强直性脊柱炎TH亚群激活及T细胞活化状态研究

目的 :研究强直性脊柱炎 (AS)患者TH1 TH2细胞激活状态及T细胞活化状况 ,探讨其发病机理。方法 :运用流式细胞仪 (CBA)法检测 35例AS患者TH1(INF γ、TNF α、IL 2 )、TH2 (IL 10、IL 5、IL 4 )细胞因子水平以及外周血淋巴细胞CD3 、CD4 、CD8 T细胞、B细胞 (CD19 )、NK细胞 (CD16 5 6 )和CD3 HLA DR 、CD4 HLA DR 、CD8 HLA DR T细胞百分率 ,并与健康对照组比较。结果 :AS患者血浆TNF α水平、IL 2水平均显著低于健康对照组 (P <0 0 1,P <0 0 5 ) ,IL 10水平显著高于健康对照组 (P <0 0 5 )。CD3 、CD3 CD8 T细胞百分率显著低于健康对照。CD8 HLA DR T细胞百分率均显著低于健康对照 (P <0 0 5 ) ,CD4 HLA DR T显著高于健康对照 (P <0 0 5 )。结论 :AS患者血浆低水平的TNF α、IL 2和高水平的IL 10提示其体内存在着TH1 TH2平衡的偏移 ;TH1激活程度低下 ,而TH2激活程度增强 ,细胞因子水平的改变尤以TH1细胞因子TNF α改变特别明显。AS患者在多个层面存在细胞免疫功能紊乱     

尖锐湿疣患者外周血T淋巴细胞上活化抗原的表达

目的 :探讨尖锐湿疣 (CA)患者外周血CD6 9和HLA DR分子在T淋巴细胞上表达的变化及其意义。方法 :采用免疫荧光三标记流式细胞术检测 30例CA患者外周血T细胞CD6 9和HLA DR抗原的表达 ,并以 31例正常人作为对照。结果 :CA患者外周血CD3 T细胞CD6 9的表达 (6 6 3%± 3 13% )与正常人对照组 (5 12 %± 1 6 4 % )相比 ,差异有显著性 (P <0 0 5 ) ,CD4 T细胞CD6 9的表达与正常人对照组相比 ,差异无显著性 (P >0 0 5 ) ,CD8 T细胞表达CD6 9水平 (4 6 1%± 3 0 9% )明显高于对照组 (2 6 7%± 1 31% ,P <0 0 1) ;患者组CD3 T细胞中HLA DR 细胞 (2 1 6 5 %± 8 84 % )比对照组 (13 5 6 %± 5 15 % )显著增高 (P <0 0 0 1)。结论 :CA患者外周血T淋巴细胞的激活以CD8 T细胞为主 ,其免疫激活状态在抗病毒感染中起着重要作用。     

恢复期SARS患者淋巴细胞亚群分布及其与T淋巴细胞Ag-NOR含量相关性的初步分析

目的　通过检测恢复期SARS患者外周血淋巴细胞亚群分布及其与T淋巴细胞核仁形成区嗜银蛋白 (Ag NOR)含量的相关性分析 ,探讨恢复期SARS患者的免疫状态及淋巴细胞亚群与活性状态改变的关系。方法　以流式细胞术检测患者外周血淋巴细胞亚群 ,用KL型免疫图像分析系统检测外周血T淋巴细胞Ag NOR ,对两者进行相关分析。结果　恢复期患者T淋巴细胞总数及CD4 +亚群基本正常 ,而CD8+亚群偏高 ,CD4 +/CD8+降低 ,CD4 +/CD8+<1的比例占 37.4 % ,激活T细胞中以CD8+细胞为主。B细胞比例正常。NK细胞明显低于对照组。重症患者CD4 +、CD4 +/CD8+、CD1 9+CD5+降低及CD8+、CD3+HLA DR+升高。 50岁以上及使用大剂量激素患者CD3+HLA DR+升高。淋巴细胞Ag NOR的含量 (IS % )在正常范围 ,但患者IS值呈偏态分布 ,低于正常范围者占 4 4.99%。淋巴细胞Ag NOR的含量与CD3+、NK、CD3+HLA DR+、CD3+CD2 5+在统计学上具有相关性。结论　恢复期SARS患者免疫功能趋于恢复正常 ,但部分患者的淋巴细胞亚群数量及淋巴细胞活性仍未恢复正常 ,这些病人在临床症状得到改善之后 ,尚需一定时间的观察随访 ,以了解SARS病毒对人体免疫机能的长期影响。     

