Anti-Idiotypic Regulatory Responses Induced by Vaccination with DNA Encoding Murine TCR Vα5 and Vβ2

There is evidence suggesting that anti-idiotypic regulation against T cells plays a role in maintaining homeostasis in the immune system, although its mechanism is not fully understood. By using DNA constructs encoding the TCR Vα5.2 and Vβ2.1 chains derived from ：in ovalbumin （OVA）-specific T cdl clone （OVA-T）, we herein demonstrated that vaccination with TCR-DNA effectively induced anti-idiotypic cellular as well as humoral responses. Serum samples from the TCR-DNA-vaccinated BALB/c mice were able to stain T cells in an idiotype-specific manner. CD4^＋ T cells from the TCR-DNA-vaccinated mice proliferated in response to stimulation with irradiated syngeneic OVA-T calls and secreted interferon-y but very little IL-4. Splenocytes from the TCR-DNA-vaccinated mice showed strong idiotype-specific CTL activity against the OVA-T cdls. Furthermore, adoptive transfer of the CD4^＋ or CD8^＋ T cells from the TCR-DNA-vaccinated mice resulted in hyporesponsiveness of syngeneic recipients. These results demonstrated that vaccination with DNA encoding TCR can effectively activate anti-idiotypic regulatory responses in vivo and thus providing a useful way for immunological intervention.     

Vδ2 T cell deficiency in granulomatosis with polyangiitis (Wegener's granulomatosis)

Previous studies have characterized phenotypic and functional alterations within T-cell receptor αβ-expressing T cells in patients with granulomatosis with polyangiitis (GPA). We analyzed the frequency, subset composition and in vitro activation of blood γδ T cells in GPA patients. We observed a significant reduction of γδ T cell numbers, due to the selective depletion of the Vδ2 subset which remained consistent over time upon repeated analysis. The loss of Vδ2 T cells was not due to migration into the inflamed lesions as very few γδ T cells were detected in inflammatory infiltrates. The memory subset distribution did not differ among Vδ2 T cells from healthy donors and GPA patients. Importantly, the remaining Vδ2 T cells were capable of responding to phosphoantigen stimulation in vitro. The marked depletion of blood Vδ2 T cells in GPA suggests cellular exhaustion, possibly due to chronic exposure to and continuous overstimulation by microbial phosphoantigens.     

Live attenuated measles vaccine expressing HIV-1 Gag virus like particles covered with gp160ΔV1V2 is strongly immunogenic

Although a live attenuated HIV vaccine is not currently considered for safety reasons, a strategy inducing both T cells and neutralizing antibodies to native assembled HIV-1 particles expressed by a replicating virus might mimic the advantageous characteristics of live attenuated vaccine. To this aim, we generated a live attenuated recombinant measles vaccine expressing HIV-1 Gag virus-like particles (VLPs) covered with gp160ΔV1V2 Env protein. The measles-HIV virus replicated efficiently in cell culture and induced the intense budding of HIV particles covered with Env. In mice sensitive to MV infection, this recombinant vaccine stimulated high levels of cellular and humoral immunity to both MV and HIV with neutralizing activity. The measles-HIV virus infected human professional antigen-presenting cells, such as dendritic cells and B cells, and induced efficient presentation of HIV-1 epitopes and subsequent activation of human HIV-1 Gag-specific T cell clones. This candidate vaccine will be next tested in non-human primates. As a pediatric vaccine, it might protect children and adolescents simultaneously from measles and HIV.     

