蚯蚓肽对小鼠非特异性免疫功能的影响

通过用不同剂量的蚯蚓肽(EP)对健康和免疫抑制小鼠灌胃15 d,观察EP对小鼠淋巴细胞增殖反应,巨噬细胞细胞毒效应以及巨噬细胞和脾细胞产生NO的影响,以探讨EP对小鼠非特异性免疫功能的影响。试验发现0.1,0.5 mg/mL组明显提高淋巴细胞增殖率,增强巨噬细胞细胞毒效应,提高巨噬细胞和脾细胞分泌NO的水平(P<0.0l),0.5 mg/mL明显提高免疫抑制小鼠的免疫功能(P<0.01)。结果表明:一定剂量下,EP具有调节免疫功能、拮抗环磷酰胺引起的免疫抑制的作用。     

Interactions of 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) with immune responses of rainbow trout

Chlorinated dioxins, as typified by the most potent isomer, TCDD, are immunosuppressive in mammalian species and can enhance the susceptibility to a number of diseases. In recent years chlorinated dioxins have been detected in fish in many freshwater and marine habitats. Thus far, the effects of these chemicals on the immune responses of fish have not been examined. We studied the influence of TCDD on the defense mechanisms of rainbow trout. Yearling trout were injected intraperitoneally with the vehicle, 0.1, 1.0 or 10 micrograms/kg of TCDD. Interactions with the humoral immune response to sheep red blood cells (SRBC) were assessed by the Jerne plaque assay using head kidney and spleen leukocytes. Serum antibody was measured by complement-mediated lysis of SRBC in a chromium release assay. Effects of TCDD on the cellular immune responses were evaluated by the response of thymic and splenic lymphocytes to Con A and PWM. In addition, the phagocytic activity of peritoneal macrophages was examined in vitro. Trout which received 0.1 or 1.0 micrograms/kg TCDD remained clinically normal, and defense mechanisms were unaltered in these fish. Trout which received 10 micrograms/kg of TCDD became hypophagic and exhibited fin necrosis, ascites and suppression of hematopoiesis. In this treatment group, Con A-induced blastogenesis of thymic and splenic lymphocytes was not significantly changed, however, suppression of the PWM-induced response of splenic lymphocytes occurred. No statistically significant alterations occurred in humoral immune responses, and phagocytic activity of peritoneal macrophages was not decreased. The dose-response curve for various biologic effects of TCDD in the rainbow trout appears different from that in sensitive mouse strains. The 30-day, single-dose, parenteral LD50 for TCDD in the C57BL mouse is 100 micrograms/kg, and TCDD suppresses both cell-mediated and humoral immune responses at 1-2 micrograms/kg in this mouse strain. In the rainbow trout, however, immunosuppression was evident only at doses of TCDD approaching the 80-day, single-dose, parenteral LD50 of 20 micrograms/kg.     

维生素A对小白鼠免疫机能的影响

本试验通过在普通日粮中成倍添加维生素A,研究了维生素A对小白鼠免疫机能的影响。结果表明,成倍添加维生素A后小白鼠外周血液中T细胞相对含量与绝对含量均增加(P<0.05),体脾比减小(P<0.05),脾脏巨噬细胞增数(P<0.01),腹腔巨噬细胞吞噬百分率增加(P<0.05)。从而提示在普通日粮中添加大剂量维生素A可以增强小白鼠免疫机能,尤其是细胞免疫机能和非特异性免疫机能。     

金线莲多糖对免疫抑制小鼠脾淋巴细胞增殖及免疫器官的影响

目的：探讨金线莲多糖对免疫抑制小鼠脾淋巴细胞增殖及免疫器官的影响。方法：按大、中、小剂量给小鼠灌胃金线莲多糖,腹腔注射环磷酰胺（CY）建立免疫抑制模型;测定小鼠体重及胸腺、脾脏重量,计算胸腺、脾指数;采用MTT法检测脾淋巴细胞的增殖。结果：3个金线莲多糖剂量组小鼠的体重及免疫器官指数均低于空白组而高于模型组,脾淋巴细胞增殖均显著（P〈O．05）高于模型组。结论：金线莲多糖能提高免疫抑制小鼠体重及免疫器官指数,促进脾淋巴细胞增殖。     

Anterior chamber-associated immune deviation.

