TKI治疗晚期非小细胞肺癌患者免疫功能的变化及意义

目的:通过观察晚期非小细胞肺癌患者TKI治疗前后外周血IgG、IgM、IgA、C3、C4、C-反应蛋白及CD3+、CD4+、CD8+、CD4+/CD8+细胞的表达变化,探讨TKI治疗对晚期非小细胞肺癌患者免疫功能的影响及意义。方法:检测TKI组30例非小细胞肺癌患者TKI治疗前、治疗一个月后外周血IgG、IgM、IgA、C3、C4、C-反应蛋白及CD3+、CD4+、CD8+、CD4+/CD8+细胞表达水平,分析表达变化及与疗效的关系。30例非小细胞肺癌患者作为对照组。结果:治疗前,TKI组与对照组IgG、IgM、IgA、C3、C4、C-反应蛋白水平基本正常,但CD4+细胞数量减低、CD4+/CD8+比值较低、CD8+细胞数量增高,两组相比IgG、IgM、IgA、C3、C4、C-反应蛋白、CD3+、CD4+、CD8+、CD4+/CD8+差异均无统计学意义(P0.05);TKI治疗一个月后,TKI组与对照组IgG、IgM、IgA、C3、C4、C-反应蛋白水平无明显变化,而CD4+细胞数量增多、CD4+/CD8+较前增高,CD8+细胞数量较前减低,两组相比CD3+、IgG、IgM、IgA、C3、C4、C-反应蛋白差异无统计学意义(P0.05),而CD4+、CD4+/CD8+、CD8+差异有统计学意义(P0.01)。结论:TKI治疗后,晚期非小细胞肺癌患者细胞免疫功能得到改善,体现在CD4+、CD8+细胞数量的变化上,且TKI治疗的疗效可通过比较外周血CD4+、CD4+/CD8+、CD8+细胞表达变化体现。     

Regulatory function for murine intraepithelial lymphocytes. Two subsets of CD3+, T cell receptor-1+ intraepithelial lymphocyte T cells abrogate oral tolerance

The murine intraepithelial lymphocyte (IEL) population is enriched in T cells that express the gamma delta-TCR, however, the biologic function served by these T cells remains obscure. IEL are considered to be major effector cells in mucosal immunity, and we have investigated whether IEL subsets could reverse orally induced systemic unresponsiveness (oral tolerance; OT) and support secondary type responses when adoptively transferred to mice orally tolerized with SRBC. When purified CD3+ IEL from mice orally primed with SRBC were transferred to adoptive hosts and challenged with SRBC, splenic IgM, IgG1, IgG2b, and IgA anti-SRBC plaque-forming cell responses were observed. However, CD3+ IEL from HRBC orally primed mice did not abrogate SRBC induced OT. Further, HRBC-primed CD3+, IEL converted HRBC-specific OT but not SRBC-specific OT. CD3+ IEL could be separated into four subsets based on expression of CD4 and CD8. CD3+, CD4-, 8+ T cells were the major subset (74.5%), with smaller numbers of CD4- and CD8- (double negatives, DN) (7.8%), CD4+, 8- (7.6%) and CD4+, CD8+ (double positives) (10.1%) T cells. Interestingly, both the CD3+, CD8+, and the CD3+, DN IEL subsets abrogated OT, resulting in significant IgM, IgG1, IgG2b, and IgA anti-SRBC plaque-forming cell responses when adoptively transferred to mice with OT. However, neither CD3+, CD4+, CD8-, nor double positive T cells affected OT when studied in this system. The CD3+, CD8+ IEL subset could be further separated into Thy-1+ (16.6%) and Thy-1- (83.4%) cells; adoptive transfer of Thy-1- cells abrogated oral tolerance whereas the Thy-1+ subset was without effect. When the expression of TCR on IEL with this biologic function was determined by use of monoclonal anti-alpha beta TCR (H57.597), TCR2-, CD3+ IEL possessed immunoregulatory function whereas the alpha beta-TCR+ (TCR2+) fraction did not abrogate OT. Immunoprecipitation of membrane fractions obtained from purified CD3+, CD4-, CD8+, Thy-1- IEL with polyclonal anti-delta peptide (Tyr-Ala-Asn-Ser-Phe-Asn-Asn-Glu-Lys-Leu) antibody revealed bands of 45 and 35 kDa, corresponding to the delta- and gamma-chains, respectively. These results suggest that gamma delta-TCR+ IEL possess a regulatory function, namely the restoration of immune responses in a state of oral tolerance. Further, both CD3+, CD4-, CD8+, Thy-1-, and CD3+, DN IEL T cells exhibit this effector contrasuppressor function.     

