OBJECTIVETo explore the diagnostic values of interleukin-12 (IL-12) and interferon-γ (IFN-γ) for the patients with acute leukemia (AL).

METHODSA total of 76 cases of AL were enrolled in this study, and the 40 healthy persons were used as control group. The levels of IL-12 and IFN-γ were determined by enzyme linked immunosorbent assay (ELISA). The results were analyzed.

RESULTSThe levels of IL-12 and IFN-γ in the untreated AL group, ALL and ANLL groups were lower significantly than those in the control group (P<0.05), there was no significant difference between untreated AL and ANLL groups (P>0.05). The levels of IL-12 and IFN-γ in CR patients of AL group after treatment obviously higher than that of patients before treatment (P<0.05), but there was no significant difference as campared with that in control. The levels of IL-12 and IFN-γ in NR patients of AL group after treatment were obviously lower than that in control group (P<0.05), but there was no significant difference in comparision with patients before treatment (P>0.05). The levels of IL-12 and IFN-γ of AL-CR and AL-NR patients before treatment were not significant difference before treatment (P>0.05). The levels of IL-12 and IFN-γ of AL-CR patients obviously higher than that in AL-NR patients (P<0.05). According to immure classification, the levels of IL-12 and IFN-γ of patients in untreated group were not significant difference. In regard to the clinical risk degree, the level of IL-12 of patients in untreated group was not obvious difference (P>0.05), but the level of IFN-γ of patients in untreated group was obvious different (P<0.05). The level of IL-12 of patients in untreated group positively correlated with level of IFN-γ (r=0.735, P<0.05), but the level of IL-12 did not significantly correlated with the level of IFN-γ (r=0.292, P>0.05).

CONCLUSIONThe serum levels of both IL-12 and IFN-γ are lower, but the changes of both serum levels may be helpful to diagnose and treatment of AL patients.

Acute Disease ; Case-Control Studies ; Enzyme-Linked Immunosorbent Assay ; Humans ; Interferon-gamma ; blood ; Interleukin-12 ; blood ; Interleukin-4 ; blood ; Leukemia ; blood ; diagnosis ; Remission Induction

BACKGROUNDChronic obstructive pulmonary disease (COPD) is characterized by progressive loss of lung function and local and systemic inflammation, in which CD8+ T-cells are believed to play a key role. Activated CD8+ T-cells differentiate into distinct subpopulations, including interferon-γ (IFN-γ)-producing Tc1 and interleukin (IL)-17-producing Tc17 cells. Recent evidence indicates that Tc17 cells exhibit considerable plasticity and may convert into IL-17/IFN-γ-double producing (Tc17/IFN-γ) cells when driven by inflammatory conditions. The aim of this study was to investigate the Tc17/IFN-γ subpopulation in peripheral blood of patients with COPD and to evaluate their potential roles in this disease.

METHODSPeripheral blood samples were collected from 15 never-smokers, 23 smokers with normal lung function, and 25 patients with COPD (Global Initiative for Chronic Obstructive Lung Disease 2-4). Proportions of the IL-17/IFN-γ-double expressing subpopulation were assessed using flow cytometry. Plasma concentrations of cytokines favoring Tc17/IFN-γ differentiation were measured by enzyme-linked immunosorbent assay.

RESULTSPatients with COPD had higher proportions of Tc17 cells and Tc17/IFN-γ cells in the peripheral blood than smokers and never-smokers. The plasticity of Tc17 cells was higher than that of Th17 cells. The percentages of Tc17 cells and Tc17/IFN-γ cells showed negative correlations with forced expiratory volume in 1 s % predicted value (r = -0.418, P = 0.03; r = -0.596, P = 0.002, respectively). The plasma concentrations of IL-6, transforming growth factor-β1, and IL-12 were significantly higher in patients with COPD compared with smokers and never-smokers.

CONCLUSIONSPeripheral Tc17 cells are increased and more likely to convert to Tc17/IFN-γ cells in COPD, suggesting that Tc17 cell plasticity may be involved in persistent inflammation of the disease.

