Distribution of Th1, Th2, and Th0 and the Th1/Th2 cell ratios in human peripheral and endometrial T cells

PROBLEM: To examine whether normal pregnancy involves type 2 T-helper (Th2) immune condition or not. METHOD OF STUDY: We measured the percentage of Th0, Th1, and Th2 and the Th1/Th2 cell ratios of human peripheral blood and endometrial T cells using flow cytometry, which can analyze both the surface marker CD3, and intracellular cytokines, interleukin 4 (IL-4) and interferon gamma (IFNgamma). RESULTS: No significant differences were found in the percentages of Th1, Th2, and Th0 and the Th1/Th2 cell ratios in the peripheral blood T cells of nonpregnant women and women in early pregnancy. On the other hand, the percentage of Th1 cells was highest during the proliferative phase of the endometrium, followed by the secretory phase and early pregnancy decidua. The percentage of Th2 cells was highest in early pregnancy decidua and lowest during the proliferative phase of the endometrium. The Th1/Th2 ratio was 147.48+/-96.68 during the proliferative phase of the endometrium, 37.74+/-21.33 during the secretory phase, and 1.31+/-0.48 in the early pregnancy decidua. CONCLUSIONS: These data indicate that Th1 cells predominate in the nonpregnant endometrium, especially during the proliferative phase, while Th2 cells predominate in early pregnancy decidua.     

Monocytes are primed to produce the Th1 type cytokine IL-12 in normal human pregnancy: an intracellular flow cytometric analysis of peripheral blood mononuclear cells

This paper considers both monocytes and peripheral blood lymphocytes as potential targets for maternal immunological modulation in pregnancy. Peripheral blood mononuclear cells (PBMCs) from non-pregnant and normal pregnant donors were stimulated in vitro, and cytokine production detected intracellularly by flow cytometry. It was found that monocyte production of TNF-alpha was unaltered in pregnancy, while production of IL-12 was significantly enhanced. In contrast, production of the Th1 type cytokine IFN-gamma was suppressed in the lymphocyte subsets: CD4+ T helper cells and CD56+ NK cells. Production of the Th2 type cytokine IL-4 in CD4+ cells was not significantly altered in pregnancy. These data suggest that the concept that pregnancy is a 'Th2 phenomenon' cannot be generalized to the function of all aspects of maternal cellular immunity as, paradoxically, circulating monocytes are 'primed' to produce the Th1 cytokine IL-12. Furthermore, these data support the hypothesis that components of maternal innate immunity are activated in normal pregnancy.     

Intravenous immunoglobulin treatment in women with recurrent abortions: increased cytokine levels and reduced Th1/Th2 lymphocyte ratio in peripheral blood

PROBLEM: The aim of this study was to investigate changes in peripheral blood Th1/Th2 cytokine levels and lymphocyte ratios after massive intravenous immunoglobulin (MIVIg) treatment for women with recurrent spontaneous abortion (RSA) of unexplained etiology. METHOD OF STUDY: Serum Th1 (IFN-gamma, TNF-alpha) and Th2 cytokine (IL-4, IL-10) levels were assessed by ELISA methods (n = 9) and peripheral blood Th1/Th2 lymphocyte ratios (n = 4) by flow cytometry before and after MIVIg treatments in women with four or more consecutive RSA. RESULTS: Pre-treatment serum IFN-gamma (0.06 +/- 0.09 pg/mL, mean +/- SD), TNF-alpha (0.21 +/- 0.45 pg/mL), IL-4 (0.70 +/- 1.16 pg/mL), and IL-10 (1.12 +/- 1.67 pg/mL) increased to 0.17 +/- 0.16 pg/mL, 0.77 +/- 0.28 pg/mL, 1.82 +/- 0.89 pg/mL, and 3.44 +/- 0.48 pg/mL, respectively, after MIVIg treatments (P < 0.05). CD4-positive IFN-gamma/IL-4 lymphocyte ratios (17.3 +/- 9.1) were reduced to 11.5 +/- 7.1 after treatment (P < 0.05). CONCLUSIONS: Massive intravenous immunoglobulin treatments increased peripheral blood cytokine levels and decreased Th1/Th2 lymphocyte ratios; thus, MIVIg treatments modify the peripheral Th1/Th2 balance.     

