• BMB Reports
• /
• v.29 no.2
• /
• pp.127-132
• /
• 1996

Germinal centers (GCs) are formed in peripheral lymphoid tissues in response to protein antigens. In order to see if immunoglobulin isotype switching takes place in GC B-cells, we isolated GC B-cells (PNA positive cells) from mouse popliteal lymph nodes by a flow cytometer after the staining of lymph node cells with PNA-FITC and anti-B220-PE, and determined the expression of ${\gamma}1$ germline transcript and ${\gamma}1$ mRNA by RT-PCR. ${\gamma}1$ germline transcript and ${\gamma}1$ mRNA were amplified specifically in cDNAs from hybridoma expressing IgG1 or splenocytes stimulated LPS plus IL-4. Germinal center B-cells formed in popliteal lymph nodes of mice immunized with chicken ovalbumin were isolated 7 days after immunization. We sorted GC B-cells five times. Immunoglobulin ${\gamma}1$ germline transcripts were expressed in germinal center B-cells in three out of five sorts whereas two out of five sorts did not express ${\gamma}1$ germline transcripts in GC B-cells. The contents of GC B-cells ranged from 5 to 7% of total lymph node cells in most flow cytometric analyses but those of two sorted cells which did not express ${\gamma}1$ germline transcripts were out of normal range. These results imply that isotype switching of immunoglobulins may take place in GCs.

• BMB Reports
• /
• v.41 no.12
• /
• pp.863-867
• /
• 2008

CD320 has been recently discovered and reported as a follicular dendritic cell (FDC) protein. Although CD320 is known to enhance proliferation of germinal center (GC) B cells, little other information is available. In this study, we investigated its cellular distribution in the GC. Confocal microscopy of human tonsil sections revealed co-localization of CD320 with CD19 and CD38 but not with CD3 indicating that GC B cells expressed CD320 in addition to FDC. In purified GC B cells, CD320 expression was inhibited in the nucleus, membrane and cytoplasm. Reverse transcriptase-polymerase chain reaction confirmed CD320 mRNA expression in B cells. These finding indicate that CD320 is expressed in B cells in addition to FDC, and that its GC activity may be more complicated than previously thought.

• Microbiology and Biotechnology Letters
• /
• v.50 no.1
• /
• pp.63-70
• /
• 2022

Two small cryptic plasmids, pHG1 and pHG2, were isolated from Levilactobacillus zymae (formerly Lactobacillus zymae) GU240 and characterized. pHG1 is 1,814 bp in size with a GC content of 37.4% and contains two open reading frames. orf1 can potentially encode a protein of 101 amino acids (aa) with 99% identity with the copy number control protein of Lacticaseibacillus paracasei. orf2 can potentially encode a protein of 230 aa with 99% identity with a replication protein from multiple species. Six inverted repeats (IR I-VI) and six direct repeats (DR I-VI) were found in pHG1. pHG2 is 2,864 bp in size, with a GC content of 39.6%. pHG2 has two orfs. orf1 might encode a protein with 99% identity with the TrsL transmembrane protein. orf2 might encode a protein with 99% identity with plasmid recombination proteins from lactic acid bacteria. Both pHG1 and pHG2 may be useful as frames for constructing lactic acid bacteria-Escherichia coli shuttle vectors.

• Asian Pacific Journal of Cancer Prevention
• /
• v.15 no.3
• /
• pp.1375-1379
• /
• 2014

Background: Differences in clinicopathological characteristics of gastric cancer (GC) between young and older patients are controversial and a matter of debate. Determining the statistical significance of clinicopathological information with respect to age might provide clues for better management and treatment ofGC. Materials and Methods: A total ofl03 Indiao GC patients were enrolled for study and specimens were classified according to the AjCC-TNM system. Patients were grouped into two age-wise categories, young patients (<40 years; n=13) and older patients (${\geq}40$ years, n=90). The clinicopathological features of both groups were retrospectively examined and compared. p53 alterations were analyzed by polymerase chain reaction-single strand conformational polymorphism and immunohistochemistry methods at gene and protein levels respectively. The cases were considered p53 over-expressed if it was present in more than 25% of the tumor cells and p53 alterations was correlated with the clinicopathological characteristics of the patients as well as etiological factors for GC in both groups. Results: We found significant association of young patients with cancer stage (p=0.01), and very strong association with histology grade (p=0.064) and poorly differentiated (p=0.051) state of GC. However, neither young nor elderly patients showed associations with location, gender, etiological factors and p53 expression and alteration. Overall the male-to-female ratio of GC patients was 3.12 and the value was higher in the young (5.5) than in the older group (2.91). Conclusions: Clinicopathological features of GC like caocer stage, cell differentiation and histological grades were significantly different among young and old age cohorts. We observed a male predominance among the young group that decreased significantly with advancing age. More awareness of GC onset is required to detect cancer at an early stage for successful treatment.