The eye possesses a critical method of self-preservation in response to intraocular antigen presentation. Instead of conventional immunity by means of delayed-type hypersensitivity (DTH), the eye participates in a systemic immune response involving the thymus and spleen, ultimately leading to suppression of cell-mediated (T helper 1) immunity. The immune response begins with intraocular capture of antigen by specialized ocular antigen-presenting cells (APCs). These activated APCs then migrate preferentially to the marginal zone of the spleen, where they become part of an intricate and highly specific cluster of immune cells. The end result is the emergence of a population of antigen-specific T-regulatory lymphocytes that return to the eye and suppress DTH.     

乌鸡转移因子对小鼠T,B淋巴细胞及体重增长的影响

乌鸡脾转因子的提取及免疫活性的研究已有几年，并认为它可以提高细胞免疫活性，这已用鸡做了研究，而它对哺乳动物的免疫活性及生长发育有无影响？这一工作无人进行对照研究，为此，笔者用双重对照的方法（用人转移因子组和正常对照组〈生理盐水组〉进行了乌鸡转移因子对小鼠Ｔ、Ｂ淋巴细胞的影响及生长发育的影响）进行了研究。     

Impact of fresh or used litter on the posthatch immune system of commercial broilers

This study was carried out to investigate the effects of exposure of growing broiler chickens of commercial origin to used poultry litter on intestinal and systemic immune responses. The litter types evaluated were fresh wood shavings or used litter obtained from commercial poultry farms with or without a history of gangrenous dermatitis (GD). Immune parameters measured were serum nitric oxide (NO) levels, serum antibody titers against Eimeria or Clostridium perfringens, mitogen-induced spleen cell proliferation, and intestinal intraepithelial lymphocyte or splenic lymphocyte subpopulations. At 43 days posthatch, birds raised on used litter from a GD farm had higher serum NO levels and greater Eimeria or C. perfringens antibody levels compared with chickens raised on fresh litter or used, non-GD litter. Birds raised on non-GD and GD used litter had greater spleen cell mitogenic responses compared with chickens raised on fresh litter. Finally, spleen and intestinal lymphocyte subpopulations were increased or decreased depending on the litter type and the surface marker analyzed. Although it is likely that the presence of Eimeria oocysts and endemic viruses varies qualitatively and quantitatively between flocks and, by extension, varies between different used litter types, we believe that these data provide evidence that exposure of growing chicks to used poultry litter stimulates humoral and cell-mediated immune responses, presumably due to contact with contaminating enteric pathogens.     

感染鸡传染性贫血病毒（CIAV）雏鸡免疫器官抗体生成细胞的动态变化

对１日龄雏鸡感染鸡传染性贫血病毒（ＣＩＡＶ）后免疫器官法氏囊、脾脏和胸腺的ＩｇＧ、ＩｇＭ、ＩｇＡ抗体生成细胞数量的动态变化进行了检测。结果发现,感染雏鸡法氏囊、脾脏和胸腺的３种抗体生成细胞数量均程度不同地低于未感染对照雏鸡,其中法氏囊的ＩｇＧ、ＩｇＭ抗体生成细胞和ＩｇＡ抗体生成细胞分别在感染后７～３５ｄ和１４～３５ｄ明显减少;脾脏红髓、白髓和淋巴小结的ＩｇＧ抗体生成细胞分别于７～３５ｄ、１４～３５ｄ和１４～２１ｄ明显减少,ＩｇＭ抗体生成细胞分别在７～４２ｄ、２８～３５ｄ和１４ｄ时明显减少,ＩｇＡ抗体生成细胞仅在红髓中（７～２８ｄ）明显减少;胸腺髓质的ＩｇＧ、ＩｇＭ、ＩｇＡ抗体生成细胞分别在１４～２８ｄ、７～２１ｄ和２１ｄ时明显减少。结果表明,ＣＩＡＶ感染雏鸡免疫器官的体液免疫功能明显降低。     

