The nonopsonic allogeneic cell phagocytosis of macrophages detected by flow cytometry and two photon fluorescence microscope

Phagocytosis, one of the apparent functions for macrophages, represents an early and crucial event in triggering host defenses against invading pathogens as well as allo- or xenogeneic rejection. Now, some methods have been used in detecting the opsonic phagocytosis of macrophages in xenogeneic settings. Efficient nonopsonic phagocytosis analysis method has not been established yet. In the present studies, allogeneic lymphocytes pre-labeled with 5-(and-6)-carboxyfluorescein diacetate succinimidyl ester (CFSE) or derived from green fluorescent protein transgeneic B6 mice (GFP-B6 mice) were co-incubated with primary murine peritoneal macrophages (PEMs) for 1–2 h or were injected into murine peritoneal cavity for 30 to 240 min. Assays by flow cytometry (FCM) and two photon laser scanning microscope (TPM) showed an efficient uptake of both allogeneic lymphocytes and xenogeneic chicken red blood cells. The continuing process of nonopsonic phagocytosis of allogeneic lymphocytes by PEMs was recorded by TPM. Furthermore, the phenotype differences of PEMs with or without phagocytosis of allogeneic cells were determined by three-color FCMs. Significantly upregulated expressions of CD11b, CD44, TLR2 and TLR4 on PEMs were observed as early as 6 h after phagocytosis of allogeneic cells. Our present data indicated that the FCM and TPM combined method is a practical approach to detect macrophage nonopsonic phagocytosis of allogeneic lymphocytes and to identify the phenotype alteration of macrophages after phagocytosis.     

Lipopolysaccharide-mediated enhancement of zymosan phagocytosis by RAW 264.7 macrophages is independent of opsonins,laminarin, mannan,and complement receptor 3

Background

Fungal and bacterial coinfections are common in surgical settings; however, little is known about the effects of polymicrobial interactions on the cellular mechanisms involved in innate immune recognition and phagocytosis.

Materials and methods

Zymosan particles, cell wall derivatives of the yeast Saccharomyces cerevisiae, are used to model fungal interactions with host immune cells since they display carbohydrates, including beta-glucan, that are characteristic of fungal pathogens. Using in vitro cell culture, RAW 264.7 macrophages were challenged with zymosan, and phagocytosis determined via light microscopy. The effects of different concentrations of lipopolysaccharide (LPS) on zymosan phagocytosis were assessed. In addition, the transfer of supernatant from LPS-treated cells to naïve cells, the effects of soluble carbohydrates laminarin, mannan, or galactomannan, and the impact of complement receptor 3 (CR3) inhibition on phagocytosis were also determined.

Results

LPS enhanced phagocytosis of zymosan in a dose-dependent manner. Transfer of supernatants from LPS-primed cells to naïve cells had no effect on phagocytosis. Laminarin inhibited zymosan phagocytosis in naïve cells but not in LPS-primed cells. Neither mannan, galactomannan, nor CR3 inhibition had a significant effect on ingestion of unopsonized zymosan in naïve or LPS-treated cells.

Conclusions

Zymosan recognition by naïve cells is inhibited by laminarin, but not mannan, galactomannan, or CR3 inhibition. LPS enhancement of phagocytosis is laminarin insensitive and not mediated by supernatant factors or zymosan engagement by the mannose or CR3 receptors. Our data suggest alternative mechanisms of zymosan recognition in the presence and absence of LPS.     

