% CI 0.15-0.69) in curatively treated patients with R0 resection/distant metastasis-free disease. CONCLUSION: High PIGR expression independently predicts a decreased risk of recurrence and an improved survival in patients with adenocarcinoma of the upper gastrointestinal tract. These findings are of potential clinical relevance and merit further validation.

Dataset Information

Immunoglobulin G Expression in Human Sperm and Possible Functional Significance.

Publications

Immunoglobulin G Expression in Human Sperm and Possible Functional Significance.

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