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SARS-CoV-2 specific IgG responses play critical roles for patients to recover from COVID-19, in-depth dissecting of the IgG responses on systems level is of great interest. Herein, we adopted a newly developed high-throughput epitope mapping technology (AbMap), analyzed 55 COVID-19 convalescent sera and 226 antibody samples enriched by specific proteins or peptides from these sera. We revealed three areas that are rich of IgG epitopes, two are on Spike protein but outside of RBD, and one is on Nucleocapsid protein. We identified 29 significant epitopes on Spike protein, from two of these significant epitopes, two critical epitope residues were found, i. e., D936 and P1263, which are highly related to the infectivity of SARS-CoV-2 In summary, we provided the first global map of IgG binding epitopes for SARS-CoV-2 at single amino acid resolution. This map will facilitate the precise development of therapeutic antibodies and vaccines. HIGHLIGHTSO_LIA map of SARS-CoV-2 specific IgG binding epitopes at single amino acid resolution C_LIO_LITwo areas outside of RBD that are rich of significant epitopes were identified C_LIO_LIOne area rich of significant epitopes was determined on Nucleocapsid protein C_LIO_LITwo critical epitope residues (D936 and P1263) on Spike protein are highly related to the infectivity of SARS-CoV-2 C_LI
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OBJECTIVE@#To investigate the prevalence of respiratory viral infections in patients with primary immunodeficiency disease (PID) during hematopoietic stem cell transplantation.@*METHODS@#108 specimens of nasopharyngeal aspirate were collected from 22 PID patients before and after hematopoietic stem cell transplantation from July 2016 to July 2018 in the Department of Hematology. The TR-PCR was used to detect for respiratory viruses including respiratory syncytial virus(RSV),human metapneumoviros(hMPV),coronavirus(CoV) and parainfluenza 1-3 (PIV1-3). And the clinical characteristics and co-infection were analyzed.@*RESULTS@#Among the total 108 specimens, viral pathogens were identified in 41 (37.96%) specimens. Among which the pathogens of highest detection rate was RSV (25.9%). Different types of PID showed different virus infection rates, among which the highest infection rate was severe combined immunodeficiency disease (SCID) patients, with the virus detection rate was 57.9%. The incidence of co-infection with two or more than two viruses was 19.5%.@*CONCLUSION@#Patients with PID who undergo hematopoietic stem cell transplantation are more susceptible to respiratory viruses. RSV is an important respiratory tract virus pathogen after hematopoietic stem cell transplantation.
Subject(s)
Humans , Hematopoietic Stem Cell Transplantation , Hematopoietic Stem Cells , Metapneumovirus , Primary Immunodeficiency Diseases , Therapeutics , Respiratory Syncytial Virus, Human , Respiratory Tract InfectionsABSTRACT
Objective To analyze the epidemiological characteristics of malaria reported in the malaria surveillance system in Jinan City from 2012 to 2016,so as to provide the evidence for improving the ability of diagnosis,treatment,prevention and control of malaria.Methods The data of all reported malaria cases in the malaria surveillance system were retrospectively ana-lyzed.Results From 2012 to 2016,91 malaria cases were found in Jinan City,of which one patient died.All the cases were imported and 95.60%(87 cases)of them came from Africa(note:most of the people were Chinese residents who had been in Af-rica for the export of labor service and came back China).Falciparum malaria accounted for the most(82.42%,75 cases).All the cases were adult males,and were mainly migrant workers.The median time from onset to being confirmedly diagnosed was 5 days,and the median time from seeing a doctor to being diagnosed was 1 day.The medical institutions where the patients first visited were mainly municipal medical institutions(42 cases,46.15%).The misdiagnosis rate was 100% in village clinics and township health centers(8/8 and 2/2,respectively). The misdiagnosis rate was lowest in the municipal medical institutions (3/42,7.14%).There were 41 malaria patients(45.05%)with complications.Conclusions The situation of overseas imported malaria in Jinan City is becoming more and more serious.It is necessary to further strengthen the related professional training for doctors and strengthen the multi-sectoral cooperation for health education,etc.in order to find the cases in time and conduct the active standardized treatment,so as to prevent the second generation cases.