恢复期SARS患者T淋巴细胞凋亡相关基因蛋白表达

目的 :探讨SARS的免疫发病机理 ,研究恢复期SARS患者T淋巴细胞凋亡相关基因蛋白表达。方法 :流式细胞仪测定 15例恢复期SARS患者的T淋巴细胞及其亚群和凋亡相关基因蛋白———CD95、Bcl 2、DR5及人冠状病毒 2 2 9E受体—CD13表达 ;并以 10例高危、健康的医务人员为对照。结果 :恢复期SARS患者外周血T淋巴细胞亚群 (CD3、CD4、CD8)均恢复正常 ;CD4 DR5、CD13,CD8 DR5、CD13表达均阴性 ,与对照组无明显差异。但是 ,恢复期SARS患者CD4、CD8阳性细胞仍明显表达Fas和Bcl 2 ,其阳性细胞的百分数较对照组明显增多 (P <0 0 1)。结论 :Fas FasL途径导致T淋巴细胞凋亡可能是SARS患者的免疫发病机理之一 ,而恢复期Bcl 2表达增多对抑制T淋巴细胞的进一步凋亡、T淋巴细胞亚群恢复正常可能具有重要作用。SARS患者T淋巴细胞凋亡不通过Trail DR5 细胞凋亡途径。     

CRTH2在特发性血小板减少性紫癜T细胞亚群表达的检测

目的　研究CRTH2在特发性血小板减少性紫癜 (ITP)患儿外周血T淋巴细胞亚群的表达及其意义。方法　收集ITP患儿及健康儿童外周抗凝静脉血 ,分离纯化T细胞 ,以PE标记的抗CRTH2单抗和Cy5标记的抗CD4、CD8单抗作双色流式细胞术 ,分析CRTH2在ITP患儿CD4 、CD8 T细胞表达的水平。结果　ITP患儿外周血T细胞与健康儿童相比 ,CD4 细胞百分率及CD4 /CD8 比例明显下降 (P <0 .0 5 ) ,CD8 细胞百分率无明显变化 (P >0 .0 5 ) ;CD4 CRTH2 -及CD4 CRTH2 细胞百分率明显下降 (P <0 .0 5 ) ,CD8 CRTH2 -及CD8 CRTH2 细胞百分率无明显变化 (P >0 .0 5 ) ,CD4 CRTH2 -/CD4 CRTH2 及CD8 CRTH2 -/CD8 CRTH2 比例明显升高 (P <0 .0 5 )。结论　ITP患儿外周血存在T细胞亚群比例失衡 ,呈明显TH1类细胞优势 ,T细胞亚群比例失衡与该病的免疫发病机制有关。     

重症肌无力患者外周血中CD4~+CD25~+调节性T细胞的变化及意义

研究CD4 + CD2 5 + 调节性T细胞在重症肌无力 (MG )发病中的作用。本文采用三色流式细胞术对 2 9例MG患者和 2 3例健康对照者外周血中CD4 + CD2 5 + T细胞 (CD3+ CD4 + CD2 5 + )的百分率进行测定。结果显示病情未能很好控制的MG患者外周血CD4 + CD2 5 + T细胞比率略低于健康对照组 (分别为 3 79%± 1 4 0 %、 4 5 3%± 0 96 % ,P =0 12 ) ,病情稳定或缓解的MG患者CD4 + CD2 5 + T细胞比率 (8 4 5 %± 1 96 % )显著高于健康对照组 (P =0 0 0 0 1) ;胸腺切除的MG患者CD4 + CD2 5 + T细胞比率 (8 4 4 %± 2 39% )显著高于非胸腺切除的MG患者 (5 88%± 2 89% ,P =0 0 38)和健康对照组 (4 5 3%± 0 96 % ,P =0 0 0 3)。提示MG患者外周血中存在异常比例的CD4 + CD2 5 + 调节性T细胞 ,可能参与疾病的发生与发展。     

