S-O_2-1菌苗对小鼠免疫细胞的作用

本文探讨 S-O_2-1菌苗对小鼠淋巴细胞和单核吞噬系统的影响。正常小鼠接种菌苗后,脾脏T细胞百分率剧烈下降、胸腺显著缩小。脾脏细胞的抗 SRBC抗体应答和 GVH反应性明显降低。经 ALS处理的免疫低下小鼠接种菌苗后,抗 SRBC应答和GVH反应性的抑制更加明显。这些结果表明该菌苗具有抑制T细胞功能活性。另一方面,接种菌苗的小鼠,无论对无机碳粒还是对鸡红细胞,都显著加快了廓清速率,其腹腔细胞形成 EA花环能力也成倍上升,提示菌苗对单核吞噬系统有强烈刺激作用。以上资料证明,S-O_2-1菌苗的免疫刺激效应不在T细胞方面,而是针对单核吞噬系统。     

紫色杆菌LPS对小鼠脾细胞免疫活性的抑制作用

细菌内毒素或脂多糖(LPS)是机体内强烈的免疫调节剂。天然低毒性紫色杆菌LPS 体内处理小鼠,能促进脾细胞的分化、增殖,但降低脾细胞对 Con A 和同种细菌 LPS 的反应性,抑制混合淋巴细胞反应(MLR)和自然杀伤(NK)细胞的活性。LPS 处理供体小鼠还抑制其脾细胞在 F_1 小鼠内诱导的移植物抗宿主反应。应用混合培养方法,在 Con A和 LPS 诱导的淋巴细胞转化反应中分别检测到非特异性抑制细胞活性,但在MLR和NK 活性测定中未发现抑制细胞的存在。上述结果说明 LPS体内抑制T、B 淋巴细胞功能和 NK细胞活性,而这种抑制作用除由抑制细胞介导外,还存在其它尚未明瞭的机理。     

S-O_2-1菌苗的免疫效应和抗肿瘤活性

本文报告我们自己分离的革兰氏阴性杆菌(S-O_2-1)菌苗对小鼠的免疫效应和抗肿瘤活性的研究。实验证明该菌苗主要作用于单核巨噬细胞系统,还能激发免疫淋巴细胞释放淋巴因子,对移植艾氏腹水癌的小鼠有较高的保护力,并能抑制癌细胞在小鼠腹腔内增殖,表明该菌苗有一定程度的佐剂性能。     

S-O_2-1菌苗诱生干扰素的研究 Ⅱ小鼠脾脏细胞体外诱生

本文表明S-O_2-1菌苗具有较强的诱导小鼠脾细胞产生干扰素的能力。将事先用菌苗免疫的C57BL/6小鼠脾细胞体外培养时,即使不加菌苗也能产生较高滴度的干扰素活性,该活性在免疫后4天制备的脾细胞培养中开始出现,免疫后7天则达高峰(7.47±0.31 log_2U/ml),以后逐渐下降。若培养时添加菌苗,干扰素高峰出现稍晚些,于免疫后10天达高峰,但滴度明显上升(9.10±0.26 log_2U/ml)。另外,C57BL/6正常小鼠的脾细胞(5×10~6细胞/ml)添加菌苗(0.1亿菌/ml)体外培养12小时后可开始测出干扰活性,培养48小时后活性达高峰(8.23±0.30log_2U/ml)。该干扰素具有小鼠Ⅱ-型干扰素相同的特性。我们在实验中还发现,将S-O_2-1菌苗与刀豆素A(Con A)合用诱生的干扰素滴度明显高于两者单独使用时诱生滴度的累加值。     

S-O_2-1细菌核糖体制剂对小鼠淋巴细胞的作用

<正> 我们实验室曾报道了 S-O_2-1细菌核糖体制剂是低毒、高效的免疫增强剂,能激活小鼠的单核巨噬细胞系统,增强小鼠腹腔粘附细胞对S_(180)肿瘤细胞的体外细胞毒作用,还能促进小鼠NK细胞活性,并能在体外诱生Ⅰ型干扰素和抑瘤因子。但关于该核糖体对淋巴细胞的作用还不清楚,本文观察S-O_2-1菌核糖体制剂对淋巴细胞的作用,从中探讨其免疫调节与抑瘤作用机理。     

