A Novel Retrograde AAV Variant for Functional Manipulation of Cortical Projection Neurons in Mice and Monkeys / 神经科学通报·英文版

Retrograde adeno-associated viruses (AAVs) are capable of infecting the axons of projection neurons and serve as a powerful tool for the anatomical and functional characterization of neural networks. However, few retrograde AAV capsids have been shown to offer access to cortical projection neurons across different species and enable the manipulation of neural function in non-human primates (NHPs). Here, we report the development of a novel retrograde AAV capsid, AAV-DJ8R, which efficiently labeled cortical projection neurons after local administration into the striatum of mice and macaques. In addition, intrastriatally injected AAV-DJ8R mediated opsin expression in the mouse motor cortex and induced robust behavioral alterations. Moreover, AAV-DJ8R markedly increased motor cortical neuron firing upon optogenetic light stimulation after viral delivery into the macaque putamen. These data demonstrate the usefulness of AAV-DJ8R as an efficient retrograde tracer for cortical projection neurons in rodents and NHPs and indicate its suitability for use in conducting functional interrogations.

Culture of human sapovirus GⅠ.1 in vitro and preparation of polyclonal antibody against capsid protein VP1 / 中国生物制品学杂志

@#Objective To culture human sapovirus(HuSaV) GⅠ.1 in vitro and prepare polyclonal antibody against the capsid protein VP1.Methods HuSaV GⅠ.1 positive stool specimens preserved in diarrhea department of National Institute for Viral Disease Control and Prevention were inoculated with HuTu-80 cells supplemented with different bile acid salts[glycine chenodeoxycholic acid(GCDCA) and glycine cholic acid(GCA)],and the infection,proliferation and passage of the virus were determined by PCR and RT-qPCR.The VP1 gene was amplified by PCR and cloned into prokaryotic expression vector pGEX-6P-1.The constructed recombinant expression plasmid pGEX-6P-1-VP1 was transformed into E.coli BL21(DE3) and induced by IPTG.Two female New Zealand white rabbits were immunized with the purified recombinant VP1 protein for 4 times.The blood samples were collected 18,28,38 and 48 d after immunization,and the serum titers were detected by ELISA.Results HuTu-80 cells were effectively infected by HuSaV GⅠ.1 in the presence of bile acid salt GCA,and the proliferated virus were stably and continuously transmitted for three generations in HuTu-80 cells.The expressed recom-binant GST-VP1 protein showed a relative molecular mass of about 86 000,and about 60 000 after purification(GST tag excision).The titer of polyclonal antibody against HuSaV VP1 protein was over 1:12 800.Conclusion HuSaV was successfully isolated and cultured in vitro using HuTu-80 cells supplemented with bile acid salt,and polyclonal antibody with high titer against HuSaV VP1 protein was prepared,which laid a foundation of in-depth research of HuSaV identification,infection and pathogenesis.

Prokaryotic expression and immunogenicity of hepatitis A virus capsid proteins VP1 and VP3 / 中国生物制品学杂志

@#Objective To express the capsid proteins VP1 and VP3 of hepatitis A virus(HAV) in prokaryotic cells and evaluate their immunogenicity.Methods VP1 and VP3 gene fragments were amplified by PCR,cloned into vector pETG28a to construct recombinant expression plasmids pET-G28a-VP1 and pET-G28a-VP3,which were transformed into competent E.coli BL21(DE3),induced by IPTG,and then analyzed by 12% SDS-PAGE.The target protein was purified by ion exchange chromatography,renatured and combined with several adjuvants to immunize mice.The mice were divided into VP1-MF59,VP1-AH,VP1-AP,VP1-AP-10CpG,VP1-AP-50CpG,VP3-AP,VP3-VP1-AP and PBS control groups,five for each group.Serum IgG antibody titers of mice in various groups were detected by ELISA,and serum neutralizing antibody titers of mice in VP1-AP,VP3-VP1-AP and PBS control groups were detected by rapid fluorescence focus immunosuppression experiment.Results Colony PCR and sequencing showed that the recombinant plasmids pET-G28a-VP1 and pET-G28a-VP3were constructed correctly.The recombinant proteins VP1 and VP3,with relative molecular masses of about 37 000 and26 000 respectively,mainly existed in the form of inclusion bodies,the expression levels were 18.6% and 32.4%,and the purity was 86.3% and 84.7%,respectively.The recombinant proteins VP1 and VP3 reacted specifically with rabbit anti-HAV antiserum and mouse anti-HAV-VP3 antiserum respectively.The serum IgG antibody titer of mice in VP1-AP-50CpG group was significantly higher than that in VP1-AP group(q=22.05,P <0.01),and the serum IgG antibody titer of mice in VP3-VP1-AP group was significantly higher than that in VP1-AP group and VP3-AP group(q=22.05 and 22.49 respectively,each P <0.01).Compared with PBS control group,the serum neutralizing antibody titer of mice in VP1-AP and VP3-VP1-AP group increased significantly(q=7.79 and 25.11 respectively,P<0.01) Conclusion Prokaryotic HAV capsid proteins VP1 and VP3 showed high purity and good immunogenicity,and the addition of CpG in the preparation was beneficial to enhance the immunogenicity of antigens.This study laid a foundation of the development of HAV recombinant subunit vaccine.

