从基因水平对新型佐剂在旋毛虫疫苗中免疫调节作用的探讨

目的 从细胞因子的基因转录水平探讨两种新型佐剂霍乱毒素B亚单位(CT-B)和皂素(Saponin)在旋毛虫疫苗中的免疫调节作用。方法 36只NIH小鼠随机分为CT-B佐剂免疫组、Saponin佐剂免疫组和对照组。将CT-B和saponin分别与旋毛虫肌幼虫可溶性抗原混匀后，以口服或皮下注射途径免疫小鼠，隔周1次。第3次免疫后1周，经口感染旋毛虫感染期肌幼虫。感染后第8d，采用RT-PCR技术分析各组小鼠脾细胞在体外旋毛虫肌幼虫抗原刺激下，转录细胞因子IFN-γ、IL-2、IL-4及IL-5 mRNA的水平。结果 各组小鼠的脾细胞在体外抗原诱导下均未能转录IL-2和IFN-γ mRNA，但均有不同程度的IL-4和IL-5特异扩增，两免疫组的IL-4及IL-5 mRNA转录水平明显高于对照组。结论 Th2细胞及其分泌的细胞因子在机体抗旋毛虫的保护性免疫中发挥着重要作用；从佐剂角度提高旋毛虫疫苗的保护性是可行的。     

结核分支杆菌Ag85B DNA疫苗的构建及其免疫作用的研究

目的 研究pcDNA3-Ag85B质粒DNA疫苗的免疫原性及其诱生细胞免疫应答的作用。方法 将结核分支杆菌Ag85B基因插入载体pcDNA3中,制备pcDNA3-Ag85B DNA疫苗。分别以生理盐水、pcDNA3及pcDNA3-Ag85B免疫BALB／c小鼠,检测小鼠抗Ag85B抗体水平（ELISA）和体外循异抗原刺激诱导的免疫小鼠脾细胞IL－2、IL－4、IL－10、IFN－γ表达水平。结果 pcDNA3-Ag85B诱生的抗Ag85B效介明显高于生理盐水组和pcDNA3组;pcDNA3-Ag85B免疫小鼠脾细胞在Ag85B抗原刺激下,IL－2和IFN－γ mRNA转录水平明显高于对照组,而IL－4和IL－10 mRNA转录水平在3组中无明显变化。结论 pcDNA3-Ag85B质粒DNA疫苗不仅能增强体液免疫,亦可诱导Th1型细胞免疫。     

CD40和白介素-12缺陷型小鼠辅助T细胞应答变化揭示了Th1应答在肥头绦虫幼虫清除中的关键作用

为了研究IL 12和CD4 0在肥头绦虫幼虫引起的鼠囊虫病中对Th应答的调节作用 ,实验通过腹腔注射肥头绦虫囊尾蚴的方法感染IL 12 p35 / 和CD4 0 / 型、野生型BALB/c小鼠以及作为抵抗对照的STAT6 / 型小鼠 ,分析了血清抗体状况和脾细胞、巨噬细胞的细胞反应和细胞因子的分泌情况。实验发现IL 12 p35 / 小鼠感染后无法诱导Th1应答 ,而是诱导强烈的Th2应答 ,产生的针对肥头绦虫幼虫抗原的Th2相关的IgG1、IgE等抗体水平和IL 4、IL 5、IL 13等细胞因子水平均高于野生型小鼠 ;而Th1相关的IgG2a水平和IFN γ水平明显低于野生型小…     

旋毛虫排泄分泌抗原对小鼠的保护性免疫

目的比较旋毛虫成虫排泄分泌抗原(ES抗原)、肌幼虫ES抗原、成虫和肌幼虫ES混合抗原对小鼠的免疫保护作用。方法用生理盐水培养法从培养液中提取成虫ES抗原、肌幼虫ES抗原,分别用成虫ES抗原、肌幼虫ES抗原、成虫和肌幼虫ES混合抗原免疫小鼠,同时设佐剂组和对照组,间隔7d共免疫3次。末次免疫后7天,每只小鼠用200条旋毛虫感染期幼虫经口进行攻击感染。感染后7天和30天检查各组小鼠肠道成虫数和肌幼虫数。结果旋毛虫成虫ES抗原组、肌幼虫ES抗原组、成虫和肌幼虫ES混合抗原组的成虫减虫率分别为87.95%、69.48%、84.34%,肌幼虫减虫率分别为74.79%、87.97%、86.87%。成虫ES抗原组、成虫与肌幼虫ES抗原混合组的成虫减虫率均高于肌幼虫ES抗原组(P均<0.05)。肌幼虫ES抗原组、成虫与肌幼虫ES抗原混合组的肌幼虫减虫率均高于成虫ES抗原组(P均<0.01)。结论旋毛虫成虫和肌幼虫ES混合抗原均能诱导小鼠产生抗成虫及肌幼虫较强的免疫力。     