田七总皂苷的制备及其免疫活性试验

用700mL/L乙醇回流提取、大孔吸附树脂分离纯化得到田七总皂苷(PNS),进行薄层色谱鉴别和含量测定。为评价PNS的免疫调节作用,用环磷酰胺(CTX)为免疫抑制剂建立小鼠免疫抑制模型,分别以4种剂量的PNS(5、25、50、75mg/kg)腹腔注射给药,测定胸腺和脾脏指数以及血清中IL-2、IFN-γ和TNF-α水平,并从组织形态上观察胸腺与脾脏的结构变化。结果表明,PNS能使小鼠胸腺和脾脏指数有所提高,但无统计学意义(P>0.05);25mg/kg剂量能显著升高血清中IFN-γ和TNF-α水平(P<0.05),50mg/kg剂量能显著升高血清中TNF-α水平(P<0.05);PNS处理组能不同程度地对抗模型所致小鼠胸腺和脾脏的萎缩,增加胸腺皮质厚度和细胞数量,增加脾小结大小和淋巴细胞数量,在高剂量组尤为明显。     

Mesenteric lymph node cells from neonates present a prominent IL-12 response to CpG oligodeoxynucleotide via an IL-15 feedback loop of amplification

ABSTRACT: At birth, the immune system is still in development making neonates more susceptible to infections. The recognition of microbial ligands is a key step in the initiation of immune responses. It can be mimicked to stimulate the immune system by the use of synthetic ligands recognising pattern recognition receptors. In human and mouse, it has been found that neonatal cytokine responses to toll-like receptor (TLR) ligands differ in many ways from those of adults but the relevant studies have been limited to cord blood and spleen cells. In this study, we compared the responses in neonate and adult sheep to CpG oligodeoxynucleotides (ODN), a TLR9 ligand, in both a mucosal and a systemic organ. We observed that in response to CpG-ODN more IL-12 was produced by neonatal than adult sheep cells from mesenteric lymph nodes (MLN) and spleen. This higher IL-12 response was limited to the first 20 days after birth for MLN cells but persisted for a longer period for spleen cells. The major IL-12-producing cells were identified as CD14+CD11b+. These cells were poor producers of IL-12 in response to direct stimulation with CpG-ODN and required the cooperation of other MLN cells. The difference in response to CpG-ODN between neonates and adults can be attributed to both a higher proportion of CD14+CD11b+ cells in neonate lambs and their higher capacity to produce IL-15. The IL-15 increases IL-12 production by an amplifying feedback loop involving CD40.     

Quantitative Analysis of Interleukin-6 Expression in Porcine Spleen Cells and Alveolar Macrophages using Real-Time PCR

Interleukin-6 (IL-6), a multifocal cytokine produced by lymphoid and non-lymphoid cells, regulates immune responses, acute-phase reactions against bacterial infections, and haematopoiesis. After cloning and sequencing of porcine IL-6, the expression pattern of porcine IL-6 mRNA was evaluated through real-time RT-PCR using porcine immune cells (spleen cells and alveolar macrophages) following stimulation with LPS. The sequence has been reported to GenBank with Accession no. AF 518322. The nucleotide sequence was different at the 89th and 205th positions in comparison with M80258, but only at the 205th with M86722. Comparison of porcine IL-6, Accession no. AF 518322, with IL-6 of human, canine, ovine, and mouse showed homologies of 78%, 81%, 82% and 73% in nucleotide sequence and 42%, 69%, 61% and 42% in amino acids. Expression of IL-6 mRNA was induced by stimulation with LPS. IL-6 mRNA expression in alveolar macrophages peaked at 2 h and decreased sharply to control levels at 4 h, whereas it peaked at 14 h and decreased at 24 h in spleen cells after stimulation with LPS (1 microg/ml). These results suggest that IL-6 mRNA expression in porcine immune cells is cell-type specific and the results of this study could be used as the basis for research on the porcine immune system.     