门静脉高压症脾亢脾巨噬细胞数量及其吞噬功能的观察

目的观察门静脉高压症(PH)脾亢脾脏巨噬细胞(MΦ)数量及其吞噬功能的变化,为进一步探讨PH脾功能亢进(脾亢)的发病机制提供依据。方法取20例PH脾亢患者的手术脾脏作为PH组,按脾脏肿大程度的分级标准,再分为中度肿大组(11例)和重度肿大组(9例)。6例正常脾外伤性破裂患者的手术脾脏作为对照组。称重后贴壁法分离培养脾脏MΦ,用细胞计数板在显微镜下计数MΦ的相对数(每g脾脏组织中的MΦ数)并计算MΦ绝对数(整个脾脏内MΦ的总数),鸡红细胞吞噬法计算MΦ的吞噬率和吞噬指数。结果对照组、PH中度和重度脾大组MΦ的相对数分别为(13．13±3．72)×10~5/g、(8．80±0．97)×10~5/g、(7．29±1．33)×10~5/g,PH组比对照组均显著减少(P＜0．01),但PH的两组之间差异无统计学意义(P＞0．05)；PH组脾脏的重量明显增加(P＜0．01),可达正常脾重的3．5～6．0倍(放血后)；各组MΦ的绝对数分别为(1．94±0．55)×10~8、(4．91±1．12)×10~8、(6．86±0．77)×10~8,PH组比对照组均显著增多(P＜0．01),且PH重度肿大组也显著多于中度肿大组(P＜0．01)；各组MΦ的吞噬率和吞噬指数分别为(6,33±0．58)%和(0．07±0．01)、(11．25±2．19)%和(0．14±0．03)、(13．00±2．38)%和(0．16±0．04),PH组比对照组均显著增强(P＜0．01),但PH的两组之间比较差异无统计学意义(P＞0．05)。结论PH脾亢时,脾脏MΦ的相对数虽然减少,但绝对数增多,MΦ的吞噬功能增强。MΦ在PH脾亢的形成中可能扮演着非常重要的角色。进一步支持了脾亢发病中的“脾内阻留学说”。     

梗阻性黄疸大鼠腹腔巨噬细胞凋亡和吞噬功能的改变

目的　探讨梗阻性黄疸大鼠腹腔巨噬细胞功能的改变。方法　将 66只成年Wistar大鼠随机分为正常 (A)组、假手术 (B)组和胆总管结扎 (CBDL ,C)组 ,每组术后又分为 1、3、7、10、14d五个时相点 ;应用流式细胞术检测两组腹腔巨噬细胞 (PMs)的凋亡、吞噬凋亡细胞及主要组织相容性复合物 (MHC)Ⅱ表达能力的变化。结果　C组大鼠术后各时相点PMs的凋亡指数 (AI)显著增高、吞噬能力和MHCⅡ (I A)类抗原表达能力下降 ,与A组和B组均有明显差异 (P <0 .0 1) ,且C组各时相点PMs的AI与吞噬能力和MHCⅡ类抗原表达能力改变有明显相关性。结论　CBDL大鼠PMs的AI明显增高 ,吞噬能力和MHCⅡ类抗原表达能力降低 ,可能导致梗阻性黄疸机体免疫机能的紊乱。     

The effect of macrophages posttrauma on T cell functions

Objective:To study molecular mechanism of suppressive effect of macrophages posttrauma on T cell functions.Methods:A murine closed trauma model was used,macrophages were harvested from the abdominal cavity and added into the culture system of T cells,which were separated from splenocytes in normal mice using nylon column.t cell functions and intracellular messenger molecules were determined.In addition,the effect of macrophages‘ removal from splenocytes of traumatized mice on T cell functions and intracellular messenger molecules was investigated.Results:Macrophages posttrauma in vitro could obviously suppress ConA stimulated normal t cell functions such as T lymphocyte transformation,interleukin 2(IL-2) production,Il-2 receptor α (IL-2Rα) expression,IL-2 mRNA and Il-2Rα mRNA levels,and elevate cAMP contents of activated normal t cells while decreasing cGMP contents,intracellular free calcium ([Ca^2 ]i)concentration and protein kinase C (PKC) activity.Removal of macrophages from splenocytes of traumatized mice could at certain degree reverse the suppression of T cell functions,decrease cAMP contents while increasing cGMP contents,[Ca^2 ]i concentration and PKC activity. Conclusions:Macrophages posttrauma may suppress T cell functions via altering messenger molecule levels in activated T cells.     