两种新型免疫佐剂CT-B、Saponin制备的旋毛虫疫苗对小鼠免疫保护作用的比较研究

目的　比较两种新型免疫佐剂CT -B(霍乱毒素B亚单位 )、saponin(皂素 )制备的旋毛虫疫苗对小鼠的免疫保护作用。方法　将旋毛虫肌幼虫可溶性抗原分别与CT -B、saponin混合 ,以口服或皮下注射途径免疫NIH小鼠 ,间隔 1周共免疫 3次 ,末次免疫后 1周 ,与对照组同时予 2 0 0条旋毛虫感染期肌幼虫攻击 ,比较三组小鼠肠道成虫数、雌虫生殖力及肌幼虫数。结果　与对照组比较 ,CT -B组成虫减虫率、新生幼虫减虫率及肌幼虫减虫率分别为 91 5 9%、6 1 74 %和 90 32 % ,saponin组成虫减虫率、新生幼虫减虫率及肌幼虫减虫率分别为 79 2 1%、6 7 4 4 %和 88 39%。 结论　CT -B和saponin均能有效提高机体对旋毛虫的保护性免疫力 ,CT -B对肠道成虫的影响更显著     

旋毛虫肌幼虫抗原免疫小鼠抗感染能力及其机制的研究

目的探讨旋毛虫肌幼虫抗原免疫后小鼠产生的抗感染能力及其机制。方法制备旋毛虫肌幼虫抗原,免疫小鼠,再用旋毛虫感染性幼虫对免疫小鼠实施攻击感染,检查感染后小鼠肠道成虫和肌肉幼虫数并计算减虫率;同时采用酶联免疫吸附试验(ELISA)分别检测旋毛虫肌幼虫抗原免疫后7、14、284、2 d小鼠外周血单个核细胞(PBMC)培养上清中IL-4和IFN-γ水平。结果肌幼虫抗原免疫组小鼠的肠道成虫数减虫率为38.54%,肌幼虫减虫率为35.31%;ELISA检测免疫小鼠PBMC培养上清IL-4和IFN-γ水平均升高,直至免疫后42 d,以免疫后14、28、42 d的IL-4水平升高更显著(P0.01)。结论旋毛虫肌幼虫抗原免疫能诱导小鼠产生有效的抗旋毛虫感染作用,其抗感染机制以Th2细胞介导的体液免疫为主。     

旋毛虫肌幼虫排泄分泌抗原诱导的肠道免疫保护作用研究

目的 研究旋毛虫肌幼虫排泄分泌抗原(Trichinella spiralis muscle larvae excretory-secretory, Ts-Es)诱导小鼠肠道保护性免疫能力以及免疫调节作用。方法 54只BALB/c小鼠随机分为3组，空白对照组、旋毛虫感染组(Ts)、旋毛虫Ts-Es抗原免疫组(Ts-Es)。空白组小鼠腹腔注射PBS;抗原免疫组小鼠腹腔注射0.1 mL的Ts-Es抗原与等量弗氏完全佐剂；旋毛虫感染组腹腔注射PBS和弗氏完全佐剂，每隔7 d注射1次，共3次，末次注射后7 d旋毛虫感染组和Ts-Es抗原免疫组用400条/mL旋毛虫感染性幼虫灌胃攻击感染。解剖取材，检查肠道成虫及肌肉幼虫减虫率，用HE染色法观察肠道病理变化，RT-qPCR法检测小肠中目的基因mRNA表达水平。结果 与旋毛虫感染组相比Ts-Es抗原免疫组成虫和肌幼虫减虫率分别为49%(t=8.109,P<0.05)和67%(t=8.090,P<0.05);与空白对照组相比旋毛虫感染组小肠T-bet(t=5.25,P<0.05)、GATA3(t=2.50,P<0.05)、ROR...     