卡介菌多糖核酸提高复发性生殖器疱疹患者CD8+ T细胞的细胞因子表达

目的　研究卡介菌多糖核酸 (BCG PSN)对复发性生殖器疱疹 (RGH)患者外周血CD8 T细胞IL 12、IFN γ、IL 4表达的影响。方法　应用流式细胞仪对 4 5例RGH患者和 15名健康志愿者外周血CD8 T细胞IL 12、IFN γ和IL 4的表达进行检测。结果　治疗前RGH患者外周血IFN γ和IL 12阳性CD8 T细胞百分率均显著下降 (P <0 .0 5 ) ,Tc1 Tc2平衡失调 (P <0 .0 5 )。治疗后BCG PSN组外周血IL 12和IFN γ阳性CD8 T细胞百分率显著升高 (P <0 .0 5 ) ,Tc1 Tc2恢复平衡。病例对照组IL 12、IFN γ及IL 4阳性CD8 T细胞百分率于治疗前后变化差异均无显著性 (P >0 .0 5 )。BCG PSN组与病例对照组治疗前后IL 12、IFN γ阳性CD8 T细胞百分率间差异均有显著性 (P <0 .0 5 )。BCG PSN组复发率较低 ,复发病情较轻。结论　RGH患者存在以Tc1水平低下为主的Tc1 Tc2比例失衡和IL 12表达水平低下 ;BCG PSN通过上调外周血CD8 T细胞IFN γ、IL 12的表达 ,纠正机体细胞因子失衡状态而减少疾病的复发     

Th1/Th2、Tc1/Tc2亚群在乙肝肝硬化患者中的作用

目的 :探讨乙肝肝硬化患者外周血 (PBMC)中CD4 和CD8 T细胞内Th1和Th2类细胞的平衡状态 ,探明Th1、Th2类细胞在乙肝肝硬化中的作用。方法 :乙肝肝硬化患者CD4 T细胞和CD8 T细胞中IFN γ 和IL 4 细胞的百分率 ,观察乙肝肝硬化患者Th1 Th2、Tc1 Tc2比例的变化。结果 :乙肝肝硬化患者PBMC中CD4 ,CD8细胞 ,CD4 CD8比值与健康对照者相比无统计学差异 (P >0 0 5 ) ,Th1细胞及Tc1细胞百分率为 8 8% ,9 0 % ,较健康对照者 7 5 % ,7 7%升高 (P <0 0 5 )。结论 :乙肝肝硬化患者外周血T细胞亚群发生Th1类偏移 ,在乙肝肝硬化的发生和发展中可能起重要作用     

促肝细胞生长素及其S4组份对人PBMC表型的影响

本文采用生物素 链霉亲和素 (BSA )免疫细胞化学法及流式细胞仪技术检测CD3+、CD4+、CD8+、HLA DR+及CD2 5 +细胞百分率 ,藉以研究促肝细胞生长素 (pHGF )及其组分S4对免疫活性细胞的作用。结果表明 ,pHGF可使脐血中CD4+、CD8+及HLA DR+细胞数有明显增长 ;对正常人及肿瘤患者PBMC的HLA DR+细胞数亦有促进作用 ,与PHA的刺激作用相近似。用FPLC及HPLC等方法分离的单一成分S4对成人PBMC表面CD2 5抗原表达有明显促进作用。     