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<p><b>OBJECTIVE</b>To evaluate the association between the use of antibacterial agents in the first years of life and childhood asthma.</p><p><b>METHODS</b>The Chinese and English databases CNKI, Wanfang Data, VIP, PubMed, and EBSCO were searched for prospective cohort studies on the association between the use of antibacterial agents in the first years of life and childhood asthma. Stata12.0 software was used to analyze the association through a Meta analysis.</p><p><b>RESULTS</b>The articles with a high quality score and adjusted effective values for factors for lower respiratory tract infection were pooled, and a total of 8 studies were included. The results of the Meta analysis showed that the use of antibacterial agents in the first years of life increased the risk of childhood asthma (OR=1.14, 95%CI: 1.10-1.17, P<0.05). Compared with the children who used antibacterial agents 0-1 times in the first years of life, those who used more than 4 times had an increased risk of asthma (OR=1.28, 95%CI: 1.19-1.38, P<0.05). High-risk children (at least one immediate family member had asthma) who used antibacterial agents had an increased risk of asthma (OR=1.47, 95%CI: 1.20-1.81, P<0.05).</p><p><b>CONCLUSIONS</b>The use of antibacterial agents in the first years of life increases the risk of childhood asthma. High-risk children who use antibacterial agents have an increased risk of asthma. The increased frequency of use of antibacterial agents in the first years of life is associated with an increased risk of childhood asthma, but the detailed dose relationship needs further investigation.</p>
Subject(s)
Humans , Infant , Infant, Newborn , Anti-Bacterial Agents , Asthma , History, Ancient , RiskABSTRACT
<p><b>OBJECTIVE</b>To perform a genome-wide alternative polyadenylation (APA) profiling in both mouse female germline stem cells (FGSCs) and embryonic stem cells (ESCs) and explore the role of germline-specific APA in the biological behaviors of FGSCs.</p><p><b>METHODS</b>We used a high-throughput sequencing-based method 3T-Seq to profile the genome-wide 3' termini of the transcripts and delineate all the APA sites in mouse FGSCs and ESCs. The genes with altered APA sites in FGSCs compared with ESCs were analyzed with DAVID Gene Ontology tool for their biological roles.</p><p><b>RESULTS</b>We identified a total of 50243 APA sites in 16973 genes. In FGSCs, 1148 genes were shown to have alterations in 3'UTR length, among which 795 ( 66%) genes had shortened and 353 (34%) had lengthened 3'UTR. Some of the genes with shortened 3'UTR were involved in germ cell development.</p><p><b>CONCLUSIONS</b>Our genome-wide APA profiling analysis reveals a cell type-specific APA alternation in FGSCs, and APA-mediated 3'UTR alteration contributes to germline-related biological process. This study provides a framework for understanding the post-transcriptional regulation mechanisms in FGSCs.</p>
Subject(s)
Animals , Female , Mice , 3' Untranslated Regions , Cell Differentiation , Embryonic Germ Cells , Metabolism , Embryonic Stem Cells , Metabolism , Gene Expression Regulation , Genome , PolyadenylationABSTRACT
Trauma has become a common cause of death, and its complication posttraumatic acute respiratory distress syndrome (ARDS) is one significant factor for patient death during several days to several weeks after trauma. At present, emergency surgery is still insufficient in treating ARDS after trauma care: on one hand, early recognition of it is not favorable, on the other hand, the care measures after recognition are also improper. Therefore, emergency surgeons must master the regular pattern of ARDS development after trauma for early recognition of ARDS, and implement care principle for combination of timeliness and integrity to increase care rate.
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<p><b>OBJECTIVE</b>To improve the management of the early neurogenic pulmonary edema(NPE)in patients with non-traumatic cerebral hemorrhage.</p><p><b>METHODS</b>Totally 140 eligible patients with non-traumatic cerebral hemorrhage who were treated in the emergency department of our hospital from October 2008 to October 2014 were divided into two groups:NPE group(n=25)and non-NPE group(n=115). The clinical data were analyzed and compared.</p><p><b>RESULTS</b>Although the mean arterial pressure was similar between these two groups,the median pH and the bicarbonate ion(HCO(3)(-))were significantly lower in the NPE group than in the non-NPE group(pH:7.32 vs.7.39,P=0.002;HCO(3)(-),20.6 mmol/L vs.22.7 mmol/L,P=0.01). Multivariate regression analysis indicated that younger age and higher glucose level were significantly correlated with the early onset of NPE in the NPE group than in the non-NPE group(age:50.1 years vs.65.1 years,P=0.0008;glucose,15.4 mmol/L vs.10.78 mmol/L,P=0.001).There were only 3 patients in all with non-traumatic cerebral hemorrhage happened the fulminant NPE in 1 hour. Within 24 hours after patients visited the emergency room,the condition was improved in 20 of 25 patients in the NPE group. However,5 patients died,among whom 3 patients with fulminant NPE(onset within 1 hour)died due to acute respiratory distress syndrome and complicated with multiple organ failure,and 2 died of cerebral hernia.</p><p><b>CONCLUSIONS</b>NPE is a rare and severe complication in patients with non traumatic cerebral hemorrhage. The possibility of NPE should be considered in relatively young patients with higher glucose and lower blood pH value. Timely prevention and treatment can improve the outcomes.</p>
