Bioengineered hydrogels enhance ex vivo preservation of patient-derived tumor explants for drug evaluation.

Ex vivo patient-derived tumor slices (PDTS) are currently limited by short-term viability in culture. Here, we show how bioengineered hydrogels enable the identification of key matrix parameters that significantly enhance PDTS viability compared to conventional culture systems. As demonstrated using single-cell RNA sequencing and high-dimensional flow cytometry, hydrogel-embedded PDTS tightly preserved cancer, cancer-associated fibroblast, and various immune cell populations and subpopulations in the corresponding original tumor. Cell-cell communication networks within the tumor microenvironment, including immune checkpoint ligand-receptor interactions, were also maintained. Remarkably, our results from a co-clinical trial suggest hydrogel-embedded PDTS may predict sensitivity to immune checkpoint inhibitors (ICIs) in head and neck cancer patients. Further, we show how these longer term-cultured tumor explants uniquely enable the sampling and detection of temporal evolution in molecular readouts when treated with ICIs. By preserving the compositional heterogeneity and complexity of patient tumors, hydrogel-embedded PDTS provide a valuable tool to facilitate experiments targeting the tumor microenvironment.

Application of spherical lens with 0.05 D intervals in optometry for small incision lenticule extraction in myopic eyes / 中华实验眼科杂志

Objective:To investigate the clinical value of spherical lens with 0.05 D intervals in optometry for small incision lenticule extraction (SMILE) in myopic eyes.Methods:A randomized controlled clinical study was conducted.Sixty patients (120 eyes) with low to moderate myopia and myopic astigmatism who underwent SMILE in the 989th Hospital of the PLA from June 2021 to February 2022 were enrolled.The patients were randomly divided into 0.05 D interval group (optometry with spherical lens at 0.05 D interval) and 0.25 D interval group (optometry with spherical lens at 0.25 D interval), with 30 cases (60 eyes) in each group.There was no significant difference in matched age, sphericity, cylindricity, and best corrected visual acuity (BCVA) (all at P>0.05). The preoperative monocular red-green balance, 1- and 3-month postoperative monocular red-green balance, uncorrected visual acuity and spherical equivalent of both groups were compared.This study adhered to the Declaration of Helsinki.The study protocol was approved by the Ethics Committee of the 989th Hospital of the PLA (No.WZLL-2021-034). Written informed consent was obtained from each subject before any medical examination. Results:The preoperative red-green balance rate in 0.05 D interval group was 95.00%(57/60), which was higher than 35.00%(21/60) in 0.25 D interval group, showing a statistically significant difference ( Wald χ2=17.642, P<0.001). The 1- and 3-month postoperative red-green balance rates in 0.05 D interval group were 63.33%(38/60) and 56.67%(34/60), which were higher than 23.33%(14/60) and 21.67%(13/60) in 0.25 D interval group respectively, showing statistically significant differences ( Wald χ2=9.137, P=0.003; Wald χ2=7.483, P=0.006). The 1- and 3-month postoperative visual acuity in 0.05 D interval group were -0.1(-0.1, -0.1) and -0.1(-0.1, -0.1), which were higher than 0.0(-0.1, 0.0) and -0.1(-0.1, 0.0) in 0.25 D interval group respectively, showing statistically significant differences ( Wald χ2=11.624, P=0.001; Wald χ2=12.841, P<0.001). The 1- and 3-month postoperative spherical equivalent were -0.07(-0.25, 0.13)D and -0.13(-0.25, 0.13)D in 0.05 D interval group, which were higher than -0.13(-0.38, 0.25)D and -0.13(-0.38, 0.25)D in 0.25 D interval group respectively, showing no statistically significant difference between the two groups ( Wald χ2=0.029, P=0.866; Wald χ2=0.189, P=0.664). Conclusions:Compared with spherical lens at 0.25 D interval, 0.05 D interval can improve the accuracy of preoperative and postoperative red-green balance rate and postoperative visual acuity in patients with low to moderate myopia who undergo SMILE.

Notch-dependent cooperativity between myeloid lineages promotes Langerhans cell histiocytosis pathology.