Aged ; Female ; Forced Expiratory Volume ; Humans ; Interferon-gamma ; biosynthesis ; Interleukin-12 ; blood ; Interleukin-6 ; blood ; Lung ; physiopathology ; Male ; Middle Aged ; Pulmonary Disease, Chronic Obstructive ; immunology ; physiopathology ; Th17 Cells ; immunology

OBJECTIVEThis study aimed to conduct a preliminary study on the possible role and significance of interleukin (IL)-12 and IL-27 in the pathogeneses of oral lichen planus (OLP).

METHODSThirty cases of patients with OLP (fifteen cases of reticular OLP and fifteen cases of erosive OLP) were enrolled in this study, and twenty cases of healthy people served as controls. Lymphocyte subsets CD3+, CD4+, CD8+, CD19+, and CD16+56 [natural killer cell (NK)] were tested using flow cytometry, and humoral immunity [immunoglobulin (Ig)G, IgA, IgM, C3, C4] were examined using nephelometry assays. IL-12 and IL-27 contents in serum of patients with OLP and normal controls were detected through enzyme linked immunosorbent assay. The correlations between the levels of IL-12, IL-27, immune status, and clinical characteristics of patients with OLP were analyzed, respectively.

RESULTSCD3+, CD4+, and CD8+in patients with OLP were markedly lower than the normal value, whereas CD 19+ of OLP in patients was significantly higher than the normal value (P<0.05). IgM inpatients with OLP was increased, whereas C4 was declined (P<0.05). IL-12 and IL-27 levels showed significant upregulation or ULF patients compared with control groups (P<0.05). Meanwhile, positive correlations existed between IL-12 andIL-27 levels in the serum of patients with OLP (r=0.912, P<0.01). No significant correlations of IL-12 and IL-27 epressions with clinical characteristics of OLP were found (P>0.05). Negative correlations of IL-12 and IL-27 levels with CD16+56(NK) cells were observed (r1 = -0.416, P1 = 0.022; r2 = -0.392, P2=0.032, respectively), whereas a positive correlation existed for IgG (r1=0.445, P1=0.014; n=0.549, P2=0.002, respectively).

CONCLUSIONA cellular immune dysfunction mainly dominate in patients with OLP, accompanied by some degree of humoral-immunity-function disorder. The abnormally high expressions of IL-12 and IL-27 are possibly synergized and promoted inflammation development in OLP. Its promotion takes place through the negatie feedback regulation of humoral immune responses, which are involved in the regulation of immune mechanisms of OLP.

Flow Cytometry ; Humans ; Immunoglobulins ; blood ; Interleukin-12 ; blood ; Interleukin-12 Subunit p35 ; metabolism ; Interleukin-27 ; blood ; Interleukins ; metabolism ; Killer Cells, Natural ; Lichen Planus, Oral ; blood ; immunology

OBJECTIVETo investigate the effects of intragastric administration of human interferon-α (hIFN-α)-transformed Bifidobacterium on immune functions of mice.

METHODSThe E.coli-Bifidobacterium shuttle expression vector containing hIFN-α gene was constructed and transformed into Bifidobacterium. The hIFN-α-transformed Bifidobacterium suspension (1010 /ml) was prepared after induction with 0.2% L-arabinose for hIFN-α expression and administered intragastrically in male Balb/C mice at the dose of 0.1 ml every other day for 2 weeks, with the mice receiving empty vector-transformed Bifidobacteria as the negative control and those having an equal volume of saline as the blank control. The percentages of mononuclear cell subsets in the thymus, spleen and blood were detected in the mice by flow cytometry, and the serum levels of IL-4, IL-12, IFN-γ and TNF-α were assayed using mouse cytokine FlowCytomix Kit.

RESULTSThe percentages of CD3⁺CD8⁺ and CD4⁺CD8⁺ cells in the thymus, CD3⁺CD4⁺, CD3⁺CD8⁺ and CD4⁺CD8⁺ cells in the spleen, and CD3⁺CD8⁺ cells in the blood all increased significantly in IFN group as compared with those in the negative and blank control groups (P<0.01 or 0.05). The serum level of IFN-γ also increased significantly (P<0.05) while IL-4 level remained unchanged in IFN group compared with those in the two groups.

CONCLUSIONIntragastric administration of hIFN-α-transformed Bifidobacterium promotes lymphocyte proliferation and maturation and increases the serum levels of Th1 cytokines in mice.