喘可治注射液对人外周血单个核细胞Th1/Th2细胞因子谱的影响

目的：通过研究喘可治注射液对人外周血单个核细胞（PBMCs）Th1／Th2细胞因子谱的影响，探讨喘可治注射液的免疫调节作用机制。方法：以流式微球分析（CBA）法检测不同处理情况下，人外周血单个核细胞分泌Th1（IFN-γTNF-α、IL-2）和Th2（IL4、IL-6、IL-10）细胞因子水平。结果：健康人PBMCs体外培养12小时后，上清中细胞因子主要为TNF-α和IL-6，喘可治使Th1和Th2细胞因子全面升高；喘可治对PDB加离子霉素诱导的PBMCs分泌Th1和Th2细胞因子具有抑制作用，并能抑制流感病人异常升高的INF-γ、TNF-α、IL-6和IL-10分泌。结论：喘可治注射液上调健康人PBMCs分泌TH1和Th2细胞因子，对异常活化的PBMCs分泌的Th1和Th2细胞因子则具有下调作用。     

Relationship of Th1/Th2 cell balance with the immune response to influenza vaccine during pregnancy

To determine the optimal timing for influenza vaccination in pregnant women, we measured alterations in the types 1 and 2 T helper cell (Th1/Th2) balance during pregnancy, monitored specific immunity to inoculated antigens after vaccination with inactivated influenza vaccine, evaluated the relevance of the Th1/Th2 ratio and immune responses to the vaccination, monitored the maintenance of high antibody titers until delivery and measured the transplacental antibody transfer rate. No significant alterations of the Th1/Th2 balance were noted in the 65% of pregnant women among whom the Th1/Th2 ratio was lower than 9.9% in the first trimester. In those groups with a ratio higher than 10% in the first trimester, there was a tendency for the ratio to decrease as gestation advanced. The efficiency of immunization was not influenced by the Th1/Th2 status or by the stage of gestation. The antibody titer decreased steadily with time from 1 month after vaccination to the time of delivery. Conversely, the transfer rate of antibodies from maternal to fetal blood at the time of delivery increased with the duration of gestation after vaccination. Nevertheless, the antibody titers in both maternal and fetal blood were sufficient to afford protection against infection. Thus, efficient influenza vaccination can be undertaken at any stage of pregnancy. J. Med. Virol. 81:1923–1928, 2009. © 2009 Wiley‐Liss, Inc.     

乙型肝炎患者外周血细胞中Th1/Th2型细胞因子的表达及其临床意义

目的 :探讨Th1 Th2型细胞因子在HBV感染的临床多种表现形式中的表达特征及其临床意义。方法 :采用四色法流式细胞术检测乙型肝炎患者和正常人外周血细胞中CD3+CD8+、CD3+CD8-细胞内的IFN γ和IL 4的表达情况。结果 :与正常对照组相比 ,急性乙肝患者的Tc2细胞百分比增高 ,慢性乙肝患者的Th1、Tc1细胞百分比减低 ,也低于急性乙肝患者 ,但Th1、Tc1细胞百分比随着慢性乙型肝炎肝脏炎症活动的加剧而逐渐增多。分泌IL 4的Th、Tc细胞在正常人和各组病例中无显著性差异。结论 :急慢性乙肝患者的Th、Tc细胞均以分泌Th1型细胞因子占优势 ,但数量的差异可能是HBV感染者不同结局的原因之一。     

Th1 and Th2 in human IVF pregnancy with allogenic fetus

PROBLEM: Positive obstetric outcome of allogenic in vitro fertilization (IVF) pregnancies makes interesting the subject of additional regulatory mechanisms after oocyte donation. METHOD OF STUDY: Eighty eight women: 23 donation of oocytes (DO), 33 IVF, 32 natural conception (NC) were studied in first trimester of pregnancy. Intracellular production of cytokines and chemokine receptors expression were studied by flow cytometry. RESULTS: Intracellular production of interferon-gamma (IFN-gamma), interleukin (IL)-4, tumor necrosis factor-alpha by CD4 T lymphocytes in DO women was higher than in IVF and NC women. Ratio IFN-gamma/IL-4 in DO was lower than in IVF. We found higher expression of chemokine receptor CCR4 but not CXCR3 on CD4 T cells in DO compared with IVF and NC. Ratio CXCR3/CCR4 in DO was lower than in NC. CONCLUSION: Hyperactivation of T helper 1 (Th1) and T helper 2 (Th2) by allogenic fetus is specific for DO pregnancy. Preferable activation of Th2 and relative suppression of Th1 chemokine expression reflect additional regulatory counteractive mechanism(s).     