S-O_2-1菌苗对S_(180)肿瘤细胞DNA合成的抑制作用

用小鼠 S_(180)肿瘤细胞作为靶细胞,以~3H-TdR 的掺入抑制百分率作为抗肿瘤活性的指标,在体外培养条件下,S-O_2-1 菌苗能刺激正常小鼠及带瘤小鼠的免疫活性细胞,释放对 S_(180)肿瘤细胞 DNA 合成有明显抑制作用的可溶性物质。实验提示,该类免疫因子的产生能增强免疫细胞对肿瘤细胞的细胞毒效应。同时发现 S-O_2-1菌苗本身对 S_(180)肿瘤细胞还具有直接杀伤作用。     

肺癌DC融合细胞诱导的抗肿瘤免疫应答

目的 研究肺癌DC融合细胞FLD－A11体内外诱导免疫应答的能力,方法 用MTT法测定FLD－A11细胞诱导淋巴细胞增殖的能力;ELISA法检测其诱导淋巴细胞分泌IL－2的水平;LDH释放法测定FLD－A11细胞免疫后小鼠脾淋巴细胞的特异性CTL活性。结果 FLD－A11有有效刺激淋巴细胞的增殖反应（SC：RC＝1：50时,SI＝2.38）,诱导淋巴细胞分泌IL－2。FLD－A11细胞免疫后,小鼠的胸腺和脾脏重量均高于对照组（P＜0.05）,脾淋巴细胞对Lewis肺癌细胞的杀伤活性显著高于对照组。结论 FLD－A11细胞具有诱导初次抗肿瘤免疫应答的能力,不仅能在体外诱导淋巴细胞的增殖和IL－2的分泌,而且体内免疫接种也能引起小鼠免疫器官的增生反应,产生针对Lewis肺癌的特异性CTL杀伤作用。     

铜绿假单胞菌外毒素A的抗原性及其与辅助细胞的关系

目的　研究铜绿假单胞菌外毒素A(PE)的某些抗原性及其与辅助细胞的关系。方法用MTT法观察PE在有无辅助细胞参与条件下对鼠脾淋巴细胞的刺激作用 ,并用鼠细胞毒性T细胞株CTLL进行证实。结果　PE在 0 0 1～ 10 0ng/ml浓度范围能刺激鼠脾淋巴细胞增殖 ,PE的丝裂原作用无需粘附细胞的辅助 ,但粘附细胞及其粘附细胞的PE刺激上清能协同PE对鼠脾淋巴细胞的刺激作用。PE在丝裂原作用范围内能刺激CTLL细胞生长 ,并与IL 2有协同作用。PE在体内能使小鼠脾淋巴细胞自发淋转率增高 ,对ConA刺激的增殖反应增强 ,但对淋巴细胞自发分泌IL 2和ConA诱导IL 2的产生水平无影响。结论　PE在无辅助细胞参与条件下也能刺激淋巴细胞增殖     

肝,脾iRNA的细胞免疫活性研究

本文应用了白细胞粘附抑制试验和IL_2活性检测试验分别探讨了不同脏器来源的抗大肠杆菌iRNA在肌体内诱导的细胞免疫活性的异同,正常小鼠分别经肝、脾iRNA致敏后,取腹腔渗出细胞(PEC)测其粘附抑制情况和取脾测其产生IL_2的活性。结果表明:脾-iRNA能特异地诱导正常小鼠PEC对其相应抗原出现显著的粘附抑制,肝—iRNA无此活性;肝—iRNA和牌—iRNA都能特异地诱导正常小鼠脾细胞在其相应抗原刺激下释放IL_2,二者活性无显著差异。本文提示,肝—iRNA和脾—iRNA都具有在动物体内特异地诱导细胞免疫的的活性,但二者间存在着差异。     