Subject(s)
Humans , Emergency Service, Hospital , Glucose , Pulmonary EdemaABSTRACT
<p><b>OBJECTIVE</b>To study the effect of inflammatory factors on cell proliferation and apoptosis in insulin-like grown factor 1 (IGF1)-slienced human coronary artery smooth muscle cells (hCASMCs).</p><p><b>METHODS</b>We silenced the expression of IGF1 in hCASMCs using the lentivirus-mediated RNA interference technology. Blank control group and negative control group were set using the hCASMCs without the infection of a virus vector and the hCASMCs with the infection of a negative control virus vector, respectively. After the treatment of these cells with both tumor necrosis factor-α 50 ng/ml and interleukin-1β 40 ng/ml, the concentration of IGF1 in cell-culture medium was detected by enzyme-linked immunosorbent assay, and the proliferation and apoptosis were evaluated by MTT assay and flow cytometry.</p><p><b>RESULTS</b>After the simulation with inflammatory factors, the concentration of IGF1 in the supernatant fluid of cultured IGF1-slienced hCASMCs was significantly lower than those in the blank control group and negative control group [(426.35±120.96) vs. (1 030.69±54.69) and (992.82±26.90)pg/ml, P=0.000). The proliferation of IGF1-slienced hCASMCs was substantially much less than the two control groups (0.302±0.011 vs. 0.401±0.028 and 0.302±0.011, F=37.628, P=0.000), and the apoptosis rate of IGF1-slienced hCASMCs was significant increased compared with the other two groups [(10.57±0.99)% vs. (0.19±0.13)% and (1.31±0.30)%, P=0.001].</p><p><b>CONCLUSION</b>Inflammatory factors can inhibit the cell proliferation and promote apoptosis after the knock-down of IGF1 in hCASMCs.</p>
Subject(s)
Humans , Apoptosis , Cell Proliferation , Coronary Vessels , Cell Biology , Insulin-Like Growth Factor I , Genetics , Metabolism , Interleukin-1beta , Pharmacology , Myocytes, Smooth Muscle , RNA Interference , Tumor Necrosis Factor-alpha , PharmacologyABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of Helicobacter pylori (Hp) eradication therapy on prognosis in children with Henoch-Schonlein purpura (HSP).</p><p><b>METHODS</b>A total of 153 children with HSP were divided into Hp infection treatment group (n=22), Hp infection control group (n=21), and Hp infection-negative group (n=110). The Hp infection treatment group received one-week triple therapy for Hp eradication in addition to conventional treatment, while the Hp infection control group and Hp infection-negative group received conventional treatment. All patients were followed up for prognostic evaluation.</p><p><b>RESULTS</b>The response rates of the Hp infection treatment, control, and negative groups were 86% (19/22), 90% (19/21) and 85% (94/110), respectively (P>0.05). The recurrence rates of HSP in the Hp infection treatment, control, and negative groups were 14% (3/22), 24% (5/21) and 31% (34/110), respectively (P>0.05). The incidence of Henoch-Schonlein purpura nephritis (HSPN) in the Hp infection-negative group (36%, 40/110) and control group (33%, 7/21) was significantly higher than that in the Hp infection treatment group (5%, 1/22) (P<0.05 for both), but no significant difference in the incidence of HSPN was found between the control and negative groups (P>0.05).</p><p><b>CONCLUSIONS</b>One-week triple therapy for Hp eradication may be useful to reduce the incidence of HSPN in children with HSP infected with Hp.</p>
Subject(s)
Child , Child, Preschool , Female , Humans , Male , Helicobacter Infections , Drug Therapy , Helicobacter pylori , Incidence , Prognosis , IgA Vasculitis , Epidemiology , RecurrenceABSTRACT
MonoMAC syndrome is a newly discovered immune deficiency syndrome caused by GATA-2 mutation, which is an autosomal dominant genetic disease. MonoMAC syndrome has typical immune cell abnormalities, with severe infection and is prone to develop into a hematological disease. Therapeutics for this disease mainly relies on symptomatic treatment and hematopoietic stem cell transplantation. In this paper, the research advances in clinical manifestations, laboratory tests, pathogenesis, diagnosis and treatment of MonoMAC syndrome are reviewed.
