巨细胞病毒感染对新生豚鼠胸腺的影响

新生豚鼠皮下接种豚鼠巨细胞病毒(GPCMV)后,导致动物胸腺急性感染。感染豚鼠胸腺在接种后第五天开始出现病毒,第十天达高峰。此外,感染动物胸腺的发育受到抑制,细胞总数和T淋巴细胞数随朐腺中病毒滴度的增高而进行性下降,至接种GPCMV后第十天最显著。由于病毒对T细胞的作用,细胞表面红细胞受体的丧失导致胸腺Null细胞百分比高于对照动物。     

体内外感染PRRSV处理下仔猪脾淋巴细胞增殖活性的变化

为了探讨体内或体外感染猪繁殖与呼吸综合征病毒(porcine reproductive and respiratory syndrome virus,PRRSV)的仔猪脾淋巴细胞在体外培养的增殖能力,本研究将PRRSV-GXA分离株经Marc-145细胞大量增殖培养,然后体内感染仔猪,接种病毒后分别于第11天、第14天和第21天从活体猪收获脾脏,分离脾淋巴细胞后用ConA和LPS于体外进行诱导增殖.研究结果显示第11天收获的T淋巴细胞增殖能力高于对照组,而B淋巴细胞增殖能力明显下降;第14天收获的T淋巴细胞和B淋巴细胞的增殖能力均极显著低于对照组;第21天收获的B淋巴细胞增殖能力略高于对照组,T淋巴细胞增殖能力略低于对照组.同时,用不同滴度的PRRSV体外感染PRRSV阴性的猪脾淋巴细胞,经ConA和LPS诱导增殖后,发现病毒滴度在103~106 TCID50/mL范围内,能明显提高脾淋巴细胞的体外增殖能力;而病毒滴度在100~102 TCID50/mL范围时,脾淋巴细胞增殖能力低于对照组.本研究结果说明PRRSV体内或体外感染对仔猪脾淋巴细胞增殖活性均有显著的影响,将为临床上PRRS的综合防治提供理论依据.     

新生豚鼠巨细胞病毒感染后唾液腺和脾脏的病理改变

豚鼠皮下接种巨细胞病毒(CMV)后,出现发育迟缓、唾液腺和脾脏肿大,唾液腺中出现高滴度的CMV。镜检唾液腺和脾脏发现有唾液腺导管细胞变性和导管细胞核内嗜酸性包涵体;脾脏中性粒细胞浸润、散在的灶状出血、大量网状内皮细胞增生、脾小结增大甚至急性窦卡他性炎症。     

BN大鼠和豚鼠对卵清白蛋白致主动过敏反应的免疫学特性比较

目的 比较BN大鼠和豚鼠对卵清白蛋白(OVA)致敏前后机体免疫学特性的变化.方法 BN大鼠和豚鼠分别用OVA(每只1 mg)隔日致敏(i.p.),共5次;于末次致敏第10天以OVA(每只2 mg)激发致敏(i.v.);分别设正常对照组和OVA致敏组.于激发致敏后1h处死动物,分离腹腔肥大细胞、脾脏和骨髓,并制备脾脏和骨髓淋巴细胞.以annexin-V作为标志检测肥大细胞活性,同时以Fluo-3/AM标记胞内钙离子,检测钙离子水平;以PHA和LPS作为有丝分裂原,分别检测脾脏和骨髓T、B淋巴细胞活性.结果 ①致敏BN大鼠和豚鼠脾脏及骨髓T、B淋巴细胞活性均升高,其中骨髓淋巴细胞活性BN大鼠显著高于豚鼠,脾脏淋巴细胞活性两种属间差异无显著性;②致敏后,腹腔肥大细胞活性两种属间差异无显著性,但BN大鼠致敏后是致敏前的6倍,豚鼠是3倍;③肥大细胞内钙离子水平两种属致敏后均升高,豚鼠致敏前后钙离子水平具有统计学意义.结论 OVA致敏后,BN大鼠骨髓淋巴细胞活性明显高于豚鼠,豚鼠肥大细胞内钙离子较BN大鼠升高明显,肥大细胞活性两者无明显差异.因此,在实验中可以根据两种属在过敏反应中的特点以及具体的实验要求选择动物模型.     

