腺病毒编码人TRP2修饰DC诱发抗小鼠黑色素瘤免疫的研究

目的比较腺病毒载体(Ad)介导人和小鼠酪氨酸酶相关蛋白2(tyrosinase-relatedpro-tein2,TRP2)修饰小鼠骨髓来源的树突状细胞(BM-DC)诱发抗小鼠黑色素瘤免疫的差异。方法Ad编码人或小鼠TRP2(AdhTRP2或AdmTRP2)体外感染小鼠BM-DC并体内皮下免疫C57BL/6小鼠,7d后取出被免疫小鼠脾细胞行体内细胞毒性T淋巴细胞杀伤试验(invivoCTL)和细胞内IFN-γ染色(ICS)分析CTL的杀伤活性和IFN-γ的产生;或给免疫后小鼠皮下接种小鼠B16.F10黑色素瘤细胞,观察荷瘤小鼠的成活情况。结果invivoCTL和ICS分析显示,AdhTRP2/BM-DC免疫小鼠,其6hCTL杀伤率为(98.7±1.2)%,IFN-γ产生的CD8+T细胞占总CD8+T细胞的(1.25±0.21)%;而AdmTRP2/BM-DC免疫的小鼠,其6hCTL杀伤率和产生IFN-γ的CD8+T细胞比例分别为(28.6±6.3)%和(0.24±0.06)%。荷瘤试验表明,AdhTRP2/BM-DC免疫小鼠后1周皮下接种106B16.F10细胞,观察3个月100%的小鼠无瘤生长;而接种5×104B16.F10细胞至AdmTRP2/BM-DC免疫1周的小鼠,3个月后小鼠成活率仅为40%。结论Ad介导异种(人)TRP2较自身(小鼠)TRP2修饰的BM-DC更为有效地打破肿瘤免疫耐受、诱导强烈的抗黑色素瘤免疫反应,是一种高效的以DC为基础的肿瘤疫苗。     

Immune responses to DNA in normal and aberrant immunity

Because of structural microheterogeneity, DNA can exert powerful effects that lead to immune system activation as well as antibody induction. These activating effects resemble those of endotoxin and result from sequences that occur much more commonly in bacterial DNA than in mammalian DNA. In contrast, mammalian DNA can inhibit the response to bacterial DNA as well as other stimuli and may serve a counterregulatory role during infection. The recognition of the immune effects of DNA is relevant to the pathogenesis of a variety of infectious and inflammatory diseases including systemic lupus erythematosus, a prototypic autoimmune disease characterized by anti-DNA autoantibodies.     

Operation Everest II: Alterations in the immune system at high altitudes

We investigated the effects on immune function after progressive hypobaric hypoxia simulating an ascent to 25,000 ft (7620 m) over 4 weeks. Multiple simultaneousin vitro andin vivo immunologic variables were obtained from subjects at sea level, 7500 ft (2286 m), and 25,000 ft during a decompression chamber exposure. Phytohemag-glutinin-stimulated thymidine uptake and protein synthesis in mononuclear cells were reduced at extreme altitudes. Mononuclear-cell subset analysis by flow cytometry disclosed an increase in monocytes without changes in B cells or T-cell subsets. Plasma IgM and IgA but not IgG levels were increased at altitudes, whereas pokeweed mitogen-stimulatedin vitro IgG, IgA, and IgM secretion was unchanged. During exposure to 25,000 ft,in vitro phytohemagglutinin-stimulated interferon production and natural killer-cell cytotoxicity did not change statistically, but larger intersubject differences occurred. IgA and lysozyme levels (nasal wash) and serum antibodies to nuclear antigens were not influenced by altitude exposure. These results suggest that T-cell activation is blunted during exposure to severe hypoxemia, whereas B-cell function and mucosal immunity are not. Although the mechanism of alteredin vitro immune responsiveness after exposure to various environmental stressors has not been elucidated in humans, hypoxia may induce alterations in immune regulation as suggested byin vitro immune assays of effector-cell function.Some of this study's results were presented as an abstract at the FASEB meeting in St. Louis, Missouri, 1986.     