Subject(s)
Humans , GATA2 Transcription Factor , Genetics , Immunologic Deficiency Syndromes , Genetics , Monocytes , Pathology , Mutation , Mycobacterium Infections , SyndromeABSTRACT
Numerous studies have indicated that human metapneumovirus (hMPV) is an important viral pathogen in acute respiratory infections in children, presenting similar manifestations with respiratory syncytial virus (RSV). HMPV infection peaks in the winter-spring season and is more prevalent in younger ages, especially in children less than 1 year old. Host innate immune response has been implicated in recognition of pathogen-associated molecular patterns (PAMPs) of the virus. This recognition occurs through host pattern recognition receptors (PRRs). Toll like receptors (TLRs) are one of the largest class of PRRs which initiate and regulate adaptive immune responses. Some studies have indicated that TLR 3 and TLR 4 may play critical roles in hMPV infection. Construction of recombinant mutant viruses lacking one or two N-linked glycosylation sites in the F protein by using site-directed mutagenesis and reverse genetics may be helpful for developing attenuated live vaccines.
Subject(s)
Humans , Metapneumovirus , Allergy and Immunology , Paramyxoviridae Infections , Vaccines, Attenuated , Allergy and Immunology , Vaccines, Synthetic , Allergy and Immunology , Viral Vaccines , Allergy and ImmunologyABSTRACT
The influence of early-stage intensive insulin therapy on the plasma levels of vascular endothelial growth factor (VEGF) and the related parameters in patients with severe trauma and the clinical implication were investigated. Sixty-four cases of severe trauma (injury severity score ≥20) with stress hyperglycemia (blood glucose >9 mmol/L) were randomly divided into intensive insulin therapy group and conventional therapy group. ELISA method, radioimmunoassay and density gradient gradation one-step process were used to determine plasma VEGF, endothelin-1 (ET-1), and the number of circulating endothelial cells (CECs) at the day of 0, 2, 3, 5 and 7 after admission. Simultaneously, the changes of CRP concentration in plasma were monitored to evaluate inflammatory response. The results showed that plasma levels of observational indexes in patients receiving early-stage intensive insulin therapy were all significantly lower than those in conventional therapy groups 2, 3, 5 and 7 days after admission [for VEGF (ng/L), 122.2±23.8 vs. 135.9±26.5, 109.6±27.3 vs. 129.0±18.4, 88.7±18.2 vs. 102.6±27.3, 54.2±26.4 vs. 85.7±35.2, P<0.05, 0.01, 0.05, 0.05 respectively; for ET-1 (ng/L), 162.8±23.5 vs. 173.7±13.2, 128.6±17.5 vs. 148.8±22.4, 96.5±14.8 vs. 125.7±14.8, 90.7±16.9 vs. 104.9±22.5, P<0.05, 0.01, 0.01, 0.01 respectively; for CRP (mg/L), 23.2±13.8 vs. 31.9±16.5, 13.6±17.3 vs. 23.5±18.4, 8.7±10.2 vs. 15.6±13.3, 5.2±9.4 vs. 10.7±11.2, all P<0.05; for CECs (/0.9 μL), 10.9±5.6 vs. 13.9±6.2, 8.5±4.9 vs. 11.3±5.3, 6.3±6.4 vs. 9.4±5.7, 4.8±7.1 vs. 7.8±4.8, all P<0.05]. It was concluded that intensive insulin therapy could antagonize the endothelium injury after trauma and reduce inflammation response quickly, which was one of important mechanisms by which intensive insulin therapy improves the prognosis of trauma patients.