甲状旁腺细胞与睾丸Sertoli细胞共同移植产生免疫赦免的研究

目的　研究甲状旁腺细胞与睾丸细胞共同移植是否产生免疫赦免。方法　随机将SD大鼠分为以下四组 :1组为单纯甲状旁腺细胞移植组 ;2组为甲状旁腺细胞 1× 10 6个睾丸Sertoli细胞移植组 ;3组为甲状旁腺细胞 2× 10 6个睾丸Sertoli细胞移植组 ;4组为甲状旁腺细胞 4× 10 6个睾丸Sertoli细胞移植组。观察移植物的存活情况 ;体外检测移植物内细胞成分及淋巴细胞凋亡情况。结果　 1组平均存活期为 (17.2 2± 3.6 3)d ;2、3、4组存活时间延长 ,分别为 (19.4 4± 4 .6 4 )d、(32 .2 2± 6 .6 7)d和 (48.80± 3.33)d。 4组在 5 0d观察期内 ,多数鼠的血清钙及甲状旁腺激素 (PTH)值维持在正常范围内 (P <0 .0 1)。移植物内可见表达FasL的睾丸细胞、甲状旁腺细胞及凋亡的淋巴细胞。FasL的表达及凋亡指数随着移植的Sertoli细胞数增加而增加。结论　睾丸Sertoli细胞FasL的表达诱导了浸润的活化淋巴细胞凋亡 ,使共同移植的甲状旁腺细胞存活期延长。     

大鼠脊髓损伤后NgR在巨噬细胞中的表达对其吞噬作用及神经再生的影响

【摘要】 目的 体外研究大鼠脊髓损伤后表达Nogo蛋白受体（Nogo receptor, NgR）的巨噬细胞的吞噬作用及其对损伤神经元再生的作用。 方法 将髓鞘碱性蛋白（myelin basic protein, MBP）分别加入表达NgR的巨噬细胞、空载巨噬细胞及正常巨噬细胞中,应用Western blot比较这3组巨噬细胞的吞噬作用。将3组巨噬细胞加入损伤神经元中培养,采用MTT法及测定乳酸脱氢酶（lactate dehydrogenase, LDH）的含量,比较3组巨噬细胞对损伤神经元轴突再生的作用。 结果 Western blot结果显示表达NgR的巨噬细胞组吞噬作用与其他2组比较差异有统计学意义(P＜0.05)。MTT比色法及LDH含量测定结果显示表达NgR的巨噬细胞组吞噬作用与其他2组比较差异有统计学意义 (P＜0.05)。 结论 NgR的表达在体外可增强巨噬细胞的吞噬作用,表达NgR的巨噬细胞可促进神经损伤后的轴突再生。     

大鼠脊髓损伤后NgR在巨噬细胞中的表达对其吞噬作用及神经再生的影响

目的体外研究大鼠脊髓损伤后表达Nogo蛋白受体（Nogo receptor,NgR）的巨噬细胞的吞噬作用及其对损伤神经元再生的作用。方法将髓鞘碱性蛋白（myelin basic protein,MBP）分别加入表达NgR的巨噬细胞、空载巨噬细胞及正常巨噬细胞中,应用Western blot比较这3组巨噬细胞的吞噬作用。将3组巨噬细胞加入损伤神经元中培养,采用MTT法及测定乳酸脱氢酶（lactate dehydrogenase,LDH）的含量,比较3组巨噬细胞对损伤神经元轴突再生的作用。结果Western blot结果显示表达NgR的巨噬细胞组吞噬作用与其他2组比较差异有统计学意义（P〈0．05）。MTT比色法及LDH含量测定结果显示表达NgR的巨噬细胞组吞噬作用与其他2组比较差异有统计学意义（P〈0．05）。结论NgR的表达在体外可增强巨噬细胞的吞噬作用,表达NgR的巨噬细胞可促进神经损伤后的轴突再生。     