脾切断流术围术期患者外周血淋巴细胞的变化及免疫功能的研究

目的:本实验旨在探讨脾切除门奇静脉断流术后患者外周血T淋巴细胞、B淋巴细胞、NK细胞的变化,评估患者的抗肿瘤免疫能力是否有所影响,及机体的免疫系统有何变化.方法:选择择期行脾切除门奇静脉断流术的肝硬化患者20名,给予全凭静脉麻醉.分别于麻醉前(T0)、切皮前(T1)、脾切除即刻(T2)、脾切除后1h(T3)、手术完毕(T4)、手术后1d(T5)、手术后7d(T6)时抽取病人外周静脉血2mL,采用流式细胞仪测定CD3+、CD4+、CD8+、CD19+B细胞、CD3-56+(NK细胞)绝对数量.结果:与麻醉前相比,切皮前CD3+T细胞、CD3+CD4+T细胞、CD3+CD8+T细胞、CD19+B细胞、NK细胞均明显的降低,而在脾脏切除即刻各系细胞又明显恢复,基本与麻醉前水平相当.然而随着手术继续,在脾脏切除后1h,仅B细胞低于术前,一直持续到手术完毕,但是,此时B细胞与麻醉前比已没有统计学差异.手术完毕时T、B细胞和NK细胞再次降低,但仍明显高于切皮前水平.手术后1d时,CD4+T细胞与NK细胞仍然低于麻醉前,CD3T细胞、CD3+CD8+T细胞和B细胞已经恢复到麻醉前水平.术后7d时,CD3+T细胞、CD3+CD4+T细胞、CD3+CD8+T细胞及B细胞不仅得到恢复,而且还比麻醉前明显升高,但是NK细胞仍与麻醉前的水平相当.结论:异丙酚联合瑞芬太尼麻醉对门脉高压患者行脾切除门奇静脉断流术患者的T、B淋巴细胞和NK细胞有快速、短期的降低作用,术后7d人体淋巴细胞数量不仅得到恢复,并且反馈性地升高,提示脾脏切除手术能够有效提升患者的免疫细胞数量.     

连翘酯苷对小鼠脾脏淋巴细胞体外增殖与分泌功能的影响

目的分析连翘酯苷（FS）对小鼠脾脏T和B淋巴细胞增殖、分泌NO和TNF-α的影响,初步探讨其免疫调节作用机制。方法无菌操作分离小鼠脾脏,制备脾脏细胞并用含10%胎牛血清的RPMI 1640培养,在培养液中分别加入刺激剂刀豆蛋白（ConA）和脂多糖（LPS）以及不同浓度40、80、160μg/mL的FS共培养不同时间,采用MTT法检测T和B淋巴细胞的吸光度变化,ELISA和Griess法分别检测细胞分泌TNF-α和NO的水平。结果低浓度和中浓度FS对ConA诱导T淋巴细胞24 h和48 h后细胞增殖和存活率明显提高,诱导时间延长至72 h后FS明显抑制细胞转化;低浓度FS对LPS诱导脾脏B淋巴细胞24 h后细胞增殖和生存率显著提高;FS促进小鼠脾脏T和B淋巴细胞分泌NO;FS促进B淋巴细胞分泌TNF-α,中浓度FS促进T淋巴细胞分泌TNF-α而高浓度反而抑制其分泌。此外,FS对环磷酰胺（CY）处理小鼠的脾脏淋巴细胞体外增殖有明显影响,对细胞NO分泌影响不显著。结论结果提示FS可能通过影响小淋巴细胞增殖和细胞因子分泌而调节免疫细胞功能。     