Characterization of the immunophenotype and the metastatic properties of a murine T-lymphoma cell line. Unexpected expression of cytoplasmatic CD4

We report the first characterization of a mouse T-lymphoma cell line that surprisingly expresses cytoplasmatic (cy) yCD4. Phenotypically, LBC cells are CD5+, CD8+, CD16+, CD24+, CD25+, CD2-/dim, CD3-/dim, TCRbeta-/dim, TCRgammadelta, CD154 , CD40-, and CD45R. Coexpress cyTCRbeta, cyCD3, cyCD4, and yet lack surface CD4 expression. Transplantation of LBC cells into mice resulted in an aggressive T-lymphoblastic lymphoma that infiltrated lymph nodes, thymus, spleen, liver, ovary, and uterus but not peripheral blood or bone marrow. LBC cells display a modal chromosome number of 39 and a near-diploid karyotype. Based on the characterization data, we demonstrated that the LBC cell line was derived from an early T-cell lymphocyte precursor. We propose that the malignant cell transformation of LBC cells could coincide with the transition stage from late double-negative, DN3 (CD4- CD8 CD44-/low, CD25+) or DN4 (CD4-low, CD8-/low, CD44-, CD25-) to double-positive (DP: CD4+CD8+) stage of T-cell development. LBC cells provide a T-lymphoblastic lymphoma model derived from a malignant early T-lymphocyte that can be potentially useful as a model to study both cellular regulation and differentiation of T-cells. In addition, LBC tumor provides a short latency neoplasm to study cellular regulation and to perform preclinical trials of lymphoma-relatel clisorders.     

变应性鼻炎患者肠道微生态变化及其与T淋巴细胞亚群的关系

探讨变应性鼻炎患者肠道微生态变化及其与T淋巴细胞亚群的关系。

回顾性分析2019年4月至2020年4月医院收治的64例变应性鼻炎患者的资料, 记为A组; 另回顾性分析同期在该院体检的58例健康者的资料, 记为B组。对比A组和B组研究对象肠道菌群多样性和丰富度的变化, 对比A组和B组研究对象血清T淋巴细胞亚群水平, 分析变应性鼻炎患者肠道菌群相对丰度与血清T淋巴细胞亚群水平的相关性。