Animals ; Bifidobacterium ; Cell Proliferation ; Genetic Vectors ; Humans ; Interferon-alpha ; pharmacology ; Interferon-gamma ; blood ; Interleukin-12 ; blood ; Interleukin-4 ; blood ; Lymphocyte Activation ; drug effects ; Male ; Mice ; Mice, Inbred BALB C ; Recombinant Proteins ; pharmacology ; Spleen ; cytology ; Th1 Cells ; cytology ; Thymus Gland ; cytology ; Tumor Necrosis Factor-alpha ; blood

The present study was aimed to investigate the effects of exercise and nutrition intervention on rat immune function. Flor-Essence is a kind of health food produced by FLORA company in Canada and certified by Quality Assurance International (QAI). Its main components are burdock root, cress leaves of grass, kelp, Turkish rhubarb root, et al. Flor-Essence has been shown to activate the body detoxification path, improve the physical environment, and inhibit cancer cell growth and proliferation. Sixty male Sprague-Dawley (SD) rats were randomly divided into six groups: control, NS + training, low-dose Flor-Essence + training, low-dose Flor-Essence, high-dose Flor-Essence + training, high-dose Flor-Essence groups. The rats in NS + training, low-dose Flor-Essence + training, high-dose Flor-Essence + training groups swam 35 min per day in the water tank for 6 days a week. One hour before exercise, the rats were given low- (2.5 mg/mL) or high-dose (5 mg/mL) Flor-Essence daily by intragastric administration, and the rats in NS + training group were given equivalent volume of NS. On the last day of four training weeks, all rats took part in a bout of exhaustive exercise, and then were sacrificed immediately. Arterial blood serum samples were taken for the assays of IL-6, IL-12 and TNF-α contents, spleens for natural killer (NK) cells activity. The results showed that serum IL-6 content in NS + training group was decreased compared with that in control group. Low- and high-dose Flor-Essence groups showed decreased IL-6, IL-12 and TNF-α serum contents, as well as longer exhaustive time, compared with control group. The improving effects of high-dose Flor- Essence on IL-6, TNF-α and exhaustive time were greater than those of low dose. Compared with NS + training, low- and high-dose Flor-Essence + training reduced serum contents of IL-6 and TNF-α, and prolonged exhaustive time; only high-dose Flor-Essence + training decreased serum IL-12 content and enhanced NK cells activity. These results suggest Flor-Essence in combination with exercise can improve rat immune function and sports performance, and the effect of high-dose Flor-Essence is better than that of low dose.
Animals ; Interleukin-12 ; blood ; Interleukin-6 ; blood ; Killer Cells, Natural ; drug effects ; Male ; Physical Conditioning, Animal ; Plant Extracts ; pharmacology ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Swimming ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVETo study the effect of Fuzheng Sanjie recipe in regulating tumor-associated macrophages (TAMs) in Lewis lung cancer mice.

METHODEfforts were made to establish the Lewis lung cancer mouse model, weigh tumors and calculate the anti-tumor rate. The immunohistochemical method was used to examine the infiltration degree of CD68 + in tumor tissues in each group. ELISA was used to examine the content of IFN-γ, TGF-β, IL-4, IL-13, IL-6, IL-10, IL-12, TNF-α in mice serum.

RESULTCompared with the tumor-bearing model group, all of the other groups showed higher tumor inhibition rates, i. e. 50.28% for the DDP group, 34.37% for the TCM-preventing group and 66.76% for the Chinese and western medicine group, with statistical difference (P < 0.05), but without statistical difference in the infiltration degree of CD68+. The expressions of the IFN-γ, IL-6, IL-12 in tumor-bearing groups were lower than that in the blank control group, but with higher contents of IL-4, IL-13, TGF-β. Intervened with different drugs, there were significant differences in content among some relevant cytokines (P < 0.05), as well as statistical differences among the TCM prevention group, the Chinese and western medicine group and the tumor-bearing control group (P <0. 05) , but without statistical difference in TNF-α and IL-10 content from the tumor-bearing control group (P < 0.05).