Subject(s)
Adult , Female , Humans , Male , Endothelin-1 , Blood , Hyperglycemia , Blood , Diagnosis , Drug Therapy , Hypoglycemic Agents , Therapeutic Uses , Insulin , Therapeutic Uses , Reproducibility of Results , Sensitivity and Specificity , Treatment Outcome , Vascular Endothelial Growth Factor A , Blood , Wounds and Injuries , Blood , Diagnosis , Drug TherapyABSTRACT
<p><b>OBJECTIVE</b>To discuss the influence of intensive insulin therapy on insulin resistance of patients with severe burn or trauma.</p><p><b>METHODS</b>Sixty patients with severe burn or trauma hospitalized in the Third People's Hospital of Chongqing or Southwest Hospital of the Third Military Medical University from January 2010 to December 2011 were randomly divided into intensive insulin therapy group (IT, treated with intensive insulin therapy to control the blood glucose to the level of 6.0-8.0 mmol/L) and control group (C, treated with routine therapy) according to the paired grouping method, with 30 patients in each group. Before treatment and on post treatment day (PTD) 1, 3, 7, 10, 14, the levels of fasting blood glucose and fasting plasma insulin were determined. Insulin resistance index and β-cell function index were calculated using homeostasis model assessment. Data were processed with t test, analysis of variance, and LSD test.</p><p><b>RESULTS</b>On PTD 1, 3, 7, 10, levels of fasting blood glucose in group IT [(6.8 ± 1.4), (6.7 ± 1.3), (5.8 ± 1.9), (5.4 ± 1.6) mmol/L] were significantly lower than those of group C [(14.8 ± 4.9), (12.7 ± 3.7), (7.7 ± 1.9), (6.6 ± 1.3) mmol/L, with t values respectively 12.453, 11.386, 5.563, 4.731, P < 0.05 or P < 0.01]. On PTD 3, 7, levels of fasting insulin in group IT [(14 ± 5), (10 ± 3) mU/L] were significantly lower than those of group C [(16 ± 4), (13 ± 4) mU/L, with t values respectively 4.212, 4.364, P values below 0.05]. Levels of fasting blood glucose and fasting insulin in the two groups at each time point were statistically significantly different from those before treatment (with P values below 0.01), except for the level of fasting blood glucose on PTD 3. On PTD 1, 3, 7, 10, levels of insulin resistance index in group IT (1.60 ± 0.80, 1.46 ± 0.70, 0.96 ± 0.21, 0.90 ± 0.23) were significantly lower than those in group C (2.15 ± 1.35, 2.21 ± 1.21, 1.50 ± 0.95, 1.17 ± 0.66, with t values respectively 8.316, 10.607, 7.825, 5.217, P < 0.05 or P < 0.01). Levels of insulin resistance index of patients in the two groups at each time point after treatment were significantly lower than those before treatment (with P values below 0.01). On PTD 1, 3, 7, levels of β-cell function index in group IT (4.6 ± 2.9, 4.5 ± 3.3, 4.5 ± 3.6) were significantly higher than those in group C (3.4 ± 2.5, 3.6 ± 2.2, 4.2 ± 2.5, with t values respectively 8.243, 7.914, 4.338, P < 0.05 or P < 0.01). Levels of β-cell function index in group C on PTD 1 and 3 were significantly lower than that before therapy (with P values below 0.05).</p><p><b>CONCLUSIONS</b>Intensive insulin therapy can alleviate insulin resistance of patients with severe burn or trauma.</p>
Subject(s)
Adolescent , Adult , Aged , Female , Humans , Male , Middle Aged , Young Adult , Burns , Insulin , Therapeutic Uses , Insulin ResistanceABSTRACT
The influence of early-stage intensive insulin therapy on the plasma levels of vascular endothelial growth factor (VEGF) and the related parameters in patients with severe trauma and the clinical implication were investigated. Sixty-four cases of severe trauma (injury severity score ≥20) with stress hyperglycemia (blood glucose >9 mmol/L) were randomly divided into intensive insulin therapy group and conventional therapy group. ELISA method, radioimmunoassay and density gradient gradation one-step process were used to determine plasma VEGF, endothelin-1 (ET-1), and the number of circulating endothelial cells (CECs) at the day of 0, 2, 3, 5 and 7 after admission. Simultaneously, the changes of CRP concentration in plasma were monitored to evaluate inflammatory response. The results showed that plasma levels of observational indexes in patients receiving early-stage intensive insulin therapy were all significantly lower than those in conventional therapy groups 2, 3, 5 and 7 days after admission [for VEGF (ng/L), 122.2±23.8 vs. 135.9±26.5, 109.6±27.3 vs. 129.0±18.4, 88.7±18.2 vs. 102.6±27.3, 54.2±26.4 vs. 85.7±35.2, P<0.05, 0.01, 0.05, 0.05 respectively; for ET-1 (ng/L), 162.8±23.5 vs. 173.7±13.2, 128.6±17.5 vs. 148.8±22.4, 96.5±14.8 vs. 125.7±14.8, 90.7±16.9 vs. 104.9±22.5, P<0.05, 0.01, 0.01, 0.01 respectively; for CRP (mg/L), 23.2±13.8 vs. 31.9±16.5, 13.6±17.3 vs. 23.5±18.4, 8.7±10.2 vs. 15.6±13.3, 5.2±9.4 vs. 10.7±11.2, all P<0.05; for CECs (/0.9 μL), 10.9±5.6 vs. 13.9±6.2, 8.5±4.9 vs. 11.3±5.3, 6.3±6.4 vs. 9.4±5.7, 4.8±7.1 vs. 7.8±4.8, all P<0.05]. It was concluded that intensive insulin therapy could antagonize the endothelium injury after trauma and reduce inflammation response quickly, which was one of important mechanisms by which intensive insulin therapy improves the prognosis of trauma patients.
