Enhancement of Immune Responses Elicited by Nanovaccines through a Cross-Presentation Pathway

Numerous studies have aimed to develop novel advanced vaccines, in part because traditional vaccines have been unsuccessful in preventing rapidly emerging and reemerging viral and bacterial infections. There is a need for an advanced vaccine delivery system to ensure the successful induction of humoral and cellular immune responses. In particular, the ability of nanovaccines to modulate intracellular antigen delivery by inducing exogenous antigens (loaded onto major histocompatibility complex class 1 molecules) in CD8+ T cells, the so-called cross-presentation pathway, has attracted a great deal of attention. Protection against viral and intracellular bacterial infections relies on cross-presentation.This review discusses the advantages, requirements, and preparation of nanovaccines, the cross-presentation mechanism, the several parameters affecting cross-presentation by nanovaccines, and future perspectives.

Molecular basis of broad neutralization against SARS-CoV-2 variants including Omicron by a human antibody

Omicron, a newly emerging SARS-CoV-2 variant, carried a large number of mutations in the spike protein leading to an unprecedented evasion from many neutralizing antibodies (nAbs). Here, we performed a head-to-head comparison of Omicron with other existing highly evasive variants in terms of their reduced sensitivities to antibodies, and found that Omicron variant is significantly more evasive than Beta and Mu variants. Of note, some key mutations occur in the conserved epitopes identified previously, especially in the binding sites of Class 4 nAbs, contributing to the increased Ab evasion. We also reported a broadly nAb (bnAb), VacW-209, which effectively neutralized all tested SARS-CoV-2 variants and even SARS-CoV. Finally, we determined six cryo-electron microscopy structures of VacW-209 complexed with the spike ectodomains of wild-type, Delta, Mu, C.1.2, Omicron, and SARS-CoV, and revealed the molecular basis of the broadly neutralizing activities of VacW-209 against SARS-CoV-2 variants. Overall, Omicron has once again raised the alarm over virus variation with significantly compromised neutralization. BnAbs targeting more conserved epitopes among variants will continue to play a key role in pandemic control and prevention. One sentence summaryStructural and functional analyses reveal that a human antibody named VacW-209 confers broad neutralization against SARS-CoV-2 variants including Omicron by recognizing a highly conserved epitope.

Study on the age composition of blood donors in some areas of China / 中国输血杂志

【Objective】 To explore the recruitment and retention strategy of blood donors by investigating the age composition of blood donors in some areas of China, so as to promote blood donation and enhance clinical blood supply. 【Methods】 Through the working platform of Practice Comparison Working Group of China’s Mainland Blood Collection and Supply Institutions, the average age and age composition of blood donors from 22 blood centers were collected, and statistical analysis was conducted after eliminating invalid data. 【Results】 The median average age of blood donors during the survey year was 30.02.The median age in 2.89% of the blood centers was lower than 25. The average age of different genders was statistically significant only in 2018(P<0.05). Fot first-time blood donors, the median constituent ratio of donors <25 and ≥25 years old was 54.53% and 44.28%, with median retention rate at 10.30% and 9.61%, respectively. The median overall participation rate of blood donors was 2.7%, with median participation rate of blood donors <25 years old at 5.1%. 【Conclusion】 The recruitment and retention of blood donor is crucial to enhance clinical blood supply. Blood donors <25 years old, with a longer period for future donation, should be the main target of blood donation recruitment. Meanwhile, the revision of upper age limit for blood donation is another important initiative to grow the blood donor pool.

Spike mutations in SARS-CoV-2 variants confer resistance to antibody neutralization