A组研究对象肠道菌群Chao1指数和Shannon指数均低于B组(P < 0.05), A组肠道菌群门水平拟杆菌门、变形菌门、厚壁菌门的相对丰度均高于B组(P < 0.05), 放线菌门的相对丰度低于B组(P < 0.05), 属水平毛螺菌属、链球菌属的相对丰度均高于B组(P < 0.05), 肠球菌属、双歧杆菌属、棒状杆菌属的相对丰度均低于B组(P < 0.05)。主成分分析显示2组菌群样品均能明显分开, 横纵坐标的贡献率分别为48.9%和16.7%, 两坐标轴总共解释了不同细菌群落差异的65.6%, 2组群落结构差异明显(P < 0.05)。A组CD3⁺比例、CD4⁺比例和CD4⁺/CD8⁺比值高于B组(P < 0.05), CD8⁺比例低于B组(P < 0.05);采用Pearson相关性检验, 变应性鼻炎患者肠道菌群门水平拟杆菌门、变形菌门和厚壁菌门的相对丰度分别与CD3⁺、CD4⁺比例和CD4⁺/CD8⁺比值呈正相关(P < 0.05), 分别与CD8⁺比例呈负相关(P < 0.05), 放线菌门的相对丰度分别与CD3⁺、CD4⁺比例和CD4⁺/CD8⁺比值呈负相关(P < 0.05), 与CD8⁺比例呈正相关(P < 0.05);属水平毛螺菌属和链球菌属的相对丰度分别与CD3⁺、CD4⁺比例和CD4⁺/CD8⁺比值呈正相关(P < 0.05), 分别与CD8⁺比例呈负相关(P < 0.05), 肠球菌属、双歧杆菌属和棒状杆菌属的相对丰度分别与CD3⁺、CD4⁺比例和CD4⁺/CD8⁺比值呈负相关(P < 0.05), 分别与CD8⁺比例呈正相关(P < 0.05)。

变应性鼻炎患者肠道菌群多样性降低, 门、属水平菌群比例均发生改变, 菌群结构差异明显, 血清CD3⁺、CD4⁺水平和CD4⁺/CD8⁺比值升高, CD8⁺水平降低, 变应性鼻炎患者肠道菌群相对丰度与血清T淋巴细胞亚群水平相关。

     

Characterization of interleukin 2-expanded human peripheral blood lymphocytes: not only NKH-1+-NK cells but also NKH-1+ and NKH-1- T cells are LAK effectors

In vitro culture of human peripheral blood mononuclear cells (PBMC) with interleukin 2 (IL-2) results in the expansion of lymphocytes including lymphokine-activated killer (LAK) cells. Using flow cytometry, studies were undertaken to determine the phenotype and LAK activity of each subset of lymphocytes expanded in vitro as a result of incubation for 2 weeks with 2500 U/ml of recombinant IL-2. Such expanded PBMC, when examined by two-color staining with various combinations of anti-CD3, 4, 8, 16, and NKH-1 monoclonal antibodies, consisted of the following six subgroups of cells: (1) CD3+4+8-, (2) CD3+4-8+, (3) CD3+4-8-, (4) CD3-16+NKH-1+, (5) CD3-16-NKH-1+, and (6) CD3-16-NKH-1-. Of the six subgroups, all five subgroups that could be tested, i.e., CD3+ T cells (CD3+4+8-, CD3+4-8+, CD3+4-8-), CD16+ natural killer (NK) cells (CD3-16+NKH-1+), and CD3-16-NKH-1- non-T non-NK cells, possessed LAK activity. Both NKH-1- as well as NKH-1+ T and non-T cells possessed LAK activity.     

Predominant CD4 T-lymphocyte tropism of human herpesvirus 6-related virus.

Human herpesvirus 6 (HHV-6)-related virus was isolated from CD4+ CD8- and CD3+ CD4+ mature T lymphocytes but could not be isolated from CD4- CD8+, CD4- CD8-, and CD3- T cells in the peripheral blood of exanthem subitum patients. HHV-6-related virus predominantly infected CD4+ CD8+, CD4+ CD8-, and CD3+ CD4+ cells with mature phenotypes and rarely infected CD4- CD8+ cells from cord blood mononuclear cells, which suggested predominant CD4 mature T-lymphocyte tropism of HHV-6-related virus.     

