IL-8 and IFN-γ as Preoperative Predictors of the Outcome of Tonsillectomy.

OBJECTIVES: Tonsillectomy (TE) and tonsillotomy (TO) due to recurrent episodes of acute tonsillitis (RAT) belong to the most frequent surgical procedures. However, an adequate objective marker predicting the outcome of TE/TO preoperatively is missing. METHODS: Patients with RAT who underwent TE/TO (n = 31) were included in this pilot study. A panel of cytokines and chemokines in serum and saliva were determined preoperatively. Health-related quality of life was assessed pre- and postoperatively by the Tonsillectomy Outcome Inventory-14. RESULTS: Health-related quality of life improved significantly after surgery. Increased serum levels of interleukin-8 (IL-8) and interferon gamma (IFN-γ) are associated with a less successful outcome. No correlation between the number of acute tonsillitis episodes and the health-related quality of life after TE or TO could be observed. CONCLUSIONS: Tonsillectomy and TO improve health-related quality of life independently from the number of past acute tonsillitis episodes. Interleukin-8 and IFN-γ in serum may serve as promising markers, predicting the benefit of TE or TO for patients preoperatively.

Significance of IL28RA in diagnosis of early pancreatic cancer and its regulation to pancreatic cancer cells by JAK/STAT signaling pathway - effects of IL28RA on pancreatic cancer.

OBJECTIVE: To explore the significance of IL28RA in diagnosis of early pancreatic cancer and its regulation to pancreatic cancer cells by the Janus kinase/signal transducer and activator of transcription (JAK/STAT) signaling pathway. PATIENTS AND METHODS: A total of 81 patients with early pancreatic cancer were enrolled as a pancreatic cancer group, and 81 patients with benign pancreatic diseases were enrolled as a benign disease group. Western blot was adopted to analyze the serum IL28RA expression of the two groups and its diagnostic value in early pancreatic cancer. A pancreatic cancer cell model was constructed, and the IL28RA expression in pancreatic cancer cells, PANC-1 and BXPC-3, was up-regulated to explore the biological function of pancreatic cancer cells after up-regulation of IL28RA and the effects on JAK-STAT signaling pathway. RESULTS: Lowly expressed in serum of patients with pancreatic cancer, IL28RA showed a sensitivity of 80.25%, specificity of 75.31%, and area under the curve (AUC) of 0.846 in diagnosis of early pancreatic cancer. It was found that up-regulation of IL28RA expression in pancreatic cancer cells inhibited proliferation and invasion abilities of pancreatic cancer cells, increased apoptosis rate and expression of pro-apoptotic protein bax, decreased expression of anti-apoptosis protein bcl-2, and significantly inhibited phosphorylation level of JAK2 and STAT3 proteins. CONCLUSIONS: IL28RA is lowly expressed in pancreatic cancer patients, and has certain diagnostic value for early pancreatic cancer. Its up-regulated expression can inhibit the proliferation and invasion of pancreatic cancer cells, and promote their apoptosis by inhibiting the activation of JAK-STAT signaling pathway.

Granulocyte colony-stimulating factor downregulates interferon-gamma receptor expression and stimulates interleukin-6 production in activated human macrophages.

We studied direct effects of human granulocyte colony-stimulating factor (G-CSF) on phenotypical properties of human macrophage cells in vitro. CD14+ monocyte/macrophages (Mc/Mphs) were isolated from blood of healthy donors by positive magnetic separation. G-CSF (0.01-1.0 ng/mL), when added to Mc/Mphs along with lipopolysaccharide (LPS, 1.0 µg/mL), was able to noticeably reduce proportions of CD119 (interferon-γ receptor 1)-positive cells, with no stable effects on CD16 (FcγRIII)+ and СD124 (IL-4 receptor subunit alpha)-positive cells. In addition, G-CSF markedly upregulated IL-6 production by LPS-activated Mph cells, without significantly affecting IL-1ß, IL-10 and tumor necrosis factor-α (TNF-α) secretion. Our data suggests that G-CSF could restrain Mph polarization to pro-inflammatory (M1) phenotype, thus potentially supporting pro-regenerative Mph activity with implications for immunotherapeutic interventions.

[Lymphocyte subpopulations, levels of interferon, and expression of their receptors in patients with chronic hepatitis B and C: Correlation with the species of viruses and the degree of liver fibrosis]. / Subpopuliatsii limfotsitov, uroven' interferonov i ékspressiia ikh retseptorov u bol'nykh khronicheskimi gepatitami V i S: zavisimost' ot vida virusov i stepeni fibroza pecheni.