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<p><b>OBJECTIVE</b>To construct a replication-defective recombinant adenovirus expressing the fusion gene of neuraminidase (NA) gene in influenza virus A/FM/1/47 and C3d and to evaluate the induced immune efficacy.</p><p><b>METHODS</b>NA-C3d was cloned into shutter vector pAdTrack-CMV, which was cotransformated with adenovirus DNA into E. coli BJ5183. The recombinant adenovirus genomic DNA was generated through homological recombination. The recombinant adenovirus was produced by transfecting 293 cell line with the genomic DNA and the induced immune efficacy in mice were analyzed.</p><p><b>RESULTS</b>The integration of NA-C3d in the adenovirus genomic DNA and its expression were confirmed by PCR and Western-Blot assays respectively. After intranasal immunization, the serum IgG was induced at a titer of 1: 1000 and 1:100 000 in BALB/c mice at primary and secondary immunization respectively. The vaccinated mice were completely survived when challenged with wide influenza virus.</p><p><b>CONCLUSION</b>recombinant adenovirus expressing NA-C3d was successfully constructed and it could induce desired immune efficacy.</p>
Subject(s)
Animals , Mice , Adenoviridae , Genetics , Metabolism , Physiology , Cloning, Molecular , Complement C3d , Genetics , Genetic Vectors , Genetics , Immunoglobulin G , Allergy and Immunology , Alphainfluenzavirus , Genetics , Mice, Inbred BALB C , Neuraminidase , Genetics , Recombinant Fusion Proteins , Genetics , Transfection , Methods , Virus ReplicationABSTRACT
<p><b>OBJECTIVE</b>To establish a novel flow cytometry-based assay for measuring the expression of lysosomal-associated membrane protein 1 (LAMP-1, CD107α) on the cell surface of natural killer (NK) cells and cytotoxic T lymphocyte (CTL) and evaluate the screening value of this assay for cytotoxic defects-related diseases such as familial hemophagocytic lymphopro-liferative (FHL) syndrome.</p><p><b>METHOD</b>Three suspected Chediak-Higashi Syndrome (CHS) patients, three suspected FHL patients and 10 healthy children were enrolled in the study from October 2010 to June 2011. Their PBMCs were separated and activated overnight with IL-2. After the granule release of NK cells activated by phytohemagglutinin (PHA) and CD8+T cells by anti-CD3, the CD107α expression were analyzed by flow cytometry. The peripheral blood DNA and RNA of the patients were extracted to analyze the pathogenic genes via DNA-PCR/RT-PCR and direct sequencing.</p><p><b>RESULT</b>The CD107α expression on CTL in the ten healthy children significantly increased after activation by anti-CD3 [(0.18 ± 0.07)% vs. (4.47 ± 2.36)%, P < 0.05] and NK cells after activation by PHA [(0.27 ± 0.07)% vs. (5.80 ± 2.83)%, P < 0.05]. The frequency of CD107α-expression NK cells in three suspected CHS after activation was significantly elevated when compared with the healthy control [0.5%, 0.6% vs. (5.80 ± 2.83)%] except patient 2. After the anti-CD3 activation, the frequency of CD107α expression on CTL cells also showed no significant difference [0.3%, 0.9%, 0.2% vs. (4.47 ± 2.36)%] in three patients. All of their mean fluorescence intensity (MFI) showed the same trend. Patient 1 and 3 were identified to have LYST mutations (Patient 1: c.5411-5414 del TTTC, L1741fsX1758 and c.7975 C > T, R2596X; Patient 3: c.4863G > A, R1563H and c.5392-5393delAA, E1739fsX1756). There was no mutation identified in the LYST gene for patient 2. CD107α expression of NK cells and CTL in the suspected FHL patients and in mirror of these findings, no underlying gene variation of PRF, MUNC13-4 and STX11 were identified.</p><p><b>CONCLUSION</b>We developed a method to quantitatively assess cytotoxicity of the NK cells and CTL by measuring the expression of CD107α on the cell membrane, which appeared to be an effective and rapid screening test for cytotoxic defects-related diseases such as FHL and other HLH secondary to primary immunodeficiency.</p>
Subject(s)