Human airway and peripheral blood eosinophils enhance Th1 and Th2 cytokine secretion

BACKGROUND: The effector function of eosinophils involves their release of toxic granule proteins, reactive oxygen species, cytokines, and lipid mediators. Murine studies have demonstrated that eosinophils can also enhance T cell function. Whether human eosinophils, in particular, airway eosinophils, have similar immunoregulatory activity has not been fully investigated. The aim of this study was to determine whether human blood and airway eosinophils can contribute to Th1 and Th2 cytokine generation from CD4+ T cells stimulated with superantigen. METHODS: Eosinophils were obtained from blood or bronchoalveolar lavage fluid 48 h after segmental allergen bronchoprovocation. Purified eosinophils were co-cultured with autologous CD4+ blood T cells in the presence of staphylococcal enterotoxin B (SEB). Cytokine levels in the supernatant fluid were determined by enzyme-linked immunosorbent assay (ELISA). Eosinophil expression of major histocompatibility complex (MHC) class II and co-stimulatory molecules was assessed by flow cytometry before culture, 24 h after granulocyte-macrophage colony-stimulating factor (GM-CSF) stimulation, and 24 h after co-culture with CD4+ T cells and SEB. RESULTS: Interleukin (IL)-5, IL-13, and interferon (IFN)-gamma generation increased when CD4+ T cells were co-cultured with either blood or airway eosinophils in the presence of SEB. The ability of eosinophils to enhance cytokine generation was independent of their source (blood vs airway), activation by GM-CSF, or detectable expression of human leukocyte antigen (HLA)-DR, CD80, or CD86. CONCLUSION: Our data demonstrate that SEB-induced generation of Th1 and Th2 cytokines is increased in the presence of human blood and airway eosinophils. Thus, eosinophils can have an immunoregulatory function in pathogen-associated allergic diseases such as atopic dermatitis, chronic sinusitis, and asthma exacerbations.     

合并HBV感染的晚期血吸虫病患者外周血Th1/Th2类细胞因子的表达及临床意义

目的 研究合并HBV感染的晚期血吸虫病患者外周血Th1/Th2类细胞因子的表达及临床意义.方法 选择合并HBV感染的晚期血吸虫病患者、乙型肝炎后肝硬化患者、单纯晚期血吸虫病患者各30例,分别设为A、B和C组,分别采用日立7600全自动生化分析仪检测肝功能,lightcycle基因荧光定量分析仪检测HBV DNA,流式细胞检测技术检测外周血Th1和Th2类细胞因子,并进行比较.结果 A组丙氨酸转氨酶水平、天冬氨酸氨基转移酶水平、总胆红素水平高于B组、C组,差异均有统计学意义,(A组与B组比较t=4.725、3.292、3.012;A组与C组比较t=4.568、3.126、3.217,P均＜0.05);A组HBV DNA水平低于B组,差异有统计学意义(t =3.916,P＜0.05).A组外周血IFN-γ、TFN-α表达水平较B组、C组增加,A组与C组之间差异有统计学意义(t =3.652、3.413,P均＜0.05).A组外周血IL-4、IL-13表达水平高于B组,低于C组,但与B组和C组之间结果比较差异均无统计学意义(A组与B组比较t=0.721、0.452、A组与C组比较t=0.625、0.632,P均＞0.05).结论 合并HBV感染的晚期血吸虫病患者,在疾病的发生发展中Th1类细胞因子可能起了重要作用.     

外周血Th1/Th2细胞在大鼠肝移植早期的变化

目的观察外周血Th1/Th2细胞在大鼠肝移植早期不同免疫状态下的变化趋势是否和免疫状态有关。方法大鼠分3组：A组：同基因移植组（BN→BN）;B组：异基因移植（Lewis→BN）＋环孢素A（CsA）组;C组：异基因移植组。均36只,另外各设观察组观察生存期。于移植后1、3、5、7和14d应用流式细胞分析外周血CD4^＋CD45RC^&#177;的百分数,观察移植肝排斥反应病理分级及受体存活时间。结果（1）A组和B组的生存期平均大于100d,明显长于C组（34.3&#177;3.6）d。（2）除第1天外,C组和B组各时间段的排斥分级明显高于A组（P〈0.05或〈0.01）;C组亦明显高于B组（P〈0.05或〈0.01）。（3）移植后3d起CD4^＋CD45RC^＋%/CD4^＋CD45RC^-%在C组明显高于A组和B组（P〈0.05或〈0.01）。（4）CD4^＋CD45RC^＋%/CD4^＋CD45RC^-%在B组和排斥分级存在负相关（r=-0.565,P〈0.01）,而在C组CD4^＋CD45RC^＋%/CD4^＋CD45RC^-%和排斥分级存在正相关（r=0.745,P〈0.01）。结论外周血Th1/Th2细胞的变化趋势和大鼠肝移植早期不同免疫状态有关。     

Effect of glutamine on Th1 and Th2 cytokine responses of human peripheral blood mononuclear cells