The functional state of follicular dendritic cells in severe combined immunodeficient (SCID) mice: Role of the lymphocytes

In the present study, we have investigated the capacity of follicular dendritic cells (FDC) to trap immune complexes (IC) in the splenic white pulp of severe combined immunodeficient (SCID) mice and the influence of lymphocyte transfer on FDC function. FDC are absent in the splenic white pulp of naive SCID mice as revealed by in vitro IC trapping assay. One week after transfer of syngeneic lymphocytes, functional FDC with complement receptors appeared in the primary follicles coincident with B cell segregation, and IC were trapped on those FDC in a complement-dependent manner. Next, we immunized the reconstituted SCID mouse to see whether another type of FDC could be induced in the secondary follicle. Antigenic stimulation induced FDC with an additional capacity to capture IC via FcR γ II. As seen in immunocompetent mice, this type of FDC was located only in the light zone of the secondary follicle. The newly generated FDC did not carry H-2 antigen of transferred lymphocytes from F₁ mice. In SCID mice, in which normally no functional FDC are detectable, the microenvironments of the splenic white pulp have a capacity to develop and differentiate normally after transfer of lymphocytes. Apparently, the generation of functional IC-trapping FDC causes the induction of complement receptor(s) and Fc receptor on meshwork cells, which requires the presence of the lymphocytes.     

几种肿瘤患者红细胞免疫粘附功能的初步观察

本文报道了30例食管癌、20例胃癌、15例乳腺癌、8例肝癌、10例淋巴瘤等患者红细胞免疫粘附功能的初步结果,并与67例献血员值进行了比较。几种肿瘤病人的RBC C_(3b)RR低下,RBC-ICR较高,两者与献血员值比较都有明显差别(P<0.01),几种肿瘤患者CIC阳性率在60～83.4％之间。对食管癌患者部分病例进行跟踪检测,并对其临床意义进行了初步探讨。     

Toll样受体与树突状细胞

Toll样受体(TLR)在介导固有免疫和适应性免疫应答中有重要作用,可以表达于多种免疫细胞,包括树突状细胞(DC).了解Toll样受体的免疫学基础、与DC之间的联系以及其在免疫耐受干预方面的作用很有必要.     

Strong cellular and humoral anti-HIV Env immune responses induced by a heterologous rhabdoviral prime-boost approach

Recombinant rhabdovirus vectors expressing human immunodeficiency virus (HIV) and/or simian immunodeficiency virus (SIV) proteins have been shown to induce strong immune responses in mice and rhesus macaques. However, the finding that such responses protect rhesus macaques from AIDS-like disease but not from infection indicates that further improvements for these vectors are needed. Here, we designed a prime-boost schedule consisting of a rabies virus (RV) vaccine strain and a recombinant vesicular stomatitis virus (VSV) both expressing HIV Envelope (Env). Mice were primed and boosted with the two vaccine vehicles by different routes and in different combinations. Mucosal and systemic humoral responses were assessed using enzyme linked immunosorbent assay (ELISA) while the cellular immune response was determined by an IFN-gamma ELISPOT assay. We found that an immunization combination of RV and VSV elicited the highest titers of anti-Env antibodies and the greatest amount of Env-specific IFN-gamma secreting cells pre- and post-challenge with a recombinant vaccinia virus expressing HIV(89.6) Env. Furthermore, intramuscular immunization did not induce antigen-specific mucosal antibodies while intranasal inoculation stimulated vector-specific IgA antibodies in vaginal washings and serum. Our results show that it is feasible to elicit robust cellular and humoral anti-HIV responses using two different live attenuated Rhabdovirus vectors to sequentially prime and boost.     