New SARS-CoV-2 variants continue to emerge from the current global pandemic, some of which can replicate faster and with greater transmissibility and pathogenicity. In particular, UK501Y.V1 identified in UK, SA501Y.V2 in South Africa, and BR501Y.V3 in Brazil are raising serious concerns as they spread quickly and contain spike protein mutations that may facilitate escape from current antibody therapies and vaccine protection. Here, we constructed a panel of 28 SARS-CoV-2 pseudoviruses bearing single or combined mutations found in the spike protein of these three variants, as well as additional nine mutations that within or close by the major antigenic sites in the spike protein identified in the GISAID database. These pseudoviruses were tested against a panel of monoclonal antibodies (mAbs), including some approved for emergency use to treat SARS-CoV-2 infection, and convalescent patient plasma collected early in the pandemic. SA501Y.V2 pseudovirus was the most resistant, in magnitude and breadth, against mAbs and convalescent plasma, followed by BR501Y.V3, and then UK501Y.V1. This resistance hierarchy corresponds with Y144del and 242-244del mutations in the N-terminal domain as well as K417N/T, E484K and N501Y mutations in the receptor binding domain (RBD). Crystal structural analysis of RBD carrying triple K417N-E484K-N501Y mutations found in SA501Y.V2 bound with mAb P2C-1F11 revealed a molecular basis for antibody neutralization and escape. SA501Y.V2 and BR501Y.V3 also acquired substantial ability to use mouse and mink ACE2 for entry. Taken together, our results clearly demonstrate major antigenic shifts and potentially broadening the host range of SA501Y.V2 and BR501Y.V3, which pose serious challenges to our current antibody therapies and vaccine protection.

Mucosal Vaccine Delivery Using Mucoadhesive Polymer Particulate Systems

Vaccination has been recently attracted as one of the most successful medical treatments of the prevalence of many infectious diseases. Mucosal vaccination has been interested in many researchers because mucosal immune responses play part in the first line of defense against pathogens. However, mucosal vaccination should find out an efficient antigen delivery system because the antigen should be protected from degradation and clearance, it should be targeted to mucosal sites, and it should stimulate mucosal and systemic immunity. Accordingly, mucoadhesive polymeric particles among the polymeric particles have gained much attention because they can protect the antigen from degradation, prolong the residence time of the antigen at the target site, and control the release of the loaded vaccine, and results in induction of mucosal and systemic immune responses. In this review, we discuss advances in the development of several kinds of mucoadhesive polymeric particles for mucosal vaccine delivery.

Mucosal Vaccine Delivery Using Mucoadhesive Polymer Particulate Systems

Vaccination has been recently attracted as one of the most successful medical treatments of the prevalence of many infectious diseases. Mucosal vaccination has been interested in many researchers because mucosal immune responses play part in the first line of defense against pathogens. However, mucosal vaccination should find out an efficient antigen delivery system because the antigen should be protected from degradation and clearance, it should be targeted to mucosal sites, and it should stimulate mucosal and systemic immunity. Accordingly, mucoadhesive polymeric particles among the polymeric particles have gained much attention because they can protect the antigen from degradation, prolong the residence time of the antigen at the target site, and control the release of the loaded vaccine, and results in induction of mucosal and systemic immune responses. In this review, we discuss advances in the development of several kinds of mucoadhesive polymeric particles for mucosal vaccine delivery.

Structural basis for bivalent binding and inhibition of SARS-CoV-2 infection by human potent neutralizing antibodies

Neutralizing monoclonal antibodies (nAbs) to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) represent promising candidates for clinical intervention against coronavirus virus diseases 2019 (COVID-19). We isolated a large number of nAbs from SARS-CoV-2 infected individuals capable of disrupting proper interaction between the receptor binding domain (RBD) of the viral spike (S) protein and the receptor angiotensin converting enzyme 2 (ACE2). In order to understand the mechanism of these nAbs on neutralizing SARS-CoV-2 virus infections, we have performed cryo-EM analysis and here report cryo-EM structures of the ten most potent nAbs in their native full-length IgG or Fab forms bound to the trimeric S protein of SARS-CoV-2. The bivalent binding of the full-length IgG is found to associate with more RBD in the "up" conformation than the monovalent binding of Fab, perhaps contributing to the enhanced neutralizing activity of IgG and triggering more shedding of the S1 subunit from the S protein. Comparison of large number of nAbs identified common and unique structural features associated with their potent neutralizing activities. This work provides structural basis for further understanding the mechanism of nAbs, especially through revealing the bivalent binding and their correlation with more potent neutralization and the shedding of S1 subunit.