Langerhans cell histiocytosis (LCH) is a potentially fatal neoplasm characterized by the aberrant differentiation of mononuclear phagocytes, driven by mitogen-activated protein kinase (MAPK) pathway activation. LCH cells may trigger destructive pathology yet remain in a precarious state finely balanced between apoptosis and survival, supported by a unique inflammatory milieu. The interactions that maintain this state are not well known and may offer targets for intervention. Here, we used single-cell RNA-seq and protein analysis to dissect LCH lesions, assessing LCH cell heterogeneity and comparing LCH cells with normal mononuclear phagocytes within lesions. We found LCH discriminatory signatures pointing to senescence and escape from tumor immune surveillance. We also uncovered two major lineages of LCH with DC2- and DC3/monocyte-like phenotypes and validated them in multiple pathological tissue sites by high-content imaging. Receptor-ligand analyses and lineage tracing in vitro revealed Notch-dependent cooperativity between DC2 and DC3/monocyte lineages during expression of the pathognomonic LCH program. Our results present a convergent dual origin model of LCH with MAPK pathway activation occurring before fate commitment to DC2 and DC3/monocyte lineages and Notch-dependent cooperativity between lineages driving the development of LCH cells.

Application of extended trochanter osteotomy wire fixation combined with autologous bone graft in revision of total hip arthroplasty / 中国骨伤

OBJECTIVE@#To investigate the clinical effect of using lengthened trochanteric osteotomy wire fixation combined with autologous bone graft in patients undergoing revision total hip arthroplasty.@*METHODS@#From December 2010 to December 2018, 18 patients underwent revision of total hip arthroplasty with extended trochanteric osteotomy wire fixation and autogenous bone graft, including 8 males and 10 females with an average age of (78.89±3.32) years old ranging from 68 to 82 years. The time from the initial replacement to the revision was 9 to 22 (16.33±2.93) years. The patients were followed up regularly after operation. The healing time of osteotomy, the time of full weight-bearing activity, Harris score of hip joint and complications were recorded.@*RESULTS@#All 18 patients were followed up for 16 to 38 months with an average of (25.78±6.65) months. The incision length was 16 to 21 cm with an average of (18.89±1.32) cm; the operation time was 105 to 128 min with an average of (115.44±6.59) min, the bleeding volume was 240 to 285 ml with an average of (267.44±13.77) ml. The healing time of osteotomy was 12 to 18 weeks with an average of (15.61±1.75) weeks. Harris score of hip joint was (47.11±5.04) before operation, (76.39±3.85) during full weight-bearing activities, and (82.22±2.76) at the final follow-up(P<0.05). During the follow-up period, there were no complications such as limb shortening, infection, poor incision healing, prosthesis loosening and sinking, and periprosthetic fracture.@*CONCLUSION@#In revision total hip arthroplasty, the use of extended trochanteric osteotomy wire fixation combined with autologous bone graft can achieve satisfactory clinical results, but the surgeon needs to make a systematic plan for the pre-revision, intraoperative and postoperative recovery.

Data-Driven Analysis of COVID-19 Reveals Persistent Immune Abnormalities in Convalescent Severe Individuals.

Severe SARS-CoV-2 infection can trigger uncontrolled innate and adaptive immune responses, which are commonly associated with lymphopenia and increased neutrophil counts. However, whether the immune abnormalities observed in mild to severely infected patients persist into convalescence remains unclear. Herein, comparisons were drawn between the immune responses of COVID-19 infected and convalescent adults. Strikingly, survivors of severe COVID-19 had decreased proportions of NKT and Vδ2 T cells, and increased proportions of low-density neutrophils, IgA+/CD86+/CD123+ non-classical monocytes and hyperactivated HLADR+CD38+ CD8+ T cells, and elevated levels of pro-inflammatory cytokines such as hepatocyte growth factor and vascular endothelial growth factor A, long after virus clearance. Our study suggests potential immune correlates of "long COVID-19", and defines key cells and cytokines that delineate true and quasi-convalescent states.