Mast cells in the seminal plasma of infertile men as detected by flow cytometry

Increased numbers of mast cells (MCs) in the testis have been associated with testicular dysfunction, where accumulation of MCs occurs. Furthermore, it has been reported that MCs might affect sperm function as it has been demonstrated that MC-derived tryptase in the seminal fluid might reduce sperm motility. Although MCs have been detected in rat epididymis, only little is known about the presence of MCs in human seminal plasma. Thus, we analysed MC numbers in the ejaculate of men during routine semen analysis of male patients suspected for infertility ( n  = 100). MCs were detected by c-kit (CD117) expression using flow cytometry. Thereby, we detected significant numbers of MCs in the ejaculate of most patients (559 ± 525 MCs ml⁻¹, mean ± SD). However, we could neither detect a correlation with respect to MCs and sperm count, motility or morphology nor to the seminal inflammatory markers like polymorphonuclear elastase. Nevertheless, a significant correlation of MCs to spermatozoa-bound IgA ( r  = 0.5; P  = 0.03; n  = 21) was observed. It is concluded that significant numbers of MCs can be detected in the human ejaculate without necessarily influencing sperm function. A potential role of MCs in seminal plasma as well as the association between MCs and IgA on spermatozoa remains to be elucidated.     

Pretransplantation minimal residual disease monitoring by multiparameter flow cytometry predicts outcomes of AML patients receiving allogeneic hematopoietic stem cell transplantation

Background and purposeIs minimal residual disease (MRD) monitoring by multiparameter flow cytometry (MFC) prognostic for acute myeloid leukemia (AML) patients before allogeneic hemopoietic stem cell transplantation (allo-HSCT)? And if so, what level of MRD eradication can be used to help guide the timing of HSCT? Can haplo-HSCT improve the prognosis of AML patients with MRD positive? To figure out these questions, we initiated this retrospective study.Methods96 AML patients were included retrospectively and divided into 5 groups, according to pre-transplantation MRD levels (from 5 × 10⁻² to <1 × 10⁻⁴), to analyze the overall survival (OS), disease-free survival (DFS) and cumulative incidence of relapse (CIR). Secondly, we compared the prognosis of MRD-negative (MRD^neg) and MRD-positive (MRD^pos) AML patients (cutoff value = 1 × 10⁻³) who underwent allo-HSCT, and further analyzed the prognosis of MRD^pos patients after received different transplantation modalities.ResultsIt is found that the 2-year OS and DFS of MRD negative group were better than the MRD positive group, and that the deeper the eradication of MRD before transplantation, the better the prognosis of patients. The CIR in patients received HLA-identical transplantation, was higher in the MRD^pos than in the MRD^neg. Haploid transplantation reduced the CIR disparity between MRD^pos and MRD^neg group. Subsequently, in AML patients who remain MRD positive before HSCT, we show that haplo-HSCT offered a better prognosis than HLA-identical transplantation (MSDT and MUDT).ConclusionIt is suggested that achieving MFC-MRD <10⁻³ (10⁻⁴ or even better) before allo-HSCT could reduce the relapse of AML and improve OS and DFS significantly, while haplo-HSCT may be preferred for patients not achieving MRD negativity.     

Analysis and sorting of prostate cancer cell types by flow cytometry.