SARS患者康复期T细胞亚群、活化状态及TCR Vβ表达格局的分析

目的：研究中西医结合治疗后，SARS患者康复期淋巴细胞功能状态和T细胞受体(TCR)Vβ24个亚家族表达的格局。方法：应用流式细胞分析技术，观察我院76例康复期患者T细胞亚群，T、B细胞的活化状态及CD3^ 和TCR Vβ1、2、3、4、5．1、5．2、5．3、7．1、7．2、8、9、11、12、13．1、13．2、13．6、14、16、17、18、20、21．3、22及23等24个亚家族的表达。结果：T细胞亚群中，CD3^ 和CD4^ T细胞的百分率均数低于参考值；CD8^ T细胞的百分率高于参考值。CD8^ ／CD28^-T细胞(Ts)的百分率较参考值增高；而CD8^ ／CD28^ T细胞(Tc)的百分率较参考值降低，其中有39例Tc细胞的百分率降低，有48例Ts细胞的百分率升高。CD3^ HLA-DR^ 和CD3^-／HLA-DR^ 细胞的百分率高于正常参考值，增加的例数分别是36例和30例。TCR Vβ24个亚家族中，TCR Vβ14的表达最高，TCR Vβ5．3和23的表达次之，表现出TCR Vβ呈优势表达。正常对照组TCR Vβ24个亚家族中，只TCR Vβ14的表达最高，与SARS患者的结果相一致；但TCR Vβ24个亚家族的分布格局不一样。两组比较，Vβ1、Vβ5．2、Vβ5．3、Vβ7．2、Vβ9、Vβ11、Vβ13．1、Vβ13．2、Vβ17、Vβ18、Vβ22和Vβ23表达有显著性差异，康复期SARS患者组均高于正常对照组。同时，SARS患者激素用量大于1000U组，TCR Vβ4、22和23的表达均高于未用激素组。结论：康复期患者CD8^ ／CD28^-T细胞数的升高可导致CD8^ T细胞数升高；Ts细胞数的增加，因其分泌的抑制因子增多，从而造成CD3^ 和CD4^ T细胞数降低。SARS患者康复期TCRVB24个亚家族在T细胞上的表达不同：CD3^ ／HLA-DR^ 和CD3^ ／HLA-DR^ T细胞数的升高，可能与T、B细胞的活化较晚有关。     

Activated T lymphocytes in pre-eclampsia

PROBLEM: The aim of our study was to investigate the activation markets of T CD3(+), T helper CD4(+) and T cytotoxic CD8(+) cells, as well as, the populations of T na?ve CD4(+) CD45RA(+), T memory CD4(+) CD45RO(+) and T regulatory lymphocytes in PE and healthy pregnant women. METHOD OF STUDY: Twenty-five patients with PE and thirty healthy third trimester pregnant women were included in the study. Peripheral blood mononuclear cells were isolated from peripheral blood, stained with monoclonal antibodies and estimated using the flow cytometric method. RESULTS: The percentages of CD4(+)CD25(+), CD4(+)CD25(dim), CD3(+)HLA-DR(+), CD4(+)HLA-DR(+) and CD8(+)HLA-DR(+) cells did not differ between study groups. The population of T regulatory CD4(+)CD25(bright) lymphocytes was significantly lower in the group of patients with PE when compared with the controls (P < 0.01). The percentages of CD3(+)CD25(+) (P < 0.05), CD8(+)CD25(+) (P < 0.05), CD4(+)45RO(+) (P < 0.01) lymphocytes were significantly higher, while CD4(+)CD45RA(+) (P < 0.01) cells--significantly lower in peripheral blood of patients with PE when compared with the control group. CONCLUSION: The increased levels of T CD4(+)45RO(+) and T CD8(+) CD25(+) cells can suggest the activation of CD4(+) and CD8(+) T lymphocytes in pre-eclampsia. It seems possible that the activation of T lymphocytes is associated with the deficiency of T regulatory cells in PE.     

Immunophenotype of peripheral T lymphocytes, NK cells and expression of CD69 activation marker in patients with recurrent spontaneous abortions, during the mid-luteal phase

PROBLEM: Determination of subpopulations of T lymphocytes, natural killer (NK) and activation status, in peripheral blood during the mid-luteal phase from patients with unexplained recurrent spontaneous abortion (RSA). METHOD OF STUDY: Peripheral blood samples from non-pregnant women with RSA and normal multiparous were taken and evaluated for subpopulations of T lymphocytes: CD4, CD8, ('naive-like' and 'memory-like'), TCR receptor (alphabeta and gammadelta), activation status by CD69(+surface or intracellular)/CD3(+), and NK cells (CD16(+)/CD56(dim)/CD3(-), CD16(+)/CD56 (bright)/CD3(-), CD69(+surface or intracellular)/CD56(+)/CD3(-) cells). RESULTS: The evaluation of T lymphocytes only showed an increase in the expression of CD69 (surface and intracellular) in the RSA group. Additionally, we observed an increase in the total NK cells, CD56(+) NK cells percentages, CD56(dim) NK cells and CD69 NK cells in RSA group. CONCLUSION: These observations support the concept that immunological activation of T lymphocytes and NK cells could be involved in peripheral blood during the mid-luteal phase in patients with unexplained RSA.     

