scholarly journals Study on the immunogenicity of pcDNA3.1(+)-cagT recombinant vector against Helicobacter pylori in BALB/c mice

2020 ◽  
Vol 22 (4) ◽  
pp. 159-166
Author(s):  
Armita Balash ◽  
Abbas Doosti

Background and aims: The role of Helicobacter pylori in the development of gastric ulcer and gastrointestinal cancer was identified in this study. More precisely, the study focused on the creation of a DNA vaccine based on the cagT gene of this bacterium and the investigation of its immunogenicity against H. pylori in infused BALB/c mice. Materials and Methods: To this end, the pcDNA3.1(+)-cagT was prepared and transformed into Escherichia coli. Then, animals were injected with recombinant pcDNA3.1(+)-cagT plasmid, pcDNA3.1(+)-cagT + nanoparticles, and pcDNA3.1(+). After the plasmid purification and confirmation of the transformation by digestion and polymerase chain reaction (PCR), chitosan nanoparticles were synthesized using the ionic gelation method. Next, the animals were classified into three groups each including 21 mice. The injectable solutions including pcDNA3.1(+)-cagT, pcDNA3.1(+)-cagT + nanoparticles, or empty pCDNA3.1 (as a control group) were injected into the quadriceps muscle of mice, separately. Finally, the blood and tissue samples of each mouse were collected 15, 30, and 45 days after the last injection, and the expression levels of transforming growth factor-beta (TGF-β1), interleukin-4 (IL-4), and interferon-gamma (IFNγ) were evaluated by real-time PCR. Results: The IFNγ and TGF-β1 expression increased in the infused mice (P<0.01) while the IL4 expression represented a significant decrease (P<0.01). Moreover, the IFNγ and IL4 expression level in pcDNA3.1(+)-cagT + nanoparticle significantly altered (P<0.01) compared to the pcDNA3.1(+)-cagT group although the TGF-β1 expression was not significantly different (P=0.075). Contrarily, the cagT gene expression in the tissue samples of both groups was significantly different 15, 30, and 45 days after the last injection (P<0.01). Eventually, the expression of the cagT gene in the infused mice by pcDNA3.1(+)-cagT and in the nanoparticle group was not significantly different 45 days after the last injection (P=0.105). Conclusion: In general, the decrease of IL-4 expression was observed in the injected mice by pcDNA3.1(+)-cagT and indicated that the immune system work by a Th1 pattern. The findings showed that a pcDNA3.1(+)-cagT construct combined with chitosan nanoparticles can increase the stimulation of the immune system in an animal model and thus it can be used as an appropriate method for controlling H. pylori infection.

2003 ◽  
Vol 10 (4) ◽  
pp. 542-545 ◽  
Author(s):  
Peer R. E. Mittl ◽  
Lucas Lüthy ◽  
Christoph Reinhardt ◽  
Hellen Joller

ABSTRACT The family of Helicobacter cysteine-rich proteins (Hcp) constitutes one of the largest protein families that are specific for proteobacteria from the delta/epsilon subgroup. Most of the proteins belonging to this family have so far only been recognized on the genome level. To investigate the expression of Hcp proteins in vivo we analyzed titers of antibody against HcpA (HP0211), HcpB (HP0336), HcpC (HP1098), and HcpE (HP0235) in sera from 30 Helicobacter pylori-positive individuals and in a control group of six H. pylori-negative individuals. Significantly higher titers of antibody were observed for H. pylori-positive individuals (P < 0.00005). The highest and lowest titers were observed for HcpC (Δ mean = 1.06) and HcpB (Δ mean = 0.333), respectively. There is a clear correlation among anti-HcpA, -HcpC, and -HcpE immunoglobulin G titers in H. pylori-positive individuals (correlation > 0.7), but there is only a weak correlation for HcpB (correlation < 0.4). These results confirm that Hcp proteins are expressed by H. pylori under natural environmental conditions and that these proteins are recognized by the immune system of the host. The observed correlations are in agreement with the expected distribution of Hcp proteins among H. pylori strains. HcpA, HcpC, and HcpE are present in the genomes of strains 26695 and J99, whereas HcpB is absent from most strains. Since Hcp proteins are specific for H. pylori, immunological assays including Hcp proteins might be of value to detect H. pylori infection and perhaps to distinguish among different groups of H. pylori-positive patients.