BACKGROUND: Prostate tumor heterogeneity as manifested by differential expression of markers can be attributed to multiple types of cancer cells populating a tumor. Does the composition differ between primary tumor and metastasis? How can one isolate the different cancer cell types to study? What is the relationship among cancer cell types? METHODS: Flow cytometry keying on the prostate epithelial cell surface markers CD57 and CD44 was applied to analyze and sort single cells prepared from tumor tissue samples by collagenase digestion. In normal tissue, CD57 is found on luminal cells and CD44 on basal cells. RESULTS: CD57(+) and CD44(+) cells were sorted from various prostate tumor tissue specimens. The CD57(+) cancer cell type was found to predominate in primary tumors, while the CD44(+) cancer cell type was found to predominate in two visceral metastases. All tumors could be characterized by a ratio of CD57(+) and CD44(+) cancer cells. CONCLUSIONS: Two types of prostate cancer cells, CD57(+) and CD44(+), were identified. The finding that most primary tumors contain a predominantly CD57(+) cancer cell population agrees with the argument that cancer cells arise from the transformation of CD57(+) luminal cells. However, CD44(+) cancer cells are also present in some primary tumors; and in some metastases, they, and not CD57(+) cells, constitute a predominant population.     

流式细胞仪检测基底细胞癌的T细胞亚群

目的：研究基底细胞癌的细胞免疫功能。方法：用流式细胞仪技术检测了32例基底细胞癌患者外周血的T细胞亚群。结果：基底细胞癌患者CD4细胞百分率下降，CD8细胞百分率增高，与对照组相比差异均有显著性。结论：基底细胞癌患者存在细胞免疫功能缺陷。     

Relationship between sperm quality and chromatin condensation measured by sperm DNA fluorescence using flow cytometry

DNA flow cytometry of sperm from 100 randomly chosen men undergoing fertility investigation revealed a general association between reduced sperm quality, as judged by conventional parameters, and the appearance of sperm with lower degrees of chromatin condensation in the ejaculate as measured by DNA fluorescence intensity. Chromatin hypocondensation, as measured by increased fluorescence, was manifested to different degrees in different samples. In many cases of more extreme sperm pathology, such as oligoasthenoteratozoospermia (OAT), the whole population of spermatozoa appeared to be affected. Significant numbers of hypercondensed spermatozoa were present in both normozoospermic men and men with different degrees of disturbance in sperm quality. All of the different parameters of sperm quality could be correlated significantly with certain of the flow parameters, although not one in particular could be used to predict deviations from the normal flow profile. In several asthenoteratozoospermic men and a small proportion of men with OAT, the DNA profiles were normal, implying that in these cases the disturbance may not be so fundamental. The presence of leucocytes in the ejaculate was associated with a general increase in the preponderance of hypocondensed subpopulations of spermatozoa in men with OAT as well as in normozoospermic subjects, emphasizing the effect of inflammatory conditions in the reproductive tract on sperm quality.     

同种异体冻干小块骨的临床应用

目的探讨临床应用辐照冻干同种异体小块骨移植的成骨效果、并发症及影响因素。方法自1995年12月-2000年12月使用同种异体小块骨移植治疗患者778例，男590例，女188例；年龄7-76岁，平均37-2岁。其中骨肿瘤84例，骨折551例，骨关节融合143例。植入前用自体新鲜血液或温生理盐水充分浸泡，根据病种及骨缺损情况，将其植入骨缺损处或需融合的关节周围。术后引流2-3d，常规应用抗生素预防感染。植骨量为4-64g，平均24g。结果术后随访36～96个月，平均59．7个月。术后早期体温、引流量、肢体肿胀等一般情况与自体骨移植同类手术相比差异无显著性。722例(92．80％)在3～8个月达到骨性愈合，平均4．5个月。并发症中伤口渗出44例，占5．65％；伤口延期愈合29例，占3．72％；伤口大量积液2例，占0．26％；骨折不愈合43例，占5．52％；肿瘤复发4例，占0．51％；关节融合失败9例，占1．15％；再骨折6例，占0．77％。全组按Mankin及Komender标准评定，满意688例，占88．44％；不满意90例，占11．56％。结论冻干同种异体小块骨具有良好的组织相容性及成骨作用，是骨移植术中良好的植骨材料，植骨区的稳定性、局部血运情况及外科操作技术是影响骨愈合的重要因素，骨折不愈合、晚期感染及再骨折是其主要并发症。     