Data-driven analysis of COVID-19 reveals specific severity patterns distinct from the temporal immune response

Key immune signatures of SARS-CoV-2 infection may associate with either adverse immune reactions (severity) or simply an ongoing anti-viral response (temporality); how immune signatures contribute to severe manifestations and/or temporal progression of disease and whether longer disease duration correlates with severity remain unknown. Patient blood was comprehensively immunophenotyped via mass cytometry and multiplex cytokine arrays, leading to the identification of 327 basic subsets that were further stratified into more than 5000 immunotypes and correlated with 28 plasma cytokines. Low-density neutrophil abundance was closely correlated with hepatocyte growth factor levels, which in turn correlated with disease severity. Deep analysis also revealed additional players, namely conventional type 2 dendritic cells, natural killer T cells, plasmablasts and CD16+ monocytes, that can influence COVID-19 severity independent of temporal progression. Herein, we provide interactive network analysis and data visualization tools to facilitate data mining and hypothesis generation for elucidating COVID-19 pathogenesis.

A follow up study on the treatment of tibial fractures with intramedullary interlocking nail through the suprapatellar approach / 中国骨伤

OBJECTIVE@#To investigate the short-term effect of suprapatellar interlocking intramedullary nail in the treatment of tibial fractures.@*METHODS@#Eighty patients with tibial fractures treated from January 2016 to June 2018 were treated with interlocking intramedullary nail, who were divided into observation group (suprapatellar approach) and control group (patellar ligament approach) according to different surgical approaches. There were 40 cases in the observation group, including 28 males and 12 females, aged 28 to 67 years with a mean of (46.70±10.34) years. There were 40 cases in the control group, including 30 males and 10 females, aged 31 to 69 years with a mean of(49.38±10.74) years. The operation time, incision length, intraoperative C-arm X-ray fluoroscopy times, intraoperative blood loss, fracture healing time, postoperative active straight leg raise (SLR) time, hospital stay, visual analogue scale (VAS), knee pain rate and postoperative Hospital for Special Surgery (HSS) score were recorded and compared between two groups.@*RESULTS@#All the patients were followed up, and the duration ranged from 19 to 38 months, with an average of(24.60±4.52) months. In the observation group, the operation time was(53.83± 7.01) min;the incision length was (3.98±0.83) cm;the number of intraoperative C-arm X-ray fluoroscopy was (18.90±1.75) times;the fracture healing time was (10.03±0.89) weeks;the postoperative active SLR time was (1.19±0.25) days;and the hospital stay was(6.73±1.06) days. The above indexes were better than those in the control group (@*CONCLUSION@#The treatment of tibial fractures with suprapatellar interlocking intramedullary nail has the advantages of less trauma and better recovery of knee function. It can obtain more satisfactory clinical results and can be further widely used.

Analysis of related risk factors for the refracture of adjacent vertebral body after percutaneous kyphoplasty / 中国骨伤

OBJECTIVE@#To investigate the risk factors of vertebral refracture after percutaneous kyphoplasty (PKP) for osteoprotic vertebral compression fractures (OVCFs), and to provide reference for clinical prevention.@*METHODS@#A retrospective analysis of 228 OVCFs patients who met the inclusion criteria admitted from November 6, 2013 to December 14, 2018. There were 35 males and 193 females, with a male-to-female ratio of 3∶20, and aged 58 to 91 years with an average of (69.70±7.03) years. All patients were treated with PKP and had complete clinical data. According to whether refracture occurred after operation, they were divided into refracture group (24 cases) and non refracture group (204 cases). Factors that may be related to refracture (including gender, age, surgical segment, number of vertebral bodies in the surgical segment, whether combined with degenerative scoliosis, whether anti-osteoporosis treatment) were included in the univariate analyses, and the single factor analysis of statistically significant risk factors was carried out with multiple Logistic regression analysis to further clarify the independent risk factors for vertebral body refracture after PKP. Survival analysis was performed using the time of vertebral refracture after PKP as the end time of follow up, the occurrence of refracture after PKP as the endpoint event, and the presence or absence of degenerative lateral curvature as a variable factor.@*RESULTS@#All 228 patients were followed up for 1.8 to 63.6 months with an average of (28.8±15.6) months, and the refracture rate was 10.5%(24/228). There were statistically significant differences between two groups in age, number of operative vertebral bodies, whether combinedwith degenerative scoliosis and whether anti osteoporosis treatment (@*CONCLUSION@#Combined scoliosis is an independent risk factor for refracture after OVCFs vertebroplasty, and it is also a possible high-risk factor for refracture after surgery.

