Transcriptome analysis of Procambarus clarkii infected with infectious hypodermal and haematopoietic necrosis virus.

Infectious hypodermal and haematopoietic necrosis virus (IHHNV) is a major viral pathogen in cultured penaeid shrimp. IHHNV has many hosts, mainly including crustaceans. It has recently been reported that Procambarus clarkii can be infected by IHHNV. In the present study, we studied the hepatopancreas of P. clarkii by transcriptome high-throughput sequencing to analyze the response of P. clarkii to IHHNV infection. After de novo assembly, there were 400,340,760 clean reads. A total of 237 differentially expressed genes (DEGs) were obtained, including 77 significantly up-regulated unigenes and 160 significantly down-regulated ones. The expression levels of 12 immune-related DEGs were validated by qRT-PCR, substantiating the reliability of RNA-Seq results. The enrichment analysis of DEGs showed that the immune-related pathways were closely related to apoptosis and phagocytosis. Moreover, a large number of pathways related to metabolic function were down-regulated, suggesting that IHHNV infection might affect the growth of P. clarkii.

A novel real-time PCR approach for detection of infectious hypodermal and haematopoietic necrosis virus (IHHNV) in the freshwater crayfish Procambarus clarkii.

Infectious hypodermal and haematopoietic necrosis virus (IHHNV) infects many crustacean hosts, including cultured penaeid shrimp. In the present study, we aimed to develop a novel sensitive SYBR Green-based real-time PCR method to specifically amplify DNA fragments of IHHNV. Our newly developed real-time PCR method with a 195-bp amplicon specifically detected IHHNV and showed no cross reaction with white spot syndrome virus (WSSV), hepatopancreatic parvovirus (HPV), Enterocytozoon hepatopenaei (EHP), infectious myonecrosis virus (IMNV) and yellow-head virus (YHV). This method could detect as low as one single copy of IHHNV plasmid DNA, more sensitive than other SYBR Green-based real-time PCR methods and less expensive and more convenient than the TaqMan probe-based real-time PCR. Moreover, our data using the newly designed method showed that 80% of IHHNV-fed Procambarus clarkii samples were IHHNV positive. Our findings further confirmed that P. clarkii can be infected by IHHNV.

Identification of Mycobacterium tuberculosis PPE68-specific HLA-A*0201-restricted epitopes for tuberculosis diagnosis.

PPE68 is a Mycobacterium tuberculosis-specific protein which is absent from the vaccine strains of BCG. A panel of 14 PPE68-derived peptides predicted to bind to HLA-A*0201 was synthesized. The HLA-A*0201 restriction of these peptides was determined in T2 cell line and HLA-A*0201 transgenic mice. The specificity of peptides was assessed in pulmonary tuberculosis (TB) patients using IFN-γ enzyme-linked immunospot (ELISPOT) assay, and immunodominant peptides were further used to evaluate their diagnostic potential in HLA-A*0201-positive pulmonary TB patients. 13 out of 14 peptides were identified as high-affinity binders. Of these peptides, 12 peptides induced significant IFN-γ-secreting T cell response in transgenic mice and 9 peptides were efficiently recognized by peripheral blood mononuclear cells of 10 HLA-A*0201-positive TB patients. Four immunodominant HLA-A*0201-restricted epitopes (PPE68126-134, PPE68133-141, PPE68140-148, and PPE68148-156) were recognized by the most of 80 HLA-A*0201-positive TB patients (81, 86, 74, and 84 %, respectively). These epitopes may be used for a potential diagnosis of M. tuberculosis infection.

Construction of photo-responsive lignin as a broad-spectrum sunscreen agent.