2020 ◽  
Vol 4 (1) ◽  
pp. 21-25
Author(s):  
Noor Alhuda K. Ibrahim ◽  
Wasnaa J. Mohammad ◽  
Sanan T. Abdawahab

Polycystic ovary syndrome (PCOS) is one of the most common causes of infertility in women of reproductive age. The aim of the study was to determine the level of apelin, insulin resistance (IR), transforming growth factor beta 1 (TGF-β1), and endoglin in women with polycystic ovary syndrome. Fifty PCOS patients and 40 non-PCOS infertile patients were recruited. The fasting serum levels of folliclestimulating hormone (FSH), luteinizing hormone (LH), testosterone (T), prolactin, fasting blood glucose, insulin, and apelin at the early follicular phase were measured. Levels of apelin, LH, LH/FSH, T, and fasting insulin, as well as homeostatic model assessment of IR (HOMA-IR) in PCOS patients, were significantly higher than in the control group. Correlation analysis showed that apelin level was positively correlated with body mass index and HOMA-IR. Apelin levels and TGF-β1 were significantly increased in PCOS patients while show decrease levels of endoglin.


Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1844
Author(s):  
Marta Rybska ◽  
Magdalena Woźna-Wysocka ◽  
Barbara Wąsowska ◽  
Marek Skrzypski ◽  
Magdalena Kubiak ◽  
...  

Cystic endometrial hyperplasia (CEH) and pyometra are the most frequently diagnosed uterine diseases affecting bitches of different ages. Transforming growth factor beta (TGF-β) has been classified in females as a potential regulator of many endometrial changes during the estrous cycle or may be involved in pathological disorders. The aim of this study was to determine the expression of TGF-β1, -β2 and -β3 in the endometrium of bitches suffering from CEH or a CEH–pyometra complex compared to clinically healthy females (control group; CG). A significantly increased level of TGF-β1 mRNA expression was observed in the endometrium with CEH–pyometra compared to CEH and CG. Protein production of TGF-β1 was identified only in the endometrium of bitches with CEH–pyometra. An increase in TGF-β3 mRNA expression was observed in all the studied groups compared to CG. The expression of TGF-β2 mRNA was significantly higher in CEH and lower in CEH–pyometra uteri. The results indicate the presence of TGF-β cytokines in canine endometrial tissues affected by proliferative and degenerative changes. However, among all TGF-β isoforms, TGF-β1 could potentially be a key factor involved in the regulation of the endometrium in bitches with CEH–pyometra complex.


2018 ◽  
Vol 12 (03) ◽  
pp. 358-362
Author(s):  
Nike Hendrijantini ◽  
Tuti Kusumaningsih ◽  
Rostiny Rostiny ◽  
Pungky Mulawardhana ◽  
Coen Pramono Danudiningrat ◽  
...  

ABSTRACTObjective: The aim of this study is to prove that human umbilical cord mesenchymal stem cell (hUCMSC) therapy on mandibular osteoporotic model is able to increase transforming growth factor-beta-1 (TGF)-β1 expression, Runx2, and osteoblasts. Materials and Methods: This research is true experimental posttest control group design. Thirty female Wistar rats were divided into 6 groups randomly, which consisted of sham surgery for control (T1), ovariectomy as osteoporotic group (T2), osteoporotic group injected with gelatine for 4 weeks (T3), 8 weeks (T4) injected with hUCMSC-gelatine for 4 weeks (T5) and 8 weeks (T6). All mice were presented for immunohistochemistry examination for TGF-β1, Runx2, and histology for osteoblasts. Results: The lowest level of osteoblast was osteoporotic group injected with gelatine in 4 weeks compared to other groups. There were increases of TGF-β1, Runx2, and osteoblasts from osteoporotic group compared to osteoporotic post-hUCMSC-gelatine injection group. Conclusion: The hUCMSC has a high osteogenic effect and increases the osteoporotic mandibular bone regeneration on the animal model that is showed by the increase of the level of TGF-β1, Runx2, and osteoblasts.


2017 ◽  
Vol 131 (6) ◽  
pp. 549-556 ◽  
Author(s):  
N Siupsinskiene ◽  
I Katutiene ◽  
V Jonikiene ◽  
D Janciauskas ◽  
S Vaitkus

AbstractObjective:To identify Helicobacter pylori infection in tonsillar tissue samples from patients undergoing tonsillectomy for chronic tonsillitis versus tonsillar hypertrophy, and to assess the possible relationships between H pylori and patients’ sociodemographic data and laryngopharyngeal reflux.Methods:In this prospective study, 97 patients who underwent tonsillectomy were divided into the following 2 groups: patients with chronic tonsillitis (n = 62) and patients with tonsillar hypertrophy (control group; n = 35). H pylori infection in the tonsillar biopsy samples was identified using histochemical and rapid urease tests.Results:The incidence of H pylori infection was significantly higher in the chronic tonsillitis group (56.5 per cent) compared to the control group (31.4 per cent). Similar findings were obtained for both subgroups of adults (68.6 vs 42.3 per cent) and children (40.7 vs 0.0 per cent). Significant relationships between a positive H pylori finding and laryngopharyngeal reflux related signs of vocal fold oedema, diffuse laryngeal oedema and hypertrophy of the posterior commissure were revealed.Conclusion:H pylori infection may be related to chronic tonsillitis and laryngopharyngeal reflux.