pH modulates conducting and gating behaviour of single calcium release channels

Intracellular pH changes affect excitation-contraction coupling in skeletal, and cardiac muscles. However the proton implication in modulating the sarcoplasmic reticulum Ca²⁺ release channel activity has never been visualized at single channel level. A large conducting Ca²⁺ release pathway has previously been characterized after incorporation of skeletal and cardiac sarcoplasmic reticulum vesicles into planar lipid bilayers. This channel has been activated by micromolar and millimolar concentrations of Ca²⁺ and ATP, respectively. The pH was independently varied on each side of the channels. Acidification of the cis-chamber (7.4 to 6.6) induced a modification of the gating behaviour, resulting in a decrease of the open probability. This effect was completely reversible. On the other hand, acidification of the trans-chamber (7.4 to 6.8) induced a reduction of the unitary conductance of the sarcoplasmic reticulum Ca²⁺ release channel.     

Circadian and age-related modulation of thermoreception and temperature regulation: mechanisms and functional implications

At older ages, the circadian rhythm of body temperature shows a decreased amplitude, an advanced phase, and decreased stability. The present review evaluates to what extent these changes may result from age-related deficiencies at several levels of the thermoregulatory system, including thermoreception, thermogenesis and conservation, heat loss, and central regulation. Whereas some changes are related to the aging process per se, others appear to be secondary to other factors, for which the risk increases with aging, notably a decreased level of fitness and physical activity. Moreover, functional implications of the body temperature rhythm are discussed. For example, the relation between circadian rhythm and thermoregulation has hardly been investigated, while evidence showed that sleep quality is dependent on both aspects. It is proposed that the circadian rhythm in temperature in homeotherms should not be regarded as a leftover of ectothermy in early evolution, but appears to be of functional significance for physiology from the level of molecules to cognition. A new view on the functional significance of the circadian rhythm in peripheral vasodilation and the consequent out-of-phase rhythms in skin and core temperature is presented. It is unlikely that the strong, daily occurring, peripheral vasodilation primarily represents heat loss in response to a lowering of set point, since behavioral measures are simultaneously taken in order to prevent heat loss. Several indications rather point towards a supportive role in immunological host defense mechanisms. Given the functional significance of the temperature rhythm, research should focus on the feasibility and effectiveness of methods that can in principle be applied in order to enhance the weakened circadian temperature rhythm in the elderly.     

壳聚糖纳米颗粒包裹对鼠白细胞介素2基因表达和调节小鼠免疫效应的影响

本实验制备壳聚糖纳米颗粒 (CNP) ,包裹小鼠白细胞介素 2基因 (mIL 2 )真核表达质粒 (VRMIL 2 ) ,肌注接种 2 1d昆明小鼠 ,观察mIL 2基因体内表达及其对免疫应答和免疫保护的影响。实验结果发现 :CNP包裹VRMIL 2注射小鼠血液中IgG、IgM和IgA不同程度地增多 ,均显著高于CNP包裹空白质粒组 (P <0 .0 5 ) ;其血清中IL 2、IL 4和IL 6的含量明显升高 ,与对照组差异显著 (P <0 .0 5 ) ;外周血液的白细胞和淋巴细胞数量也较对照组显著增加。免疫后 35d以大肠杆菌口服攻毒实验小鼠 ,检测发现 :CNP包裹VRMIL 2组小鼠的上述免疫指标除中性粒细胞外均显著多于对照小鼠 ,VRMIL 2接种小鼠均健康存活 ,而对照小鼠均发病 ;尽管CNP包裹VRMIL 2接种小鼠的体液和细胞免疫指标与未包裹VRMIL 2免疫鼠差异不显著 (P >0 .0 5 ) ,但剂量仅为后者的 1/ 5。这些结果表明 :壳聚糖纳米颗粒包裹VRMIL 2可显著提高外源IL 2基因体内表达水平 ,明显增强体液和细胞免疫水平的效应 ,增强对大肠杆菌的抗病力 ,提示壳聚糖纳米颗粒包裹IL 2基因可明显增强动物的体液和细胞免疫 ,可作为有效的抗感染免疫调节剂。