Effect of double plate technique combined with autogenous iliac bone graft in the treatment of femoral nonunion after intramedullary nailing / 中国骨伤

OBJECTIVE@#To investigate the clinical effect of double plate combined with iliac bone graft in the treatment of femoral nonunion after intramedullary nailing.@*METHODS@#From December 2008 to December 2017, double plate combined with autogenous iliac bone graft was used to treat femoral nonunion after intramedullary nailing. There were 11 cases, including 10 males and 1 female, aged 35 to 62 years, and the time from fracture to nonunion was 12 to 20 months. According to Judet classification, there were 8 cases of atrophic nonunion and 3 cases of proliferative nonunion. Regular follow-up was conducted after operation to record the fracture healing time, load-bearing activity time and complications, and to observe the repair effect of double plate fixation combined with iliac bone graft on nonunion after femoral shaft fracture operation.@*RESULTS@#All patients were followed up for 12 to 22 months. The operation time was 70 to 130 min and the blood loss was 180 to 350 ml. After operation, 2 cases had knee stiffness, which recovered after passive exercise with CPM machine for 2 weeks;1 case had pain in iliac bone donor area, which was relieved after 3 months. The time of fracture healing was 24 to 40 weeks, and the time of complete weight-bearing activity was 14 to 32 weeks. SF-36 quality of life score at the final follow-up:body pain 70 to 82, activty 70 to 82, social function 72 to 83, the overall health 72 to 82. At the end of the follow-up, there were no complications such as limb shortening, infection, poor wound healing, internal fixation failure (fracture, loosening).@*CONCLUSION@#It is an effective method to treat nonunion of femur after intramedullary nailing by using double plate combined with autogenous iliac bone graft.

Frequent Import and Multiple Sources of Dengue Fever have Changed the Epidemic Situation of the Disease in Fujian Province, China / 生物医学与环境科学（英文）

Objective@#The aim of this study was to update the epidemic situation of dengue fever (DF) and provide new insights for the consideration of disease control in Fujian province, China.@*Methods@#Details about DF cases in Fujian reported during 2004-2017 were collected and analyzed. The envelope (E) genes of isolates of dengue virus (DENV) were sequenced for phylogenetic analysis.@*Results@#The number of imported DF cases had increased dramatically since 2013, and the source regions expanded from Southeast Asia to South Asia, America, Oceania, and Africa, as well as the surrounding provinces. This resulted in local outbreaks and indigenous cases of DF that occurred more frequently, with 10 of 13 local outbreaks and 85.9% (1,252/1,458) of indigenous cases reported in 2013-2017. Compared with only two coastal cities before 2013, four coastal and one inland city in 2013-2017 experienced the local DF outbreaks. The phylogenetic analysis of E genes confirmed that the import of DENV, not only from abroad but also from the surrounding provinces, played an important role in dissemination and local outbreaks of DF in Fujian.@*Conclusions@#The frequent import of DF cases from not only abroad but also the surrounding provinces resulted in increased incidence, frequent local outbreaks, and expansion of distribution in Fujian in recent years. There is a need for urgent measures to improve disease control in this province.