The use of melittin to enhance transgene expression mediated by recombinant adeno-associated virus serotype 2 vectors both in vitro and in vivo / 中西医结合学报

OBJECTIVE@#Melittin, a cell-penetrating peptide, improves the efficiency of many non-viral gene delivery vectors, yet its application in viral vectors has not been well studied. The non-pathogenic recombinant adeno-associated virus (rAAV) vector is an ideal in vivo gene delivery vector. However, its full potential will only be achieved after improvement of its transduction efficiency. To improve the transduction efficiency of rAAV2 vectors, we attempted to develop a melittin-based rAAV2 vector delivery strategy.@*METHODS@#The melittin peptide was inserted into the rAAV2 capsid either in the loop VIII of all viral proteins (VPs) or at the N terminus of VP2. Various rAAV2-gfp or -fluc vectors were subjected to quantitative real-time polymerase chain reaction and Western blot assays to determine their titers and integrity of capsid proteins, respectively. Alternatively, the vectors based on wild-type capsid were pre-incubated with melittin, followed by transduction of cultured cells or tail vein administration of the mixture to C57BL/6 and BALB/c nude mice. In vivo bioluminescence imaging was performed to evaluate the transgene expression.@*RESULTS@#rAAV2 vectors with melittin peptide inserted in the loop VIII of VPs had low transduction efficiency, probably due to dramatically reduced ability to bind to the target cells. Fusing the melittin peptide at the N-terminus of VP2 produced vectors without the VP2 subunit. Interestingly, among the commonly used rAAV vectors, pre-incubation of rAAV2 and rAAV6 vectors with melittin significantly enhanced their transduction efficiency in HEK293 and Huh7 cells in vitro. Melittin also had the ability to increase the rAAV2-mediated transgene expression in mouse liver in vivo. Mechanistically, melittin did not change the vector-receptor interaction. Moreover, cell counting kit-8 assays of cultured cells and serum transaminase levels indicated melittin had little cytotoxicity.@*CONCLUSION@#Pre-incubation with melittin, but not insertion of melittin into the rAAV2 capsid, significantly enhanced rAAV2-mediated transgene expression. Although further in vivo evaluations are required, this research not only expands the pharmacological potential of melittin, but also provides a new strategy to improve gene therapy mediated by rAAV vectors.

Single-shot AAV-vectored vaccine against SARS-CoV-2 with fast and long-lasting immunity

Due to the insufficient long-term protection and significant efficacy reduction to new variants of current COVID-19 vaccines, the epidemic prevention and control are still challenging. Here, we employ a capsid and antigen structure engineering (CASE) strategy to manufacture an adeno-associated viral serotype 6-based vaccine (S663V-RBD), which expresses trimeric receptor binding domain (RBD) of spike protein fused with a biological adjuvant RS09. Impressively, the engineered S663V-RBD could rapidly induce a satisfactory RBD-specific IgG titer within 2 weeks and maintain the titer for more than 4 months. Compared to the licensed BBIBP-CorV (Sinopharm, China), a single-dose S663V-RBD induced more endurable and robust immune responses in mice and elicited superior neutralizing antibodies against three typical SARS-CoV-2 pseudoviruses including wild type, C.37 (Lambda) and B.1.617.2 (Delta). More interestingly, the intramuscular injection of S663V-RBD could overcome pre-existing immunity against the capsid. Given its effectiveness, the CASE-based S663V-RBD may provide a new solution for the current and next pandemic.