Influence of HLA-DR2 on perforin-positive cells in pulmonary tuberculosis

Perforin is one of the key effector molecules of cytotoxic T cells and natural killer cells. The influence of HLA-DRB1 alleles on peripheral blood perforin-positive CD4, CD8, CD16 and CD 56 cells was studied by flow cytometry. HLA-DRB1 typing was done in normal healthy subjects (NHS: n = 156) and patients with pulmonary tuberculosis (PTB: n = 102) by polymerase chain reaction-based sequence-specific oligonucleotide hybridization method. We observed a significantly decreased percentage of total perforin-positive cells (per(+)) (P = 0.0004); CD8(+)/Per(+) (P = 0.0005); CD16(+)/Per(+) (P = 0.05) and CD 56(+)/Per(+) cells (P = 0.001) in HLA-DR2-positive PTB patients compared to non-DR2 patients. Subtyping of HLA-DR2-positive subjects at the allelic level revealed that the percentage of CD8(+)/Per(+) cells did not differ among DRB1*1501 and DRB1*1502 patients while a trend towards a decreased percentage of CD16(+)/Per(+) and CD 56(+)/Per(+) cells was noticed in DRB1*1501 patients compared to DRB1*1502 patients. The present study suggests that HLA-DR2 may be associated with down-regulation of perforin-positive cytotoxic lymphocytes and natural killer cells in pulmonary tuberculosis.     

HLA-DQB1 alleles may influence the surface expression of DQ molecules in lymphomononuclear cells of type 1 diabetes mellitus patients

To evaluate the expression of human leucocyte antigen (HLA) class II (DR and DQ) molecules on lymphomononuclear cells involved in the pathogenesis of type 1 diabetes, we studied 20 patients and 20 controls matched to patients for age, sex and HLA class II profile. The coexpression of HLA and CD3, CD4, CD8, CD19 and CD14 molecules was evaluated by flow cytometry. HLA-DRB1, -DQA1 and -DQB1 alleles were assigned using amplified DNA hybridized with sequence-specific primers. The fluorescence intensity of HLA-DR and -DQ molecules observed on the surface of the lymphomononuclear cells of patients did not differ significantly from controls. Patients presented decreased percentage of double-positive CD4(+)/DQ(+) cells and increased percentage of CD19(+)/DR(+) cells, irrespective of the HLA class II profile; however, the more dramatic alteration of the lymphomononuclear phenotype profile was observed for patients possessing the HLA-DQB1*0201 allele. These patients exhibited decreased percentage of CD3(+), CD4(+), CD8(+), CD19(+) and CD14(+) cells bearing HLA-DQ molecules and decreased fluorescence intensity for HLA-DQ molecules on CD19(+) cells compared to patients without the DQB1*0201 allele. Although type 1 diabetes patients shared CD4/DQ or CD19/DR phenotype abnormalities, patients typed as DQB1*0201 presented additional abnormalities in terms of DQ expression and cell phenotypes bearing DQ molecules.     