• IMMUNE NETWORK
• /
• v.20 no.5
• /
• pp.42.1-42.10
• /
• 2020

Long-lasting post-switched plasma cells (PCs) arise mainly from germinal center (GC) reactions, but little is known about the mechanism by which GC B cells differentiate into PCs. Based on our observation that the expression of the transcription factor CCAAT/enhancer binding protein β (C/EPBβ) is associated with the emergence of post-switched PCs, we enquired whether a cell-autonomous function of C/EPBβ is involved in the program for PC development. To address this, we generated C/EPBβ-deficient mice in which the Cebpb locus was specifically deleted in B cells after transcription of the Ig γ1 constant gene segment (Cγ1). In response to in vitro stimulation, B cells from these Cebpb^fl/flCγ1^Cre/+ mice had defects in the induction of B lymphocyte-induced maturation protein 1 (Blimp1) and the formation of IgG1⁺ PCs, but not in proliferation and survival. At steady state, the Cebpb^fl/flCγ1^Cre/+ mice had reduced serum IgG1 titers but normal IgG2c and IgM titers. Moreover, upon immunization with T-dependent Ag, the mice produced reduced levels of Ag-specific IgG1 Ab, and were defective in the production of Ag-specific IgG1 Ab-secreting cells. These results suggest that a cell-autonomous function of C/EPBβ is crucial for differentiation of post-switched GC B cells into PCs through a Blimp1-dependent pathway.

• Asian-Australasian Journal of Animal Sciences
• /
• v.22 no.11
• /
• pp.1513-1520
• /
• 2009

Forty-six Holstein heifers were used in a completely randomized design and assigned to 1 of 2 treatments to evaluate the effects of 2 diets varying in ruminal fermentable carbohydrate sources, namely ground corn (GC) and rolled wheat (RW), on metabolism and performance of primiparous cows in the periparturient period. The heifers were fed diets as a total mixed ration (TMR) with similar energy and crude protein content including i) 18.57% GC, or ii) 18.57% RW from -24.13${\pm}$7.73 d relative to expected calving until calving. After calving, all animals received the same lactation diet until 28 d. Animals were group fed from the beginning of the study to -7 d relative to expected calving, fed individually from d -7 to 7 days in milk (DIM), and again group fed to 28 DIM. The pre-partum diets affected (p<0.05) dry matter intake (DMI), energy intake, energy balance (EB) and urinary pH during the last week pre-partum. There was no effect of pre-partum carbohydrate source on overall plasma concentration of glucose, nonesterified fatty acid (NEFA), $\beta$-hydroxybutyrate (BHBA), albumin, triglyceride (TG), cholesterol, aspartate aminotransferase (AST), insulin, and cortisol during the periparturient period. Cows fed the RW diet during the pre-partum period had greater calcium for the first week (p<0.05) and during 28 d (p = 0.08) of lactation compared with heifers fed the GC diet. Primiparous cows fed the RW diet produced greater milk protein content and yield (p<0.05). Primiparous cows fed the RW diet had lower milk urea nitrogen (MUN) and somatic cell count (SCC) than cows fed the GC diet (p<0.05). The results of this study show that feeding pre-partum diets with a rapidly fermentable source of starch but low energy content can improve animal metabolism and performance and smooth the transition of primiparous Holstein cows from gestation to lactation.

• Asian Pacific Journal of Cancer Prevention
• /
• v.15 no.6
• /
• pp.2663-2667
• /
• 2014

Background: Single nucleotide polymorphisms of C-reactive protein (CRP) have been shown to be related to circulating CRP level, risk and prognosis in cancer patients. However, accumulating evidence of rs1800947 involvement in risk of cancer is inconsistent. Thus, a meta-analysis was performed to obtain a more precise relationship. Materials and Methods: The pooled odds ratio (OR) and its 95% confidence interval were assessed in 10 eligible articles with 12 studies containing 5,601 cancer cases and 8,669 cancer-free controls. Results: No significant association was observed overall and in subgroups in comparison of genotype GC vs GG ($P_H$=0.847, OR=0.939, 95%CI=0.810-1.087), GC/CC vs GG ($P_H$=0.941, OR=1.021, 95%CI=0.901-1.157) and allele C vs G ($P_H$=0.933, OR=1.026, 95%CI=0.909-1.159). However, statistically significance was evident in comparison of genotype CC vs GG in cancer risk ($P_H$=0.586, OR=2.854, 95%CI= 1.413-5.763), especially in colorectal cancer ($P_H$=0.481, OR=4.527, 95%CI= 1.664- 12.315). Conclusions: Genotype CC of rs1800947 in the CRP gene is strongly associated with increased cancer risk, particularly in colorectal cancer.