A canonical Vγ4Vδ4+ γδ T cell population with distinct stimulation requirements which promotes the Th17 response

We previously reported a subset of γδ T cells in mice which preferentially responds following intradermal immunization with collagen in complete Freund’s adjuvant (CFA). These cells express a nearly invariant “canonical” Vγ4Vδ4+ TCR. They are potent producers of IL-17A and promote the development of collagen-induced arthritis. In this study, we report that CFA emulsified with PBS alone (without collagen) is sufficient to induce a strong response of Vγ4Vδ4+ cells in the draining lymph nodes of DBA/1 and C57BL/6 mice and that the TCRs of the elicited Vγ4Vδ4+ cells in both strains heavily favor the canonical sequence. However, although both CFA and incomplete Freund’s adjuvant (which lacks the killed mycobacteria present in CFA) induced Vγ4Vδ4+ γδ T cell to expand, only CFA stimulated them to express IL-17A. The route of immunization was also critical, since intraperitoneal CFA induced only a weak response by these cells, whereas intradermal or subcutaneous CFA strongly stimulated them, suggesting that the canonical CFA-elicited Vγ4Vδ4+ cells are recruited from Vγ4+ γδ T cells normally found in the dermis. Their IL-17A response requires the toll-like receptor adapter protein MyD88, and their activation is enhanced by IFNγ, although αβ T cells need not be present. The CFA-elicited Vγ4Vδ4+ γδ T cells show a cytokine profile different from that of other previously described IL-17-producing γδ T cells. Finally, the Vγ4Vδ4+ subset appears to promote the Th17 αβ T cell response, suggesting its importance in mounting an effective immune response against certain pathogens.     

Sphingosine Kinase-1 Associates with Integrin αVβ3 to Mediate Endothelial Cell Survival

Sphingosine kinase (SK)-1 promotes endothelial cell (EC) survival through the cell junction molecule CD31 (platelet endothelial cell adhesion molecule-1). The integrin α_vβ₃ is also essential for EC survival; inhibition of α_vβ₃ ligation promotes apoptosis. Herein we demonstrate that under basal conditions, SK-1, α_vβ₃, and CD31 exist as a heterotrimeric complex. Under conditions that affect EC survival such as loss of contact with the extracellular matrix or growth factor activation, more of this heterotrimeric complex forms. Overexpression studies demonstrate a requirement for SK-1 phosphorylation at serine 225 for increased heterotrimeric complex formation, activation of α_vβ₃, and EC survival signals, including Bcl-X and nuclear factor-κB pathways. Moreover, β₃ integrin depletion confirmed the requirement for this heterotrimeric complex in SK-1-mediated EC survival. Thus, with α_vβ₃ integrin being identifiable primarily on angiogenic ECs and SK-1 being highly expressed in tumors, targeting SK-1 may affect multiple survival pathways, and its inhibition may be highly efficacious in controlling pathological EC survival.Survival of vascular endothelial cells (ECs) demands three specific conditions: attachment to an extracellular matrix (ECM),¹ cell-cell interactions (reviewed in Ref. ²), and exposure to growth factors (GF) (for example, vascular endothelial growth factor [VEGF]) and fibroblast growth factor [FGF]).³ ECM and GF-induced survival signals involve integrins, a family of transmembrane heterodimeric cell surface proteins that are formed by two noncovalently associated subunits, α and β. Integrins exist in two distinct functional states, an inactive nonadhesion-promoting state and a ligand-bound, active, and adhesion-promoting conformation.