Functional expression of a bovine major histocompatibility complex class I gene in transgenic mice

Major histocompatibility complex (MHC) class I restricted cellular immune responses play an important role in immunity to intracellular pathogens. By binding antigenic peptides and presenting them to T cells, class I molecules impose significant selection on the targets of immune responses. Candidate vaccine antigens for cellular immune responses should therefore be analysed in the context of MHC class I antigen presentation. Transgenic mice expressing human MHC (HLA) genes provide a useful model for the identification of potential cytotoxic T lymphocyte (CTL) antigens. To facilitate the analysis of candidate CTL vaccines in cattle, we have produced transgenic mice expressing a common bovine MHC (BoLA) class I allele.The functional BoLA-A11 gene, carried on a 7 kb genomic DNA fragment, was used to make transgenic mice by pronuclear microinjection. Three transgenic mouse lines carrying the BoLA-A11 gene were established. Expression of the BoLA-A11 gene was found in RNA and the A11 product could be detected on the surface of spleen and blood cells. Functional analysis of the A11 transgene product, and its ability to act as an antigen presenting molecules in the mouse host will be discussed.     

T cell-mediated immunity to Cowdria ruminantium in mice: the protective role of Lyt-2+ T cells

The inability of athymic nude mice to make a drug-aided recovery from infection with either the Kümm or the Welgevonden stocks of Cowdria ruminantium and their inability to mount an immune response, suggest that immunity in heartwater is cell-mediated. The adoptive transfer of immunity with the spleen cells of mice immune to the Welgevonden stock is supportive evidence. Immune spleen cells depleted of Lyt-2+ T cells are unable to confer resistance to challenge to recipient mice, whereas the depletion of L3T4+ T cells had no effect on the protection conferred by immune spleen cells. This is conclusive evidence that immunity in heartwater is largely cell-mediated. Immune serum, C. ruminantium and complement incubated in the presence of mouse peritoneal macrophages, inhibits the infectivity of the heartwater agent, but not in the absence of macrophages. The decreased resistance to challenge of immune mice treated with gloxazone adds further support to the concept that in heartwater persistence of C. ruminantium in the host is associated with immunity.     

Changes in mouse circulating leukocyte numbers in C57BL/6 mice immunosuppressed with dexamethasone for Cryptosporidium parvum oocyst production

The Iowa strain of Cryptosporidium parvum will not propagate in immunocompetent mice, but will successfully infect genetically immunocompromised nude or SCID mice as well as immunocompetent mice which have been immunosuppressed with glucocorticoids. Using dexamethasone-tetracycline is one published method for immunosuppressing mice for the production of C. parvum oocysts. However, dexamethasone-induced immunosuppression is variable, because it is dependent on the total daily water consumption of each individual mouse. The changes in circulating leukocytes and other immune system associated organs before, during and after dexamethasone suppression were analyzed for comparison with a new single injection methylprednisolone acetate (MPA) suppression model. The dexamethasone-induced immunocompromised state was associated with a greater than 90% sustained drop in circulating T-lymphocytes, a greater than 700% increase in circulating mature segmented neutrophils and a severe depletion of circulating monocytes. The thymus and spleen decreased in size by over 80%. Oocyst shedding in suppressed mice started within 4 days of oocyst inoculation and persisted for 6 days post-dexamethasone treatment. Seven days after dexamethasone withdrawal, circulating neutrophils still were 549% higher than controls. Circulating CD3 and CD4 lymphocytes remained depressed by 85-90% while on dexamethasone and for 7 days after discontinuing dexamethasone. CD8 lymphocyte numbers initially decreased by 90%, but rose even while on dexamethasone and even with severe thymic involution. At day 7 post-dexamethasone treatment, the spleen was 119 mm(3), approximating the same size as controls. Fourteen days post-dexamethasone treatment, which was 8 days after oocyst shedding had ceased, the CD8 counts per 5000 events were only 1.6% below controls, while the CD3 and CD4 counts were still depressed by 66%. The thymus now was about one quarter smaller than the controls. The rise in circulating CD8 lymphocytes, when oocyst production stopped, suggests that CD8 positive lymphocytes may play a significant role in vivo in clearing the parasite. The overall pattern of immunosuppression was nearly identical to that observed with the methylprednisolone acetate immunosuppression model.     

