Preparation and identification of polyclonal antibody against human cytomegalovirus encoding protein UL23

Chai, Huizi; Wu, Songbin; Deng, Jiaming; Feng, Linyuan; Yang, Xiaoping; Ran, Yanhong; Li, Hongjian

doi:10.1016/j.pep.2019.04.008

Cited by 6 publications

(3 citation statements)

References 10 publications

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“…Mouse antitubulin (66031-1-Ig), anti-STAT2 (16674-1-AP), anti-IRF9 (14167-1-AP), and rabbit anti-Jak1 (66466-1-Ig) were provided by (Manchester, United Kingdom) Rabbit anti-Tyk2 (A2128) was purchased from (Wuhan, China), rabbit antiphosphor-Tyk2 (ab138394) was purchased from Abcam, Inc. (Cambridge, United Kingdom), and rabbit anti-phosphor-Jak1 (AF5857) was purchased from (Shanghai, China). Mouse anti-UL23 HCMV monoclonal antibodies and rabbit anti-UL23 HCMV polyclonal antibodies utilized in the present study are consistent with those described previously (Chai et al, 2019;Li et al, 2020).…”

Section: Cells Viruses Reagents and Antibodiessupporting

confidence: 90%

Human Cytomegalovirus UL23 Attenuates Signal Transducer and Activator of Transcription 1 Phosphorylation and Type I Interferon Response

Feng

et al. 2021

Front. Microbiol.

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Human cytomegalovirus (HCMV), the human beta-herpesvirus, can cause severe syndromes among both immunocompromised adult patients and newborns. Type I interferon (IFN-I) exerts an important effect to resist infections caused by viruses such as HCMV, while IFN evasion may serve as a key determining factor for viral dissemination and disease occurrence within hosts. In this study, UL23, a tegument protein of HCMV, was confirmed to be a key factor for negatively regulating the type I IFN immune response. A detailed analysis indicated that the viral UL23 protein increases the IFN-I antiviral resistance during HCMV infections. Furthermore, UL23 was shown to significantly reduce the levels of IFN-stimulated genes (ISGs) and promoter activity of IFN-I-stimulated response element. Mechanically, UL23 was discovered to impair the signal transducer and activator of transcription 1 (STAT1) phosphorylation, although it was not found to affect phosphorylation and expression of STAT2, Janus activated kinase 1, or tyrosine kinase 2, which are associated with IFN-I signal transduction pathway. Additionally, a significantly reduced nuclear expression of STAT1 but not of IFN regulatory factor 9 or STAT2 was observed. Findings of this study indicate that HCMV UL23 is a viral antagonist that acts against the cellular innate immunity and reveal a possible novel effect of UL23 on IFN-I signaling.

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Section: Cells Viruses Reagents and Antibodiessupporting

confidence: 90%

Human Cytomegalovirus UL23 Attenuates Signal Transducer and Activator of Transcription 1 Phosphorylation and Type I Interferon Response

Feng

et al. 2021

Front. Microbiol.

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“…At present, Ni column affinity chromatography is one of the most effective methods for protein purification. Therefore, Ni column affinity chromatography was used to purify the inducible INHα protein in this experiment as the procedure can both remove inclusion body impurities as far as possible and assist the correct folding of the protein in subsequent renaturation (Chai et al, 2019). The purified INHα protein was analyzed by SDS-PAGE and a single band of 36 kDa with a concentration of 1.5 mg/ml was obtained.…”