CONCLUSIONFuzheng Sanjie recipe could reverse the immune remodeling effect and control the tumor growth by down-regulating the expressions of IL-4, IL-13, TGF-α in lung cancer immune microenvironment and up-regulating the expression of IFN-γ.

Animals ; Cell Line, Tumor ; Disease Models, Animal ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Interleukin-10 ; blood ; Interleukin-12 ; blood ; Interleukin-13 ; blood ; Lung Neoplasms ; blood ; drug therapy ; immunology ; Macrophages ; drug effects ; immunology ; Male ; Mice ; Mice, Inbred C57BL ; Transforming Growth Factor beta ; blood ; Tumor Necrosis Factor-alpha ; blood

Toxoplasma gondii is an intracellular protozoan parasite that causes a Th1 cellular immunity. Our previous study showed that T. gondii lysate antigen (TLA) treatment in S180 tumor-bearing mice resulted in tumor reduction by suppressing CD31 expression, a marker of angiogenesis. In the present study, to investigate tumor suppressive effect of TLA under the absence of T lymphocytes, athymic nude mice were compared with euthymic mice in the anti-tumorigenic effect triggered by TLA in CT26 tumors. According to the results, intratumorally injected TLA reduced tumor growth and TIMP-1 level, a metastatic marker, in both euthymic and athymic mice. TLA treatment led to a sharp increase in IL-12 expression in serum cytokine profiling of athymic mice, and increased MyD88 signals in macrophages derived from the bone marrow, implying the activation of innate immunity. The selective induction of IL-12 by TLA treatment had an anti-tumorigenic effect.
Animals ; Antigens, Protozoan/*immunology ; Disease Models, Animal ; Immunity, Innate ; Immunotherapy/*methods ; Interleukin-12/*blood ; Macrophages/immunology ; Mice, Inbred BALB C ; Mice, Nude ; Myeloid Differentiation Factor 88/analysis ; Neoplasms/pathology/*therapy ; Toxoplasma/*immunology ; Treatment Outcome

The relationship between T cell immunoglobulin domain and mucin domain protein 3 (Tim-3)/Galectin (Gal)-9 pathway and recurrent spontaneous abortion (RSA) was studied. Thirty-one pregnant women with RSA and 27 normal early gravidas were investigated to detect the levels of Tim-3 and Gal-9 in villi and deciduas by Western blotting. Meanwhile, the concentration of interleukin (IL)-4 and IL-12 in peripheral blood plasma was determined by ELISA in 25 healthy fertile non-pregnant controls, the normal early gravidas and pregnant women with RSA mentioned above, respectively. It was found that the relative expression levels of Tim-3 and Gal-9 in villi and deciduas were significantly increased in pregnant women with RSA as compared with those in the normal early gravidas. The concentration of IL-4 in peripheral blood plasma of pregnant women with RSA was lower than that of the normal early gravidas (P<0.05) and healthy fertile non-pregnant controls (P<0.05), but that of IL-2 in pregnant women with RSA was significantly higher than that of the normal early gravidas (P<0.05) and healthy fertile non-pregnant controls (P<0.05). It was suggested that the overexpression of Tim-3/Gal-9 pathway may be related to the pathogenesis of RSA.
Abortion, Spontaneous ; metabolism ; pathology ; Adolescent ; Adult ; Chorionic Villi ; metabolism ; pathology ; Female ; Galectins ; biosynthesis ; Hepatitis A Virus Cellular Receptor 2 ; Humans ; Interleukin-12 ; blood ; Interleukin-4 ; blood ; Membrane Proteins ; biosynthesis ; Pregnancy ; Pregnancy Proteins ; biosynthesis ; Up-Regulation

OBJECTIVETo study the infiltration of macrophages and their phenotype in the healing process of full-thickness wound in rat.