Guiding significance of lumbar quantitative computed tomography in percutaneous vertebroplasty for osteoporotic vertebral compression fractures / 中国骨伤

OBJECTIVE@#To investigate the guiding significance of lumbar quantitative computed tomography (QCT) in percutaneous vertebroplasty (PKP) for osteoporotic vertebral compression fractures (OVCF).@*METHODS@#The clinical data of 90 patients with OVCF underwent PKP from December 2017 to December 2018 were retrospectively analyzed. There were 24 femalesand66males, withanaverage agedof (74.47±6.60) yearsold. Allpatientswere received QCT examination before surgery, andaccording to the QCT value oflumbarspine, the patientswere dividedinto osteopenia decrease group (80 to 120 g/L, 17 cases, 30 vertebrae), osteoporosis group (40 to 80 g/L, 44 cases, 66 vertebrae) and severe osteoporosis group (<40 g/L, 29 cases, 39 vertebrae). Bone cement was injected into vertebral body, AP and lateral X-rays were done during operation. The diffusion and leakage of bone cement in injured vertebrae of patients with different QCT values were observed. Unilateral approach was used for patients whose bone cement diffused beyond the midline of the vertebral body, otherwise, and bilateral approach was adopted, and guiding significance of QCT in PKP for OVCF was analyzed.@*RESULTS@#In 90 cases of 135 vertebrae, 72 cases of 98 vertebral bone cement diffused beyond the midline, accounting for 72.59%. Unilateral approach was used for the 72 patients whose bone cement diffused beyond the midline of the vertebral body, among them, there were 5 cases with 8 vertebrae in osteopenia group, 40 cases with 55 vertebrae in osteoporosis group and 27 cases with 35 vertebrae in severe osteoporosis group. There was significant difference in the bone cement dispersion between three groups (=41.397, =0.000). Moreover, no bone cement leakage occurred in osteopenia group, 3 cases of 4 vertebrae occurred in osteoporosis group and 2 cases of 3 vertebrae in severe osteoporosis group. However, none of the patients with bone cement leakage caused nerve injury and other symptoms, and there was no significant difference in bone cement leakage between the three groups (=2.242, =0.326).@*CONCLUSION@#According to the QCT examination of lumbar spine, defining the degree of osteoporosis and guiding the puncture method can shorten the operation time, reduce the number of fluoroscopy, and effectively improve the safety of vertebroplasty.

Systematic review on the curative effect of total hip arthroplasty through direct anterior approach versus posterolateral Approach / 中国医师杂志

Objective To systematic evaluate the curative effect of total hip arthroplasty (THA) through direct anterior approach versus (DAA) posterolateral approach (PSA).Methods All the randomized controlled trial (RCT) articles and observational research articles about the curative effect of THA through direct anterior approach (DAA group) versus posterolateral approach (PSA group)for treatment of hip disease that published at home and abroad included from database establishing to December 2017 were retrieved from Cochrane Library,Pubmed,Web of Science,CBM and China national knowledge internet (CNKI) through computer.The articles were screened and the information was extracted independently by two authors.A Meta-analysis was performed utilizing the RevMan 5.3 software.Results Two hundred and forty-nine articles were searched out in the initial stage.After screening,7 Chinese articles and 2 English articles (942 cases)were included in the final analysis,including 8 RCT articles and 1 observational research article,and 463 patients in direct anterior approach and 479 patients in posterolateral approach.The results of Meta-analysis demonstrated that the incision length,operation time,intraoperative blood less,Harris hip function scores at 1 month after surgery,visual analogue scale (VAS) scores at 24 hours after surgery,postoperative bed rest time and the hospital stay were better in DAA group compared to PSA group (SMD =-2.51,95％ CI:-4.84,-0.19,P=0.03);(SMD=0.84,95％ CI:0.10,1.59,P=0.03);(SMD=-1.52,95％ CI:-2.98,-0.05,P=0.04);(SMD=2.58,95％ CI:0.65,4.51,P=0.009);(SMD=-0.65,95％ CI:-0.91,-0.38,P＜0.01);(SMD=-1.85,95％ CI:-3.00,-0.70,P=0.002);(SMD =-1.12,95％ CI:-1.62,-0.61,P ＜0.01).There were no statistical difference in drainage (SMD =-1.45,95％ CI:-3.40,0.49,P =0.14),Harris hip function scores at last follow-up (SMD =0.91,95％ CI:-0.31,2.14,P =0.15),VAS scores at 48 hours after surgery (SMD =-1.21,95％ CI:-3.03,0.61,P =0.19),acetabular anteversion angles (SMD =-0.50,95％ CI:-1.06,0.05,P =0.07),acetabular abduction angles (SMD =-0.01,95％ CI:-0.32,0.30,P =0.97) and incidence of complications (OR =1.10,95％ CI:0.59,2.07,P =0.77) between the 2 groups.The result of analysis of publication bias according to Harris hip function scores at last follow-up and incidence of complications showed that the funnel-plot was essentially bilateral symmetry.Conclusions THA through DAA can significantly reduce the soft-tissue damage,and it is beneficial for patient's quick recovery,also it conforms to the current concept of minimally invasive treatment that worthy of promotion.