Potent human neutralizing antibodies elicited by SARS-CoV-2 infection

The pandemic caused by emerging coronavirus SARS-CoV-2 presents a serious global public health emergency in urgent need of prophylactic and therapeutic interventions. SARS-CoV-2 cellular entry depends on binding between the viral Spike protein receptor-binding domain (RBD) and the angiotensin converting enzyme 2 (ACE2) target cell receptor. Here, we report on the isolation and characterization of 206 RBD-specific monoclonal antibodies (mAbs) derived from single B cells of eight SARS-CoV-2 infected individuals. These mAbs come from diverse families of antibody heavy and light chains without apparent enrichment for particular families in the repertoire. In samples from one patient selected for further analyses, we found coexistence of germline and germline divergent clones. Both clone types demonstrated impressive binding and neutralizing activity against pseudovirus and live SARS-CoV-2. However, the antibody neutralizing potency is determined by competition with ACE2 receptor for RBD binding. Surprisingly, none of the SARS-CoV-2 antibodies nor the infected plasma cross-reacted with RBDs from either SARS-CoV or MERS-CoV although substantial plasma cross-reactivity to the trimeric Spike proteins from SARS-CoV and MERS-CoV was found. These results suggest that antibody response to RBDs is viral species-specific while that cross-recognition target regions outside the RBD. The specificity and neutralizing characteristics of this plasma cross-reactivity requires further investigation. Nevertheless, the diverse and potent neutralizing antibodies identified here are promising candidates for prophylactic and therapeutic SARS-CoV-2 interventions.

Associations Among Plasma Stress Markers and Symptoms of Anxiety and Depression in Patients with Breast Cancer Following Surgery

OBJECTIVE: The objective of present study is to analyze the prevalence of depression and anxiety following breast cancer surgery and to assess the factors that affect postoperative psychological symptoms. METHODS: The Hamilton Rating Scale for Depression (HAM-D), Hamilton Anxiety Rating Scale (HAM-A), Body Image Scale (BIS), and Rosenberg Self Esteem Scale (RSES) were used to assess the psychological states of patients who had been diagnosed with and had undergone surgery for breast cancer. Blood concentrations of the stress markers adrenocorticotropic hormone, cortisol, arginine-vasopressin, and angiotensin-converting enzyme were measured. Pearson’s correlation analysis and multilinear regression analysis were used to analyse the data. RESULTS: At least mild depressive symptoms were noted in 50.5% of patients, while 42.4% of patients exhibited at least mild anxiety symptoms. HAM-D score was positively correlated with HAM-A (r=0.83, p < 0.001) and BIS (r=0.29, p < 0.001) scores and negatively correlated with RSES score (r=-0.41, p < 0.001). HAM-A score was positively correlated with BIS score (r=0.32, p < 0.001) and negatively correlated with RSES score (r=-0.27, p < 0.001). There were no statistically significant associations between stress markers and depression/anxiety. CONCLUSION: Patients with breast cancer frequently exhibit postoperative depression and anxiety, which are related to low levels of self-esteem and distorted body image.

Single B cell sorting-based amplification and functional identification of HIV-1 monoclonal antibody genes / 中华微生物学和免疫学杂志

Objective@#To amplify and identify monoclonal antibody genes from HIV-1-infected patients.@*Methods@#Single cell sorting was used to isolate antigen-specific single B cells. Sequence Identity Matrix and the international ImMunoGeneTics information system were used to analyze antibody variable region genes. Binding abilities were detected by enzyme linked immunosorbent assay. Neutralizing activities were tested by TZM-bl/pseudovirus assay.@*Results@#The heavy and light chain genes of four, seven, and eleven antibodies were amplified and sequenced from three HIV-1-infected patients, respectively. They were derived from various germline genes with flexible CDR3 lengths and somatic mutations. A1 and B3 antibodies bound to HIV-1 clade B, CRF01_AE, and CRF07_BC antigens. The half maximal inhibitory concentration values of A1 and B3 against MW965 virus were 0.04 μg/ml and 37.34 μg/ml.@*Conclusion@#In this study, we acquired a lot of monoclonal antibody genes and two HIV-1 monoclonal binding and neutralizing antibodies, which would provide basic data for further research on monoclonal antibody identification.

