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1.
Zhang M Wang Z Graner MW Yang L Liao M Yang Q Gou J Zhu Y Wu C Liu H Zhou B Chen X 《Cellular immunology》2011,(2):217-223
Although it has been recognized that ectopic follicle-like B cell aggregate formation is common in the lungs of patients with tuberculosis, the role of infiltrated B cells in human tuberculosis remains to be elucidated. In the present study, we showed that ectopic B cell aggregate formation was associated with containment of Mycobacterium tuberculosis. The area ratio of ectopic B cell aggregates was correlated with localized IL-17 mRNA expression and peripheral TGF-β and IL-6 mRNA expression. Depletion of B cells from pleural fluid mononuclear cells resulted in significantly diminished M. tuberculosis antigen-specific IL-17 and IL-22 production, but not in IFN-γ secretion. Therefore, ectopic lung B cell formation is important for containment of M. tuberculosis, and up-regulation of IL-17 and IL-22 responses may be an important mechanism underlying the protective role B cells in human tuberculosis. 相似文献
2.
The S gene region of the hepatitis B virus (HBV) is responsible for the expression of surface antigens and includes the ‘a’-determinant
region. Thus, mutation(s) in this region would afford HBV variants a distinct survival advantage, permitting the mutant virus
to escape from the immune system. The aim of this study was to search for mutations of the S gene region in different patient groups infected with genotype D variants of HBV, and to analyse the biological significance of these mutations. Moreover, we investigated S gene mutation inductance among family members. Forty HBV-DNA-positive patients were determined among 132 hepatitis B surface
antigen (HbsAg) carriers by the first stage of seminested PCR. Genotypes and subtypes were established by sequencing of the
amplified S gene regions. Variants were compared with original sequences of these serotypes, and mutations were identified.
All variants were designated as genotype D and subtype ayw3. Ten kinds of point mutations were identified within the S region. The highest rates of mutation were found in chronic hepatitis patients and their family members. The amino acid mutations
125 (M → T) and 127 (T → P) were found on the first loop of ‘a’-determinant. The other consequence was mutation inductance
in a family member. We found some mutations in the S gene region known to be stable and observed that some of these mutations
affected S gene expression. 相似文献
3.
With a view to diagnosing tuberculosis in populations in endemic areas, excretory-secretory antigen fraction(Mtb EST-6) of purifiedMycobacterium tuberculosis H37Ra and affinity purified polyclonal antibodies againstMtb EST were used to detect both antibodies and circulating antigen in the sera of patients and disease-free individuals. Indirect
stick penicillinase ELISA system usingMtb EST-6 detected antigen-specific IgG antibody in 84% of sputum positive, 77% of sputum negative pulmonary tuberculosis patients
and 7% of healthy and 11% of subjects with nontub~rculosis diseases. Similarly, a sandwich penicillinase ELISA system using
affinity purified antiMtb EST antibodies detected circulating antigen in 83% and 61% of sputum positive and negative pulmonary tuberculosis subjects.
In contrast only 24% of healthy and 18% of disease controls showed seropositivity. Antibody assay showed higher sensitivity
and specificity (83% and 91% respectively) compared to antigen detection (sensitivity of 79% and specificity of 79%). However,
by concomitant use of both assays it was possible to enhance the specificity of detection to 98%, though sensitivity was reduced
marginally to 70%. The present study confirms the presence of both antigen and specific antibodies in the circulation during
clinical disease and draws attention to the utility ofMtb EST-6 as a diagnostic marker of pulmonary tuberculosis. 相似文献
4.
Grewal J.S. Reddy M.P. Singh Arvinderjit Singh Mohinder 《World journal of microbiology & biotechnology》2002,18(8):721-725
Secretion of a low molecular weight (24 kDa) Mycobacterium tuberculosis-specific antigen was analysed in the urine samples of tuberculosis patients under antimycobacterial therapy regime. The urine samples of sputum-positive and culture-positive tuberculosis patients under therapy regime were collected after 2, 3, 4, 5 and 6 months of antimycobacterial therapy. After concentration of the samples by ultrafiltration the proteins were resolved by SDS–PAGE. The antibodies raised in rabbits against M. tuberculosis strain H37Ra were used to check specificity of the bands by Western blotting. It was found that the tuberculosis patients secreted a 24 kDa M. tuberculosis-specific antigen in their urine. This band was present in the samples taken upto 5 months of drug therapy but was absent in the samples taken after 6 months of antimycobacterial therapy. This study gives evidence in support of the continuation of chemotherapy of tuberculosis patients for at least 6 months. Also, the 24 kDa excreted/secreted antigen can be used as a marker for monitoring the drug therapy as well as for the diagnosis of tuberculosis patients. Moreover, the urine sample taken in the study is a non-invasive and safer sampling method as compared to sampling blood or sputum which is the usual procedure in these patients. 相似文献
5.