• Molecules and Cells
• /
• v.41 no.7
• /
• pp.653-664
• /
• 2018

The RNA-binding protein tristetraprolin (TTP) binds to adenosine-uridine AU-rich elements in the 3'-untranslated region of messenger RNAs and facilitates rapid degradation of the target mRNAs. Therefore, it regulates the expression of multiple cancer and immunity-associated transcripts. Furthermore, a lack of TTP in cancer cells influences cancer progression and predicts poor survival. Although the functions of TTP on cancer cells have previously been researched, the mechanism of TTP on the interaction between cancer cells with their micro-environment remains undiscovered. In this study, we admed to determine the role of cancer cell TTP during the interaction between tumor and immune cells, specifically regulatory T cells (Tregs). We evaluate the capability of TTP to modulate the antitumor immunity of GC and explored the underlying mechanism. The overexpression of TTP in GC cells dramatically increased peripheral blood mononuclear lymphocyte (PBML) -mediated cytotoxicity against GC cells. Increased cytotoxicity against TTP-overexpressed GC cells by PBMLs was determined by Treg development and infiltration. Surprisingly, we found the stabilization of programmed death-ligand 1 (PD-L1) mRNA was declining while TTP was elevated. The PD-L1 protein level was reduced in TTP-abundant GC cells. PD-L1 gas been found to play a pivotal role in Treg development and functional maintenance in immune system. Taken together, our results suggest the overexpression of TTP in GC cells not only affects cell survival and apoptosis but also increases PBMLs -mediated cytotoxicity against GC cells to decelerate tumor progression. Moreover, we identified PD-L1 as a critical TTP-regulated factor that contributes to inhibiting antitumor immunity.

• The Korean Journal of Physiology and Pharmacology
• /
• v.9 no.5
• /
• pp.275-282
• /
• 2005

By using differential display, we identified one of the genes encoding the multi-subunit complex protein V-ATPase, c subunit gene (ATP6L), and showed alterations of the gene expression by oxidative stresses. Expression of the ATP6L gene in Neuro-2A cells was increased by the treatment with $H_2O_2$ and incubation in hypoxic chamber, implying that the expression of the ATP6L gene is regulated by oxidative stresses. To examine mechanisms involved in the regulation of the gene expression by oxidative stresses, the transcriptional activity of the rat ATP6L promoter was studied. Transcription initiation site was determined by primer extension analysis and DNA sequencing, and promoter of the rat ATP6L and its deletion clones were constructed in reporter assay vector. Significant changes of the promoter activities in Neuro-2A cells were observed in two regions within the proximal 1 kbp promoter, and one containing a suppressor was in -195 to -220, which contains GC box that is activated by binding of Sp1 protein. The suppression of promoter activity was lost in mutants of the GC box. We confirmed by electrophoretic mobility shift and supershift assays that Sp1 protein specifically binds to the GC box. The promoter activity was not changed by the $H_2O_2$ treatment and incubation in hypoxic chamber, however, $H_2O_2$ increased the stability of ATP6L mRNA. These data suggest that the expression of the ATP6L gene by oxidative stresses is regulated at posttranscriptional level, whereas the GC box is important in basal activities of the promoter.

• Journal of Ginseng Research
• /
• v.40 no.2
• /
• pp.185-195
• /
• 2016

Background: To investigate the effect of pectinase-treated Panax ginseng (GINST) in cellular and male subfertility animal models. Methods: Hydrogen peroxide ($H_2O_2$)-induced mouse spermatocyte GC-2spd cells were used as an in vitro model. Cell viability was measured using MTT assay. For the in vivo study, GINST (200 mg/kg) mixed with a regular pellet diet was administered orally for 4 mo, and the changes in the mRNA and protein expression level of antioxidative and spermatogenic genes in young and aged control rats were compared using real-time reverse transcription polymerase chain reaction and western blotting. Results: GINST treatment ($50{\mu}g/mL$, $100{\mu}g/mL$, and $200{\mu}g/mL$) significantly (p < 0.05) inhibited the $H_2O_2$-induced ($200{\mu}M$) cytotoxicity in GC-2spd cells. Furthermore, GINST ($50{\mu}g/mL$ and $100{\mu}g/mL$) significantly (p < 0.05) ameliorated the $H_2O_2$-induced decrease in the expression level of antioxidant enzymes (peroxiredoxin 3 and 4, glutathione S-transferase m5, and glutathione peroxidase 4), spermatogenesis-related protein such as inhibin-${\alpha}$, and specific sex hormone receptors (androgen receptor, luteinizing hormone receptor, and follicle-stimulating hormone receptor) in GC-2spd cells. Similarly, the altered expression level of the above mentioned genes and of spermatogenesis-related nectin-2 and cAMP response element-binding protein in aged rat testes was ameliorated with GINST (200 mg/kg) treatment. Taken together, GINST attenuated $H_2O_2$-induced oxidative stress in GC-2 cells and modulated the expression of antioxidant-related genes and of spermatogenic-related proteins and sex hormone receptors in aged rats. Conclusion: GINST may be a potential natural agent for the protection against or treatment of oxidative stress-induced male subfertility and aging-induced male subfertility.