肿瘤宿主的免疫状态 Ⅲ.带Ehrlich实体瘤小鼠脾脏细胞在体外对肿瘤细胞的细胞静止效应的动态观察

应用5-~(125)碘-2’-脱氧尿嘧啶核苷(~(125)IUdR)参入抑制试验,证明带Ehrlich实体瘤的C57BL 小鼠脾脏细胞可在体外对同一移植肿瘤细胞的DNA 合成产生显著的抑制。随着肿瘤的增长,带瘤小鼠的脾脏进行性增大,脾细胞对肿瘤细胞的抑制效应也随之增强。这种对肿瘤细胞DNA合成的抑制作用是非特异的。肿瘤切除后,抑制效应也随之下降,到术后21天,细胞静止效应已不能测得。本文还应用~(125)IUdR技术同时测定了带瘤小鼠脾脏的T 杀伤细胞,脾脏淋巴细胞对致分裂原Con A 与PWM 的转化反应的动态变化,实验表明带瘤小鼠脾脏T、B 淋巴细胞的功能是受抑制的。组织病理学观察提示,带瘤动物脾脏的网织细胞和巨噬细胞的增多可能与细胞静止效应的增强有关。     

与豚鼠红细胞形成自然花环的狗淋巴细胞类别的探讨

本文研究了豚鼠E花环形成狗淋巴细胞(E-RFC)的类别。狗淋巴细胞于37℃孵育后,可提高E花环形成率。E-RFC与总淋巴细胞的SIg阳性率相近。E-EAC(豚鼠红细胞—抗体和补体包被的绵羊红细胞)混合花环实验表明一部分狗淋巴细胞同时具有E和补体二种受体,总淋巴细胞与补体受体淋巴细胞的E花环阳性率无显著差异。这些结果说明E受体不仅表现于一部分T细胞上,而且见于某些B细胞上。因此,不宜用E花环试验作为检测狗T细胞的方法。     

用Epstein-Barr病毒转化法获得分泌肾综合征出血热病毒抗体的B淋巴细胞系

人B淋巴细胞膜上带有Epstein-Barr(EB)病毒受体,用EB病毒转化人B淋巴细胞是获得人源性单克隆抗体的重要方法之一。最近David等报道,先用EB病毒转化感染巨细胞病毒(CMV)患者的脾脏B淋巴细胞,再将转化了的B淋巴细胞和人骨髓瘤细胞WL-L_2-727融合,获得迄今为止分泌抗体滴度最高的人—人杂交瘤312A和914,能稳定分泌抗体达12     

海蛇乙醇浸出物对小鼠免疫系统的影响

目的 探讨海蛇乙醇浸出物（AEBFSS）在小鼠体内、体外对免疫系统的影响。方法 体外试验：采用AEBFSS与小鼠脾细胞共孵育72h,测定T、B淋巴细胞转化功能;体内试验：采用每天灌胃给予小鼠1次AEBFSS,共6天。测定脾脏中T淋巴细胞转化功能和溶血素抗体生成水平。结果 在体外条件下低浓度AEBFSS促进T、B淋巴细胞增殖,高浓度时则作用相反;在体内条件下低浓度AEBFSS能增加B细胞生成溶血素抗体,对T细胞增殖无明显影响,高浓度则抑制T、B细胞功能。结论 AEBFSS对小鼠免疫系统有一定的双向调节作用。     

Characterization of a monoclonal antibody to guinea pig T cells that inhibits rosette formation of the cells with rabbit erythrocytes: similarity of the antigen to E-receptor on human T cells

A monoclonal antibody, GPT-1, was prepared by fusion of the splenic cells of mice immunized with guinea pig thymocytes with a mouse myeloma cell line. GPT-1 completely inhibited spontaneous rosette formation of T cells with papain-treated rabbit erythrocytes. GPT-1 reacted with 90% of thymocytes, 70% of peripheral blood lymphocytes, and 45% of splenic lymphocytes, but not with B cells. These results indicate that GPT-1 has pan-T reactivity. The antibody specifically bound to a single polypeptide chain with a molecular size of 50-65 kD. The surface density of the antigen was higher on thymocytes than on peripheral T cells, suggesting that the antigen is a certain differentiation antigen on T cells. Phytohemagglutinin-activated T cells expressed more antigen molecules than resting T cells. In addition, GPT-1 suppressed the proliferation of T cells induced by the mitogen, indicating that GPT-1 recognizes a T cell-specific surface antigen which is associated with T cell activation. Based on these results, it was concluded that GPT-1 reacts with a guinea pig T cell surface antigen which is similar to the E-receptor protein on human T cells (CD2 molecule).     