Immunobiology of the reproductive tract in a female baboon

The aim of this study was to assess the suitability of various antihuman antibodies directed against immunocomponent cells to identify components involved in cellular and humoral immune responses in the immune organs of a female baboon, and to use these reagents to analyze the immunobiology of its reproductive tract. A female baboon of reproductive age was euthanized in the luteal phase of the menstrual cycle, and samples of spleen, intestines, tonsil, lymph nodes, Fallopian tube, uterus, cervix, and vagina were removed. Tissues were either fixed in 10% unbuffered formaldehyde, Bouin's fluid, or 95% ethanol containing 5% glacial acetic acid, and embedded in paraffin, or frozen unfixed. Frozen sections were then fixed in 100% acetone. Subsequently, tissue sections were reacted with the following antihuman antibodies directed against CD3, CD45RA, CD45RO, CD4, CD8, CD20, CD68, HLA-DR, CD57, CD103, CD15, and TIA-1: IgA, IgG, IgM, J-chain, secretory component, and neutrophil elastase, using routine immunohistology techniques. Human tissues (spleen, small intestine, lymph node, and tonsil) were used as positive controls. All antihuman antibodies crossreacted with baboon tissues, except neutrophil elastase, CD15, CD45RO, CD57, and CD1A. The distribution of immune cells in the reproductive tract of the female baboon was comparable to that in the human and offers the potential for this primate to be used as a model for the study of human reproductive immunology.     

Phenotypic and functional analysis of murine CD3+,CD4-,CD8- TCR-gamma delta-expressing peripheral T cells

Murine CD3+,CD4-,CD8- peripheral T cells, which express various forms of the TCR-gamma delta on their cell surface, have been characterized in terms of their cell-surface phenotype, proliferative and lytic potential, and lymphokine-producing capabilities. Three-color flow cytofluorometric analysis demonstrated that freshly isolated CD3+,CD4-, CD8- TCR-gamma delta lymph node cells were predominantly Thy-1+,CD5dull,IL-2R-,HSA-,B220-, and approximately 70% Ly-6C+ and 70% Pgp-1+. After CD3+,CD4-,CD8-splenocytes were expanded for 7 days in vitro with anti-CD3-epsilon mAb (145-2C11) and IL-2, the majority of the TCR-gamma delta cells expressed B220 and IL-2R, and 10 to 20% were CD8+. In comparison to CD8+ TCR-alpha beta T cells, the population of CD8+ TCR-gamma delta-bearing T cells exhibited reduced levels of CD8, and about 70% of the CD8+ TCR-gamma delta cells did not express Lyt-3 on the cell surface. Functional studies demonstrated that splenic TCR-gamma delta cells proliferated when stimulated with mAb directed against CD3-epsilon, Thy-1, and Ly-6C, but not when incubated with an anti-TCR V beta 8 mAb, consistent with the lack of TCR-alpha beta expression. In addition, activated CD3+,CD4-,CD8- peripheral murine TCR-gamma delta cells were capable of lysing syngeneic FcR-bearing targets in the presence of anti-CD3-epsilon mAb and the NK-sensitive cell line, YAC-1, in the absence of anti-CD3-epsilon mAb. Finally, activated CD3+, CD4-,CD8-,TCR-gamma delta+ splenocytes were also capable of producing IL-2, IL-3, IFN-gamma, and TNF when stimulated in vitro with anti-CD3-epsilon mAb.     

化疗在中晚期非小细胞肺癌患者中对淋巴细胞亚群影响分析

目的:探讨化疗在中晚期非小细胞肺癌患者中对淋巴细胞亚群的影响。方法:随机抽取本院收治的60例中晚期非小细胞肺癌患者编为实验组进行化疗,另选取同期体检的50例健康志愿者作为对照组。随访12月-15月,采用流式细胞仪技术分别对两组外周血淋巴细胞亚群进行检测计数。结果:两组间相比,实验组患者的CD3+、CD4+、NK细胞的数量以及CD4+/CD8+比值均低于对照组(P<0.05),而CD8+细胞的比例却高于对照组。化疗后CD3+、CD4+、CD4+/CD8+、NK均较化疗前升高(P<0.05),但CD8+不变(P>0.05)。结论:应用化疗治疗中晚期非小细胞肺癌,可明显改善患者的免疫功能。     

Extrathymic T cell maturation. Phenotypic analysis of T cell subsets in nude mice as a function of age.

