Identification and optimization of peptide inhibitors to block VISTA/PSGL-1 interaction for cancer immunotherapy

Developing new therapeutic agents for cancer immunotherapy is highly demanding due to the low response ratio of PD-1/PD-L1 blockade in cancer patients. Here, we discovered that the novel immune checkpoint VISTA is highly expressed on a variety of tumor-infiltrating immune cells, especially myeloid derived suppressor cells (MDSCs) and CD8+ T cells. Then, peptide C1 with binding affinity to VISTA was developed by phage displayed bio-panning technique, and its mutant peptide VS3 was obtained by molecular docking based mutation. Peptide VS3 could bind VISTA with high affinity and block its interaction with ligand PSGL-1 under acidic condition, and elicit anti-tumor activity in vivo. The peptide DVS3-Pal was further designed by d-amino acid substitution and fatty acid modification, which exhibited strong proteolytic stability and significant anti-tumor activity through enhancing CD8+ T cell function and decreasing MDSCs infiltration. This is the first study to develop peptides to block VISTA/PSGL-1 interaction, which could act as promising candidates for cancer immunotherapy.

Perfil clínico dos pacientes atendidos em uma unidade especializada em queimados: estudo transversal / Clinical profile of patients treated at a burns unit: a cross-sectional study / Perfil clínico de pacientes atendidos en una unidad especializada en quemados: estudio transversal

Objetivo: descrever o perfil clínico do paciente admitido terapêutica em um Centro de Tratamento de Queimados. Método: estudo transversal e retrospectivo realizado em um hospital militar do Rio de Janeiro da cidade do Rio de Janeiro. Amostra composta por 43 prontuários. Análise de variáveis de admissão e desfecho óbito. Protocolo de pesquisa aprovados pelos Comitês de Ética em Pesquisa das instituições envolvidas. Resultados: homens de até 39 anos foi o público mais afetado com lesões prevalentes de segundo e terceiro graus, com maior acometimento na região torácica, por agente causador de origem física, apresentando como principal complicação a sepse e desfecho óbito relacionada com a maior taxa de superfície corpórea queimada. Conclusão: os achados demonstram semelhança aos dados do Ministério de Saúde do Brasil, e representam informações adicionais como agente, extensão e grau de queimadura, que podem contribuir na prática assistencial e no gerenciamento de recursos humanos.

Objective: to describe the clinical profile of patients admitted to therapy at a Burn Treatment Center. Method: this retrospective, cross-sectional study was conducted at a military hospital in Rio de Janeiro city on a sample of 43 medical records to examine admission variables and death as outcome. The research protocol was approved by the research ethics committees of the institutions involved. Results: the group most affected was males up to 39 years old, with prevalent second- and third-degree injuries, mostly involving the thoracic region, caused by agents of physical origin, with sepsis as the main complication, and death as outcome related to greater burn surface area. Conclusion: the findings proved similar to data from Brazil's Ministry of Health, and constitute additional information, such as agent, extent, and degree of burns, which can contribute to care practice and human resource management.

Objetivo: describir el perfil clínico del paciente ingresado a terapia en un Centro de Tratamiento de Quemados. Método: estudio transversal y retrospectivo realizado en un hospital militar de Río de Janeiro, en la ciudad de Río de Janeiro. Muestra compuesta por 43 historias clínicas. Análisis de las variables de ingreso y desenlace de muerte. Protocolo de investigación aprobado por los Comités de Ética en Investigación de las instituciones involucradas. Resultados: hombres de hasta 39 años fue el público más afectado, con prevalencia de lesiones de segundo y tercero grados, con mayor número de lesiones en la región torácica, por agente causal de origen físico, siendo la principal complicación la sepsis y la muerte relacionada con la tasa más alta superficie corporal quemada. Conclusión: los hallazgos demuestran similitud con los datos del Ministerio de Salud de Brasil y representan información adicional, como el agente, la extensión y el grado de la quemadura, que pueden contribuir para la práctica asistencial y la gestión de recursos humanos.

Screening and identification of cell-penetrating peptides by display technology / 国际药学研究杂志

Screening and identification of cell-penetrating peptides, CPP which have different functions by display technology are hot topics in recent years. Currently the most widely used display technology is phage display and mRNA display technology. This paper describes the principle and application of display technology such as phage display and mRNA display and reviews in detail the advance in study on the screening and identification of CPP by display technology in recent years.

Screening and identification of cell-penetrating peptides by display technology / 国际药学研究杂志

Screening and identification of cell-penetrating peptides(CPP)which have different functions by display technolo?gy are hot topics in recent years. Currently the most widely used display technology is phage display and mRNA display technology. This paper describes the principle and application of display technology such as phage display and mRNA display and reviews in detail the advance in study on the screening and identification of CPP by display technology in recent years.