IL-12在抗伯氏疟原虫感染中的免疫佐剂作用

目的　探讨IL - 12在抗伯氏疟原虫感染中的免疫佐剂作用。方法　BALB/c小鼠分别皮下用抗原sAg、IL - 12、抗原sAg联合IL - 12或抗原sAg联合弗氏佐剂进行免疫 ,共免疫 3次 ,每次间隔 2wk ,末次免疫后 1wk以 5× 10 5个感染P berghei的RBC攻击感染 ,观察各组小鼠原虫血症变化及存活时间。攻击感染后 1wk收集各组小鼠血清 ,采用ELISA检测血清中IFN -γ、IL - 4、IL - 10及IgG水平。结果　抗原sAg联合IL - 12免疫可较其它各免疫组及未免疫对照组显著延长小鼠存活时间 ,且全部小鼠在原虫血症达 5 0 %时开始死亡 ;而单独抗原sAg、单独IL - 12或抗原sAg联合弗氏佐剂免疫与未免疫比较 ,对小鼠原虫血症及存活时间均无明显影响 ,且半数以上小鼠均在原虫血症小于 2 5 %时死亡。抗原联合IL - 12免疫组小鼠血清中IFN -γ及IgG水平较单独抗原sAg、单独IL - 12及未免疫组显著升高。 结论　结果表明 ,IL - 12与sAg抗原共同免疫小鼠可诱导对P berghei攻击感染的部分抵抗力 ,具有一定的免疫佐剂活性。     

刺激原诱导的华支睾吸虫感染大鼠淋巴细胞体外增殖和细胞因子的产生

目的　研究感染华支睾吸虫的大鼠T淋巴细胞对寄生虫抗原或促有丝分裂原植物血凝素刺激后的体外增殖及细胞因子产生的免疫效应。　方法　观察感染华支睾吸虫的大鼠脾淋巴细胞和肠系膜淋巴结细胞 ,在促有丝分裂原PHA ,华支睾吸虫分泌排泄抗原 ,华支睾吸虫成虫粗抗原和异尖线虫幼虫粗抗原刺激下 ,体外淋巴细胞增殖及细胞因子的水平 ,包括IFN γ ,IL 2 ,IL 4,IL 1 0。　结果　肠系膜淋巴结的淋巴细胞增殖高于脾淋巴细胞。淋巴细胞浓度为 3× 1 0 6或 9× 1 0 6 细胞/孔时 ,脾脏和肠系膜淋巴细胞增殖均显著高于对照组 (P <0 .0 1 )。淋巴细胞数为 5× 1 0 6 细胞 /孔 ,刺激原浓度为 5或 1 0 μg/ml时 ,淋巴细胞活化明显。肠系膜淋巴结细胞的细胞因子 ,IFN γ和IL 1 0的产生水平显著增加。　 结论　淋巴细胞浓度 5×1 0 6 细胞/孔和刺激原浓度 1 0 μg/ml的条件为激活T淋巴细胞增殖和细胞因子产生的最适培养条件。华支睾吸虫感染大鼠的成虫分泌排泄抗原在体内可能刺激淋巴细胞增殖和细胞因子产生     

HBcAg核酸疫苗与白细胞介素表达质粒联合接种小鼠的免疫应答观察

目的观察HBcAg核酸疫苗与鼠白细胞介素12(IL12)和白细胞介素18(IL18)表达质粒联合免疫小鼠所诱导的特异性免疫应答。方法小鼠随机分为载体质粒组、HBcAg核酸疫苗组(核酸疫苗组)、HBcAg核酸疫苗 IL12组(C IL12组)、HBcAg核酸疫苗 IL18组(C IL18组)和HBcAg核酸疫苗 IL12/IL18组(C IL12/IL18组)。载体质粒、HBcAg核酸疫苗、IL12及IL18表达质粒经肌内注射法免疫各组小鼠。采用酶联免疫吸附试验检测小鼠血清HBcAg特异性抗体、IgG亚类(IgG1,IgG2a)以及小鼠脾淋巴细胞培养上清液干扰素(IFN)γ含量。采用乳酸脱氢酶(LDH)释放法检测免疫小鼠特异杀伤性T淋巴细胞(CTL)活性。结果除对照质粒组外,核酸疫苗免疫的各组小鼠均能检出血清抗HBc,C IL12组、C IL18组和C IL12/IL18组的抗HBc终点滴度与C组相比均明显增高(P<0.05)。各组小鼠抗HBcIgG亚类均以IgG2a占优。核酸疫苗免疫组除C IL12 IL18组外,小鼠脾细胞培养上清液IFNγ水平均显著高于对照质粒组(P<0.01)。C IL18组和C IL12/IL18组小鼠脾细胞HBcAg特异性CTL活性强于其他各组。结论IL12和(或)IL18表达质粒与HBcAg核酸疫苗联合免疫,具有增强HBcAg核酸疫苗所激发的免疫应答特别是细胞免疫应答的作用。     