T cell maturation in an extrathymic environment has been studied using as a model the congenitally athymic nude mouse. Phenotypic analyses as a function of age were conducted on lymphocytes obtained from the spleens and lymph nodes of nude mice through use of mAb recognizing T cell surface markers and multiparameter flow cytometry. The data show that nude mice accumulate increasing numbers of lymphocytes bearing Thy-1, CD3, CD4, and CD8 with age characterized by a progression from heterogeneous dim to more homogeneous bright expression. In contrast, the expression of heat-stable Ag (HSA), a marker of immature thymocytes, decreases with age. By analogy to intrathymic maturation, spleens and lymph nodes in nude mice contain T cells defined as immature, transitional, and mature based on the expression of these markers. Although the proportion of CD4+ and CD8+ T cells associated with bright CD3 expression increases with age, at no age are significant numbers of CD4+8+ cells observed, in contrast to intrathymic T cell maturation. In addition to the frequently observed inversion in the ratio of CD4 to CD8, the CD8 T cell subpopulation in older nude mice contains mainly mature cells (CD8+, CD3+, HSA-) whereas only 50% of CD4+ T cells express the mature (CD4+, CD3+, HSA-) phenotype. At any age, the spectrum of phenotypes observed indicates that lymph nodes contain more mature T cells than spleen, suggesting a role for environmental Ag in driving extrathymic maturation, a process occurring most efficiently among CD8+ T cells. Because extrathymic maturation mirrors some but not all aspects of the intrathymic pathway, we propose that the nude mouse may be a useful model for further dissecting those interactions crucial to establishing the T cell repertoire in euthymic individuals as well as elucidating the contribution of extrathymically derived T cells to the peripheral immune system.     

Peripheral murine CD3+, CD4-, CD8- T lymphocytes express novel T cell receptor gamma delta structures

A mAb directed against the CD3 molecule was used to identify a subset of CD3+, CD4-, CD8- T cells previously undefined in the peripheral lymphoid organs of the mouse. Biochemical analysis of CD3+, CD4-, CD8- splenocytes revealed that the vast majority of these cells express one of at least two distinct CD3-associated TCR gamma delta heterodimeric structures, but no detectable TCR alpha beta. One disulfide-linked heterodimer (77 kDa) is composed of two chains of 45 to 46 and 32 kDa. The latter chain was immunoprecipitated with an anti-TCR C gamma 1/C gamma 2 antiserum and was not glycosylated. An antiserum produced against a peptide corresponding to the C-terminal region of the predicted C gamma 4 gene product immunoprecipitated additional heterodimers (80 to 90 kDa). One heterodimer, composed of disulfide-linked 41- to 45-kDa protein (including a V gamma/C gamma 4 component), is expressed on a T cell hybridoma, DN-1.21, which was derived from fused splenic CD3+, CD4-, CD8- T cells. Another V gamma/C gamma 4-containing heterodimer is composed of disulfide-linked 46- to 47-kDa glycoproteins. These findings demonstrate that CD3+, CD4-, CD8- T cells present in the peripheral lymphoid organs express a variety of paired TCR gamma delta proteins. Unlike CD3+, CD4-, CD8- thymocytes, these cells express high levels of C gamma 4, but little, if any TCR alpha beta.     