Child, Preschool , Female , Humans , Infant , Male , Case-Control Studies , Cell Degranulation , Allergy and Immunology , Cell Membrane , Metabolism , Chediak-Higashi Syndrome , Diagnosis , Genetics , Allergy and Immunology , Metabolism , Cytotoxicity, Immunologic , Flow Cytometry , Methods , Interleukin-2 , Metabolism , Killer Cells, Natural , Allergy and Immunology , Metabolism , Lymphohistiocytosis, Hemophagocytic , Diagnosis , Genetics , Allergy and Immunology , Metabolism , Lysosomal-Associated Membrane Protein 1 , Metabolism , Mutation , Phytohemagglutinins , Metabolism , T-Lymphocytes, Cytotoxic , Allergy and Immunology , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To investigate the value of gene analysis of amniotic fluid exfoliated cells and WASP detection from cord blood in prenatal diagnosis of high-risk fetus with Wiskott-Aldrich syndrome.</p><p><b>METHOD</b>Seven patients with Wiskott-Aldrich syndrome were diagnosed by gene analysis and WASP detected by flow cytometry from 2008 to 2010. After detailed inquiry for medical history and gene analysis of related family members, seven pedigree trees were drawn, including 15 carriers of abnormal genes. From 2008 to 2011, seven samples of amniotic cell gotten by amniocentesis were collected from seven high-risk pregnant women with abnormal gene during 18 to 20 gestational weeks. WASP gene was amplified by polymerase chain reaction (PCR) from DNA of amniotic cell gotten and sequencing was performed directly on the PCR products forward and reversely. Embryo blood sample was collected from one high-risk fetus by needle puncture of umbilical blood vessel and WASP expression was detected by flow cytometry. Karyotyping was performed in amniotic cell gotten cultivated by orthotopic slice and G band staining. Gene analysis of WASP, WASP expression detected by flow cytometry and evaluation of immune function were reexamined in high-risk fetus after delivery.</p><p><b>RESULT</b>Amniocentesis and culture of amniotic cell succeeded in all the seven fetuses. Gene analysis and karyotyping showed that one male fetus and four female fetuses were normal and two female fetuses were carriers. WASP expression detected from embryo blood sample of the patient was normal. After delivery, the result of gene analysis, WASP detection and evaluation of immune function was the same as that of prenatal diagnosis.</p><p><b>CONCLUSION</b>Karyotyping, gene analysis and WASP detection of cord blood can provide reliable service of prenatal diagnosis for high-risk pregnant women with Wiskott-Aldrich syndrome.</p>
Subject(s)
Female , Humans , Male , Pregnancy , Amniocentesis , Fetal Diseases , Diagnosis , Flow Cytometry , Prenatal Diagnosis , Wiskott-Aldrich Syndrome , Diagnosis , Genetics , Wiskott-Aldrich Syndrome Protein , Blood , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To understand the clinical characteristics and outcome associated with respiratory syncytial virus (RSV) infection in hematopoietic stem cell transplantation (HSCT) recipients with primary immunodeficiencies (PIDs).</p><p><b>METHOD</b>Nasopharyngeal aspirate samples were collected consecutively before and after HSCT from 9 recipients from Apr. 2009 to Sep. 2010 and analyzed for the presence of RSV using real-time polymerase chain reaction assay. To further verify the presence of the virus, positive samples for PCR were isolated for RSV. RSV G gene was amplified, sequenced and used for phylogenetic analysis.</p><p><b>RESULT</b>The presence of RSV was detected in 3 out of 9 children. The viral replication in all the patients was prolonged for months. All the 3 patients with RSV infection were treated with intravenous immune globulin (IVIG) and one was treated with antiviral medication. All patients survived and achieved successful immune reconstitution.</p><p><b>CONCLUSION</b>This study indicates that the HSCT recipients with PID are at increased risk for RSV infection. RSV can shed for months after the initial infection and the patients recover with the course of immune reconstitution.</p>
Subject(s)
Child, Preschool , Female , Humans , Infant , Hematopoietic Stem Cell Transplantation , Immunologic Deficiency Syndromes , General Surgery , Virology , Prognosis , Respiratory Syncytial Virus Infections , Diagnosis , Drug Therapy , Respiratory Syncytial Viruses , Genetics , Physiology , Virus Replication , Virus SheddingABSTRACT
<p><b>OBJECTIVE</b>X-linked lymphoproliferative disease (XLP), a genetic disorder characterized by immunodeficiency to Epstein-Barr virus (EBV) infection, has been linked to mutations in the SH2D1A gene. XLP patient displays EBV associated fulminant infectious mononucleosis or hemophagocytic lymphohistocytosis, hypogammaglobulinemia or malignant lymphoma. Here we report the clinical features, gene mutation and SAP expression on PBMCs of a Chinese patient with XLP and potential carriers.</p><p><b>METHOD</b>A 6 years old male patient and his maternal relatives were enrolled in this study. The patient was found to have with a renal Burkitt lymphoma on the right waist at 5 years of age by accident. His elder brother and a maternally related cousin both died of multiple systemic organ dysfunction syndrome (MODS) due to fulminant infectious mononucleosis (FIM) at the age of one year. The patient and his maternal relatives were subjected to detection of SAP expression on the PBMCs by flow cytometry and gene mutation analysis of SH2D1A by using PCR based on genomic DNA.