The immunoneuroendocrine role of melatonin

Abstract: A tight, physiological link between the pineal gland and the immune system is emerging from a series of experimental studies. This link might reflect the evolutionary connection between self-recognition and reproduction. Pinealectomy or other experimental methods which inhibit melatonin synthesis and secretion induce a state of immunodepression which is counteracted by melatonin. In general, melatonin seems to have an immunoenhancing effect that is particularly apparent in immunodepressive states. The negative effect of acute stress or immunosuppressive pharmacological treatments on various immune parameters are counteracted by melatonin. It seems important to note that one of the main targets of melatonin is the thymus, i.e., the central organ of the immune system. The clinical use of melatonin as an immunotherapeutic agent seems promising in primary and secondary immunodeficiencies as well as in cancer immunotherapy. The immunoenhancing action of melatonin seems to be mediated by T-helper cell-derived opioid peptides as well as by lymphokines and, perhaps, by pituitary hormones. Melatonin-induced-immuno-opioids (MHO) and lymphokines imply the presence of specific binding sites or melatonin receptors on cells of the immune system. On the other hand, lymphokines such as -γ-interferon and interleukin-2 as well as thymic hormones can modulate the synthesis of melatonin in the pineal gland. The pineal gland might thus be viewed as the crux of a sophisticated immunoneuroendocrine network which functions as an unconscious, diffuse sensory organ.     

Response of rat pineal melatonin to calcium, magnesium, and lithium is circadian stage dependent

Abstract: Herein we documented the response of pineal melatonin production to electrolytes known to be effective on pineal function in view of a possible circadian stage dependence. We studied the release of melatonin by perifused rat pineal glands at 2 different circadian stages corresponding to the middle of the light and dark periods, i.e., respectively, 7 and 19 HALO (Hours After Light Onset, L:D = 12:12). The initial efflux rates were, as expected, much higher in the perifusates of glands removed from rats sacrificed during the dark phase than of those removed during the light phase. After 3 hr of perifusion, melatonin release reached similar levels which were found constant up to the 8th hr of perifusion, whatever the circadian stage. Perifusion of the glands with physiological concentrations for the rat of calcium (5.2 mmol/1) and magnesium (1.34 mmol/1) resulted in a stimulatory effect on the pineal glands removed from rats sacrificed in the middle of the dark period (19 HALO), whereas no effects were observed on the pineal glands removed from rats sacrificed during the light (7 HALO). Lithium (0.28 and 0.55 mmol/1) was ineffective on melatonin release in pineal glands removed 7 and 19 HALO. Our results show differences in the initial efflux rates of melatonin and in the response of perifused pineal glands to calcium and magnesium according to the circadian stage.     

Changes in connexin43, the gap junction protein of astrocytes, during development of the rat pineal gland

Abstract: The abundance of gap junctions between rat pineal astrocytes formed by connexin43 (Cx43) was studied during development. Levels and distribution of Cx43 were measured by immunoblotting and indirect immunofluorescence, respectively. The amount of Cx43 in cells located within the gland was low until about the 7th postnatal day and increased to adult values between the 14th and 21st days postpartum. Although astrocytes, recognized by their vimentin immunoreactivity, were scarce before birth, they were abundant by the 7th postnatal day suggesting that the low levels of Cx43 found at this age corresponded to a low expression of this protein. Localization of the immunoreactivity to Cx43 and vimentin showed a close correlation, indicating that mature or immature pineal astrocytes form gap junctions made of Cx43. Since Cx43 levels attained their adult values at about the time the innervation and the functional state of the gland reached maturity (2–3 weeks after birth), it is proposed that astrocyte gap junctions are involved in the function of the adult rat pineal gland.     

Conservative management of perforated duodenal diverticulum： A case report and review of the literature

Duodenal diverticula are a relatively common condition. They are asymptomatic, unless they become complicated, with perforation being the rarest but most severe complication. Surgical treatment is the most frequently performed approach. We report the case of a patient with a perforated duodenal diverticulum, which was diagnosed early and treated conservatively with antibiotics and percutaneous drainage of secondary retroperitoneal abscesses. We suggest this method could be an acceptable option for the management of similar cases, provided that the patient is in good general condition and without septic signs.     