Isolation and identification of neutralizing monoclonal antibodies to two Tier 2 viruses in a patient with HIV-1 infection / 中华微生物学和免疫学杂志

Objective To isolate neutralizing monoclonal antibodies to Tier 2 viruses in a Chinese patient with HIV-1 infection. Methods Monoclonal antibodies were isolated using single B cell sorting and monoclonal antibody expression technique. The international ImMunoGeneTics database (IMGT) was used to analyze antibody variable region genes. Antibody binding ability and neutralizing activity were tested by enzyme linked immunosorbent assay and TZM-bl/pseudovirus assay,respectively. Results Two monoclonal antibodies (E11 and H2) that could neutralize two Tier 2 viruses were isolated from the patient with clade B HIV-1 infection. Heavy chains of E11 and H2 were derived from IGHV4-4*08 with a somatic mutation rate of 15.79% and 14.74%,respectively. Light chains were both derived from IGKV3-15*01 with a somatic mutation rate of 8.33% and 7.95%, respectively. E11 and H2 could bind to HIV-1 clade B, CRF01_AE and CRF07_BC viruses. The half maximal inhibitory concentration(IC50) values of E11 and H2 were 18.78 μg/ml and 22.43 μg/ml against 398-F1 virus and 43.35 μg/ml and 39.45 μg/ml against 25710 virus. Conclusion In this study, two neutralizing monoclonal antibodies to two Tier 2 viruses were identified in the patient with HIV-1 infection,which might provide reference for the development of AIDS vaccines.

Analysis of Genetic Polymorphism and Genetic Distance of 19 Autosomal STR Loci in Jiangsu Han Population / 法医学杂志

Objective To investigate the distribution of alleles in 19 autosomal short tandem repeat (STR) loci in Jiangsu Han population.Methods Goldeneye20A kit was used to detect 9 025 samples.Genetic analysis was performed on typing data of 19 autosomal STR loci, and genetic distance with other 17 populations was analyzed.Results All the 19 autosomal STR loci were consistent with the Hardy-Weinberg equilibrium (P＞0.05), with the heterozygosity 0.616 1-0.916 3, probability of match 0.012 8-0.202 6, discrimination power 0.797 4-0.987 2, probability of paternity exclusion 0.310 8-0.828 8, and polymorphic information content 0.561 7-0.913 6.The cumulative discrimination power and cumulative probability of exclusion were 0.999 999 999 999 999 998 434 1 and 0.999 999 989, respectively.The Jiangsu Han population had close genetic distances with the Han population in Tianjin, Hunan and Jilin, and significant difference with Han population in Aletai region in Xinjiang (P＜0.05).Conclusion The STR allele polymorphism data and population genetic parameters of Jiangsu Han population can provide data support for the forensic application of these STR loci in Jiangsu Han population.

Genetic Polymorphism of 30 InDel Loci in Han Population from Jiangsu Province / 法医学杂志

Objective To investigate the genetic information of 30 insertion/deletion (InDel) loci in Han population from Jiangsu Province, and to explore the application values of Investigator? DIPplex kit for guiding the forensic analysis in Han population from Jiangsu Province. Methods The autosomal InDel loci of 305 unrelated healthy Han individuals from Jiangsu Province were genotyped and analysed by In-vestigator? DIPplex kit, and the allelic frequencies and forensic parameters of 30 InDel loci were statis-tically analysed. Results The distribution of 30 InDel loci in Han population from Jiangsu Province con-formed to Hardy-Weinberg equilibrium. The minor allele frequencies of 21 InDel loci were above 0.3. The polymorphism information content ranged from 0.089 to 0.375, while the discrimination power dis-tributed from 0.093 to 0.500. The paternity exclusion in duo cases and trio cases were 0.047-0.250 and 0.046-0.219, respectively. The linkage disequilibrium analysis of 30 InDel loci showed that all loci were independent from each other. The combined discrimination power was 1-7.369 ×10-8, whereas the com-bined mean exclusion chance in duo cases was 0.998933978, in trio cases was 0.997806392. The Fst values were all less than 0.06 except HLD118 and other four loci, which showed small differences be-tween groups. Conclusion The InDel loci of Investigator ? DIPplex kit can be used as complementary genetic markers for the cases associated with forensic genetics.