In vitro antibody response to the tetrapeptide repeats of Plasmodium falciparum circumsporozoite protein by splenocytes from mice infected with P. yoelii yoelii

The antibody response to the recombinant protein, R32tet32, which contained the repetitive sequence (NANP)n of Plasmodium falciparum CSP was determined in C57BL/6 mice during the course of nonlethal infection with Plasmodium yoelii 17X. Marked suppression of the IgG antibody response to R32tet32 occurred when mice were immunized at peak parasitemia (on day 16). In vitro antibody responses of spleen cells from acutely infected mice to R32tet32 were similarly suppressed. Stimulation of normal spleen cells cultured for 5 days with 100 ng/ml of R32tet32 gave an optimal IgG antibody response, but spleen cells from infected mice obtained at peak parasitemia failed to respond to a broad range of antigen concentrations. Cocultivation studies employing enriched lymphocyte populations from infected and uninfected C57BL/6 mice indicated that both T and B cells from infected mice were defective in their response to R32tet32. The response to the repetitive region was restored by the addition of recombinant mouse interleukin-2 (IL-2) at a dose of 50 U/ml to cultures of spleen cells from infected mice.     

玉屏风复合多糖对环磷酰胺免疫抑制小鼠的恢复作用

为探讨玉屏风复合多糖对免疫功能损伤的调节和保护作用,以环磷酰胺（80mg／kg）诱导的免疫低下小鼠为实验动物模型,观察不同剂量玉屏风复合多糖（150,300,600mg／kg）对免疫器官结构和功能恢复的影响。结果发现,小鼠胸腺和脾脏体积显著增大,脾脏指数显著升高,脾淋巴细胞增殖、转化能力和NK细胞活性均显著增强（P〈0．05或P〈0。01）。脾脏结构中可见2种组织学变化,脾脏固有结构受损,淋巴组织明显增生。在对玉屏风复合多糖阳性对照组（未注射环磷酰胺）的观察中发现,小鼠脾脏结构清晰,出现结构性增生和肥大。结果证实,玉屏风复合多糙可明昂但讲务痴枷制小窜脾噼垂圭柏知功能确．海茸．对正常小赢睥睦功能扣．右明昂柏增强作用     

Analysis of Immune Response Induced by Recombinant Lactobacillus reuteri Expressing Cap Protein of Porcine Circovirus Type 2 in Mice