An expression vector constructed from genes of Pichia pastoris was applied for heterologous gene expression in Saccharomyces cerevisiae. Recombinant hepatitis B surface antigen was synthesized by cloning hepatitis B virus ‘S’ gene under the control of glyceraldehyde-3-phosphate
dehydrogenase (GAP) promoter of Pichia pastoris in Saccharomyces cerevisiae. Hepatitis B surface antigen was constitutively expressed, was stable and exhibited ∼20–22 nm particle formation. Stability
and absence of toxicity to the host with the expression vector indicates the expression system can be applied for large-scale
production. 相似文献
6.
Summary
In vitro culture of hairy roots of Phyllanthus amarus induced by Agrobacterium rhizogenes was established. Their growth and ability for in vitro inactivation of hepatitis B virus surface antigen was studied and compared with adventitious roots grown in vitro. The selected hairy root clone HR-1 was capable of growing at a very fast rate, and an approximately 900-fold increase in
weight of root biomass was achieved after 4 wk of culture in hormone-free quarter-strength liquid Murashige and Skoog medium
with continuous agitation. Non-transformed roots cultured in the presence of 1.0 mg l−1 (5.71 μM) indole-3-acetic acid increased by 330-fold. The immuno-inactive property of roots was maximal in the crude extract. The
hairy roots were shown to possess 85% inhibition (in contrast to 15% in the control) in binding of hepatitis B surface antigen
(HBsAg) to its antibody (anti-HBs) after 24 h of incubation with HbsAg-positive sera in vitro at 37°C. Out of three fractions selected on the basis of molecular weight components of the extract, the Fraction III containing
comparatively lower molecular weight substances (≤3500) yielded the highest activity. The extract from non-transformed roots
was found to possess similar efficiency (87% inhibition). The levels of activity in both types of in vitro-raised roots were higher than those of naturally occurring roots and leafy shoots. The ability of P. amarus hairy root cultures to yield high biomass with the anti-viral property at high levels may provide an alternative source of
raw material for more detailed study in the field of pharmaceutical research. 相似文献
7.
Negative HBcAg in immunohistochemistry assay of liver biopsy is a predictive factor for the treatment of patients with nucleos(t)ide analogue therapy
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Mingxing Huang Jian Liu Monica Chow Xuan Zhou Zongping Han Zhenjian He Jinfang Xue Zhe Zhu Xinhua Li Jinyu Xia 《Journal of cellular and molecular medicine》2018,22(3):1675-1683
The hepatitis B core antigen (HBcAg) is an important target for antiviral response in chronic hepatitis B (CHB) patients. However, the correlation between HBcAg in the hepatocyte nucleus and nucleos(t)ide analogue (NA) therapeutic response is unclear. We sought to evaluate the role of HBcAg by analysing liver biopsies for viral response in NA‐naïve hepatitis B e antigen (HBeAg) positive (+) CHB patients via immunohistochemistry (IHC). A total of 48 HBcAg‐negative (?) patients and 48 HBcAg (+) patients with matching baseline characteristics were retrospectively analysed for up to 288 weeks. Virological response (VR) rates of patients in the HBcAg (?) group were significantly higher at week 48 and 96 than the HBcAg (+) group (77.1% versus 45.8% at week 48, respectively, P = 0.002 and 95.3% versus 83.3% at week 96, respectively, P = 0.045). The serological negative conversion rate of HBeAg was significantly higher in the HBcAg (?) than in the HBcAg (+) group from week 96 to 288 (35.4 % versus 14.6% at week 96, respectively, P = 0.018; 60.4% versus 14.6%, respectively, P < 0.001 at week 144; 72.9% versus 35.4%, respectively, P < 0.001 at week 288). The cumulative frequencies of VR and lack of HBeAg were higher in the HBcAg (?) group (both P < 0.05). Binary logistic regression analysis showed that HBcAg (?) was the predictor for the lack of HBeAg (OR 4.482, 95% CI: 1.58–12.68). In summary, the absence of HBcAg in the hepatocyte nucleus could be an independent predictor for HBeAg seroconversion rates during NA‐naïve treatment in HBeAg (+) CHB patients. 相似文献
8.