Mental Health Evaluation for Elderly in Community, Pilot Study

OBJECTIVE: With the burgeoning elderly population, mental health among the elderly has surfaced as an important medical, social, and economic issue. This study was a pilot study testing the assessment module that was developed to survey mental health among the elderly living in the city of Busan. METHODS: Basic demographic data and mental health problems were investigated using the General Health Questionnaire-12 (GHQ-12), suicide risk screening tool, subjective memory deterioration scale, sleep disorder and sleep pattern scale, health-related quality of life scale (EuroQoL-5 dimension), and alcohol abuse screening tool (Cut down, Annoyed, Guilty, Eye-opener Questionnaire). RESULTS: Four-hundred adults aged 65 years or older living in either of two districts in the Busan metropolitan city participated in the study, of whom 219 (54.8%) were female. About 26.8% of the participants were obese, while 2.3% were underweight. One hundred and twenty-five (31.3%) participants were in need of attention to or improvement of nutrition management, and 19.1% perceived themselves to be in poor health. In the GHQ-12, 68.8% were in psychological distress (≥2). About 49.8% of the participants had insomnia, 61.8% had subjective memory disturbance, and 6% were at risk of suicide. The GHQ-12 score, proportion of participants with sleep disturbance, and subjective memory deterioration was significantly higher among women than in men, while health-related quality of life was higher in men. CONCLUSION: A considerable number of older adults living in the Busan region were suffering from psychological stress, were in need of nutrition management, had low quality of life, and had poor mental health, and women were particularly more vulnerable.

Psychiatric Symptoms Among Female Adult Victims of Sexual Molestation : Comparison with Rape Victims / 정신신체의학

OBJECTIVES: According to the recent Korean National Police Agency report, rape victims were 17.1%, but sexual molestation victims were more than the rape victims by 78.0%. Despite many international reports about the occurrence of severe psychiatric symptoms in sexual molestation victims, there is no domestic research. Therefore in this study, we investigated psychiatric symptoms of sexual molestation victims, and we also compared it with psychiatric symptoms in rape victims. METHODS: 58 women who visited Busan Smile Center within 3months after sexual violence were the study subjects. Questionnaire about sociodemographic and sexual violence related characteristics were retrospectively investigated. Of the psychiatric symptoms, Beck Depression Inventory(BDI), Beck Anxiety Inventory(BAI) for severity of depression and anxiety, and Impact of Event Scale-Revised(IES-R) to check the presence of posttraumatic stress disorder(PTSD) symptoms were used. RESULTS: Of 58 sexual violence victims, sexual molestation victims were 36(62.1%) and rape victims were 22(37.9%). In sexual molestation victims, 80.6% had more than moderate severity of depression, 83.3% had more than moderate severity of anxiety, and 94.4% had significant scores at PTSD screening test. Compared with rape victims' psychiatric symptoms(each 95.5%, 95.5%, 95.5%) there were no significant difference. CONCLUSIONS: The majority of sexual molestation victims were also accompanied by depression, anxiety and PTSD symptoms as rape victims. There results suggest that appropriate assessment and early treatment for psychiatric symptom must be made in the early stage of injury in sexual molestation victims.

CD137 molecule permits single cell sorting and expansion of HIV-specific T cells in vitro / 中华微生物学和免疫学杂志

Objective To isolate HIV-specific T cell clone and to expand them in vitro through the activation-induced expression of CD137 molecule.Methods The peripheral blood mononuclear cells were isolated from HIV-infected patients and HIV Gag specific CD3+CD8+CD137+T cell subset were sorted to 96-well plate in 1 cell/well by multicolor flowcytometry and single cell sorting.After 14 days in vitro culture with feeder cells and cytokines, the numbers and phenotypes of the cultured HIV-specific T cells were calcu-lated and identified.Results The CD137 expression was low on rested T cells but up regulated by the stim-ulation with Gag peptide pool.The CD8+CD137+T cells could secret IFN-γ.The number of CD8 T cells reached to 106 after 14 days in culture and expanded to 107-108 cells after 28 days of culture in vitro 100%of the cells remained activated upon Gag stimulation.Conclusion In stead of using IFN-γ, CD137 could be utilized as a novel molecule to isolate and expand HIV specific T cells in vitro.The expanded antigen spe-cific T cell clones could maintain good activation status.