Lignin has potential to serve as promising sunscreen agents as it has good ultraviolet (UV) absorption and antioxidant properties. However, the weak absorption capacity of lignin in the long-wave UV region (UVA, 320-400 nm) limits its further development. In this work, a spiropyran-modified lignin (DLSP) with photo-responsive property was prepared by in-situ construction of spiropyran (SP) structure in the demethylated lignin (DL). Due to the presence of SP moiety, the absorption of DLSP in the UVA region was significantly improved. Under UV irradiation, its absorption peak was redshifted as unconjugated SP form isomerized to conjugated merocyanine (MC) form, and the UVA/UVB ratio increased from 0.62 to 0.74. The free-radical scavenging ability of lignin could protect SP from photodegradation, which provided DLSP excellent fatigue resistance. DLSP were blended into creams to investigate its sunscreen performance. Results indicated that DLSP exhibited radiation-enhanced sunscreen performance, the sun protection factor (SPF) of cream containing 10 wt% of DLSP improved from 20 to 67 after 8 h of UV irradiation. Moreover, DLSP showed low skin penetration and good biocompatibility. These results provide a useful guideline for the rational design of sunscreens with special functionalities.

[Regulating effects of Paecilomyces cicadae polysaccharides on immunity of aged rats].

OBJECTIVE: To investigate the effects of Paecilomyces cicadae polysaccharide (PCPS) on the immunological function of aged rats in vivo. METHOD: The young and old rats were administered with normal saline as control groups, and the rats from test group were sc given 50, 100, 200 mg x kg(-1) x d(-1) dosage of PCPS for 3 weeks. The phagocytizing rate and index of PMphi, AMphi to S. aureas were observed, and the colorimetric MTI was used to analyze the proliferative activity of spleenocytes which had been stimulated with ConA or LPS. We also inspected the ability varing of ACP, LDH, ARG of spleen, and observed the ultramicro structure of spleen under the SEM. RESULT: The phagocytosis of Mphi was lower in aged group than that in young' s group, and the proliferative activity of spleenocytes was lower too. The activities of ACP, LDH, ARG of spleen were extremely decreased (P < 0.01) in aged rats as well. The proliferative activity and phagocytotic rate were both extremely increased in PCPS groups (P < 0.01), and the mitochondrion and endoplasmic reticulum of spleen were accrementition as well (P < 0.01). CONCLUSION: PCPS could enhance the phagocytizing function of PMphi, AMphi of aged rats in vivo, and strengthen the immune function of spleen and its proliferative activity as well. Then the immunity of aged rats could be improved. The PCPS may be an anti-aging agent.

Identification of conserved and HLA-A*2402-restricted epitopes in Dengue virus serotype 2.

In this study, we set out to identify dengue virus serotype 2 (DENV-2)-specific HLA-A*2402-restricted epitopes and determine the characteristics of T cells generated to these epitopes. We screened the full-length amino-acid sequence of DENV-2 to find potential epitopes using the SYFPEITHI algorithm. Twelve putative HLA-A*2402-binding peptides conserved in hundreds of DENV-2 strains were synthesized, and the HLA restriction of peptides was tested in HLA-A*2402 transgenic mice. Nine peptides (NS4b(228-237), NS2a(73-81), E(298-306), M(141-149), NS4a(96-105), NS4b(159-168), NS5(475-484), NS1(162-171), and NS5(611-620)) induced high levels of peptide-specific IFN-γ-secreting cells in HLA-A*2402 transgenic mice. Apart from IFN-γ, NS4b(228-237-), NS2a(73-81-) and E(298-306)-specific CD8(+) cells produced TNF-α and IL-6 simultaneously, whereas M(141-149-) and NS5(475-484-) CD8(+) cells produced only IL-6. Moreover, splenic mononuclear cells (SMCs) efficiently recognized and killed peptide-pulsed splenocytes. Furthermore, each of nine peptides could be recognized by splenocytes from DENV-2-infected HLA-A*2402 transgenic mice. The SMCs from HLA-A*2402 transgenic mice immunized with nine immunogenic peptides efficiently killed DENV-2-infected splenic monocytes. The present identified epitopes have the potential to be new diagnostic tools for characterization of T-cell immunity in DENV infection and may serve as part of a universal epitope-based vaccine.