Cheong JY Cho SW Chung SG Lee JA Yeo M Wang HJ Lee JE Hahm KB Kim JH 《Biochemical genetics》2006,44(5-6):246-255
The natural history of hepatitis B virus (HBV) infection is probably related to host immune factors. Interferon-γ (IFN-γ) plays significant roles in immune defense. This study was undertaken to investigate the association between HBV infection and single nucleotide polymorphisms (SNPs) of IFN-γ, IFN-γ receptor (IFNGR)-1 and 2, and interferon regulatory factor (IRF)-1 genes. Between March 2002 and December 2002, 614 Korean patients were enrolled in two different groups: an HBV clearance group (n = 201), who were hepatitis B surface antigen (HBsAg) negative with antibodies to HBsAg and hepatitis B core antigen, and an HBV persistence group (n = 413), who were repeatedly HBsAg positive. We assessed polymorphisms in the IFN-γ gene at position +874, in the IFNGR-1 gene at positions −56 and +95, in the IFNGR-2 gene at the second position of codon 64 (Gln64Arg), and in the IRF-1 gene promoter (−410, −388), and the genotype distributions of the HBV clearance and persistence groups were compared. On the basis of unconditional logistic regression analysis with adjustment for age and sex, no statistically significant association with susceptibility to persistent HBV infection was observed with the IFN-γ, IFNGR-1 and 2, and IRF-1 gene polymorphisms under the codominant, dominant, and recessive models. 相似文献
9.
Neysi Ibarra Abel Caballero Ernesto Gonzlez Rodolfo Valds 《Journal of chromatography. B, Analytical technologies in the biomedical and life sciences》1999,735(2):314
An immunoaffinity chromatographic method was developed using a mAb immunosorbent to purify recombinant hepatitis B surface antigen (r-HBsAg) from yeast. Elution conditions using a mAb-coated ELISA were improved to select the best conditions to purify r-HBsAg. The optimum results in terms of total quantitative recovery were obtained using 20 mM Tris pH 11.6. An increase in the CB.Hep-1 mAb (anti-HBsAg) useful immunosorbents half-life and in its yield per cycle was obtained when alkaline elution conditions were used. Moreover, the basic conditions do not affect either the antigenic characteristics or the purity or the molecular integrity of r-HBsAg. 相似文献
10.
Degui Geng Yiqin Wang Peng Wang Wenbin Li Yongru Sun 《Journal of applied phycology》2003,15(6):451-456
A stable transformation system for theexpression of foreign genes in theunicellular marine green alga Dunaliella salina was established. Amongfive antibiotics, 60 g mL-1chloramphenicol completely inhibitedgrowth. Of five promoters tested, theubiquitin-$Ohgr; promoter yielded thehighest -glucuronidase (GUS) activity.The hepatitis B surface antigen (HBsAg)gene was introduced into the cells by usingelectroporation. PCR and Southern blotanalysis amd it was shown that the gene wasintegrated into the genome. The stableexpression of HBsAg protein was confirmedby enzyme-linked immunosorbent assay(ELISA) and Western blot analysis. Theintroduced DNA and HBsAg expression weremaintained stable for at least 60generations in medium devoid ofchloramphenicol. This is an important steptoward the production of useful foreignproteins in the alga. 相似文献
11.