Further evidence for heterogeneity of Fc gamma-receptors on guinea pig splenic B and T lymphocytes: analysis using monoclonal antibodies to two distinct types of Fc gamma-receptor

In our previous paper, we reported that guinea pig splenic lymphocytes expressed two distinct Fc-receptors for homologous IgG (Fc gamma Rs), one monospecific for IgG2 (Fc gamma 2R) and the other bispecific for IgG1 and IgG2 (Fc gamma 1/gamma 2R), when analyzed by EA-rosette assay. These Fc gamma Rs on the cells were further studied by using two monoclonal antibodies toward the Fc gamma Rs on guinea pig peritoneal macrophages (anti-Fc gamma 1/gamma 2R and anti-Fc gamma 2R antibody). The anti-Fc gamma 1/gamma 2R antibody completely inhibited the rosette formation of splenic lymphocytes with IgG1-sensitized sheep erythrocytes [EA(IgG1)]. On the other hand, EA(IgG2)-rosette formation was inhibited partially by anti-Fc gamma 2R but not by anti-Fc gamma 1/gamma 2R antibody. Complete inhibition of the EA (IgG2)-rosette formation was achieved by simultaneous additions of both anti-Fc gamma 2R and anti-Fc gamma 1/gamma 2R antibodies. The binding of IgG2 antibody complexed with ovalbumin to the cells was partially inhibited by either anti-Fc gamma R antibody, and complete inhibition occurred in the presence of both the antibodies, indicating that two types of Fc gamma R, Fc gamma 1/gamma 2R, and Fc gamma 2R, are expressed on the cells. The determination of these Fc gamma Rs on B and T lymphocytes by two-color flow cytometry showed that about 52% of B lymphocytes expressed Fc gamma 1/gamma 2R alone and 32% of the cells expressed both the Fc gamma Rs. On the other hand, about 12% of T lymphocytes was found to express Fc gamma 2R alone and the cells expressing Fc gamma 1/gamma 2R were in the minority (3.8%). T lymphocytes expressing both the Fc gamma Rs were not detected. These results show that guinea pig B lymphocytes bear two types of Fc gamma Rs and are heterogeneous with regard to their Fc gamma Rs and that T lymphocytes express Fc gamma 2R mainly.     

Mature B cells are required for acute splenic infection, but not for establishment of latency, by murine gammaherpesvirus 68.

Murine gammaherpesvirus 68 (gamma HV-68; also referred to as MHV-68) is a gammaherpesvirus which infects murid rodents. Previous studies showed that CD8 T cells are important for controlling gamma HV-68 replication during the first 2 weeks of infection and suggested a role for B cells in latent or persistent gamma HV-68 infection. To further define the importance of B cells and CD8 T cells during acute and chronic gamma HV-68 infection, we examined splenic infection in mice with null mutations in the transmembrane domain of the mu-heavy-chain constant region (MuMT; B-cell and antibody deficient) or in the beta2-microglobulin gene (beta2 -/-; CD8 deficient). Immunocompetent mice infected intraperitoneally with gamma HV-68 demonstrated peak splenic titers 9 to 10 days postinfection, cleared infectious virus 15 to 20 days postinfection, and harbored low levels of latent virus at 6 weeks postinfection. Beta2-/- mice showed peak splenic gamma HV-68 titers similar to those of normal mice but were unable to clear infectious virus completely from the spleen, demonstrating persistent infectious virus 6 weeks postinfection. These data indicate that CD8 T cells are important for clearing infectious gamma HV-68 from the spleen. Infected MuMT mice did not demonstrate detectable infectious gamma HV-68 in the spleen at any time after infection, indicating that mature B lymphocytes are necessary for acute splenic infection by gamma HV-68. Despite the lack of measurable acute infection, MuMT spleen cells harbored latent virus 6 weeks postinfection at a level about 100-fold higher than that in normal mice. These data demonstrate establishment of latency by a herpesvirus in an organ in the absence of acute viral replication in that organ. In addition, they demonstrate that gamma HV-68 can establish latency in a cell type other than mature B lymphocytes.     