⁴ On ligand binding, integrins are aggregated in transmembrane complexes (focal contacts) that are enriched in cytoskeletal proteins (eg, talin and vinculin) as well as signaling proteins (eg, focal adhesion kinase [FAK]) (reviewed in Refs. ^5,6). Integrin α_vβ₃ is key to EC survival because disruption of α_vβ₃ ligation inhibits blood vessel formation and initiates apoptosis⁷ and is also found at its highest levels on angiogenic ECs in wounds and tumors.⁸ GF receptors also collaborate with integrins by partitioning into common complexes within which they become activated and signal more efficiently.^9,10Sphingosine kinase (SK) is a lipid kinase that catalyzes the phosphorylation of sphingosine to form sphingosine-1-phosphate (S1P) (reviewed in Ref. ¹¹). SK has two isoforms (SK-1 and -2), and although most cells can synthesize S1P, large amounts are present in platelets¹² and recent reports have identified erythrocytes as well as vascular endothelium as major contributors of S1P in circulation.^13,14,15 S1P can act extracellularly through the G protein-coupled S1P receptors (S1P_1-5). Mature ECs express S1P receptors S1P_1-3, and these ligand/receptor interactions promote EC survival, migration, proliferation, adherens junction assembly, increased revascularization, and wound healing both in vitro and in vivo (reviewed in Ref. ¹⁶). SK-1 can also act intracellularly through as yet unknown binding partners in which the ablation of S1P receptor signaling through both chemical or genetic mechanisms does not abrogate SK-1 effects on cell proliferation, Ca²⁺ mobilization, or the survival and motility of ECs (reviewed in Ref. ¹¹). SK-1 seems to have two functional states: an intrinsic or basal state and an agonist-induced activation state that requires its phosphorylation and translocation to the plasma membrane and is responsible for its oncogenic properties.¹⁷Recently, we demonstrated that SK-1 can promote EC survival through the cell junction molecule CD31 (platelet endothelial cell adhesion molecule-1) acting specifically through phosphatidylinositol 3-kinase/Akt and not the mitogen-activated protein kinase pathway.¹⁸ With previous reports that CD31 and SK-1 can interact, at least when overexpressed in cells¹⁹ and that there is cross talk between CD31 and integrins in ECs,²⁰ we investigated whether SK-1 and integrins may be linked in their capacities to regulate EC survival. The results presented here show that enforced overexpression of SK-1 increases the levels of α_vβ₃ integrin as well as its activation state, resulting in regulation of the Bcl-X_L/Bim and nuclear factor-κB (NF-κB) survival pathways. Moreover, we demonstrate that under basal conditions, a complex of α_vβ₃, SK-1, and CD31 exists, and, on conditions of cell stress, enhanced levels of this heterotrimeric complex form. Complex formation and α_vβ₃ integrin activation requires SK-1 to be phosphorylated, thus suggesting that agonist-induced activation of SK-1 is essential for cell survival. These results, combined with our previous work showing that CD31 is an important intermediary in SK-1-induced survival, demonstrates that SK-1 is a fulcrum for the activities of these two well known key survival proteins in ECs, the integrin α_vβ₃ and CD31.     