脾细胞在CTL克隆增殖中的作用

本文证明二次MLC上清或重组IL2均不足以维持细胞毒 T细胞(CTL)克隆的正常增殖,而同种异体、同种第三者、甚至同基因的脾细胞均可增强rIL2诱导的CTL克隆增殖应答,亦可增强 CTL由 CD3∈链抗体(145-2C11)刺激的 IL-2非依赖性增殖。脾细胞的这种辅佐效应似与可溶性因子或脾粘附细胞无关。用 rIL2 预处理的脾细胞可诱导 CTL克隆的显著增殖,这种增殖可被抗IL2 受体抗体所抑制,提示脾细胞可能将其“膜结合”rIL2递给CTL而更有效地诱导增殖反应。FD18.5(抗 LFA-1),KH1.4(抗 Ly-6)及 HK2.1(抗 Thy-1)可显著抑制脾细胞对 145-2C11 激活 CTL增殖的辅佐效应,提示脾细胞对 CTL克隆 IL2非依赖性增殖的辅佐效应与一些细胞表面分子有关。     

Histamine synthesis by mouse T lymphocytes through induced histidine decarboxylase.

When spleen cells of C57BL/6 mice or mast cell-deficient W/Wv mice were cultured, their histidine decarboxylase (HDC) activity increased with increases in the histamine concentration in the cells and the medium. Addition of concanavalin A (Con A) or Escherichia coli lipopolysaccharide (LPS) enhanced the increase. The removal of adherent cells reduced both the control HDC activity and the response to the mitogens. Purified T lymphocytes responded to Con A but not to LPS. Neither Con A nor LPS had any effect on B lymphocytes. Treatment of T cells with anti-Thy-1.2 and complement completely abrogated the induction of HDC. Histamine synthesis dependent on Con A by T cells was stimulated by the addition of conditioned medium from peritoneal adherent cells activated with LPS. The addition of recombinant interleukin-1 (rIL-1) or peritoneal adherent cells fixed with paraformaldehyde significantly enhanced HDC induction dependent on Con A in T cells. These results suggest that histamine is synthesized by T lymphocytes through HDC and that the reaction was enhanced by a soluble factor(s) released from macrophages.     

乳腺癌带瘤小鼠免疫功能的动态变化

用多项免疫学指标动态检测了 615系小鼠于接种乳腺癌 Ca759后免疫功能的变化。结果表明,随着肿瘤的进行性生长,外周血淋巴细胞及ANAE 阳性细胞的百分率、脾脏淋巴细胞对ConA 的刺激反应、抗体形成细胞的功能及中性粒细胞的吞噬功能均呈进行性下降;脾脏Ea-RFC百分率及腹腔巨噬细胞的吞噬功能呈先升后降趋势;脾脏EAC-RFC百分率及血清溶菌酶水平至晚期也下降。本文对相应的免疫学指标的变化趋势进行了比较,对Ca759带瘤小鼠免疫功能的变化规律进行了分析和讨论。     

Concanavalin A-induced resistance to Listeria monocytogenes and activation of macrophages: defect in mice treated with 89Sr

⁸⁹Sr-treated mice injected with concanavalin A (Con A) 24 h prior to infection with Listeria monocytogenes (LM) could not enhance the clearance of LM from the spleen. Adoptive transfer of normal syngeneic spleen cells together with Con A rendered these animals more resistant. Spleen cells of ⁸⁹Sr-treated or age-matched control mice were stimulated with Con A for 24 h, and supernatant fluids were assessed for macrophage-activating factor (MAF), i.e. the ability to activate resident peritoneal macrophages to kill LM intracellularly in vitro. A defective MAF production by spleen cells was observed in ⁸⁹Sr-treated, 2 week-old, and athymic nude mice. Also, treatment of spleen cells with anti-Thy-1.2 antiserum plus complement inhibited MAF production. Synergism between spleen cells from ⁸⁹Sr-treated and nude mice did not occur. The cells required for MAF production were relatively resistant to gamma irradiation. Nylon wool filtration did not modify the ability of spleen cells to make MAF. ⁸⁹Sr-treated mice possess macrophages responsive to MAF derived from normal spleen cells. The data suggest that the failure of ⁸⁹Sr-treated mice to develop an anti-LM response observed in this system could be due to a defective capacity to produce protective humoral factors and/or cells in response to Con A.     