The purpose of this study was to construct a recombinant Lactobacillus reuteri (L. reuteri) expressing the cap protein of porcine circovirus type 2 (PCV2) and evaluate its effect on immune response in mice. The cap protein gene of PCV2b strain isolated and stored in the laboratory was amplified by PCR. A recombinant strain pPG-T7 g10-PPT-cap / L. reuteri expressing the cap protein was constructed using L. reuteri of pig origin as the host strain and explored the immune effect of BALB/c mice with recombinant bacteria orogastrically. Indirect ELISA was used to determine the level of antigen-specific IgG antibodies in the serum of mice after immunization, the levels of antigen-specific sIgA antibodies in stool, nasal wash, reproductive tract wash, and intestinal mucus, and the levels of various cytokines in mouse serum; MTT method was used to detect mouse spleen lymphocyte proliferation levels; flow cytometry (FCM) was used to detect the levels of CD4⁺ T cells and CD8⁺ T cells in mouse spleen lymphocytes; fluorescence quantitative PCR was used to detect the viral load of organs in challenged mice after immunization. The results showed that the serum levels of IgG antibodies in the mice of the oral immune recombinant strain group (OIG) were significantly higher than those in the control group (P<0.01); the levels of sIgA antibodies in the stool, nasal wash, reproductive tract wash, and intestinal mucus of the mice in OIG were significantly higher than those in the control group (P<0.01); Compared with the control group, the levels of cytokines in the serum of OIG were as follows: The levels of IFN-γ, IL-2, IL-4, IL-12 increased, the levels of IL-10 decreased, and the levels of IFN-α did not change significantly; Incubation of PCV2 and mouse spleen lymphocytes in vitro showed that the proliferation stimulating index of spleen lymphocytes in OIG was significantly higher than that of the control group (P<0.01); FCM results showed that CD4⁺ T cells and CD8⁺ T cells were higher than those of the control group; the results of fluorescent quantitative PCR showed that compared with the control group, the viral load in the OIG was significantly lower than that of the control group. In summary, the recombinant L. reuteri expressing the PCV2 cap protein were successfully constructed, and the constructed recombinant L. reuteri can stimulate mice to produce humoral and cellular immune responses after oragastrical immunization, and can exert a certain immune protection effect.     

表达猪圆环病毒2型cap蛋白的重组罗伊氏乳酸杆菌在小鼠诱导的免疫应答分析

旨在构建表达猪圆环病毒2型（PCV2）cap蛋白的重组罗伊氏乳酸杆菌（Lactobacillus reuteri,L.reuteri）,并评价其在小鼠体内诱导的免疫应答效果。利用PCR扩增实验室分离保存的PCV2b型毒株的cap蛋白基因,以猪源L.reuteri为宿主菌,构建表达cap蛋白的重组菌株pPG-T7 g10-PPT-cap/L.reuteri,通过口服免疫BALB/c小鼠。采用间接ELISA方法测定免疫后小鼠血清中抗原特异性IgG抗体水平,粪便、鼻腔洗液、生殖道洗液、肠黏液中抗原特异性sIgA抗体水平,小鼠血清中各细胞因子水平;MTT法检测小鼠脾淋巴细胞增殖水平;流式细胞技术检测小鼠脾淋巴细胞中CD4⁺T细胞、CD8⁺T细胞的水平;荧光定量PCR检测免疫后攻毒的小鼠体内器官的病毒载量。结果显示,口服免疫重组乳酸菌组小鼠血清IgG抗体水平显著高于对照组（P<0.01）;小鼠粪便、鼻腔洗液、生殖道洗液、肠黏液中sIgA抗体水平显著高于对照组（P<0.01）;小鼠血清中细胞因子水平和对照组相比,IFN-γ、IL-2、IL-4、IL-12水平升高,IL-10水平降低,IFN-α无显著变化;体外孵育PCV2和小鼠脾淋巴细胞结果表明,重组乳酸菌组小鼠脾淋巴细胞增殖刺激指数显著高于对照组（P<0.01）;流式细胞技术检测结果显示,口服免疫重组乳酸菌组小鼠脾细胞中CD4⁺T细胞、CD8⁺T细胞含量高于对照组;荧光定量PCR结果显示,相比于对照组,口服免疫重组乳酸菌组小鼠体内的病毒载量明显低于对照组。综上所述,本研究成功构建了表达PCV2 cap蛋白的重组罗伊氏乳酸杆菌,经口服途径免疫动物,构建的重组乳酸杆菌能够刺激小鼠产生体液免疫和细胞免疫应答,且具有一定的免疫保护效果。