Presence of immunoreactive growth hormone and prolactin in the ovine pineal gland

Abstract: The use of antisera raised against bovine growth hormone (GH) and ovine prolactin (PRL) enabled the detection of related immunoreactive (ir) sequences of proteins in ovine pineal tissue. The isolation of PRL-like ir-material was accomplished using a 0.25 M ammonium sulphate (pH 5.5) extraction followed by ethanol precipitation, whereas the resulting 2.0 M ammonium sulphate (pH 7.0) precipitate contained a GH-like immunoreactivity. Gel chromatography of the GH-like immunoreactivity (Sephadex G-100) indicated the presence of several GH-like fragments ranging in the M_r range of 7,000 to 55,000. Analyses of the PRL-like ir-material found in pineal tissue on HPLC using a TSK 545-DEAE column led to the resolution into a single peak of immunoreactivity. A single peak of activity was also observed following chromatofocusing and hydrophobic interaction chromatography of the ir-peak from the TSK 545-DEAE column. The PRL-like ir-material inhibited the binding of [¹²⁵I]ovine PRL-S14 to anti-ovine PRL antibodies without showing an affinity for binding to anti-rat PRL or anti-bovine GH antibodies. Scatchard analysis of the binding of pineal PRL-like ir-material and pituitary ovine PRL-S14 to liver membranes from day-20 pregnant rats revealed similar affinity constants (K_a of 4.7 ± 0.2 × 10⁹ M^-1). In addition, the replication of Nb 2 Node rat lymphoma cells was stimulated by pineal PRL-like ir-material, an effect known to be specific for lactogenic hormones. The pineal PRL-like immunoreactivity appeared on sodium dodecyl sulfate polyacrylamide gels as a single major band of M_r 24,000. The functional status of PRL-and GH-like ir-material in the ovine pineal remains to be determined, but evidence is presented that the overall protein synthesis rate of the rat pineal responded to circulating concentrations of PRL.     

Microbiology of human immunodeficiency virus anorectal disease

PURPOSE: Individuals who are seropositive for the human immunodeficiency virus are at high risk for opportunistic infection and anorectal disorders. Little prospective information is available regarding anorectal pathogens in these patients. METHODS: One hundred sixty-three HIV-seropositive patients presented to the colorectal clinic between 1989 and 1992. Forty-seven (29 percent) patients were thought to have an infectious process and were prospectively studied using a standardized multiculture protocol. RESULTS: Mean age was 33 (range, 19–59) years. All were male; high-risk behavior accounted for 87 percent of HIV transmissions. Presenting complaints included anorectal pain (79 percent), pus per anum (28 percent), and blood per anum (26 percent). Examination revealed perianal tenderness (60 percent), condyloma (38 percent), perianal ulcers (38 percent), and anal fissures (34 percent). Sixty-six sets of cultures were performed; 28 patients had one set, 15 had two sets, and 4 had three sets. Thirty-two of these 47 patients (68 percent) had positive cultures including herpes (50 percent), cytomegalovirus (25 percent),Neisseria gonorrhoeae (16 percent), chlamydia (16 percent), acidfast bacilli (2 percent), and others (9 percent). Six of 32 patients with positive cultures had more than one organism cultured. Sixteen (50 percent) patients with positive cultures were treated medically, 8 (25 percent) were treated surgically and 8 (25 percent) were treated with both modalities. Sixty-one procedures were performed on 17 patients for condylomata. Eighteen patients had 20 procedures for abscesses, 50 percent of whom had positive cultures for other than common bowel flora; all improved. Fourteen patients underwent 33 procedures for perianal fistulas.Mycobacterium fortuitum was cultured from one patient who required 13 procedures for abscesses and fistulas. Forty-five (96 percent) patients were followed for an average of 12.5 months ±2.9 SEM (range, 1–94 months). Symptoms were improved or resolved in 22 of 32 (69 percent) patients with positive cultures and in 11 of 13 (84 percent) with negative cultures. CONCLUSIONS: Specific pathogens may often be identified in human immunodeficiency virus-seropositive patients with anorectal disorders if aggressively sought. Although patients without specific pathogens identified may be expected to improve with planned empiric treatment, positive identification allows more directed therapy.