胎盘多肽对子宫肌瘤患者T 淋巴细胞亚群、NK细胞及炎症因子水平的影响

目的:探讨胎盘多肽对子宫肿瘤切除术后患者血清炎症因子变化和免疫功能的影响。方法:收集我院收治的子宫肿瘤切除术后患者57例,根据就诊先后顺序不同分为对照组和实验组,实验组采用胎盘多肽治疗,对照组采用常规治疗。检测两组患者CD3~+、CD4~+、CD8~+、NK细胞、CRP及IL-6水平,并比较两组的临床效果。结果:治疗后,两组患者血清CRP及IL-6水平均低于治疗前,且实验组明显低于对照组,差异具有统计学意义(P0.05)。治疗后,两组患者CD3~+、CD4~+、CD4~+/CD8~+、NK细胞均高于治疗前,而CD8~+低于治疗前,差异具有统计学意义(P0.05);实验组患者治疗后CD3~+、CD4~+、CD4~+/CD8~+、NK细胞显著高于对照组,而CD8~+低于对照组,差异具有统计学意义(P0.05)。与对照组相比,实验组患者的首次排气和排便时间明显较短,差异具有统计学意义(P0.05)。结论:胎盘多肽能够明显改善子宫肌瘤切除患者的免疫功能,降低炎症反应,有助于患者术后康复。     

急性病毒性肝炎患者外周血T 细胞亚群的检测和分析

目的:检测急性甲、乙型病毒性肝炎患者外周血T淋巴细胞亚群变化,探讨其对疗效和预后意义。方法:采用APAAP桥联酶标法检测115例急性甲、乙型病毒型肝炎患者外周血CD3+、CD4+、CD8+T细胞亚群比例,计算CD4+/CD8+值。并检测了34例患者治疗前后T淋巴细胞亚群变化。结果:115例急性甲、乙型病毒性肝炎患者外周血CD3+、CD4+T细胞比例及CD4+/CD8+值均低于正常对照组(P<0.05),而CD8+T细胞比例均高于正常对照组(P<0.01)。6例无明显疗效者,各亚群比例在治疗前后无显著差异(P>0.05)。28例有明显疗效者,治疗后各亚群比例恢复正常水平,与治疗前相比差别具有统计学意义(P<0.05)。结论:急性甲、乙型病毒性肝炎患者外周血CD4+/CD8+T细胞比值可在一定程度上反映疗效及预后。     

Effect of chronic psychoemotional stress on subpopulation spectrum of T-lymphocytes in immunocompetent organs in male mice

Subpopulation spectrum of T lymphocytes (CD3+, CD4+, CD8+, CD25+, and CD3(+)25+) in thymus, spleen and inguinal lymphatic nodes have been studied in male mice after 20 days of psychoemotional stress produced by social defeats in daily agonistic confrontations. A reduction of total number of cells, of absolute numbers of all researched subpopulations of lymphocytes and % CD3+ cells in thymus of submissive mice was shown in comparison with intact animals. Reduction of total number of splenocytes and absolute numbers of CD4+ and CD8+ lymphocytes has been observed in a spleen of submissive mice. Besides, % CD3+, CD25+, CD4+ and CD25+ cells were increased in these animals in comparison with intact mice. The absolute number of cells with CD8 phenotype was increased in inguinal lymphatic nodes. The data obtained suggest that the chronic psychoemotional stress is accompanied by serious changes of the cellular link of immunity. The effect of chronic emotional social stress on mutual interaction of the central and peripheral links of immunity has been discussed.     

Alloantigen-specific cytotoxic clones bearing the alpha,beta T cell antigen receptor but not CD4 or CD8 molecules

The vast majority of circulating lymphocytes that express the alpha,beta TCR in association with CD3 also express either CD4 or CD8 molecules, which are thought to act as important accessory structures in HLA class II- and I-restricted T cell functions, respectively. In the current study alpha,beta TCR+ clones devoid of detectable CD4 or CD8 were generated by repeated stimulation of fresh CD3+,CD4-,CD8- cells with an allogeneic lymphoblastoid cell line in the presence of conditioned medium containing IL-2. Except for the absence of CD4 and CD8, which was associated with undetectable levels of CD4 and CD8 mRNA, the clones were phenotypically indistinguishable from classical CD3+,alpha,beta TCR+ cells. Furthermore, they mediated potent cytolysis of their specific stimulator line but did not kill irrelevant LCL or NK-sensitive targets. mAb to CD3 and the alpha,beta TCR inhibited cytolysis, suggesting that the clones use the TCR/CD3 complex to recognize and respond to their targets. mAbs to CD2 and CD11a also inhibited cytolysis, indicating that the clones use these accessory molecules to interact with their targets. Finally, cytolysis was inhibited by an HLA-A,B,C framework-specific mAb (W6/32) as well as a mAb (MA2.1) specific for an HLA-A2 epitope. These results demonstrate that CD3+,alpha,beta TCR+,CD4-,CD8- cytotoxic clones can be generated from the peripheral blood of healthy adults, and use their TCR/CD3 complexes to function in an HLA class I-restricted manner.     