Urgent scientific issues to be solved in clinical trials of capsid assembly modulator combined with nucleos(t)ide analogues for the treatment of chronic hepatitis B / 临床肝胆病杂志

Chronic hepatitis B virus (HBV) infection is the main cause of viral hepatitis, liver cirrhosis, and primary liver cancer. At present, nucleos(t)ide analogues (NUC) and pegylated interferon α used in clinical practice cannot directly target covalently closed circular DNA, and it is difficult to achieve clinical cure of chronic hepatitis B patients; therefore, it is urgently needed to develop direct-acting antiviral agents targeting all stages of the HBV replication cycle. Capsid assembly modulator (CpAM) targets the assembly of viral capsids through various mechanisms, thereby exerting a direct-acting antiviral effect. Its combination with NUC should have a good synergistic antiviral effect, but the results of existing clinical trials have shown that chronic hepatitis B patients who received a limited course of antiviral therapy with CpAM and NUC all experienced off-therapy viral rebound. Based on the mechanism of action of these two types of drugs, this article provides a reasonable explanation for the above clinical trial results and points out that a longer course of antiviral therapy with CpAM and NUC may be needed in the future clinical trials with safe drug withdrawal as the end point of observation, so as to deplete or silence the pool of covalently closed circular DNA and increase the possibility of safe drug withdrawal in CHB patients. In addition, further studies are needed to explore antiviral therapeutic strategies with a combination of multiple targets.

Novel PF74-like small molecules targeting the HIV-1 capsid protein: Balance of potency and metabolic stability

Of all known small molecules targeting human immunodeficiency virus (HIV) capsid protein (CA), PF74 represents by far the best characterized chemotype, due to its ability to confer antiviral phenotypes in both early and late phases of viral replication. However, the prohibitively low metabolic stability renders PF74 a poor antiviral lead. We report herein our medicinal chemistry efforts toward identifying novel and metabolically stable small molecules targeting the PF74 binding site. Specifically, we replaced the inter-domain-interacting, electron-rich indole ring of PF74 with less electron-rich isosteres, including imidazolidine-2,4-dione, pyrimidine-2,4-dione, and benzamide, and identified four potent antiviral compounds (

Porcine circovirus 3 capsid protein induces autophagy in HEK293T cells by inhibiting phosphorylation of the mammalian target of rapamycin / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Porcine circovirus 3 (PCV3) has been detected in major pig-producing countries around the world since its first report in the US in 2016. Most current studies have focused on epidemiological investigations and detection methods of PCV3 because of lack of live virus strains for research on its pathogenesis in porcine cells or even in pigs. We constructed a recombinant plasmid pCMV-Cap carrying the PCV3 orf2 gene to investigate the effects of capsid (Cap) protein expression on autophagic response in human embryonic kidney cell line 293T (HEK293T). We demonstrate that PCV3 Cap protein induced complete autophagy shown as formation of autophagosomes and autophagosome-like vesicles as well as LC3-II conversion from LC3-I via inhibiting phosphorylation of the mammalian target of rapamycin (mTOR) in HEK293T cells. The ubiquitin-proteasome pathway is also involved in the autophagy process. These findings provide insight for further exploration of PCV3 pathogenetic mechanisms in porcine cells.

Advances in research on HBV inhibitors based on new targets (1): capsid protein inhibitors / 药学学报

Hepatitis B virus (HBV) capsid protein plays an important role in the life cycle, thus becoming an ideal target for drug design. Capsid protein inhibitors can exert a synergistic antiviral effect with nucleoside drugs by inhibiting the replication of HBV. This paper reviews the research progress of capsid protein inhibitors with different structural types from the perspective of medicinal chemistry.