In vivo normobaric oxygen exposure depresses spleen cell in vitro Con A response. Effects of 2-mercaptoethanol and peritoneal cells.

Normobaric O2 exposure decreased spleen cell (SC) response to T cell mitogen Con A. 3H-TdR incorporation of SC from O2 exposed mice (O2SC) compared to those of control mice (Air SC) decreased significantly after 72 and 87 h O2 exposure. The dose response kinetics to Con A were identical in O2SC or Air SC. Increasing SC number did not restore the response to Con A and the depressed hyperoxic effect was not related to suppressor cells in the spleen of O2 exposed mice. Response of O2SC to Con A was restored by the thiol compound 2-mercaptoethanol (2-ME), and the degree of restoration by 2-ME, was inversely proportional to the depressed response. Addition of intact peritoneal cells (PC) induced restoration within the same range as 2-ME. Restoration of the mitogenic response by 2-ME involved antioxidant properties and suggested that macrophages were functionally injured by O2 exposure. In cases where mitogen response was highly depressed, restoration was only partial; in these conditions in vivo O2 injury probably involved both macrophages and splenic T cells. The mechanisms of O2 toxicity have been discussed in terms of free radical generation under hyperoxic conditions.     

Serotonin regulation of T-cell subpopulations and of macrophage accessory function.

The role of serotonin as an immune modulator was investigated by measuring the functional competence of T cells from control mice versus from mice whose intracellular stores of serotonin had been depleted by pretreatment with p-chlorophenylalanine (PCPA). While the proportions of splenic CD4+ and CD8+ T cells isolated from control and PCPA-treated mice were similar, the level of expression of the alpha-chain interleukin-2 receptor (IL-2R) was reduced on splenic CD4+ cells but not on CD8+ cells. Culture with the T-cell mitogen concanavalin A (Con A) failed to induce expression of the IL-2R on either CD4+ or CD8+ cells of PCPA-treated mice, although IL-2R was induced on control cells. The proliferative response to Con A by these spleen cells from PCPA-treated mice was also reduced compared to that by control spleen cells. Both expression of IL-2R and proliferation in response to Con A by spleen cells from serotonin-depleted mice were increased or completely restored by supplementation of the cultures with serotonin. Studies to identify the mechanisms for the reduction in T-cell activation when serotonin levels were reduced implicated a defect in the capacity of macrophages from PCPA-treated mice to provide accessory help for T-cell activation. Splenic macrophages from control mice were able to restore the blastogenic capability of lymphocytes from PCPA-treated mice, although macrophages from PCPA-treated mice were unable to support normal lymphocyte blastogenesis unless the cultures were supplemented with serotonin. These results show the requirement of autologous serotonin for optimal T-cell activation and suggest the importance of serotonin in macrophage accessory function for T-cell activation.     

Blastogenic response of Toxoplasma-infected mouse spleen cells to T- and B-cell mitogens.

In order to differentially test the function of lymphocytes in Toxoplasma gondii-infected mice, the in vitro blastogenic response of spleen cell cultures to non-specific mitogens was studied. Phytohaemagglutinin (PHA) and concanavalin A (Con A) stimulation were used as tests of thymus-dependent lymphocyte (T cell) function and bacterial endotoxin lipopolysaccharide (LPS) was used as a probe of bursal equivalent lymphocyte (B cell) function. For the first 3 weeks following T. gondii infection, the uptake of tritiated thymidine ([3H]TdR) by spleen cells cultured with all three mitogens was markedly reduced in comparison to the uptake in spleen cells from uninfected control mice. Thereafter, the response to LPS returned to normal while stimulation by the T-cell mitogens (PHA and Con A) remained depressed. It is postulated that T. gondii infection either: (1) diluted out T cells in the spleen with unreactive cells; (2) modified T cells in such a way that they were less responsive to mitogens; (3) depleted the peripheral lymphoid tissues of T cells; (4) induced non-specific suppressor cells, which inhibited the T-cell function assays; or (5) activated macrophages which depressed T-cell function non-specifically.     