老年呼吸机相关性肺炎患者肠道菌群特征及其与疾病的相关性

探讨老年呼吸机相关性肺炎(VAP)患者肠道菌群特征及其与疾病的相关性, 为该类患者的治疗提供参考。

选择2018年1月至2020年1月在该院进行机械通气的患者141例, 根据是否发生VAP分为VAP组(n=67)和非VAP组(n=74), 同时选择同期在我院进行体检的健康者为对照组(n=50)。采用16S rRNA荧光定量PCR法检测3组研究对象肠道双歧杆菌和大肠埃希菌数量, 并计算B/E值。采用流式细胞仪检测T淋巴细胞亚群CD3⁺、CD3⁺CD4⁺、CD3⁺CD8⁺、CD4⁺/CD25⁺、CD8⁺/CD28^-以及CD8⁺/CD28⁺细胞水平。采用改良酶学分光光度法检测血浆D-乳酸水平。采用Pearson相关检验分析VAP组B/E值与免疫功能及血浆D-乳酸的相关性。

VAP组患者肠道双歧杆菌数量、B/E值显著低于非VAP组和对照组, 大肠埃希菌数量显著高于非VAP组和对照组(均P < 0.05);非VAP组和对照组比较差异亦均有统计学意义(均P < 0.05)。VAP组患者CD3⁺、CD3⁺CD4⁺、CD3⁺CD8⁺以及CD8⁺CD28⁺细胞水平显著高于非VAP组和对照组, CD8⁺CD28^-细胞水平显著低于非VAP组和对照组(均P < 0.05);非VAP组和对照组比较差异亦均有统计学意义(均P < 0.05)。VAP组患者血浆D-乳酸水平显著高于非VAP组和对照组, 非VAP组和对照组比较差异亦有统计学意义(均P < 0.05)。Pearson相关检验显示, VAP组患者B/E值与CD3⁺、CD3⁺CD4⁺、CD3⁺CD8⁺、CD8⁺/CD28⁺细胞以及血浆D-乳酸水平呈显著负相关, 与CD8⁺/CD28^-细胞呈显著正相关(均P < 0.05)。

VAP患者存在肠道菌群失衡, 且VAP与肠道菌群失衡存在相关性, 促进肠道菌群平衡有助于VAP的防治。

     

Immunological roles of Pasteurella multocida toxin (PMT) using a PMT mutant strain

The immunological role of the Pasteurella multocida toxin (PMT) in mice was examined using a PMT mutant strain. After a nasal inoculation, the mutant strain failed to induce interstitial pneumonia. Moreover, PMT had no significant effect on the populations of CD4+, CD8+, CD3+, and CD19+ immunocytes in blood or on the populations of CD4+ and CD8+ splenocytes (P<0.01). However, there was a significant increase in the total number of cells in the BAL samples obtained from the wild-type P. multocida-inoculated mice. On the other hand, the level of IL-1 expression decreased when the macrophages from the bronchio-alveolar lavage were stimulated with PMT. Overall, PMT appears to play some role (stimulating and/or inhibiting) in the immunological responses but further studies will be required to confirm this.     