Porcine circovirus 3 capsid protein induces autophagy in HEK293T cells by inhibiting phosphorylation of the mammalian target of rapamycin / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Porcine circovirus 3 (PCV3) has been detected in major pig-producing countries around the world since its first report in the US in 2016. Most current studies have focused on epidemiological investigations and detection methods of PCV3 because of lack of live virus strains for research on its pathogenesis in porcine cells or even in pigs. We constructed a recombinant plasmid pCMV-Cap carrying the PCV3 orf2 gene to investigate the effects of capsid (Cap) protein expression on autophagic response in human embryonic kidney cell line 293T (HEK293T). We demonstrate that PCV3 Cap protein induced complete autophagy shown as formation of autophagosomes and autophagosome-like vesicles as well as LC3-II conversion from LC3-I via inhibiting phosphorylation of the mammalian target of rapamycin (mTOR) in HEK293T cells. The ubiquitin-proteasome pathway is also involved in the autophagy process. These findings provide insight for further exploration of PCV3 pathogenetic mechanisms in porcine cells.

Caracterización de la proteína no capsidal 3D, del virus de la fiebre aftosa y producción de anticuerpos policlonales / Characterization of the non-capsided 3d protein, of the foot-and-mouth disease virus and the production of polyclonal antibodies

Las proteínas no capsidales del virus de la fiebre aftosa se utilizan como marcadoras en la evaluación de animales que han estado en contacto con el virus, a diferencia de los inmunizados, ya que la vacuna no debe tener estas proteínas, por lo tanto los animales no deben presentar anticuerpos contra ellas. El objetivo de esta investigación fue la caracterización de la proteína no capsidal 3D y la producción de anticuerpos policlonales in vivo. La proteína se purificó del cultivo de virus inactivo, por cromatografía de intercambio iónico. La elución de los picos fue sometida a electroforesis uni-bidimensional; demostrándose un alto grado de pureza (>90%) en el pico tres, donde se identifico la proteína 3D, por la técnica de MALDI-TOF y electroespray de trampa iónica. La proteína purificada, se inoculó en cabras y el suero hiperinmune fue precipitado y sometido a cromatografía de afinidad para la obtención de inmunoglobulinas; la reacción inmunitaria se confirmó por medio de inmunodifusión y Western blot. El proceso de purificación demostró ser eficiente y útil para la obtención de anticuerpos específicos, los cuales tendrán utilidad en la elaboración de un ensayo inmunoenzimático que mida la pureza de la vacuna frente al contenido de estas proteínas.

The noncapsid proteins of the foot and mouth disease are used as markers in the evaluation of animals that have been in contact with the virus, to discriminated the immunized animals, because the vaccine should not have these proteins, therefore animals should not present antibodies against them. The aim of this investigation was the characterization of the 3D non-capsid protein and the production of polyclonal antibodies in vivo. The protein was purified from the culture of inactivated virus, by ion exchange chromatography. The elution of the peaks were submit an one-two-dimensional electrophoresis; Demonstrated a high degree of purity (> 90%) in peak three, where the 3D protein was identified, by the MALDI-TOF technique and ion trap electrospray. The purified protein, inoculated in goats and the hyperimmune serum, was precipitate out and submitted to affinity chromatography to obtain immunoglobulins; the immune reaction was confirmed by means of immunodiffusion and Western blot. The purification process proved to be efficient and useful for obtaining specific antibodies, which will be useful in the preparation of an immunoenzymatic assay that measures the purity of the vaccine against to the content of these proteins.

Productive infection and transduction by bacteriophage P1 in the species Salmonella bongori

Background: Horizontal gene transfer (HGT) is the most important mechanism in the evolution of new genetic capabilities in bacteria, including specific degradative pathways, virulence factors, and resistance to antibiotics. Among the processes involved in HGT, transduction is noteworthy. This is a mechanism for gene transmission mediated by a bacteriophage that functions both as a reservoir and as a vector of exogenous genes, which remain protected from environmental effects in the bacteriophage's capsid. Within this context, this investigation aimed to evaluate the ability of the generalized transducing bacteriophage P1 to productively infect and transduce in the bacterial species Salmonella bongori. Results: We could establish that a derivative of bacteriophage P1, P1Cm, infects strains of S. bongori with frequencies of lysogenization in the order of ~10−2 lysogens/UFP. Through thermal induction, infective viral progeny was obtained, and we could show that P1Cm readily formed plaques on S. bongori lawns, a phenomenon thus far not reported for other members of the genus Salmonella. Finally, we showed P1Cm-mediated transduction of the model plasmid RP4 at frequencies of ~10−7 transductants/donor. Conclusion: Therefore, bacteriophage P1 can be used as a tool for the genetic manipulation in the species S. bongori.