Roles of macrophages in measles virus infection of genetically modified mice

Knowledge of the mechanisms of virus dissemination in acute measles is cursory, but cells of the monocyte/macrophage (MM) lineage appear to be early targets. We characterized the dissemination of the Edmonston B vaccine strain of measles virus (MV-Ed) in peripheral blood mononuclear cells (PBMC) of two mouse strains expressing the human MV-Ed receptor CD46 with human-like tissue specificity and efficiency. In one strain the alpha/beta interferon receptor is defective, allowing for efficient MV-Ed systemic spread. In both mouse strains the PBMC most efficiently infected were F4/80-positive MMs, regardless of the inoculation route used. Circulating B lymphocytes and CD4-positive T lymphocytes were infected at lower levels, but no infected CD8-positive T lymphocytes were detected. To elucidate the roles of MMs in infection, we depleted these cells by clodronate liposome treatment in vivo. MV-Ed infection of splenic MM-depleted mice caused strong activation and infection of splenic dendritic cells (DC), followed by enhanced virus replication in the spleen. Similarly, depletion of lung macrophages resulted in strong activation and infection of lung DC. Thus, in MV infections of genetically modified mice, blood monocytes and tissue macrophages provide functions beneficial for both the virus and the host: they support virus replication early after infection, but they also contribute to protecting other immune cells from infection. Human MM may have similar roles in acute measles.     

Thymic and splenic changes following injection of living Brucella and BCG in the Guinea pig

Summary The thymic and splenic reactions, following injections of BCG and living Brucella M. in the guinea pigs, were studied. Both microorganisms injected intravenously produced thymic involution, maximal after BCG inoculation, followed by regeneration that was complete by day 10 in Brucella M.-treated guinea pigs, and by day 15, in BCG injected guinea pigs. Increase of mitotic index was more accentuated and more persistent after Brucella M. than after BCG treatment in the thymus. After a short involution period the spleen of injected animals increased in size in both groups. The splenic enlargement was dramatic and occurred at an accelerated rate in animals given BCG. It appeared to be the result of a conspicuous involvement of the red pulp by multiple granulomas. In Brucella M. treated guinea pigs the splenic enlargement was less obvious, but the splenic white pulp was more abundant in BCG-treated guinea pigs. Granulomas were observed only in the periarterial sheaths of the white pulp.These observations provide evidence for the hypothesis that injections of both BCG and Brucella M. provoke a proliferation of B and T lymphocytes, a migration of T lymphocytes from the thymus to the T-dependent area of the spleen which seems, perhaps, more marked after injection of Brucella M., and a strong granulomatous histiocytic reaction which is more conspicuous in animals given BCG.     

Isotype Specificity and Heterogeneity of Fcγ-Receptors on Guinea Pig Splenic B and T Lymphocytes

The isotype specificity of Fc-receptors for IgG (Fcγ-Rs) on normal guinea pig splenic B and T cells was determined by a rosette assay using sheep erythrocytes sensitized with either homologous IgG1 or IgG2 anti-sheep erythrocyte antibody [EA(IgG1) or EA(IgG2)]. Approximately 70% of the lymphocytes in the highly purified B-cell fraction could form rosettes with EA(IgG2), and 55% of the cells with EA(IgG1). Inhibition experiments with soluble complexes of IgG1 or IgG2 antibody with ovalbumin demonstrated that approximately 20% of the EA(IgG2) rosette-forming B cells bore the Fcγ-R monospecific for IgG2, whereas 80% of the cells had two distinct Fcγ-Rs simultaneously; one monospecific for IgG2 and the other bispecific for IgG1 and IgG2. The existence of a B cell bearing the Fcγ-R monospecific for IgG1 was not definitively demonstrated in the B-cell fraction. In the T cell-enriched fraction, approximately 40% of the cells could form rosettes with EA(IgG2). The EA(IgG1)rosette-forming cells, however, comprised only 6% of the total cells, indicating that most of the EA(IgG2) rosette-forming T cells bear essentially the Fcγ-R monspecific for IgG2 alone. The results obtained revealed that guinea pig splenic lymphocytes bear two distinct Fcγ-Rs, which are not equally distributed on the B- and T-cell populations and also on their respective subsets.     