烧伤对小鼠腹腔巨噬细胞产生一氧化氮的影响

为探讨烧伤后巨噬细胞产生 NO 的情况及其可能机制,观察了烧伤早期不同时相点小鼠腹腔巨噬细胞(PMΦ)产生 NO 及诱导型 NO 合成酶(iNOS)活性的变化,以及单甲基精氨酸(NMMA)、放线菌素 D 及 genistein 等阻断剂对 NO 及 iNOS 活性的影响。结果显示烧伤早期 PMΦ就能产生过量的NO,且 iNOS 活性明显增强,NO 与 iNOS 两者的变化呈明显正相关。NMMA、放线菌素 D 及 genistein均能抑制 NO 的产生,并且放线菌素 D 及 genistein 还能使 iNOS 活性明显下降。提示:烧伤早期 PMΦ能产生大量的 NO,NO 是通过 iNOS 催化 L-精氨酸而产生的,而 iNOS 是经基因转录而合成的。蛋白酪氨酸激酶(PTK)信号传导系统参与了烧伤后 PMΦ iNOS 的合成从而影响 NO 的产生。     

烧伤对小鼠腹腔巨噬细胞产生一氧化氮的影响

为探讨烧伤后巨噬细胞产生ＮＯ的情况及其可能机制，观察了烧伤早期不同时相点小鼠腹腔巨噬细胞（ＰＭΦ）产生ＮＯ及诱导型ＮＯ合成酶（ｉＮＯＳ）活性的变化，以及单甲基精氨酸（ＮＭＭＡ）、放线菌素Ｄ及ｇｅｎｉｓｔｅｉｎ等阻断剂对ＮＯ及ｉＮＯＳ活性的影响。结果显示烧伤早期ＰＭΦ就能产生过量的ＮＯ，且ｉＮＯＳ活性明显增强，ＮＯ与ｉＮＯＳ两者的变化呈明显正相关。ＮＭＭＡ、放线菌素Ｄ及ｇｅｎｉｓｔｅｉｎ均能抑制ＮＯ的产生，并且放线菌素Ｄ及ｇｅｎｉｓｔｅｉｎ还能使ｉＮＯＳ活性明显下降。提示：烧伤早期ＰＭΦ能产生大量的ＮＯ，ＮＯ是通过ｉＮＯＳ催化Ｌ－精氨酸而产生的，而ｉＮＯＳ是经基因转录而合成的。蛋白酪氨酸激酶（ＰＴＫ）信号传导系统参与了烧伤后ＰＭΦｉＮＯＳ的合成从而影响ＮＯ的产生。     

DNA fragmentation assessment by flow cytometry and Sperm-Bos-Halomax (bright-field microscopy and fluorescence microscopy) in bull sperm

The aim of this study was to find the relationship between fertility (as 90-day non-return rates) and DNA fragmentation assessed by two techniques [sperm chromatin structure assay (SCSA) and Sperm-Bos-Halomax (SBH)]. Furthermore, other quality parameters were achieved (motility, morphological abnormalities, cytoplasmic droplets, viability, capacitation and acrosomal and mitochondrial status) and their correlations with fertility were analysed. Bulls were divided into three fertility groups: high [non-return rate (NRR) >or= 80], medium (80 < NRR >or= 70) and low (70 < NRR > 40). The results of this study indicate that there is a good correlation between fertility and different parameters of sperm quality (SBH and SCSA parameters, % of spermatozoa with head, neck and total abnormalities, and % of spermatozoa with proximal cytoplasmic droplets) and differences between fertility groups were observed in some of them (SBH and SCSA parameters and % of spermatozoa with head, neck and total abnormalities). In this sense, SBH parameters rendered good correlations with fertility (r = -0.42 using bright light microscope and r = -0.47 with fluorescence). Also, standard deviation of DNA fragmentation index (SD-DFI) and DFIh (cells with High DNA fragmentation index) showed good correlations with fertility (r = -0.41 and r = -0.29). No correlations were observed between SCSA and SBH parameters. A multiple regression shows that four parameters (% of proximal cytoplasmic droplets, % of intact acrosomes in total population, SD-DFI and percentage of fragmented DNA detected by bright light microscope) present a good predictive value of the fertility of sperm samples (r(2) = 0.34, p < 0.001).     