Clonación y expresión en Escherichia coli de un gen L1 completo del virus del papiloma humano 18 aislado de una paciente cubana y variantes delecionadas / Cloning and expression in Escherichia coli of the full-length and deletion variants of a human papillomavirus 18 L1 gene isolated from a Cuban patients

Introduction: Human papillomavirus (HPV) vaccines are based on the L1 major capsid protein. Objectives: To clone the HPV-18 L1 gene from a Cuban female HPV-18-infected patient and to express the full-length and deletion variants of the cloned HPV-18 L1 gene in Escherichia coli. Methods: The full-length HPV-18 L1 gene was PCR-amplified from total DNA isolated from a Cuban patient, cloned and finally subcloned into the E. coli expression vector pET26b. Three deletion mutants were constructed, which encode truncated proteins lacking 30 amino acids at the C-terminus in combination with 5, 6 or none deleted residue at the N-terminus. Production of L1 proteins in E. coli BL21(DE3) and E. coli SHuffle T7 was assessed by SDS-PAGE and Western blotting. Results: The cloned HPV-18 L1 gene was 99.9 por ciento similar to the African variant EF202152 and probably shares a common origin with the B lineage of genotype 18. The three truncated variants of HPV-18 L1 were produced at higher levels than the full-length HPV-18 L1 protein, attaining higher levels in E. coli BL21(DE3) and higher solubility in E. coli SHuffle. The C-terminus-only truncated variant, L1∆C30, was produced at similar levels to the HPV-18 L1s truncated at both termini. E. coli SHuffle produced about three times more amounts of L1∆C30 when grown under autoinduction conditions with respect to conventional induction and thus, amounts were comparable to those obtained in E. coli BL21(DE3) under conventional induction. Conclusions: Truncation of thirty amino acid residues at the carboxy-terminus of the HPV-18 L1 made a major contribution to the production and solubility of this wild-type protein in E. coli. This is the first report about soluble production of HPV-18 L1 protein in an E. coli SHuffle strain. However, higher amounts of L1 are needed to scale-up its production for developing an HPV vaccine candidate(AU)

Introducción: Las vacunas contra el virus del papiloma humano (VPH) se fundamentan en la proteína principal de la cápsida L1. Objetivo: Clonar el gen L1 del VPH-18 a partir de una paciente cubana infectada con VPH-18 y expresar las variantes de longitud completa y delecionadas del gen L1 del VPH-18 en Escherichia coli. Métodos: El gen L1 del VPH-18 de longitud completa se amplificó por PCR a partir de ADN total aislado de un paciente cubana, se clonó y finalmente se subclonó en el vector de expresión de E. coli pET26b. Se construyeron tres mutantes de deleción, que codifican para proteínas truncadas que carecen de 30 aminoácidos por el extremo carboxilo, en combinación con 5, 6 o ningún residuo delecionado por el extremo amino. La producción de las proteínas L1 en E. coli BL21(DE3) y E. coli SHuffle T7 se evaluó mediante SDS-PAGE y Western blot. Resultados: El gen L1 del VPH-18 clonado fue 99.9 percent similar a la variante africana EF202152 y probablemente comparte un origen común con el linaje B del genotipo 18. Las tres variantes truncadas de la proteína L1 del VPH-18 se produjeron a mayores niveles que la proteína L1 del VPH-18 de longitud completa, alcanzando mayores niveles en E. coli BL21(DE3) y mayor solubilidad en E. coli SHuffle. La variante truncada solo por el extremo carboxilo, L1(C30, se produjo a niveles similares a las proteínas L1 del VPH-18 truncadas por ambos extremos. E. coli SHuffle produjo aproximadamente tres veces más cantidades de L1(C30 cuando creció en condiciones de autoinducción, con respecto a la inducción convencional y, por ende, las cantidades fueron comparables a las obtenidas por E. coli BL21(DE3) bajo inducción convencional. Conclusiones: La truncación de treinta residuos de aminoácidos por el extremo carboxilo de la proteína L1 del VPH-18 tuvo una importante contribución a la producción y solubilidad de la proteína L1 nativa en E. coli. Este es el primer informe sobre la producción soluble de la proteína L1 del VPH-18 en una cepa de E. coli SHuffle. Sin embargo, se necesitan mayores cantidades de la proteína L1 para escalar su producción para desarrollar un candidato vacunal contra el VPH(AU)