</p><p><b>RESULT</b>The patient exhibited 536.9 copy/ml level of circulating EBV-DNA during remission. Sequence analysis showed that the patient harbored a nonsense mutation in exon 2 (C462T), resulting in a premature stop codon (Arg55X). His mother and some of the maternal relatives were proved to be carriers of this mutation. SAP expression from the patient was significantly reduced as compared to normal individual and the carriers.</p><p><b>CONCLUSION</b>We identified a Chinese XLP case genetically. Assessment of SAP expression on PBMCs by flow cytometry seemed to be an effective rapid diagnostic method for this disease. Absence of EBV infection does not diminish the possibility of XLP.</p>
Subject(s)
Child , Humans , Male , Carrier Proteins , Genetics , DNA, Viral , Blood , Epstein-Barr Virus Infections , Exons , Herpesvirus 4, Human , Intracellular Signaling Peptides and Proteins , Genetics , Lymphoproliferative Disorders , Genetics , Virology , Mutation , Pedigree , Signaling Lymphocytic Activation Molecule Associated ProteinABSTRACT
<p><b>OBJECTIVE</b>To investigate regulatory effect of Acheron (Achn) on proliferation and apoptosis of human vascular endothelial cell.</p><p><b>METHODS</b>(1) Eahy926 cells were cultured in serum-free DMEM medium (96-well plates) and were divided into Achn inhibition group (transfected with plasmid psi-Achn), psi4.1 group (transfected with psi4.1 empty vector), Achn induction group (transfected with pcDNA-Achn), pcDNA3.1 group (transfected with pcDNA3.1 empty vector), cotransfection group [cotransfected with pcDNA-Achn + psi-calcium/calmodulin-dependent serine protein kinase (CASK)], blank control group (treated with PBS) according to the random number table (the same method below). The cell proliferation was determined by MTT assay at post transfection hour (PTH) 1, 24, 48, 72, with expression of absorbance value. (2) Total protein of Eahy926 cells were extracted and quantitated by BCA assay, and then they were divided into Achn antibody precipitation group (100 µg protein), CASK antibody precipitation group (100 µg protein), IgG antibody group (100 µg protein), Western blot group (20 µg protein). Achn and CASK protein levels were determined by immunoprecipitation and Western blot. (3) Synchronously cultured Eahy926 cells were divided into LPS induction group (treated with 5 mol/L LPS), Achn transfection group (transfected with pcDNA-Achn), cotransfection group (cotransfected with psi-CASK and pcDNA-Achn), KCl group (treated with 5 mol/L KCl), and blank control group (treated with 5 mol/L PBS). Cells in transfection groups were stimulated by LPS for 12 hours after PTH 24. Caspase-3 protein level was detected by immunohistochemistry. (4) Synchronously cultured Eahy926 cells were divided into Achn inhibition group (transfected with psi-Achn vector), Achn induction group (transfected with pcDNA-Achn vector), and blank control group (treated with PBS). Apoptosis rate was determined by FITC/PI with flow cytometry. Data were processed with one-way analysis of variance and t test.</p><p><b>RESULTS</b>(1) The cell proliferation in Achn inhibition group was lower than that in psi4.1 group from PTH 24, and the differences were statistically significant at PTH 48, 72 (with t value respectively 10.777, 6.112, P values all below 0.05). The cell proliferation in Achn induction group during PTH 24-72 were higher that in pcDNA3.1 group (with t value respectively 5.367, 6.053, 9.831, P values all below 0.05). The cell proliferation in cotransfection group at PTH 48, 72 were significantly lower than that in Achn induction group (with t value respectively 5.481, 9.517, P values all below 0.05). (2) Achn protein was detected in CASK antibody precipitation group while CASK protein was also detected in Achn antibody precipitation group. (3) Caspase-3 level in Achn transfection group was lower [(15.6 ± 0.5)%] as compared with that in LPS induction group [(32.8 ± 2.6)%, t = 10.083, P < 0.05], and that in cotransfection group showed further inhibition [(7.0 ± 2.0)%, t = 9.827, P < 0.01]. (4) Apoptosis rate in Achn inhibition group [(45.6 ± 10.9)%] was higher than that in blank control group [(13.2 ± 4.3) %, t = 7.043, P < 0.05]; while that in Achn induction group [(5.3 ± 2.9)%] was lower than that in blank control group (t = 6.499, P < 0.05).</p><p><b>CONCLUSIONS</b>Achn can promote human vascular endothelial cell proliferation, and inhibit its apoptosis induced by LPS or burn serum, and the effect is related to CASK.</p>