METHODSThirty healthy SD rats were divided into control group (n = 6) and injury group (n = 24) according to the random number table. Two round full-thickness skin defects (11 mm diameter) were created on both sides of dorsal spine of rats in injury group with surgical scissors and homemade trephine. After injury, wound area was measured immediately. The wounds were disinfected with iodophor every day. Rats in control group received anesthesia and hair removal only. On post injury day (PID) 1, 3, 7, and 13, respectively, 6 rats of injury group were sacrificed after the measurement of wound area (wound healing rate was calculated). Wound samples were obtained by excision down to healthy fascia along wound edge. Histological study was done with HE staining. The expression of CD68 (the surface marker of macrophage) in the wound tissue was observed with immunohistochemical staining. The double positive expressions of induced nitric oxide synthase (iNOS) plus CD68 (type I macrophage) and arginase 1 (Arg-1) plus CD68 (type II macrophage) were observed with immunofluorescence staining. The levels of interferon-γ (IFN-γ), TNF-α, IL-4, IL-13, IL-10, and IL-12 in wound tissue were assayed by double-antibody sandwich ELISA, and the ratio of IL-10/IL-12 was calculated. Full-thickness skin tissues (11 mm diameter) in rats of control group were excised at the same site as rats in injury group, and the histological observation and cytokines assay were performed as well. Data were processed with one-way analysis of variance or LSD- t test.

RESULTSWound area of rats in injury group was gradually reduced after injury, and the overall difference of the wound healing rate on each PID was statistically significant (F = 358.55, P < 0.01). No abnormal appearance of skin tissue was observed in rats of control group. In injury group, inflammatory cell infiltration was obvious in wound tissue on PID 1 and 3; vascular structure and fresh collagen were observed in wound tissue on PID 7 and 13. Numbers of CD68 positive cells in skin tissue of rats in control group and wound tissue of rats in injury group on PID 1, 3, 7, and 13 were respectively (2.7 ± 1.5), (31.8 ± 3.5), (40.8 ± 4.7), (20.8 ± 2.8), (3.2 ± 2.4) per 200 times visual field (F = 180.55, P < 0.01). Compared with that in control group, the number of CD68 positive cells of rats in injury group was increased on PID 1, 3, and 7 (with t values respectively 18.81, 18.79, 14.05, P values below 0.01). No double positive expression of iNOS plus CD68 or Arg-1 plus CD68 was observed in normal tissue of rats in control group. In injury group, proportions of iNOS plus CD68 double positive cells on PID 1, 3, 7, and 13 were respectively (12.2 ± 2.8)%, (16.5 ± 2.9)%, (4.2 ± 2.3)%, (0.7 ± 0.8)% (F = 72.50, P < 0.01); proportions of Arg-1 plus CD68 double positive cells on PID 1, 3, 7, and 13 were respectively 0, (8.2 ± 1.9)%, (21.5 ± 3.4)%, (4.7 ± 2.0)% (F = 120.93, P < 0.01). In injury group, proportion of iNOS plus CD68 double positive cells on PID 3 was significantly higher than that on other PID (with t values respectively 2.65, 8.17, 12.95, P values below 0.05); proportion of Arg-1 plus CD68 double positive cells on PID 7 was higher than that on other PID (with t values respectively 15.27, 8.25, 10.38, P values below 0.01). Compared with that of Arg-1 plus CD68 double positive cells, proportion of iNOS plus CD68 double positive cells was higher on PID 1 and 3 (with t values respectively 10.71 and 5.88, P values below 0.01) and lower on PID 7 and 13 (with t values respectively 10.24 and 4.60, P values below 0.01). The overall differences of IFN-γ, TNF-α, IL-4, IL-13, and IL-10/IL-12 ratio in skin tissue of rats in control group and wound tissue of rats in injury group on every PID were statistically significant (with F values from 14.08 to 631.03, P values below 0.01). Compared with those in control group, levels of IFN-γ, TNF-α, IL-4, and IL-13 in wound tissue of rats in injury group were significantly higher on every PID (with t values from 4.58 to 9.17, P values below 0.05), while IL-10/IL-12 ratio was significantly higher on PID 1, 3, and 7 (with t values respectively 27.70, 30.51, 9.49, P values below 0.05) . In injury group, IFN-γ level on PID 1 [(61 ± 5) pg/mL] and IL-10/IL-12 ratio on PID 3 (1.647 ± 0.098) were significantly higher than those of control group and those on other PID in injury group [with IFN-γ level respectively (32 ± 4), (54 ± 6), (46 ± 7), (47 ± 4) pg/mL and IL-10/IL-12 ratio respectively 0.328 ± 0.045, 0.960 ± 0.034, 0.530 ± 0.028, 0.289 ± 0.040, with t values respectively from 3.19 to 8.20 and from 16.59 to 31.84, P values below 0.05].