Advance in human papillomavirus major capsid protein L1-based vaccines / 中华微生物学和免疫学杂志

L1 is the major capsid protein of human papillomavirus (HPV) encoded by late gene 1. Based on the fact that L1 can self-assemble into virus like particle (VLP) with good immunogenicity, it has aroused wide concern in studying the pathogenesis of and vaccines against HPV. Nevertheless, there are a few limitations of present L1-based HPV vaccines. For instance, low expression of the protein and the complexity of purification result in the relatively low yield of vaccines. Type-specific antibody induced by L1 also results in the unsatisfactory cross-protection rate. Furthermore, there is no reported therapeutic effect against HPV-related diseases because of its undefined role in virus eliminating. This review focused on the structure, immunogenicity and role in immune response of L1 and the development of and latest progress in HPV vaccines.

The expression characteristics and co-localization of HIV capsid protein p24 and TRIM22 in U-251 glioma cells / 西安交通大学学报(医学版)

Objective: To observe the expression characteristics and co-localization of exogenous TRIM22 and HIV capsid protein p24 in glioma cells. Methods: The vectors of pEGFP-N3-TRIM22 or pDsRed1-p24 were transfected into U-251 glioma cells respectively to examine the expression of TRIM22-EGFP or p24-DsRed1 by confocal microscopy. Moreover, we used a confocal z-stacking program to achieve series of optical sections and to rebuild 3-D images by ImageJ 1.50i software to detect the expression characteristics of p24-DsRed1 in U-251 cells. In the end, the vectors of pEGFP-N3-TRIM22 and pDsRed1-p24 were co-transfected into U-251 cells to detect the co-localization between TRIM22 and p24 by confocal microscopy. Results: Confocal microscopy results showed that TRIM22-EGFP or p24-Dsred1 was localized to the cell body as well as to protuberance in U-251 cells, and 3D structural reconstruction showed that p24-Dsred1 could be transferred to foot processes of U-251 cells. Simultaneously, confocal microscopy results also showed that TRIM22 and p24 could be co-localized and their combination could be released by budding in U-251 cells co-transfected with pEGFP-N3-TRIM22 and pDsRed1-p24. Conclusion: TRIM22 co-localized to HIV capsid protein p24 and their combination can be released by budding in glioma cells.

Correlation analysis between serum NRSN2 with epstein-Barr virus antibody and DNA in nasopharyngeal carcinoma / 中国医师杂志

Objective To investigate the expression of serum Neurensin 2 (NRSN2) in patients with nasopharyngeal carcinoma (NPC),and to analyze its relationships with epstein-barr virus (EBV) antibody and EBV-DNA.Methods 120 patients with NPC admitted to our hospital from April 2016 to May 2018 were selected as the study group,and 56 healthy people in the physical examination center were selected as the control group.Enzyme linked immunosorbent assay (ELISA) was used to detect the expressions of serum viral capsid antigen-IgA (VCA-IgA),nuclear related tumor antigen-IgA (EBNA-IgA) and early antigen-IgG (EA-IgG).The expression of serum NRSN2 and EBV-DNA were detected by fluorescence quantitative polymerase chain reaction (qPCR).Receiver operating characteristic (ROC) curve was used to analyze the diagnostic value of serum NRSN2 level for NPC.Spearman was used to analyze the relationship between serum NRSN2 and EBV antibody with EBV-DNA in nasopharyngeal carcinoma,multivariate logistic regression analysis was used to analyze the risk factors of NPC.Results The positive rates of serum VCA-IgA,EBNA-IgA and EA-IgG in the study group were significantly higher than those in control group (P ＜0.05);the level of serum NRSN2 and the positive expression rate of EBV-DNA in the study group was significantly higher than that in the control group (P ＜ 0.05);ROC curves showed that the area under curve (AUC) of serum NRSN2 level in diagnosing NPC disease was 0.759,with sensitivity 63.33％,specificity 80.36％;there were significant positive correlations between serum NRSN2 with VCA-IgA,EBNA-IgA,EA-IgG and EBV-DNA in NPC patients (P ＜ 0.05);multivariate regression analysis showed that VCA-IgA,EBNA-IgA,EA-IgG,EBV-DNA and NRSN2 expressions were supportive predictive value for diagnostic NPC (P ＜ 0.05).Conclusions Up-regulation of serum NRSN2 levels in NPC patients is positively correlated with serum VCA-IgA,EBNA-IgA,EA-IgG and EBV-DNA levels,which may provide a reference for the prediction,diagnosis and treatment of NPC.