RT-nPCR Assays for Amplification and Sequencing of VP1 Genes in Human Enterovirus A-D from Clinical Specimens / 生物医学与环境科学（英文）

Objective@#To develop RT-nPCR assays for amplifying partial and complete VP1 genes of human enteroviruses (HEVs) from clinical samples and to contribute to etiological surveillance of HEV-related diseases.@*Methods@#A panel of RT-nPCR assays, consisting of published combined primer pairs for VP1 genes of HEV A-C and in-house designed primers for HEV-D, was established in this study. The sensitivity of each RT-nPCR assay was evaluated with serially diluted virus stocks of five serotypes expressed as CCID @*Results@#The sensitivity of RT-nPCR assays for amplifying partial VP1 gene of HEVs was 0.1 CCID @*Conclusion@#This RT-nPCR system is capable of amplifying the partial and complete VP1 gene of HEV A-D, providing rapid, sensitive, and reliable options for molecular typing and molecular epidemiology of HEVs in clinical specimens.

Publisher Correction: Clonal analysis of Salmonella-specific effector T cells reveals serovar-specific and cross-reactive T cell responses.

In the version of this article initially published, the first affiliation lacked 'MRC'; the correct name of the institution is 'MRC Weatherall Institute of Molecular Medicine'. Two designations (SP110Y and ST110H) were incorrect in the legend to Fig. 6f,h,i. The correct text is as follows: for panel f, "...loaded with either the CdtB(105-125)SP110Y (DRB4*SP110Y) or the CdtB(105-125)ST110H (DRB4*ST110H) peptide variants..."; for panel h, "...decorated by the DRB4*SP110Y tetramer (lower-right quadrant), the DRB4*ST110H (upper-left quadrant)..."; and for panel i, "...stained ex vivo with DRB4*SP110Y, DRB4*ST110H...". In Fig. 8e, the final six residues (LTEAFF) of the sequence in the far right column of the third row of the table were missing; the correct sequence is 'CASSYRRTPPLTEAFF'. In the legend to Fig. 8d, a designation (HLyE) was incorrect; the correct text is as follows: "(HlyE?)." Portions of the Acknowledgements section were incorrect; the correct text is as follows: "This work was supported by the UK Medical Research Council (MRC) (MR/K021222/1) (G.N., M.A.G., A.S., V.C., A.J.P.),...the Oxford Biomedical Research Centre (A.J.P., V.C.),...and core funding from the Singapore Immunology Network (SIgN) (E.W.N.) and the SIgN immunomonitoring platform (E.W.N.)." Finally, a parenthetical element was phrased incorrectly in the final paragraph of the Methods subsection "T cell cloning and live fluorescence barcoding"; the correct phrasing is as follows: "...(which in all cases included HlyE, CdtB, Ty21a, Quailes, NVGH308, and LT2 strains and in volunteers T5 and T6 included PhoN)...". Also, in Figs. 3c and 4a, the right outlines of the plots were not visible; in the legend to Fig. 3, panel letter 'f' was not bold; and in Fig. 8f, 'ND' should be aligned directly beneath DRB4 in the key and 'ND' should be removed from the diagram at right, and the legend should be revised accordingly as follows: "...colors indicate the HLA class II restriction (gray indicates clones for which restriction was not determined (ND)). Clonotypes are grouped on the basis of pathogen selectivity (continuous line), protein specificity (dashed line) and epitope specificity; for ten HlyE-specific clones (pixilated squares), the epitope specificity was not determined...". The errors have been corrected in the HTML and PDF versions of the article.