Participation of Interleukins in Synergistic Effect of Bu-WSA on Concanavalin A-Induced DNA Synthesis in Mouse Splenic Lymphocytes

Butanol-extracted water-soluble adjuvant (Bu-WSA) obtained from Bacterionema matruchotii was mitogenic to murine splenic B lymphocytes, but not T lymphocytes. When murine splenic cells were cultured in the presence of Bu-WSA and concanavalin A (Con A) together, [3H]thymidine uptake of the culture cells synergically increased. The mechanism of the synergy of Con A and Bu-WSA and the participation of interleukin (IL) 1 and 2 in the synergy were studied. The proliferation cells in the synergy were Lyt-1+23- lymphocytes. Ia-positive accessory cells were required for the response. When separated cell populations and Marbrook-type culture vessels were used, a mixed cell population of T lymphocytes and B lymphocytes or macrophages (M phi) produced some active factor(s) after co-stimulation by Con A and Bu-WSA, and the factors enhanced DNA synthesis of another Con A-activated T lymphocyte population. Supernatants obtained from the spleen cell cultures or the mixed cell cultures with T lymphocytes and M phi in the presence of Con A and Bu-WSA contained greater amounts of IL-1 and IL-2 than those from cultures containing Con A or Bu-WSA alone. An addition of exogenous IL-1 or IL-2 to spleen cell cultures with Con A resulted in a proliferative response like that obtained through co-stimulation by Con A and Bu-WSA. These results suggest that the synergistic effect of Con A and Bu-WSA on the proliferative response in murine splenic cells is sustained by the enhancement of production of these T-lymphocyte growth factors.     

Further observations on the immunological induction of DNA synthesis in mouse peritoneal macrophages. Role of products of activated lymphocytes.

Subcutaneous injection of ovalbumin (OA) into mice immunized with OA in Freund's complete adjuvant was followed by an increase in the numbers of peritoneal macrophages synthesizing DNA, determined by autoradiography. The effect was immunologically specific. The increase was followed by an increase in the numbers of peritoneal macrophages; the numbers of peritoneal lymphocytes also increased. Injection of OA into immunized or normal mice was followed by a blood monocytosis. Increased DNA synthesis, determined by liquid scintillation counting, occurred in spleen or lymph node cells from immunized mice, cultured with OA. Diluted supernatants from such cultures, injected intravenously into normal mice, caused increases in the numbers of DNA-synthesizing peritoneal macrophages. Similarly, supernatants from concanavalin A stimulated spleen cells, freed of Con A, also caused an increase in DNA-synthesizing macrophages.     

Antiapoptotic and immunomodulatory effects of chlorophyllin

Chlorophyllin (CHL) was earlier shown to reduce the level of intracellular ROS and apoptosis induced by ionizing radiation and 2,2'-azobis(2-propionimidinedihydrochloride) (AAPH). In the present studies, the effect of CHL on radiation-induced immunosuppression and modulation of immune responses in mice was examined. Chlorophyllin inhibited the in vitro lymphocyte proliferation induced by concanavalin A (Con A) in a dose dependent manner at doses>or=50 microM. At lower doses (10 microM) CHL significantly inhibited activation induced cell death (AICD) in Con A stimulated spleen cells. Spleen cells obtained from CHL treated mice showed an inhibition of response to Con A depending on dose of CHL and the time after its administration. Spleen cells obtained from CHL treated mice (24 h) showed lower inhibition of response to Con A following in vitro (5 Gy) as well as whole body irradiation (2 Gy). The expression of antiapoptotic genes bcl-2 and bcl-xL was up-regulated in these cells. Chlorophyllin treatment of mice led to splenomegaly and increase in the number of peritoneal exudate cells (PEC). The numbers of T cells, B cells and macrophages in the spleen were also increased. Increased phagocytic activity was seen in PEC obtained from CHL treated mice. Most importantly, CHL administration to mice immunized with sheep red blood cells (SRBC) augmented both humoral and cell-mediated immune responses.