Section: Test Results Of Blood Biochemical Indexesmentioning

confidence: 99%

EFFECTS OF INHIBIN-α AND ANTI-INHIBIN-α IMMUNIZATION ON THE REPRODUCTIVE HORMONES IN KAZAKH SHEEP

2022

THE JAPS

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The Kazakh sheep is native to China. The breed has many advantages, including its size, hardiness, and good meat production, it has the economic disadvantage of having a low reproductive rate. Inhibin-α (INHα) regulates the synthesis and secretion of follicle-stimulating hormone (FSH) and could feasibly be used to boost reproduction in the Kazakh sheep. However, the current methods of INHα preparation are both costly and time-consuming. Here, we investigated the effects of INHα on Kazakh sheep reproductive performance by immunization with INHα and analyzing the subsequent changes in reproductive hormone levels and blood biochemical indices. An anti-INHα polyclonal antibody was raised in camel. It and a recombinant INHα protein were used to immunize groups of adult Kazakh sheep in anestrus. Reproductive hormones (FSH, luteinizing hormone [LH], progesterone [P4], and estradiol [E2]) were measured by ELISA, together with the measurement of changes in INH levels and blood physiological and biochemical indicators. The blood levels of LH and P4 in the sheep immunized with the camel anti-INHα polyclonal antibody (group A) did not differ significantly from those in the recombinant INHα protein (group B) and the control group (group C) (P ＞0.05).FSH and E2 levels in group A were significantly higher than the controls (P <0.05) and the INH concentrations were significantly lower than those in group C (P <0.05). There were no abnormalities in the blood biochemical indices in groups A, B, and C.In conclusion, immune INHα preparations significantly affected the blood reproductive hormone levels of Kazakh sheep. This technique has potential application for improving the reproductive performance in these sheep and is also relevant for future research into the development of an INHα vaccine.

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“…The bacteria were cultured in LB medium supplemented with 50 μg/mL kanamycin. When the cell density of BL21 (DE3) reached 0.4–0.6 at OD 600 nm , 1 mM isopropyl-β- d -thiogalactoside (IPTG, Sigma, USA) was added to the medium to induce the expression of His-MsrA fusion protein at 16 °C for 12 h [ 28 ]. The bacterial cells induced by IPTG were harvested, resuspended in PBS (pH 7.4) and lysed by sonication on ice (3 s on and 3 s off, total of 20 min).…”

Section: Methodsmentioning

confidence: 99%

Luteolin increases susceptibility to macrolides by inhibiting MsrA efflux pump in Trueperella pyogenes

Guo

Huang

et al. 2022

Vet Res

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Trueperella pyogenes (T. pyogenes) is an opportunistic pathogen associated with a variety of diseases in many domestic animals. Therapeutic treatment options for T. pyogenes infections are becoming limited due to antimicrobial resistance, in which efflux pumps play an important role. This study aims to evaluate the inhibitory activity of luteolin, a natural flavonoid, on the MsrA efflux pump and investigate its mechanism. The results of antimicrobial susceptibility testing indicated that the susceptibility of msrA-positive T. pyogenes isolates to six macrolides increased after luteolin treatment, while the susceptibility of msrA-negative isolates showed no change after luteolin treatment. It is suspected that luteolin may increase the susceptibility of T. pyogenes isolates by inhibiting MsrA activity. After 1/2 MIC luteolin treatment for 36 h, the transcription level of the msrA gene and the expression level of the MsrA protein decreased by 55.0–97.7% and 36.5–71.5%, respectively. The results of an affinity test showed that the equilibrium dissociation constant (KD) of luteolin and MsrA was 6.462 × 10–5 M, and hydrogen bonding was predominant in the interaction of luteolin and MsrA. Luteolin may inhibit the ATPase activity of the MsrA protein, resulting in its lack of an energy source. The current study illustrates the effect of luteolin on MsrA in T. pyogenes isolates and provides insight into the development of luteolin as an innovative agent in combating infections caused by antimicrobial-resistant bacteria.

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Preparation and identification of polyclonal antibody against human cytomegalovirus encoding protein UL23

Cited by 6 publications

References 10 publications

Human Cytomegalovirus UL23 Attenuates Signal Transducer and Activator of Transcription 1 Phosphorylation and Type I Interferon Response

Human Cytomegalovirus UL23 Attenuates Signal Transducer and Activator of Transcription 1 Phosphorylation and Type I Interferon Response

EFFECTS OF INHIBIN-α AND ANTI-INHIBIN-α IMMUNIZATION ON THE REPRODUCTIVE HORMONES IN KAZAKH SHEEP

Luteolin increases susceptibility to macrolides by inhibiting MsrA efflux pump in Trueperella pyogenes

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