2020 ◽  
Vol 8 (A) ◽  
pp. 297-301
Author(s):  
Hernita Taurustya ◽  
Mae Sri Hartati Wahyuningsih ◽  
Indwiani Astuti

BACKGROUND: Keloid is a benign fibroproliferative dermis tumor characterized by an increase in growth factors which induce fibroblast proliferation, excessive migration, and synthesis of collagen. Nerium indicum Mill. extract had been studied as a keloid therapy agent. 5α-oleandrin contained in N. indicum has antikeloid activity by inhibiting keloid fibroblast proliferation, fibroblast migration, collagen deposition, and transforming growth factor beta-1 (TGF-β1) synthesis. OBJECTIVE: This study aimed to determine the effect of administration of N. indicum extract on TGF-β1 and vascular endothelial growth factor (VEGF) expression in keloid fibroblast. METHODS: This research was a quasi-experimental research with a post-test only control group design. The research subjects were fibroblast cells passage IV-VII isolated from patients’ keloid tissue with explant techniques. Treatment groups received N. indicum extract with a serial concentration of 2 μg/ml, 1 μg/ml, and 0.5 μg/ml, and control group received medium only. The supernatant was obtained after 72 h incubation period. Examination of TGF-β1 and VEGF expressions was performed using ELISA procedure. RESULT: The expression of TGF-β1 in the treatment groups of the extract N. indicum (2 μg/ml, 1 μg/ml, and 0.5 μg/ml) was significantly lower than a control group of keloid fibroblasts (p < 0.05), according to increased concentration. VEGF expression in the treatment groups of N. indicum extract was lower compared to the control group of keloid fibroblasts. A significant decrease in keloid fibroblast VEGF levels occurred at extract concentrations of 2 μg/ml and 1 μg/ml (p < 0.05). CONCLUSION: N. indicum extract could decrease TGF-β1 and VEGF expressions compared to control medium in keloid fibroblast cultures.


2021 ◽  
Vol 11 (4) ◽  
pp. 731-735
Author(s):  
Yuchen Wang ◽  
Zhimin Huang

Gastric cancer (GC) is a common tumor with high incidence and poor prognosis. So far, the pathogenesis of GC has not been fully elucidated, which has brought great difficulty to the treatment. TGF-β regulates cell growth and differentiation. As a key member, TGF-β1 is abnormally expressed in various tumors, but its role on GC and related mechanisms have not been elucidated. Gastric cancer and adjacent tissues were collected to measure TGF-β1 level by real-time PCR. SGC-7901 cell was assigned into control group, mock group, and TGF-β1 siRNA group followed by analysis of TGF-β1 level by ELISA, cell proliferation by MTT assay, apoptosis by flow cytometry, cell migration by cell scratch test, cell invasion by Transwell chamber assay, and Bcl-2, Bax, and PTEN level by Western blot. TGF-β1 was significantly upregulated in GC tissues (P <0.05) and increased with TNM stage dependence. TGF-β1 siRNA transfection significantly decreased TGF-β1 mRNA level and secretion, inhibited cell proliferation, increased apoptosis rate, and attenuated cell migration and invasion along with downregulated Bcl-2 and elevated Bax and PTEN expression (P <0.05). Downregulation of TGF-β1 can promote gastric cancer cell apoptosis, inhibit proliferation, migration, and invasion by regulating PTEN.


2017 ◽  
Vol 85 (4) ◽  
Author(s):  
Rungtiwa Dangtakot ◽  
Somchai Pinlaor ◽  
Upsornsawan Itthitaetrakool ◽  
Apisit Chaidee ◽  
Chariya Chomvarin ◽  
...  