Decreased glutamine concentrations are found in patients with catabolic stress and are related to susceptibility to infections. In this study, we evaluated the role of glutamine in Th1/Th2 cytokine responses. Peripheral blood mononuclear cells were stimulated with phytohemagglutinin (PHA), live attenuated bacillus Calmette-Guérin (BCG), or measles virus in the presence of different glutamine concentrations. We found that glutamine at an optimal concentration (0.6 mM) significantly enhanced PHA-stimulated lymphocyte proliferation as well as Th1 [interferon-gamma (IFN-gamma) and interleukin-2 (IL-2)] and Th2 cytokine (IL-4 and IL-10) production. In the absence of glutamine, BCG and measles virus elicited minimal lymphocyte proliferation, whereas BCG enhanced Th1 cytokine response and measles virus promoted Th2 cytokine response. Interestingly, addition of glutamine promoted the BCG-elicited Th1 cytokine response (IFN-gamma), but suppressed the measles-induced Th2 cytokine response (IL-10). These results suggest that appropriate glutamine levels may influence host responses to different antigens and microorganisms. Furthermore, predominately Th1, but not Th2, cytokine responses required the presence of optimal concentrations of glutamine.     

Expansion of the fraction of Th1 cells in women with preeclampsia: inverse correlation between the percentage of Th1 cells and the plasma level of PAI-2

PROBLEM: The fractions of Th1 cells and Tcl cells may be increased in preeclamptic women compared with healthy pregnant women. METHOD OF STUDY: Eleven healthy non-pregnant women, nine healthy pregnant women (34.1+/-3.1 weeks of gestation), and 10 women with preeclampsia (32.0+/-5.4 weeks) were studied. The fractions of Th1 cells, Th2 cells, Tc1 cells, and Tc2 cells in the peripheral blood mononuclear cells were determined using a three-color flow cytometric technique. The concentrations of plasma plasminogen activator inhibitor-2 (PAI-2) were simultaneously determined. RESULTS: The fraction of Thl cells was significantly larger in women with preeclampsia (18.7+/-5.2%) than in normal pregnant women (11.0+/-5.7%), and it increased with a decrease in the PAI-2 level (r = -0.706, P = 0.002), which was significantly lower in preeclamptic women (83.4+/-46.8 ng/mL) than in normal pregnant women (225.3+/-82.0 ng/mL). The fraction of Tc1 cells increased with increases in the fraction of Th1 cells (r=0.657. P<0.001) and the ratio of Th1-to-Th2 cells (r=0.535, P=0.002). The ratio of Tc1-to-Tc2 cells also increased with an increase in the ratio of Th1-to-Th2 cells (r = 0.394, P = 0.031). CONCLUSIONS: The fraction of Th1 cells appears to be expanded in women with preeclampsia compared with healthy pregnant women.     

Th1 and Th2 cytokine profiles in recurrent aborters with successful pregnancy and with subsequent abortions

BACKGROUND: This study compared Th1-Th2 cytokine profiles in a subgroup of recurrent aborters who had an abortion with those in a subgroup of recurrent aborters who had a successful pregnancy. METHODS: Fifty-four women with a history of at least three normal pregnancies, 24 women with a history of recurrent spontaneous abortion (RSA) followed by abortion (RSA-->A) and 39 women with a history of RSA followed by normal pregnancy (RSA-->N) were studied. Blood samples and placentas were obtained at the time of delivery or abortion; peripheral blood mononuclear cells were stimulated separately with phytohaemagglutinin and with autologous placental cells, and the secreted cytokines estimated. RESULTS: Peripheral blood mononuclear cells from the RSA-->N subgroup secreted higher concentrations of Th1-type cytokines as compared with normal pregnant women, indicating a higher Th1 bias in these women. However, women in the RSA-->N subgroup had significantly higher concentrations of Th2 cytokines as compared with women in the RSA-->A subgroup. A comparison of Th1:Th2 cytokine ratios indicated a higher Th2 bias in RSA-->N women as compared with RSA-->A women. CONCLUSIONS: We conclude that abortion-prone women who proceed to have successful pregnancy are more Th2-biased than abortion-prone women who abort, and that recurrent aborters who undergo spontaneous abortion have a stronger Th1 bias than aborters who have normal pregnancy.     