CONCLUSIONSMacrophage infiltration increases in the healing process of full-thickness wound in rat with different phenotypes, among which type I macrophage appears in the inflammatory stage, and type II macrophage predominates in the proliferative stage.

Animals ; Antigens, CD ; genetics ; metabolism ; Antigens, Differentiation, Myelomonocytic ; genetics ; metabolism ; Collagen ; Enzyme-Linked Immunosorbent Assay ; Interferon-gamma ; Interleukin-10 ; Interleukin-12 ; Interleukin-13 ; Interleukin-4 ; Macrophages ; Male ; Phenotype ; Rats ; Skin ; injuries ; Tumor Necrosis Factor-alpha ; blood ; Wound Healing ; genetics

OBJECTIVETo observe the effect of Ningdong Granule (NG) on serum levels of interleukin-12 (IL-12) and tumor necrosis factor-alpha (TNF-alpha) of children patients with Tourette's syndrome (TS).

METHODSTotally 90 TS children patients were randomly assigned to the NG group, the NG + Tiapride group (abbreviated as the combined treatment group), and the Tiapride group, 30 in each group. Besides,another 30 healthy children were recruited as the healthy control group. Patients in the NG group were treated with NG (consisting of all gastrodia rhizome, Codonopsis pilosula, Ophiopogon japonicus, white peony root, Rhinocerotidae, oyster, earthworm, licorice root, etc.), one dose daily, administered by dissolving it in boiled water, taken in two portions in the morning and in the evening respectively. Patients in the Tiapride group took Tiapride Tablet, 50 -100 mg each time, twice daily. The dosage was adjusted according to individual difference and changes of pathogenic conditions. The maximal dosage was 300 mg per day. Those in the combined treatment group were treated with equal dose of NG and Tiapride Tablet in combination. The treatment course was 3 months for all. Changes of pathogenic condition before and after treatment were assessed by Yale global tic severity scale (YGTSS). Serum levels of IL-12 and TNF-alpha were detected by enzyme-labeled immunosorbent assay (ELISA) before and after treatment.

RESULTS(1) The total effective rate of the NG group, the combined treatment group, and the Tiapride group was 79.3%, 83.3%, and 67.9%, respectively. It was the lowest in the Tiapride group (P < 0.05). It was significantly higher in the combined treatment group than in the NG group (P < 0.05). (2) The post-treatment YGTSS score was obviously lower in each group after treatment than before treatment (P < 0.05). The posttreatment YGTSS score was obviously lower in the NG group and the combined treatment group than in the Tiapride group (P < 0.05), but with no statistical difference between the fromer two groups (P > 0.05).(3) Compared with the healthy control group before treatment, serum levels of IL-12 and TNF-alpha (pg/mL) were 124.95 +/- 22.78 and 209.52 +/- 21.69 in the NG group, 126.14 +/- 25.65 and 208.97 +/- 22.46 in the combined treatment group, 123.00 +/- 24.26 and 205.10 +/- 26.16 in the Tiapride group, being higher than those in the healthy control group (64.56 +/- 27.59 and 78.13 +/- 33.42; P < 0.05). After treatment, serum levels of of IL-12 and TNF-alpha were 104.67 +/- 16.84 and 183.01 +/- 24.95 in the NG group, 109.04 +/- 16.81 and 179.87 +/- 23.45 in the combined treatment group, significantly lower than before treatment (P < 0.05). But there was no statistical difference in serum levels of IL-12 or TNF-alpha in the Tiapride group between before treatment (123.00 +/- 24.26 and 205.10 +/- 26.16) and after treatment (117.75 +/- 16.79 and 199.76 +/- 33.21; P > 0.05).

CONCLUSIONNG could modulate abnormal serum levels of IL-12 and TNF-alpha in TS children patients, which might be one of its pharmacodynamic mechanisms for treating TS.

Adolescent ; Child ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Interleukin-12 ; blood ; Male ; Phytotherapy ; Tourette Syndrome ; blood ; drug therapy ; Tumor Necrosis Factor-alpha ; blood