[目的] 耐药结核分枝杆菌(drug-resistant Mycobacterium tuberculosis)的产生给结核病(tuberculosis)的治疗带来巨大困难。[方法] 使用基于全基因组测序的关联分析探究耐药强相关的单核苷酸多态性(single nucleotide polymorphism,SNP)突变,主要有GEMMA、phyc、plink。为了阐明其中最优的耐药相关SNP计算方法,本研究下载NCBI上已有的1504株结核分枝杆菌数据,并获取它们对于3种常见的一线抗结核治疗药物(isoniazid、rifampicin、ethambutol)的耐药性检验结果。并使用这3种耐药相关SNP计算方法计算与结核分枝杆菌耐药相关的SNP;并评估计算得到的耐药相关SNP在预测耐药表型的敏感性和特异性。[结果] 发现通过phyc可以预测到最多的已知耐药相关SNP和最少的耐药无关SNP,而且phyc预测的耐药相关SNP的敏感性和特异性恒定大于52.49%。[结论] phyc在预测结核分枝杆菌耐药相关SNP中结果最准确,但考虑到运行时间和表型数据的更新,GEMMA和plink的结果也应作为参考。 相似文献
12.
Lee TH Tai DI Cheng CJ Sun CS Lin CY Sheu MJ Lee WP Peng CY Wang AH Tsai SL 《Journal of biomedical science》2006,13(1):27-39
Summary Chronic infections with hepatitis B and C viruses (HBV and HCV) are etiologically linked to hepatitis, liver cirrhosis, and
hepatocellular carcinoma (HCC). Both viruses may induce activation of nuclear factor-kappa B (NF-κB) in hepatocytes that plays
a crucial role in the regulation of cell growth and apoptosis. Functional proteomics analysis of proteins associated with
NF-κB signaling complexes in both viruses-related HCC tumor and non-tumor tissues may disclose possible common mechanisms
in hepatocarcinogenesis. By functional proteomics, we analyzed proteins associated with NF-κB-signaling complexes in four-paired
human HCC tumor and non-tumor tissues from HBV- and HCV-infected patients, respectively, and in one-paired tissue with dual
viral infection. The quantity of NF-κB-associated proteins was semi-quantitatively measured by protein spot intensity on the
gels of two-dimensional polyacrylamide gel electrophoresis. The results showed that overexpression of NF-κB-associated Wnt-1 protein in tumor part was detected in the␣majority of HBV- and HCV-infected HCC samples. These data suggest that enhanced
expression of NF-κB-associated Wnt-1 protein might be a mechanism of hepatocarcinogenesis common to HBV- and HCV-infected patients. NF-κB signaling pathway and
Wnt-1 protein could be potential targets for designing highly effective therapeutic agents in treating HCC and for chemoprevention
of hepatocarcinogenesis. 相似文献
13.
High-level expression of bovine rotavirus VP4 protein in Escherichia coli using a hepatitis B core antigen vector 总被引:3,自引:0,他引:3
Six regions of the VP4 protein of bovine rotavirus strain UKtc were expressed using hepatitis B core antigen (HBcAg) as a carrier. Following induction by IPTG, the six fusion proteins, AHBcAg through FHBcAg, were expressed in Escherichia coli to a level of 20–37% of total cellular protein. Soluble fusion proteins in a particulate form were partially purified with yields ranging from 0.5–6.4 mg l–1 of culture. 相似文献
14.
Griselda Escobedo-Melendez Arturo Panduro Nora A Fierro Sonia Roman 《Memórias do Instituto Oswaldo Cruz》2014,109(6):728-737
Studies on the prevalence of infection with hepatitis B virus (HBV) among children
are scarce in Latin American countries, especially in Mexico. This study was aimed to
investigate the prevalence of HBV infection, occult hepatitis B infection (OBI) and
HBV genotypes among children with clinical hepatitis. In total, 215 children with
clinical hepatitis were evaluated for HBV infection. HBV serological markers and HBV
DNA were analysed. OBI diagnosis and HBV genotyping was performed. HBV infection was
found in 11.2% of children with clinical hepatitis. Among these HBV DNA
positive-infected children, OBI was identified in 87.5% (n = 21/24) of the cases and
12.5% (n = 3/24) were positive for both HBV DNA and hepatitis B surface antigen. OBI
was more frequent among children who had not been vaccinated against hepatitis B (p
< 0.05) than in those who had been vaccinated. HBV genotype H was prevalent in 71%
of the children followed by genotype G (8%) and genotype A (4%). In conclusion, OBI
is common among Mexican children with clinical hepatitis and is associated with HBV
genotype H. The results show the importance of the molecular diagnosis of HBV
infection in Mexican paediatric patients with clinical hepatitis and emphasise the
necessity of reinforcing hepatitis B vaccination in children. 相似文献
15.