Differences in peripheral blood lymphocyte phenotypes between Helicobacter pylori-positive children and adults with duodenal ulcer

The immunological mechanisms involved in the development of duodenal ulcer, especially in childhood, are unclear. Helicobacter pylori-positive children and adults, with and without duodenal ulcer, were therefore compared with respect to CD4(+) T-cells, and CD8(+) T-cells, B-cells and B1a-cells, as well as cell activation (CD4(+)/HLA-DR(+) and CD8(+)/HLA-DR(+)) and co-stimulatory (CD4(+)/CD28(+) and CD8(+)/CD28(+)) markers, in peripheral blood. Children with and without duodenal ulcer differed significantly. In particular, there was a phenotypic change in CD8(+) T-cells from children with ulcer that involved a 200% increase in the number of CD8(+)/HLA-DR(+) cells/mm(3) and a decrease of 34.2% in the number of CD8(+)/CD28(+) cells/mm(3). This phenotype of chronically activated memory CD8(+) T-cells, which has also been observed in patients with AIDS and tuberculosis, is associated with disease severity and progression. A lower frequency of B1a-cells was also observed in the group of children with ulcer. Conversely, no difference between infected adults with and without ulcer was observed, but the percentage of CD4(+)/HLA-DR(+) cells was lower in adults with ulcer, suggesting that a down-regulated immune response may play a role in the development of duodenal ulcer in adults. Gastric inflammation correlated positively with CD4(+) and chronically activated CD4(+) T-cells in children and adults without duodenal ulcer, respectively. These results suggest that there are differences in the immunophenotyping profile between H. pylori-positive children and adults with duodenal ulcer, indicating the possibility of distinct immune mechanisms in the development of the disease according to age.     

TCRalphabeta+/CD4+ large granular lymphocytosis: a new clonal T-cell lymphoproliferative disorder

Large granular lymphocyte (LGL) leukemia is a well-recognized disease of mature T-CD8(+) or less frequently natural killer cells; in contrast, monoclonal expansions of CD4(+) T-LGL have only been sporadically reported in the literature. In the present article we have explored throughout a period of 56 months the incidence of monoclonal expansions of CD4(+) T-LGL in a population of 2.2 million inhabitants and analyzed the immunophenotype and the pattern of cytokine production of clonal CD4(+) T cells of a series of 34 consecutive cases. Like CD8(+) T-LGL leukemias, CD4(+) T-LGL leukemia patients have an indolent disease; however, in contrast to CD8(+) T-LGL leukemias, they do not show cytopenias and autoimmune phenomena and they frequently have associated neoplasias, which is usually determining the clinical course of the disease. Monoclonal CD4(+) T-LGLshowed expression of TCRalphabeta, variable levels of CD8 (CD8(-/+dim)) and a homogeneous typical cytotoxic (granzyme B(+), CD56(+), CD57(+), CD11b(+/-)) and activated/memory T cell (CD2(+bright), CD7(-/+dim), CD11a(+bright), CD28(-), CD62L(-) HLA-DR(+)) immunophenotype. In addition, they exhibited a Th1 pattern of cytokine production [interferon-gamma(++), tumor necrosis factor-alpha(++), interleukin (IL-2)(-/+), IL-4(-), IL-10(-), IL-13(-)]. Phenotypic analysis of the TCR-Vbeta repertoire revealed large monoclonal TCR-Vbeta expansions; only a restricted number of TCR-Vbeta families were represented in the 34 cases analyzed. These findings suggest that monoclonal TCRalphabeta(+)/CD4(+)/NKa(+)/CD8(-/+dim) T-LGL represent a subgroup of monoclonal LGL lymphoproliferative disorders different from both CD8(+) T-LGL and natural killer cell-type LGL leukemias. Longer follow-up periods are necessary to determine the exact significance of this clonal disorder.     

Regeneration and tolerance factor: a correlate of human immunodeficiency virus-associated T-cell activation.