论信息科学的世界观V／V

第九章信息现象的实证思维、理论思维与哲学思维 本章以物质科学知识体系的发展脉络为参照,讨论了信息科学大致相同的发展逻辑,阐述了信息现象的实证思维、理论思维和哲学思维.笔者自1979年从数学步人计算机领域以来的学术经历,大致从计算机软件工程,经过医学信息学(及生命信息学)、理论信息学,走到了信息哲学.所以这样走过来,并不自觉,是在"跟着感觉走".笔者体会到了某种学术行进的"惯性",好像"身不由己",有时欲停不能,欲罢不忍.本章中包含了笔者与合作伙伴们近三十年来的学术经历总结,其中渗透着指导教授们的心血.     

Flow cytometric analysis of TCR Vβ repertoire in patients with 22q11.2 deletion syndrome

In 22q11.2 deletion patients, the normal decrease in T lymphocyte counts after 1–2 years is blunted such that relatively T lymphocyte numbers increase over early childhood, probably via post‐thymic expansion of peripheral lymphocytes. This may leave less T lymphocyte receptor (TCR) diversity than when derived from naive thymic emigrants. We analysed TCR Vβ repertoire on 27 22q11.2 chromosome deletion patients. No patient had infection at sampling. CD3⁺CD4⁺ recent thymic emigrants (RTEs) were identified by CD45RA and CD31 expression. TCR Vβ repertoire was determined using four‐colour flow cytometry. Patients and controls showed significant TCR Vβ family usage differences between CD3⁺CD4⁺ and CD3⁺CD4^? T lymphocyte subpopulations. Vβ family abnormalities (±3 SD of controls) were identified in 18/27 (67%) patients and 12/47 (25%) controls. In patients, the magnitude of expansions was increased, with some Vβ families representing 37% of the cells present in the subpopulations. There was a significant increase in frequency of abnormalities in CD3⁺CD4⁺ (P < 0.001) and CD3⁺CD4^? T lymphocytes (P < 0.05) in patients. A total of 11/16 patients had an abnormal CD4⁺CD25^Bright TCR Vβ repertoire. There was no difference in expansions/contractions between CD4⁺CD25^Bright and CD4⁺ T lymphocyte repertoires (P = 0.575) for individual patients but significant differences in expansions/contractions between CD4⁺CD25^Bright and CD8⁺ T lymphocytes repertoires (P = 0.011). There was bias in Vβ usage between CD3⁺CD4⁺ and CD3⁺CD4^? T lymphocyte subsets. A total of 67% patients had TCR Vβ repertoire abnormalities, with a trend towards increased repertoire abnormalities with fewer RTEs, suggesting thymic output plays an important role in TCR repertoire diversity. There was no correlation between skewed repertoire and symptoms of infection or autoimmunity.     

心电图V7、V8、V9导联Q波正常值的研究

目的:探讨心电图V_7、V_8、V_9导联Q波的正常值。方法:对1280名健康查体者,除描记常规12导联心电图外,加描V_7、V_8、V_9导联心电图。剔除其中患有或疑似患有各种心脏疾患320人,对960名入选者的V_7、V_8、V_9导联Q波、R波由专业人员进行测量、统计,并进行统计学处理。结果:测得的V_7、V_8、V_9导联Q波的正常值范围是:时间<0.04S;电压:V_7≤0.15mV,V_8≤0.20mV,V_9≤0.25mV;Q/R:V_7≤1/4,V_8≤1/2,V_9≤1。结论:此数值可作为V_7、V_8、V_9导联Q波诊断的正常值参考标准。     

Is M129V of PRNP gene associated with Alzheimer's disease? A case-control study and a meta-analysis

The methionine/valine (M/V) polymorphism at codon 129 within the prion protein gene (PRNP) represents a known risk factor for Creutzfeldt-Jakob disease (CJD). Few authors reported also the effects of this polymorphism on the risk of Alzheimer's disease (AD), although with controversial results. To better clarify this issue, we performed a novel case-control study and a meta-analysis of published association studies between PRNP and AD. Our findings argue against PRNP as a susceptibility gene for developing AD in the Italian population but support the hypothesis that the V allele influences cognitive performances. The meta-analysis, revealed that Caucasian subjects homozygous at codon 129 had a 1.3-fold increased risk [95% CI: 1.0-1.6, p = 0.05] of developing AD compared to heterozygous individuals. We also observed that MM genotype and M allele represent a risk factor for AD, independently from the ethnic background, providing a significant but modest association between this polymorphism and AD.     

Serum hormone and myocellular protein recovery after intermittent runs at the velocity associated with V˙O2max