Differential control of human Treg and effector T cells in tumor immunity by Fc-engineered anti-CTLA-4 antibody.

Anti-CTLA-4 mAb is efficacious in enhancing tumor immunity in humans. CTLA-4 is expressed by conventional T cells upon activation and by naturally occurring FOXP3+CD4+ Treg cells constitutively, raising a question of how anti-CTLA-4 mAb can differentially control these functionally opposing T cell populations in tumor immunity. Here we show that FOXP3high potently suppressive effector Treg cells were abundant in melanoma tissues, expressing CTLA-4 at higher levels than tumor-infiltrating CD8+ T cells. Upon in vitro tumor-antigen stimulation of peripheral blood mononuclear cells from healthy individuals or melanoma patients, Fc-region-modified anti-CTLA-4 mAb with high antibody-dependent cell-mediated cytotoxicity (ADCC) and cellular phagocytosis (ADCP) activity selectively depleted CTLA-4+FOXP3+ Treg cells and consequently expanded tumor-antigen-specific CD8+T cells. Importantly, the expansion occurred only when antigen stimulation was delayed several days from the antibody treatment to spare CTLA-4+ activated effector CD8+T cells from mAb-mediated killing. Similarly, in tumor-bearing mice, high-ADCC/ADCP anti-CTLA-4 mAb treatment with delayed tumor-antigen vaccination significantly prolonged their survival and markedly elevated cytokine production by tumor-infiltrating CD8+ T cells, whereas antibody treatment concurrent with vaccination did not. Anti-CTLA-4 mAb modified to exhibit a lesser or no Fc-binding activity failed to show such timing-dependent in vitro and in vivo immune enhancement. Thus, high ADCC anti-CTLA-4 mAb is able to selectively deplete effector Treg cells and evoke tumor immunity depending on the CTLA-4-expressing status of effector CD8+ T cells. These findings are instrumental in designing cancer immunotherapy with mAbs targeting the molecules commonly expressed by FOXP3+ Treg cells and tumor-reactive effector T cells.

Traditional Chinese Medicine as a Remedy for Male Infertility: A Review

Male infertility (MI) is a complex multifactorial disease, and idiopathic infertility accounts for 30% of cases of MI. At present, the evidence for the effectiveness of empirical drugs is limited, and in vitro fertilization is costly and may increase the risk of birth defects and childhood cancers. Therefore, affected individuals may feel obliged to pursue natural remedies. Traditional Chinese medicine (TCM) may represent a useful option for infertile men. It has been demonstrated that TCM can regulate the hypothalamic-pituitary-testicular axis and boost the function of Sertoli cells and Leydig cells. TCM can also alleviate inflammation, prevent oxidative stress, reduce the DNA fragmentation index, and modulate the proliferation and apoptosis of germ cells. Furthermore, TCM can supply trace elements and vitamins, ameliorate the microcirculation of the testis, decrease the levels of serum anti-sperm antibody, and modify epigenetic markers. However, the evidence in favor of TCM is not compelling, which has hindered the development of TCM. This review attempts to elucidate the underlying therapeutic mechanisms of TCM. We also explore the advantages of TCM, differences between TCM and Western medicine, and problems in existing studies. Subsequently, we propose solutions to these problems and present perspectives for the future development of TCM.

Clonal analysis of Salmonella-specific effector T cells reveals serovar-specific and cross-reactive T cell responses.

To tackle the complexity of cross-reactive and pathogen-specific T cell responses against related Salmonella serovars, we used mass cytometry, unbiased single-cell cloning, live fluorescence barcoding, and T cell-receptor sequencing to reconstruct the Salmonella-specific repertoire of circulating effector CD4+ T cells, isolated from volunteers challenged with Salmonella enterica serovar Typhi (S. Typhi) or Salmonella Paratyphi A (S. Paratyphi). We describe the expansion of cross-reactive responses against distantly related Salmonella serovars and of clonotypes recognizing immunodominant antigens uniquely expressed by S. Typhi or S. Paratyphi A. In addition, single-amino acid variations in two immunodominant proteins, CdtB and PhoN, lead to the accumulation of T cells that do not cross-react against the different serovars, thus demonstrating how minor sequence variations in a complex microorganism shape the pathogen-specific T cell repertoire. Our results identify immune-dominant, serovar-specific, and cross-reactive T cell antigens, which should aid in the design of T cell-vaccination strategies against Salmonella.