Kinetics and functional implications of Th1 and Th2 cytokine production following activation of peripheral blood mononuclear cells in primary culture

The importance of cytokine production in some disease processes is now widely recognized. To investigate temporal relationships between cytokines, we stimulated peripheral blood mononuclear cells (PBMC) in vitro using the T cell mitogen phytohemagglutinin (PHA) and various antigens chosen to induce predominantly Th1 (streptokinase: streptodornase or purified protein derivative) or Th2 (Dermatophagoides pteronyssinus, bee or wasp venom: allergens in sensitive subjects) responses. Cytokine production was measured by sensitive bioassays or enzyme-linked immunosorbent assays. Of the 30 subjects studied, 10 were normal and 20 individuals were allergic to either D. pteronyssinus (n = 10) or bee venom (n = 10) (examined before specific allergen immunotherapy). We examined the temporal profiles of a panel of cytokines produced in prmary culture. In PHA-driven cultures, cytokines were found to be sequentially produced in the order interleukin (IL)-2, IL-4, IL-5, IL-3, interferon (IFN)-γ, IL-10, IL-6, IL-12 and tumor necrosis factor (TNF)-α. The response to allergen in allergic patients was predominantly Th2 in nature, with the production of IL-4, IL-5, IL-6 and IL-10, but little or no IFN-γ. IL-2, IL-3, TNF-α and IL-12 were also produced in low amounts. The response of both atopic and normal subjects to recall bacterial antigens was predominantly Th1, with high levels of IFN-γ, IL-2 and TNF-α. The relevance of the order, amount and speed of production, characteristic kinetics (production, consumption, homeostatic regulation) and the cell source of the cytokines are discussed.     

Interleukin-12 secretion by peripheral blood mononuclear cells is decreased in normal pregnant subjects and increased in preeclamptic patients

PROBLEM: It has been reported that T-helper (Th) 2 dominance in normal pregnancy shifts to Th1 dominance in preeclampsia. Peripheral blood mononuclear cell (PBMC) production of interleukin (IL)-12, which induce Th1 responses, has not been compared between these clinical states. METHOD OF STUDY: Peripheral blood mononuclear cell from 35 non-pregnant women, 35 healthy pregnant women, 12 mildly preeclamptic patients, and 15 severely preeclamptic patients were cultured for 24 hr. IL-12 secretion was determined by enzyme-linked immunosorbent assay (ELISA). Th1/Th2 ratios in PBMC were determined flow-cytometrically, and the amounts of HLA-DR and CD14 expression on the monocytes were obtained by flow cytometry. RESULTS: Peripheral blood mononuclear cell from healthy pregnant subjects secreted less IL-12 than non-pregnant women. PBMC from severely preeclamptic patients secreted more IL-12 than those from healthy pregnant subjects, while IL-12 secretion in mild preeclampsia resembled secretion in normal pregnancy. Th1/Th2 ratios correlated were positively with IL-12. Increased HLA-DR antigens and reduced CD14 expression, suggesting monocyte activation, were observed in preeclamptic patients, although monocyte counts were unchanged. CONCLUSION: Decreased IL-12 secretion by PBMC may cause Th2 dominance in normal pregnancy, while increased IL-12 secretion by activated monocytes may cause Th1 dominance in preeclampsia.     

CD4单抗选择对于流式细胞术分析Th1/Th2极化的有效性评价

选择13B8.2和S3.5两株商品化的CD4单克隆抗体用于Th1/Th2极化的流式细胞术评价,分析PMA、Ionomycin和BFA单独或联合作用于外周血CD4抗原表达水平的变化,以确定Th1/Th2极化评价时有效的CD4设门单抗,并试图揭示CD4抗原降调的机制。结果表明,13B8.2可作为CD4的设门单抗。PMA和Ionomycin联合刺激会导致外周血CD4抗原的明显降低,而PMA单独刺激不能,且BFA与PMA和Iononmycin同时加入外周血中可部分抑制PMA和Ionomycin刺激引起的CD4抗原内吞和降解。     

A novel surface molecule of Th2- and Tc2-type cells, CRTH2 expression on human peripheral and decidual CD4+ and CD8+ T cells during the early stage of pregnancy

It has been demonstrated that pregnancy induces the immunomodulation of cytokine responses away from the Th1 paradigm and towards the Th2 paradigm. In this study, we examined the expression of CRTH2 (chemoattractant receptor-homologous molecule expressed on Th2) on decidual CD4+ and CD8+ T cells during the early stages of pregnancy. Examination of the cytokine profile revealed that CRTH2 was expressed on CD4+-type-2 T helper (Th2-type) and CD8+-type 2 T cytotoxic (Tc2-type) cells. The percentages of CRTH2+ cells in CD3+/CD4+ T cells and CD3+/CD8+ T cells were significantly higher in the decidua than in the peripheral blood. These results indicate the significance of Th2- and Tc2-type cells of the decidua in the maternal immune system, presumably through their production of cytokines which may contribute to the maintenance of pregnancy.