The responses of serum myocellular proteins and hormones to exercise were studied in ten well-trained middle-distance runners [maximal oxygen consumption (V˙O_2max)?=?69.4?(5.1)?ml?·?kg^?1?·?min^?1] during 3 recovery days and compared to various measures of physical performance. The purpose was to establish the duration of recovery from typical intermittent middle-distance running exercises. The subjects performed, in random, order two 28-min treadmill running exercises at a velocity associated with V˙O_2max: 14 bouts of 60-s runs with 60?s of rest between each run (IR₆₀) and 7 bouts of 120-s runs with 120?s of rest between each run (IR₁₂₀). Before the exercises (pre- exercise), 2?h after, and 1, 2 and 3 days after the exercises, the same series of measurements were performed, including those for serum levels of the myocellular proteins creatine kinase, myoglobin and carbonic anhydrase III (S-CK, S-Mb and S-CA III, respectively), serum hormones testosterone, Luteinizing hormone, follicle-stimulating hormone and cortisol (S-testosterone, S-LH, S-FSH and S-cortisol, respectively) and various performance parameters: maximal vertical jump height (CMJ) and stride length, heart rate and ratings of perceived exertion during an 8-min run at 15?km?·?h^?1 (SL_15?km·h?1, HR_15?km?·?h?1 and RPE_15?km?·?h?1, respectively). Two hours after the end of both exercise bouts the concentration of each measured serum protein had increased significantly (P?15?km?·?h?1 or CMJ. During the recovery days only S-CK was significantly raised (P?P?15?km?·?h?1 (P?120 the post-exercise responses returned to their pre-exercise levels within the 3 days of recovery. The present findings suggest that a single 28-min intermittent middle-distance running exercise does not induce changes in serum hormones of well-trained runners during recovery over 3 days, while changes in S-CK, CMJ and RPE_15?km?·?h?1 indicate that 2–3 days of light training may be needed before the recovery at muscle level is complete.     

V79和V798细胞周期及其调控因子的差异比较

为了解Ｖ７９和Ｖ７９－８两种细胞及调控因子的差异，用「＾３Ｈ『胸腺嘧核苷参入法检测两种细胞，发现Ｖ７９细胞无可检测到的Ｇ２期；Ｖ７９－８细胞则同时缺管可测到的Ｇ１和Ｇ２期。流式细胞光度术显示，Ｖ７９Ｇ１ＡＤＷＥ调控上存在差异。比较两者Ｇ１期周期调控因子的ｍＲＮＡ表达水平发现，Ｖ７９－８ｃｙｃｌｉｎＤ１，ｃｙｃｌｉｎＤ３和ＩＤ表达水平明显高于Ｖ７９，这可能直接导致了两者Ｇ１的差异。     

Thrombosis risk in carriers of the factor V Leiden mutation: is it associated with a defined skin color?

Factor V Leiden (FVL; G1691A) is an autosomal dominant mutation with a high risk for thrombosis. Speculation that founders of FVL lived in the Middle East is supported by a prevalence of FVL that is higher in Arabs residing in Israel, Jordan, Lebanon, and Syria (12-14%) than in other white populations like Europeans (4-5%, up to 15% in the South of Sweden). We sought to verify the appropriate use of skin color as a clinical sign by which Arab individuals in Kuwait are included or excluded from testing for FVL. After institutional approval, 200 healthy Arabs residing in Kuwait consented to participate. Skin type was distinguished for the participants by Fitzpatrick natural skin color classification: 76 (38%) skin type II (white), 96 (48%) Mediterranean skin type IV (brown), and 28 (14%) skin type VI (black). FVL was tested by real-time PCR, and the percentage of carriers was calculated in each group. FVL was positive in 17 (8.5%) of the total subjects: 8 (10.5%) skin type II, 7 (7.3%) skin type IV, and 2 (7.1%) skin type VI. Therefore, FVL shows an even distribution in Arabs, and all Arabs residing in Kuwait should be tested for FVL irrespective of skin color.     

CML中TCR Vβ2、Vβ5和Vβ17亚家族sjTRECs的检测情况

目的检测慢性粒细胞白血病（CML）患者外周血TCR Vβ2-、Vβ5-和Vβ17-Dβ1 sjTRECs的存在情况．从而了解相应3种Vβ亚家族naiveT细胞的近期胸腺输出情况．方法利用半巢式PCR扩增9例CML患者外周血单个核细胞（PBMC）、CD^4＋细胞和CD8^＋细胞DNA的Vβ2-、Vβ5-和Vβ17-Dβ1 sjTRECs，13例正常人外周血作为对照．结果TCR Vβ2-、Vβ5-和Vβ17-Dβ1 sjTRECs在部分正常人和1例CML患者CD4^＋或CD8^＋细胞可以检测到，而在CML患者PBMC中则未能检测到．结论在CML患者Vβ2-、Vβ5-和Vβ17-Dβ1 sjTRECs检出率低，与其胸腺输出相应的TCR Vβ亚家族naiveT细胞水平低下有关．     