High level expression of recombinant Mycobacterium tuberculosis culture filtrate protein CFP32 in Pichia pastoris 总被引:1,自引:0,他引:1
Benabdesselem C Barbouche MR Jarboui MA Dellagi K Ho JL Fathallah DM 《Molecular biotechnology》2007,35(1):41-49
Difficulty in obtaining large quantities of Mycobacterium tuberculosis (MTB) proteins remains a major obstacle in the development of subunit vaccines and diagnostic reagents for tuberculosis.
A major reason is because Escherichia coli has not proven to be an optimal host for the expression of MTB genes. In this article, we used the yeast Pichia pastoris to express high levels of CFP32, a culture filtrate protein restricted to the MTB complex and a potential target antigen
for serodiagnosis of tuberculosis in patients. Using shaker flasks, we generated a P. pastoris clone expressing CFP32 as a secreted protein fused to the myc-(His)6 tag, at a yield of 0.5 g of purified protein per liter of culture. Recombinant CFP32 (rCFP32) produced in P. pastoris has a molecular weight of 35 kDa, which is slightly higher than that of the native protein We identified putative acylation
and glycosylation sites in the CFP32 amino acid sequence that suggested post-translational modifications may contribute to
the size difference. The NH2-terminal peptide sequencing of rCFP32 showed that the signal peptide alpha factor is correctly excised. In addition, rCFP32
reacted with the sera of patients with tuberculosis. These data are the first to show that P. pastoris is a suitable host for high-yield production of good quality mycobacterium antigens, and especially culture filtrate proteins
that have vaccine and diagnostic potential. 相似文献
16.
Much of our understanding of the epidemiologic features of infection with Blastomyces dermatitidis has come from cluster and outbreak investigations which have established the association of human disease with recreational
pursuits and the presence of infectious microfoci in areas of moist soil with high organic content. This report describes
the clustering of eight cases of pulmonary blastomycosis without an apparent common source exposure which occurred during
a 90 day period in a 96 square mile area (population 4,450) within Oconto County, Wisconsin. We conclude that multiple high-risk
environmental foci may have existed following a sustained five-year period of diminished precipitation in the cluster area.
A case-control study which included family and community controls concluded that multiple earth-disturbing activities engaged
in by case-patients was statistically associated with illness. Lymphocyte-proliferation assays of whole blood samples detected
previously unrecognized infection withB. dermatitidis among five of 32 family controls.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
17.
Imen Ben Dhifallah Kaouther Ayouni Ghofrane Najjar Hanene Chelbi Amel Sadraoui Walid Hammami Henda Touzi Henda Triki 《Microbiology and immunology》2020,64(7):512-519
Approaches based on association studies have proven useful in identifying genetic predictors for many diseases, including susceptibility to chronic hepatitis B. In this study we were interested by the IL-1B genetic variants that have been involved in the immune response and we analyzed their role in the susceptibility to develop chronic hepatitis B in the Tunisian population. IL - 1B is a potent proinflammatory cytokine that plays an important role in inflammation of the liver. Polymorphic gene IL-1 ( − 511, +3954) was analyzed in a total of 476 individuals: 236 patients with chronic hepatitis B from different cities of Tunisia recruited in Pasteur Institute between January 2017 and December 2018 and 240 controls. Genomic DNA was obtained using the standard salting-out method and genotyping was performed by polymerase chain reaction (PCR)-restriction fragment length polymorphism. For − 511C>T polymorphism a significant association was found between patients and controls when comparing the genotypic (P = 0.007; χ2 = 9.74 and odds ratio [OR] = 0.60; confidence interval [CI] = 0.41–0.89) and allelic (P = 0.001; χ2 = 10.60) frequencies. When the viral load was taken into account a highly significant difference was found (P = 9 × 10−4; χ2 = 10.89). For +3954C>T polymorphism a significant association was found between patients and controls when comparing genotypic (P = 0.0058; χ2 = 7.60 and OR = 1.67; CI = 1.14–2.46) and allelic (P = 0.0029; χ2 = 8.81) frequencies. T allele can be used as a strong marker for hepatitis B virus disease for both polymorphisms. 相似文献
18.