ABSTRACT Persistent infection with Opisthorchis viverrini causes hepatobiliary abnormalities, predisposing infected individuals to cholangiocarcinoma (CCA). In addition, Helicobacter pylori is highly prevalent in most countries and is a possible risk factor for CCA; however, its role in enhancing hepatobiliary abnormality is unclear. Here, we investigated the effects of coinfection with H. pylori and O. viverrini on hepatobiliary abnormality. Hamsters were divided into four groups: (i) normal, (ii) H. pylori infected (HP), (iii) O. viverrini infected (OV), and (iv) O. viverrini and H. pylori infected (OV+HP). At 6 months postinfection, PCR and immunohistochemistry were used to test for the presence of H. pylori in the stomach, gallbladder, and liver. In the liver, H. pylori was detected in the following order: OV+HP, 5 of 8 (62.5%); HP, 2 of 5 (40%); OV, 2 of 8 (25%). H. pylori was not detected in normal (control) liver tissues. Coinfection induced the most severe hepatobiliary abnormalities, including periductal fibrosis, cholangitis, and bile duct hyperplasia, leading to a significantly decreased survival rate of experimental animals. The greatest thickness of periductal fibrosis was associated with a significant increase in fibrogenesis markers (expression of alpha smooth muscle actin and transforming growth factor beta). Quantitative reverse transcription-PCR revealed that the highest expression levels of genes for proinflammatory cytokines (interleukin-1 [IL-1], IL-6, and tumor necrosis factor alpha) were also observed in the OV+HP group. These results suggest that coinfection with H. pylori and O. viverrini increased the severity of hepatobiliary abnormalities to a greater extent than either single infection did.


2021 ◽  
Vol In Press (In Press) ◽  
Author(s):  
Tuan Van Nguyen ◽  
Ky Duc Ngo ◽  
Minh Hoang Thi ◽  
Lan Thi Phuong Dam ◽  
Thuan Quang Huynh

Background: The transforming growth factor-beta 1 (TGF-β1) has been demonstrated as one of the main factors in the progression of fibrosis and sclerosis glomerular damages. Glomerulonephritis is one common cause of chronic kidney disease (CKD) with the promotion of inflammatory renal damage containing fibrosis and sclerosis glomerular. Objectives: This study aimed to evaluate the TGF-β1 level in CKD patients and compare it with the healthy control group. Methods: This cross-sectional case-control study was carried out on 212 subjects admitted to the Nghe An Friendship General Hospital in Vietnam from March 2018 to February 2020. The case group included 152 patients diagnosed with CKD caused by glomerulonephritis, and the control group included 60 healthy individuals. The TGF-β1 was determined in serum by ELISA method. Results: The serum TGF-β1 concentration of the healthy control group and CKD group was 13.45 ± 7.17 and 32.35 ± 11.74, respectively. The CKD group had a significantly higher TGF-β1 level than the control group (P < 0.05). The CKD group with the eGRP ≥ 60 mL/min/1.73 m2 group had a higher TGF-β1 level than the eGRP < 60 mL/min/1.73 m2 group, and the TGF-β1 level increased from stage 1 to stage 5 (P < 0.001). The TGF-β1 had a medium correlation to urea, creatinine, and hs-CRP. Conclusions: The concentration of TGF-β1 in the CKD group was higher than the control group so that it increased early from the first stage of the disease.


2017 ◽  
Vol 3 (1) ◽  
pp. 15 ◽  
Author(s):  
Hasret Ecevit ◽  
Kubra Gunduz ◽  
Nilufer Bilgic ◽  
Muzeyyen Izmirli ◽  
Bulent Gogebakan

<p>Thymoquinone, one of the essential oil in the structure of cumin, is used for alternative therapy for many diseases from past to present. It was shown to have anti-carcinogenic and anti-inflammatory effects, as well as positive effects on fibrosis. However, there is no study on the effect of thymoquinone on cancer and fibrosis mechanism in bronchial epithelium cell line BEAS-2B. In our study, the effect of thymoquinone on cell viability and transforming growth factor-beta 1 (TGF-β1) level, which has an important role in the regulation of many biological processes including cancer and fibrosis-associated signal transduction, was evaluated. BEAS-2B cells were exposed to thymoquinone at 0–80 μmol/L concentrations for 24-, 48- and 72-hour durations. Cell viability was evaluated with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test. TGF-β1 level was determined with enzyme-linked immunosorbent assay (ELISA) method from the collected supernatant. Cell viability was found to be increased at all concentrations and durations (10–80 μmol/L; 24, 48 and 72 h) according to the control group (0 μmol/L; thymoquinone in ethanol) (p &lt; 0.0001). Moreover, thymoquinone was found to increase the level of TGF-β1 only at 80 μmol/L concentration and 24-hour exposure period (0 μmol/L, 53.41 ± 18.44 pgr/ml TGF-β1; 80 μmol/L, 174.5 ± 80.03 pgr/ml TGF-β1). As a result, thymoquinone was found to increase cell proliferation and encourage TGF-β1 release.</p>


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