Bystander CD8+ T cells are abundant and phenotypically distinct in human tumour infiltrates.

Various forms of immunotherapy, such as checkpoint blockade immunotherapy, are proving to be effective at restoring T cell-mediated immune responses that can lead to marked and sustained clinical responses, but only in some patients and cancer types1-4. Patients and tumours may respond unpredictably to immunotherapy partly owing to heterogeneity of the immune composition and phenotypic profiles of tumour-infiltrating lymphocytes (TILs) within individual tumours and between patients5,6. Although there is evidence that tumour-mutation-derived neoantigen-specific T cells play a role in tumour control2,4,7-10, in most cases the antigen specificities of phenotypically diverse tumour-infiltrating T cells are largely unknown. Here we show that human lung and colorectal cancer CD8+ TILs can not only be specific for tumour antigens (for example, neoantigens), but also recognize a wide range of epitopes unrelated to cancer (such as those from Epstein-Barr virus, human cytomegalovirus or influenza virus). We found that these bystander CD8+ TILs have diverse phenotypes that overlap with tumour-specific cells, but lack CD39 expression. In colorectal and lung tumours, the absence of CD39 in CD8+ TILs defines populations that lack hallmarks of chronic antigen stimulation at the tumour site, supporting their classification as bystanders. Expression of CD39 varied markedly between patients, with some patients having predominantly CD39- CD8+ TILs. Furthermore, frequencies of CD39 expression among CD8+ TILs correlated with several important clinical parameters, such as the mutation status of lung tumour epidermal growth factor receptors. Our results demonstrate that not all tumour-infiltrating T cells are specific for tumour antigens, and suggest that measuring CD39 expression could be a straightforward way to quantify or isolate bystander T cells.

The impact of ischemia-reperfusion injuries on skin resident murine dendritic cells.

Pressure ulcers are a chronic problem for patients or the elderly who require extended periods of bed rest. The formation of ulcers is due to repeated cycles of ischemia-reperfusion (IR), which initiates an inflammatory response. Advanced ulcers disrupt the skin barrier, resulting in further complications. To date, the immunological aspect of skin IR has been understudied, partly due to the complexity of the skin immune cells. Through a combination of mass cytometry, confocal imaging and intravital multiphoton imaging, this study establishes a workflow for multidimensionality single cell analysis of skin myeloid cell responses in the context of IR injury with high spatiotemporal resolution. The data generated has provided us with previously uncharacterized insights into the distinct cellular behavior of resident dendritic cells (DCs) and recruited neutrophils post IR. Of interest, we observed a drop in DDC numbers in the IR region, which was subsequently replenished 48h post IR. More importantly, in these cells, we observe an attenuated response to repeated injuries, which may have implications in the subsequent wound healing process.

MAIT cell clonal expansion and TCR repertoire shaping in human volunteers challenged with Salmonella Paratyphi A.

Mucosal-associated invariant T (MAIT) cells are innate-like T cells that can detect bacteria-derived metabolites presented on MR1. Here we show, using a controlled infection of humans with live Salmonella enterica serovar Paratyphi A, that MAIT cells are activated during infection, an effect maintained even after antibiotic treatment. At the peak of infection MAIT cell T-cell receptor (TCR)ß clonotypes that are over-represented prior to infection transiently contract. Select MAIT cell TCRß clonotypes that expand after infection have stronger TCR-dependent activation than do contracted clonotypes. Our results demonstrate that host exposure to antigen may drive clonal expansion of MAIT cells with increased functional avidity, suggesting a role for specific vaccination strategies to increase the frequency and potency of MAIT cells to optimize effector function.