子宫内膜异位症患者腹腔液中巨噬细胞和不育的关系

巨噬细胞是腹腔液中主要的有核细胞,是对炎症刺激发生反应的第一线细胞。子宫内膜异位症患者腹腔液增多,腹腔液中巨噬细胞浓度增加,腹腔液和血清中由巨噬细胞分泌的细胞因子也明显增多。巨噬细胞吞噬精子能力亢进,抑制精子的活动度及其功能;还可介导炎症反应引起盆腔粘连,改变盆腔正常的解剖结构,影响输卵管功能,从而导致不育。巨噬细胞分泌的一些细胞因子致胚胎毒性增加,但另一些细胞因子却对囊胚的形成具有促进作用。本综述着重从巨噬细胞及其分泌的细胞因子两方面来叙述巨噬细胞、子宫内膜异位症和不育三者之间可能的关系。     

睾丸支持细胞与共刺激通路阻断剂对异体移植肾细胞的保护作用

目的　研究表达Fas配体 (FasL)的睾丸支持细胞与共刺激通路阻断剂细胞毒性T细胞相关抗原 4免疫球蛋白 (CTLA 4Ig)对异体移植肾细胞的保护作用 ,以提高移植肾的免疫耐受性。方法　酶消化法制备睾丸支持细胞及肾细胞。取 10 6个细胞混合移植于大鼠肾包膜下 ,实验大鼠分为 3组 :对照组 (10只 )、混合细胞移植组 (混合移植组 ,16只 )、联合CTLA 4Ig组 (CTLA组 ,10只 )。分别于术后 1、7、14、2 0d检测血清白细胞介素 2 (IL 2 )水平变化。术后 2 0d取出移植物 ,以亲和素 生物素 过氧化物酶复合物技术观察肾细胞存活状况 ;MD 2 0图像分析系统测定混合移植物灰度值 ;末端脱氧核糖核酸转移酶介导的X dUTP缺口末端标记 (TUNEL)法观察移植物中凋亡的细胞。　结果对照组无移植物存活 ;混合移植组 14只移植物存活 ,灰度值为 0 36 2± 0 0 17;CTLA组 10只移植物均存活 ,灰度值为 0 4 4 5± 0 0 2 1;混合移植组与CTLA组间灰度值差异有显著意义。术后CTLA组血清IL 2水平较混合移植组低 ,差异有显著意义 ;混合移植物中可见凋亡的淋巴细胞。　结论　异体肾细胞移植中 ,睾丸支持细胞与CTLA 4Ig对移植肾细胞具有协同保护作用。     

The use of multiparameter flow cytometry to assess tumor cell heterogeneity and grade prostate cancer

Multiparametric flow cytometry has permitted the quantitative assessment of tumor cell heterogeneity within a single tumor. This technique was applied to four tumor lines with biological activity similar to the variety observed in human prostate cancer. These tumors were derived from the Dunning R3327 transplantable rat prostatic adenocarcinoma and have permitted the development of an index to assess, quantitate, and discriminate variation in the aggressive nature of these tumors. The index is based upon measurements of the mean and the variance for perpendicular light scatter and forward light scatter. Perpendicular light scatter has been shown to correlate with nuclear shape, volume, and roundness as measured by digitized image analysis. Forward light scatter increases in quantity and range as the tumors become more undifferentiated. This technique is rapid, reproducible, and applicable to the quantitative measurement of cytometric parameters on large numbers of individual tumor cells.