Predictive indicators for the therapeutic effect of OK-432 in patients with chronic active type B hepatitis 总被引:1,自引:0,他引:1
Twenty-two patients with chronic type B hepatitis were treated with OK-432. Immunological parameters were serially measured to find predictive indicators for the seroconversion from hepatitis B envelope antigen(HBe Ag) to anti-HBe. In patients who achieved the disappearance of HBe Ag associated with or without the appearance of anti-HBe, the numbers of CD8+DR+ and CD4+DR+T cells in peripheral blood increased gradually during OK-432 therapy and then reduced subsequently to the seroconversion from HBe Ag positive to anti-HBe positive. Increases of DR-positive T cells in numbers were significantly correlated with increased amounts of IFN- produced in response toin vitro OK-432 stimulation.In vitro OK-432-stimulated IFN- production and the increase of CD8+DR+T cells in number in peripheral blood could be proposed as predictive indicators for the disappearance of HBe Ag. 相似文献
19.
Jodi J. L. Rowley Valentine A. Hemingway Ross A. Alford Michelle Waycott Lee F. Skerratt Ruth Campbell Rebecca Webb 《EcoHealth》2007,4(1):31-36
Chytridiomycosis is a fatal disease of amphibians, caused by the amphibian chytrid Batrachochytrium dendrobatidis. The disease is unusual in that it may drive many amphibian species to local extinction during outbreaks. These dramatic declines
in host population numbers could be facilitated if the pathogen can grow as a saprobe or on alternative hosts, a feature common
to other chytrid species. This is also supported by in vitro work that demonstrates B. dendrobatidis can grow and reproduce in the absence of amphibian cells. In a previous study, B. dendrobatidis was detected on freshwater shrimp from rain forest streams in northern Queensland, Australia, using diagnostic PCR. We set
out to confirm and further investigate the presence of B. dendrobatidis on crustaceans by carrying out more extensive sampling of shrimp in the field, experimental B. dendrobatidis infection trials using shrimp and crayfish, and PCR verification of the presence of B. dendrobatidis from shrimp samples that previously tested positive. We could not confirm the presence of B. dendrobatidis on shrimp, and report that original positive tests in shrimp reported by Rowley et al. (2006) were likely false. Thus, we suggest that shrimp may not be an important reservoir host for B. dendrobatidis. 相似文献
20.
Antibody to hepatitis B virus core antigen (anti-HBc) is one of the most important serological markers during hepatitis B virus (HBV) infection. The quality of the hepatitis B virus core antigen (HBcAg; diagnostic antigen) is crucial to the accuracy of anti-HBc detection. In an attempt to explore the suitability of recombinant HBcAg (rHBcAg) for diagnostic purposes, HBcAg was expressed in Escherichia coli (E. coli) and Pichia pastoris (P. pastoris) and evaluated for the detection of anti-HBc. The expression level of the recombinant protein satisfied the criteria for large-scale biologic production. P. pastoris- and E. coli-derived rHBcAg were purified with gel filtration followed by sucrose gradient (reagents A and C) or with a monoclonal anti-HBc antibody binding (reagents B and D) and were utilized to detect anti-HBc in competitive inhibition enzyme-linked immunosorbent assay (ELISA) format. The ELISA using P. pastoris-derived rHBcAg had a higher specificity and sensitivity than that using E.coli-derived rHBcAg to detect the anti-HBc standard panel. Serum specimens were collected from HBV-infected patients and healthy individuals (voluntary blood donors). Anti-HBc was detected in those specimens using P. pastoris- and E. coli-derived rHBcAg. The positive rate of anti-HBc detection in HBV-infected patients' sera was 100% with reagents A and B, 96.4% with reagent C, and 93.6% with reagent D. The negative rate in healthy control sera was 100% with reagents A and B, 97.0% with reagent C, and 99.7% with reagent D. These data indicate that P. pastoris-derived rHBcAg is superior to E.coli-derived rHBcAg for the detection of anti-HBc using the diagnostic ELISA. 相似文献