Progress of screening methods in vitro for HIV-1 capsid protein inhibitors / 中国药理学通报

Human immunodeficiency virus type 1 (HIV-1) infection is highly dependent on its conical fullerene capsid, which is composed of approximately 1 500 capsid proteins. In recent years, capsid protein has been recognized as an ideal target for design and screening drugs for AIDS therapy. Screening methods are the key to develop HIV-1 capsid inhibitors. A variety of screening approaches for HIV-1 capsid protein inhibitor, which have been reported in the literature are reviewed in the article.

Advances in HBV capsid protein assembly modulators / 中国药理学通报

Hepatitis B virus (HBV) belongs to the Hepadnaviridae family. HBV infection may lead to acute and chronic hepatitis, which has become a global health issue. HBV capsid protein assembly modulators play an important role in multiple stages of HBV replication, such as encapsidation of pgRNA and synthesis of HBV DNA, thus becoming a new hotspot in the research and development of hepatitis B drugs. This paper reviews the research progress of HBV capsid protein assembly modulators.

Correlation analysis between serum NRSN2 with epstein-Barr virus antibody and DNA in nasopharyngeal carcinoma / 中国医师杂志

Objective@#To investigate the expression of serum Neurensin 2 (NRSN2) in patients with nasopharyngeal carcinoma (NPC), and to analyze its relationships with epstein-barr virus (EBV) antibody and EBV-DNA.@*Methods@#120 patients with NPC admitted to our hospital from April 2016 to May 2018 were selected as the study group, and 56 healthy people in the physical examination center were selected as the control group. Enzyme linked immunosorbent assay (ELISA) was used to detect the expressions of serum viral capsid antigen-IgA (VCA-IgA), nuclear related tumor antigen-IgA (EBNA-IgA) and early antigen-IgG (EA-IgG). The expression of serum NRSN2 and EBV-DNA were detected by fluorescence quantitative polymerase chain reaction (qPCR). Receiver operating characteristic (ROC) curve was used to analyze the diagnostic value of serum NRSN2 level for NPC. Spearman was used to analyze the relationship between serum NRSN2 and EBV antibody with EBV-DNA in nasopharyngeal carcinoma, multivariate logistic regression analysis was used to analyze the risk factors of NPC.@*Results@#The positive rates of serum VCA-IgA, EBNA-IgA and EA-IgG in the study group were significantly higher than those in control group (P<0.05); the level of serum NRSN2 and the positive expression rate of EBV-DNA in the study group was significantly higher than that in the control group (P<0.05); ROC curves showed that the area under curve (AUC) of serum NRSN2 level in diagnosing NPC disease was 0.759, with sensitivity 63.33%, specificity 80.36%; there were significant positive correlations between serum NRSN2 with VCA-IgA, EBNA-IgA, EA-IgG and EBV-DNA in NPC patients (P<0.05); multivariate regression analysis showed that VCA-IgA, EBNA-IgA, EA-IgG, EBV-DNA and NRSN2 expressions were supportive predictive value for diagnostic NPC (P<0.05).@*Conclusions@#Up-regulation of serum NRSN2 levels in NPC patients is positively correlated with serum VCA-IgA, EBNA-IgA, EA-IgG and EBV-DNA levels, which may provide a reference for the prediction, diagnosis and treatment of NPC.

Advance in human papillomavirus major capsid protein L1-based vaccines / 中华微生物学和免疫学杂志

L1 is the major capsid protein of human papillomavirus (HPV) encoded by late gene 1. Based on the fact that L1 can self-assemble into virus like particle (VLP) with good immunogenicity, it has aroused wide concern in studying the pathogenesis of and vaccines against HPV. Nevertheless, there are a few limitations of present L1-based HPV vaccines. For instance, low expression of the protein and the com-plexity of purification result in the relatively low yield of vaccines. Type-specific antibody induced by L1 also results in the unsatisfactory cross-protection rate. Furthermore, there is no reported therapeutic effect against HPV-related diseases because of its undefined role in virus eliminating. This review focused on the struc-ture, immunogenicity and role in immune response of L1 and the development of and latest progress in HPV vaccines.