Effect of dynamic articulars panning external fixation combined with finite internal fixation for comminuted fractures of distalradius / 中国基层医药

Objective To observe the effect of dynamic articulars panning external fixation combined with limited internal fixation for distal radius comminuted fracture.Methods 100 comminuted distal radius fractures patients during September 2014 and September 2014 in our hospital were selected and randomly divided into control group and research group,50 cases in each group.Patients of the control group accepted conventional manual reduc-tion combined with plaster external fixation,patients of the research group accepted dynamic articulars panning exter-nal fixation combined with limited internal fixation treatment.Curative effect,the palm inclination angle and feet were compared between the two groups.Results (1)After surgical treatment and rehabilitation therapy,15 patients in the control group achieved optimal degree,28 cases in the research group achieved optimal degree(P =0.086);9 cases in the control group were the poor degree;only 2 patients in the research group were poor degree(P =0.025).Statistical analysis showed that two differences were statistically significant,(P <0.05).(2)After 3 months,the palmar tilt of patients in the control group was (6.5 ±2.2)°,while that of the research group was (8.5 ±2.5)°(t =4.25,P <0.000 1 );The average ulnar deviation of control group was (12.3 ±4.3)°,and that of the research group was (15.6 ±6.6)°(t =2.96,P =0.003 8).In a word,the average palmar tilt and ulnar deviation of the research group were greater than those in the control group,the difference showed statistically significant.6 months later,the average palmar tilt of the control group patients was (6.2 ±2.3)°,that of the research group was (8.8 ±2.5)°(t =5.41, P <0.000 1 );the ulnar deviation of the control group was (12.1 ±4.1 )°,and that of the research group was (16.3 ±4.4)°(t =4.93,P <0.000 1),the difference was statistically significant.Conclusion Dynamic articulars panning external fixation combined with limited internal fixation for the treatment of distal radius comminuted fracture effect is significant,which is worthy of clinical promotion.

Panning and identification of humanized antinuclear antibody Fab fragment / 第二军医大学学报

Objective: To prepare humanized antinuclear antibody Fab fragment. Methods: The reconstructed humanized antinuclear antibody (ANA) Fab phage display library was enriched by 4 rounds of panning and was identified by indirect ELISA method. Phasmid DNA isolated from positive clones was deprived of g III gene. After self-ligation the recombinant plasmid was used to transform E. coli. XL1-Blue, then XL1-Blue was induced by IPTG to product soluble human antinuclear antibody Fab fragment. Finally, soluble human antinuclear antibody Fab in the supernatant was identified by indirect ELISA method and immunofluorescence. Results: The eluted phages were enriched by more than 200 folds after 4 rounds of panning. Two positive clones were isolated from the ANA Fab library. Electrophoresis after Xho I digestion proved that the self-ligation was successful after deletion of g III gene. The results of indirect ELISA indicated that the 2 positive clones of Fab had specific anti-dsDNA activity. Indirect immunofluorescence showed homogeneous fluorescence within nuclei of Hep2 and monkey hepatic cells and in the Crithidia kinetoplast. Conclusion: We have successfully prepared soluble, specific human antinuclear antibody Fab fragment, which pares a way for preparation of high affinity antinuclear antibody Fab fragment.

Preparation and Identification of Anti-lactoferrin Single Chain Fv Antibody / 医学研究杂志

Objective To prepare a specific anti-lactoferrin single chain variable fragment(ScFv).Methods Anti-lactoferrin clones were screened from a 'naive' phage antibody library against the immobilized lactoferrin antigen,then the clones were transformed to the E.coli HB2151 to give soluble expression of antibody fragments.The culture supernatant containing ScFv was purified by immobilized metal affinity chromatography,and then determined with SDS-PAGE and ELISA.Results The results demonstrated that ScFvs were specific;they did not react with transferrin,lysozyme and bovine serum albumin in ELISA.The SDS-PAGE showed that the ScFvs had high purity through affinity chromatography and the molecular weight of them was about 32 kD.Conclusions The successful generation of the ScFvs against lactoferrin provides a basis for further study and clinical applications for dry eye and other ocular diseases.

The Selection of Thrombin-specific scAbs by Alternative Panning Strategy / 微生物学通报

Specific scAbs could be obtained through biopanning from phage antibody libraries by use of antigens as target molecules. scAbs specific to thrombin were separated from mouse antibody library by the panning strategy of alternating liquid-solid phase in this paper. Thrombin was biotinylated by photobiotin at first, then avidin-coated magnetic beads were utilized to isolate specific scAbs. The eluted phages were amplified and subject to the second round panning in microtiter plate to remove the unspecific reombinant phages. 4 specific scAbs were separated from 23 phage clones after four rounds of alternative panning.

Screening the peptide high-binding VEGF receptor 3 with phage-displayed random peptide library / 第三军医大学学报

Objective To get a high affinity peptide of vascular endothelial growth factor receptor 3 (VEGFR3) as a potent carrier targeted to lymphangiogenesis of ovarian cancer via the technology of phage display. Methods Solid-phase was panned with direct VEGFR3 extracellular protein coating, then the unbound phage was washed away and the eluted phage was amplified. The positive phage clones were identified by ELISA and sequenced, and the affinity and specialty were identified by competent ELISA. Results After four-round bio-panning, the enriched positive phage clones were identified by ELISA. Eight positive phage clones were sequenced and 5 were consensus (WHGSLKQNLWWY). The short peptide displayed on screened positive phage could bind specifically to VEGFR3, and the binding could be inhibited by natural antibody VEGF-D. Conclusion The phage clone (phage-WHGSLKQNLWWY) obtained via bio-panning of peptide library has a high affinity with VEGF receptor 3. The peptide could be a potent carrier targeted to VEGFR3.