Human immunodeficiency virus (HIV) infection causes extensive phenotypic alterations in lymphocytes. Cellular markers that are normally absent or expressed at low levels on quiescent cells are upregulated throughout the disease course. The transmembrane form of regeneration and tolerance factor (RTF) is expressed at negligible levels on resting T cells but is quickly upregulated following in vitro stimulation and activation. Recently, we reported that expression of RTF was significantly higher in cells from HIV-seropositive (HIV(+)) individuals than in cells from HIV-seronegative (HIV(-)) individuals. Because T cells from HIV(+) individuals express markers reflecting chronic activation, we hypothesized that these in vivo-activated cells would coexpress RTF. Flow cytometry was used to assess RTF expression on activated (CD38(+) and HLA-DR(+)) CD4(+) and CD8(+) T cells. HIV(+) individuals had higher percentages of RTF(+) CD38(+) (P < 0.0001) or RTF(+) HLA-DR(+) (P = 0.0001) CD4(+) T cells than HIV(-) individuals. In HIV(+) individuals, increased percentages of CD4(+) T cells that were RTF(+), RTF(+) CD38(+), and RTF(+) HLA-DR(+) correlated inversely with the absolute number and percentage of CD4(+) T cells and correlated positively with plasma beta(2)-microglobulin concentrations. HIV(+) individuals had higher percentages of CD8(+) T cells that were RTF(+) CD38(+) (P = 0.0001) or RTF(+) HLA-DR(+) (P = 0.0010). In HIV(+) individuals, increased percentages of CD8(+) T cells that were RTF(+) HLA-DR(+) correlated inversely with the percentage of CD4(+) T cells, and high percentages of CD8(+) T cells that were RTF(+) CD38(+) correlated positively with plasma beta(2)-microglobulin levels. These findings strongly suggest that increased RTF expression is a correlate of HIV-associated immune system activation.     

Can immune parameters be used as predictors to distinguish between pulmonary multidrug-resistant and drug-sensitive tuberculosis?

Introduction

Despite the development and wide implementation of Directly Observed Therapy Strategies (DOTS), multidrug-resistant tuberculosis (MDR-TB) remains a serious global health threat. In this study, the role of host immune response in patients with MDR-TB is investigated and compared with that of patients with smear-positive drug-sensitive tuberculosis (SP-TB).

Material and methods

27 patients with SP-TB, 20 patients with MDR-TB, and 20 healthy controls were included in the study. Immune parameters were determined by flow cytometry using monoclonal antibodies in order to compare the percentage values of these markers in the two study groups and the control group.

Results

The levels of lymphocyte subgroups in the gate of CD45(+)/CD14(–) lymphocyte: CD45(+), CD3(+), CD4(+), NK, CD3/HLA-DR, CD 95(+) cells were significantly lower; by contrast CD23(+), CD25(+), CD19(+), CD4(+)/CD8(+), HLA-DR cells were found to be lower, but not significantly so in patients with MDR-TB, compared to levels in patients in the SP-TB and control groups. Besides these findings, the levels of NKT cells and _γδ TCR(+) cells were significantly higher in the MDR-TB than in the healthy control and SP-TB group.

Conclusions

The lower levels of CD3/ HLA-DR, CD4 (+), Fas (+), and NK, and the higher level of NKT together with _γδ T cells in patients with MDR-TB compared to those in SP-TB may indicate a profound immune suppression in MDR-TB patients and thereby may denote an accumulation in the bacterial load. Our findings may shed light on the pathogenesis and prognosis of MDR tuberculosis, and may point towards the use of flow cytometry findings as an aid to early diagnosis in MDR-TB patients.     

Lymphocyte sub-populations in gestational diabetes

PROBLEM: We hypothesize that the normal immunologic responses by the maternal immune system during pregnancy are not as well-regulated in gestational diabetes (GD) patients as in healthy pregnant women. METHOD OF STUDY: Using two-color flow cytometry we evaluated frequencies of peripheral blood lymphocytes in 20 GD patients being treated with insulin; 43 GD patients treated with dietary therapy but no insulin; 44 women experiencing normal pregnancies; and 48 non-pregnant women. RESULTS: When compared with healthy pregnant women, both GD cohorts showed higher percentages CD4(+)CD25(+) (P < 0.05), CD4(+)CD45RO(+) (P < 0.05) and CD4(+)CD29(+) (P < 0.01) but lower percentages of CD4(+)CD45RA(+) (P < 0.05). Higher percentages of the activated phenotypes CD8(+)CD25(+) and CD8(+)HLA-DR(+) cells in the diet-treated cohort and CD4(+)HLA-DR(+) cells in insulin-treated GB cohort, were observed compared with healthy pregnant subjects (P < 0.05). CONCLUSIONS: Expanded populations of activated peripheral blood T cells are associated with GD, suggesting that normal maternal immunosuppression is less effective in GD-afflicted women.