Development of antigenic heterogeneity in the splenic meshwork of severe combined immunodeficient (SCID) mice after reconstitution with T and B lymphocytes

Recently, we produced monoclonal antibodies reacting specifically with the reticular meshwork (RM) of lymphoid tissues, and demonstrated that, in the splenic white pulp of normal mouse, the antigenic heterogeneity of RM was associated with the segregation of the T and B lymphocytes. In the present study, we attempted to visualize further the interaction between splenic RM and T and B lymphocytes transferred into severe combined immunodeficient (SCID) mice. The splenic white pulp of naive SCID mice, containing a few T and B cells, showed little tendency for T-B segregation and antigenic diversity of RM. Transfer of spleen or bone marrow cells from normal mice resulted in complete recovery of lymphocyte populations, showing not only a clear segregation of T and B lymphocytes but also a remarkable antigenic diversity of RM. The same results were obtained following the transfer of spleen or bone marrow cells from the nude mouse. Next, we transferred purified T lymphocytes to one group of SCID mice and B cells to another. In mice given T cells, a few B cells were observed in the white puop; T lymphocytes lodged not only in the inner periarterial lymphatic sheath (PALS) but also in the outer PALS and follicles. In the animals to which B cells were transferred, T cells were few and the homing of B cells occurred only into their proper compartments, such as the outer PALS, follicles and marginal zone, but not in the inner PALS. Thus, B cells can home into their proper compartments of the splenic white pulp independently of T lymphocytes.     

T and B lymphocytes of irradiated, tumor-bearing mice treated with an immunostimulator, pyran.

The T and B splenic lymphocyte populations of BALB/c mice were determined in Madison lung 109 carcinoma-bearing animals. Concurrently, some groups of tumored mice were exposed to 500 rads of whole body irradiation and treated with one dose of a nonspecific immunostimulating agent, pyran. By indirect immunofluorescence, it was found that the percentage of splenic T lymphocytes were significantly depressed in the tumored irradiated mice. Mitogenic studies revealed that the PHA-sensitive T lymphocytes were more depressed in the tumored irradiated mice than in the corresponding Con A-sensitive T lymphocytes. Pyran was relatively effective in reconstituting the T cell compartment of these splenic T lymphocytes.The B cell compartment of the splenic lymphocytes of the tumored irradiated mice was found to be extremely radiosensitive. Utilizing a specific anti-F(ab')₂ serum, no B lymphocytes were detected throughout the duration of testing. Blastogenic studies using LPS as the mitogenic probe revealed that the incorporation of [³H]TdR of the tumored irradiated mice was just slightly higher than background values. Pyran proved to be relatively ineffective in reconstituting the splenic B cells of the tumored irradiated mice.     

The antigen-induced selective recruitment of specific T lymphocytes to the lung

The purpose of the present studies was to investigate the mechanisms by which specific T lymphocytes accumulate in the lung. After the intratracheal (IT) inoculation of influenza virus into guinea pigs, the detection of specific T lymphocytes in the lung coincided with the development of immunity in both hilar nodes and systemic lymphoid tissue. Animals immunized in the footpads with virus failed to develop immune responses in the lung unless rechallenged IT with immunogen. In adoptive transfer experiments, IT inoculation of influenza into nonimmune guinea pigs, followed immediately by the i.v. injection of a mixture of 3H-thymidine-labeled syngeneic T lymphocytes specific for influenza virus and 14C-thymidine-labeled syngeneic T lymphocytes specific for an irrelevant antigen resulted in the selective accumulation of the virus-specific T lymphocytes in the lung. Taken together, these studies indicate that the selective recruitment by antigen of circulating immune cells is one of the mechanisms by which specific T cells accumulate in the lung.