Phenotype of peripheral blood leukocytes and survival of patients with metastatic colorectal cancer

Immune dysfunction is prevalent in metastatic cancer. Few patients with colorectal cancer metastases are cured, and among the strategies aimed at improving the therapeutic results in patients with metastatic colorectal cancer, immunotherapy is being increasingly investigated. We evaluated retrospectively the prognostic significance of peripheral blood leukocytes in 59 patients with metastatic colorectal cancer. The relative numbers of CD3+, CD3+CD4+, CD3+CD8+, NK (CD3-CD16+CD56+), CD3+DR+, CD3+CD25+, CD3+CD69+, CD19+, CD19+CD23+, CD8+CD28+, CD8-CD28+, CD8+CD57+, CD14+DR+ and CD14+CD16+ leukocytes were analyzed by two-color flow cytometry. A three-step approach was adopted to identify predictors of prognosis using regression analysis. Based on the results of univariate survival analysis, the absolute number of white blood cells, NK/CD3+CD69+ and NK/white cell count ratios were significant indicators of prognosis. In the multivariate regression analysis a model was obtained using a single parameter, the NK/CD3+CD69+ ratio, predicting the survival with 10-15% power of regression. The present results indicate that the NK/CD3+CD69+ ratio in peripheral blood may be an independent variable in a regression model predicting the overall survival of patients with colorectal cancer metastases to be tested in prospective studies.     

氟比洛芬酯对食管癌患者围术期外周血淋巴细胞亚群的影响

目的:探究氟比洛芬酯对食管癌患者围术期外周血淋巴细胞亚群的影响。方法:选择2014年6月~2016年8月期间在我院择期行食管癌根治术患者72例为研究对象,采用随机数字法将其分为氟比洛芬酯组(39例)和对照组(33例),患者均给予常规麻醉处理,对照组患者泵入5 mg托烷司琼与20μg/kg芬太尼;氟比洛芬酯组患者给予氟比洛芬酯2 mg/kg。评价术后12 h、24 h和48 h患者疼痛情况(VAS评分),并于术前1 h、术后24 h、术后72 h检测患者血清T细胞中CD3~+、CD4~+、CD8~+及CD56~+比例。结果:两组患者术后不同时刻VAS评分比较,差异均无统计学意义(P0.05);术后24 h两组患者CD3~+、CD4~+、CD4~+/CD8~+均显著降低(P0.05),术后72 h两组患者CD3~+、CD4~+、CD4~+/CD8~+水平均升高,且氟比洛芬酯组患者恢复到术后1 h水平,而对照组患者均仍低于术后1 h,且术后72 h氟比洛芬酯组CD3~+、CD4~+、CD4~+/CD8~+均高于对照组,差异均有统计学意义(P0.05),而CD8~+与CD56~+比例在两组各个时点均没有变化(P0.05);两组患者均未发生严重不良反应。结论:食管癌患者在手术过程中均出现免疫抑制,氟比洛芬酯麻醉效果较好,且对机体免疫功能具有保护作用,促进手术患者免疫功能的恢复,具有重要的临床价值。     

Development of T cells in vitro from precursors in mouse bone marrow

Bone marrow cells from 6- to 8-week-old athymic nude mice were depleted of nylon-wool adherent cells and cultured in vitro at low cell numbers (300 cells/well) in medium supplemented with a supernatant from a thymoma cell line. About 1% of cultured cells grew. Pooled cultures contained cells expressing CD3 (52%), CD4 (37%), CD8 (11%), Thy 1.2 (72%), MAC-1 (43%) and J11d (86%) but no cells expressing sIg. They also contained cells expressing mRNA for the alpha, beta, gamma, and delta chains of the T cell receptor as assessed with C region probes using a sensitive dot blot assay. These cells appear to develop from progenitors which are CD3-. When pooled Day 10 cultures were depleted of nylon-wool adherent cells, the remaining cells were nearly all J11d+, Thy 1.2+, MAC-1-, CD3+, and either CD4+CD8+; CD4+CD8-; CD4-CD8+, or CD4-CD8-; i.e., their surface marker patterns were reminiscent of those of thymocytes. We conclude that our culture system is enabling bone marrow precursors to commence differentiation down the T cell lineage in the absence of a thymic environment.