Comparative Study of Adolescents' Mental Health between Multicultural Family and Monocultural Family in Korea

OBJECTIVES: With the observation of factors influencing depression, self-reported happiness, self-reported stress and suicidal attempt, this study compared and evaluated the mental health of adolescents from multicultural families and monocultural families in Korea. METHODS: This study compared the characteristics of adolescents from multicultural and monocultural families based on the analysis of data of The Tenth Korea Youth Risk Behavior Web-based Survey and analyzed the factors influencing adolescent depression, self-reported happiness, self-reported stress and suicidal attempt. RESULTS: There was no significant difference in the levels of depression, self-reported happiness, and self-reported stress between adolescents from multicultural and monocultural families. However, suicidal ideation and suicidal attempt were significantly higher in adolescents from multicultural families. CONCLUSION: There was no significant difference between the mental health of adolescents from multicultural and monocultural families. Because depression is the greatest factor for suicidal attempt, and violence has stronger relationship with depression of adolescents from multicultural families, further study is needed to prevent this trend.

Oral health knowledge, attitudes and practice among teachers of primary and secondary schools in Gongshu District of Hangzhou City / 中华健康管理学杂志

Objective To explore the current status and influencing factors of oral health knowledge,attitudes and practice (KAP) among the teachers of primary and secondary schools in Gongshu District of Hangzhou City.Methods A total of 652 teachers of primary and secondary schools were selected by using multi-stage stratified cluster random sampling method and completed a self-designed questionnaire,which included demographic characteristics and oral health KAP.The status and influencing factors of oral health KAP were analyzed by single and multiple-factor analysis methods.Results The questionnaire was completed by all the 652 teachers,627 of whom were qualified (96.2％).The total ratios of oral health knowledge awareness,attitudes correctness and behaviors formation were 77.1％,68.7％ and 46.3％,respectively.There was significant difference of knowledge awareness rate by age,gender,marriage status,monthly income,life status and current cigarette smoking (x2 values were 11.48,8.57,8.12,8.46,23.97 and 4.64,respectively; all P＜0.05).There was significant difference in oral health attitudes correctness rate by gender (x2=10.65,P＜0.05) and behaviors formation rate by gender,education level and life status (x2 values were 3.95,10.88 and 10.41,respectively; all P＜0.05).In logistic regression model,impact factors of oral health knowledge awareness were age,gender,life status and physical exercises; the impact factors of oral health attitudes correctness were gender and monthly income; the impact factors of oral health behaviors formation were gender,level of education and physical exercises.Conclusion The oral health awareness and attitudes correctness are acceptible among the teachers of primary and secondary schools in Gongshu District of Hangzhou City,although the formation rate of oral health behaviors is low.Oral health education should be enforced among the teachers of primary and secondary schools.

A Case of Necrotizing Fasciitis and Severe Sepsis Complicated by Emphysematous Gastritis

We report a case of necrotizing fasciitis and severe sepsis complicated by emphysematous gastritis. The patient initially presented with sepsis due to extensive necrotizing fasciitis. Although he recovered from severe sepsis after intensive treatment including amputation of involved lesion, fatal emphysematous gastritis developed later during long-term hospitalization. To the best of our knowledge, this is the first report on emphysematous gastritis associated with necrotizing fasciitis and sepsis.

[Effect of octadecyl amine (ODA) on the complex titration of magnesium and calcium ions studied with UV-visible spectrophotometry].

The effect of flotation agent octadecyl amine (ODA) on the complex titration of both magnesium and calcium ions was studied with two groups of comparative experiments: (1) Before titration, the suspension was not filtered. In this case, ODA had a great effect on the complex titration of both magnesium and calcium ions. The titration end-point of magnesium ions was difficult to be determined. Although the titration end-point of calcium ions could be determined, there was an obvious experimental error compared with the blank solution without ODA. These results were confirmed by the UV-Visible spectrum analyses of the related solutions. (2) Before titration, the suspension was filtered. In this case, the influence of ODA on the complex titration of both magnesium and calcium ions could be removed. UV-Visible spectrum studies showed that, in this case, both the spectra and time scanning curves of the tested solutions were similar to those of the blank solutions.