A polymorphism, L162V, in the peroxisome proliferator-activated receptor α (PPAR α) gene is associated with lower body mass index in patients with non-insulin-dependent diabetes mellitus

This study examined the effect a polymorphism (L162V) in the gene for peroxisome proliferator activated receptor (PPAR) alpha in the development of non-insulin-dependent diabetes mellitus (type 2 DM), obesity and hyperlipidaemia. The frequency of the L162V polymorphism in the PPARalpha gene was determined in 370 morbidly obese patients who underwent gastric banding surgery, 154 patients attending a type 2 DM clinic, 188 patients attending a lipid clinic and 199 healthy blood donors. The overall frequency of the V allele of the L162V polymorphism was 0.06. There were no significant differences in the allele frequency between patients with morbid obesity, hyperlipidaemia, type 2 DM and healthy controls, suggesting that it does not play a major role in the development of these conditions. The polymorphism was associated with a lower body mass index (BMI) in two independently recruited groups of patients with type 2 DM. There was no effect of the polymorphism on subjects without type 2 DM. Thus a polymorphism in PPARalpha protects type 2 DM patients from the overweight which is frequently associated with their condition.     

Prion proteins with different conformations accumulate in Gerstmann-Sträussler-Scheinker disease caused by A117V and F198S mutations

Gerstmann-Str?ussler-Scheinker disease (GSS) is characterized by the accumulation of proteinase K (PK)-resistant prion protein fragments (PrP(sc)) of approximately 7 to 15 kd in the brain. Purified GSS amyloid is composed primarily of approximately 7-kd PrP peptides, whose N terminus corresponds to residues W(81) and G(88) to G(90) in patients with the A117V mutation and to residue W(81) in patients with the F198S mutation. The aim of this study was to characterize PrP in brain extracts, microsomal preparations, and purified fractions from A117V patients and to determine the N terminus of PrP(sc) species in both GSS A117V and F198S. In all GSS A117V patients, the approximately 7-kd PrP(sc) fragment isolated from nondigested and PK-digested samples had the major N terminus at residue G(88) and G(90), respectively. Conversely, in all patients with GSS F198S, an approximately 8-kd PrP(sc) fragment was isolated having the major N terminus start at residue G(74). It is possible that a further degradation of this fragment generates the amyloid subunit starting at W(81). The finding that patients with GSS A117V and F198S accumulate PrP(sc) fragments of different size and N-terminal sequence, suggests that these mutations generate two distinct PrP conformers.     

Identification of a family with nonspecific mental retardation (MRX79) with the A140V mutation in the MECP2 gene: Is there a need for routine screening?

Mutations in the methyl-CpG-binding protein 2 (MECP2) cause Rett syndrome, a severe neurodevelopmental disorder occurring predominantly in females. Male patients with Rett syndrome are extremely rare, as the Rett-causing mutations in the MECP2 gene are usually lethal in hemizygous males. However, different mutations in the same gene were reported to cause mental retardation, both in sporadic non-syndromic males as well as in syndromic families with disease manifestation in carrier females. The majority of the reported MECP2 mutations in mentally retarded patients cause amino acid substitutions and, especially in isolated cases, discrimination between a disease-causing mutation and a rare polymorphism is not obvious and the significance of each individual variation should be verified. We mapped a new non-syndromic X-linked family (MRX79) to the chromosomal region Xq27.3-Xq28 and identified an A140V mutation in the MEPC2 gene in all patients with the disease haplotype. In addition to data published by others, this suggests that A140V is a recurrent mutation (and not a polymorphism) found in patients with X-linked mental retardation.