AIM: To determine whether there is a correlation of the composition of circulating lymphocyte subpopulations, the serum concentrations of interferon (IFN)-α, IFN-γ, and IFN-λ3, and the lymphocyte expression of types I and II IFN receptors with the species of a disease pathogen and the degree of liver fibrosis (LF) in patients with chronic hepatitis B (CHB) and in those with chronic hepatitis C (CHC). SUBJECTS AND METHODS: The investigation enrolled 44 patients with CHC, 9 patients with CHB, and 13 clinically healthy donors. The degree of LF in the patients was determined using transient elastography. The composition of peripheral blood lymphocyte subpopulations was examined; the concentrations of IFN-α, IFN-γ, and IL-28B were estimated. RESULTS: Lymphocyte counts were higher in patients with CHC and in those with CHB than those in healthy donors; and the number of neutrophils was lower. There were no differences between the groups in the composition of lymphocyte subpopulations with the exception of the number of CD3+CD4+ cells, which in patients with CHC was larger than in those with CHB. In CHC and CHB patients, the counts of CD118+ lymphocytes were higher than those in healthy donors. Patients with CHB and those with CHC did not differ between themselves and from healthy donors in the expression of CD119 on the lymphocytes. In CHC patients, the relative CD119+ cell counts were higher between CD4+ lymphocytes than those in healthy donors. The serum levels of IFN-α and IFN-γ in CHC and CHB patients were similar, but higher in healthy donors. The concentration of IL-28B genotype in patients with CHC was twice as high as in those with CHB, but the differences were statistically insignificant. The number of lymphocytes increased with the progression of fibrosis; that of neutrophils decreased. There was an inverse relationship between platelet counts and LF severity. Multiple comparisons of the clusters of patients with different degrees of LF revealed no differences in the number of major lymphocyte subpopulations. However, the number of CD3+CD16+CD56+ natural killer-like T (NKT) cells correlated with fibrosis severity. Patients with different degrees of LF showed no differences in the proportion of CD118- and CD119 cells between lymphocytes and in the serum levels of IFN-α, IFN-γ, and IL-28B levels. Patients with grade IV LF displayed a higher proportion of CD4+CD119+ lymphocytes between CD45+ cells than did those with grade III LF. CONCLUSION: Several new clinical and laboratory trends were identified and the nature and extent of previously described hematological and immunological changes were clarified in CHC or CHB patients with various degrees of LF. Some indicators may be used as additional criteria for the prognosis of the above forms of hepatitis, and a number of newly described facts suggest that it is necessary to revise the protective/phlogogenic value of types I, II, and III IFNs in chronic viral hepatitis C and B.

[ASSESSMENT OF INDICATORS FOR INTERFERON SYSTEM IN PATIENTS WITH COMMON FORMS OF PULMONARY TUBERCULOSIS RECEIVING COMPLEX THERAPY WITH CYCLOFERON].

We have evaluated the efficacy of cycloferon inclusion in the complex therapy of newly diagnosed patients with common forms of pulmonary tuberculosis, based on monitoring of the number of monocytes with receptors to interferon-gamma (flow cytometry) and the concentration of interferon-gamma in the serum (ELISA). For this purpose, a group of 36 patients (18 patients received standard chemotherapy, and 18 additionally received 600 mg cycloferon tablets 3 times per week) was examined for 3 months. Control group consisted of 18 apparently healthy patients. The study did not include patients with multiple or extensively drug-resistant M. tuberculosis strains. The analysis of results showed a statistically significant positive dynamics of the level of monocyte receptors to interferon-gamma in patients receiving cycloferon as manifested by an increase in their number in the first 2 months of therapy (period of clinical manifestations of the disease), followed by a decrease in the 3rd months of treatment, which corresponds to clinical improvement, in contrast to patients treated with standard chemotherapy alone.

The alteration and clinical significance of Th22/Th17/Th1 cells in patients with chronic myeloid leukemia.

T helper- (Th-) cell immunodeficiency plays important roles in tumor development and their effects in chronic myeloid leukemia (CML) remain unclear. In the present study, we mainly investigated the role of Th22, Th17, and Th1 cell and their related cytokines (IL-22, IL-17, and IFN-r) in the pathophysiology of CML. Bone marrow (BM) and peripheral blood (PB) were extracted from newly diagnosed (ND), chronic phase- (CP-) CML patients, and controls. Th subsets were examined by flow cytometry. Plasma IL-22, IL-17, and IFN-r concentrations were measured by ELISA. AHR and RORC mRNA expressions were examined by RT-PCR. The frequencies of Th22, Th17, and Th1 cells, along with the expression of specific transcription factors RORC and AHR, were significantly decreased in ND patients compared with healthy controls, while all these abnormality recovered in CP patients. In addition, there existed a significantly positive relationship between Th22 and Th17 cells in PB or BM. A significantly negative relationship was found between Th cells (Th22, Th17, or Th1) and BCR-ABL (%) IS or the number of PB white blood cells. All these results demonstrated that Th22, Th17, and Th1 cells might be important therapeutic targets in CML and could facilitate a better outcome for tumor immunotherapy.

Changes in natural killer cell subpopulations over a winter training season in elite swimmers.

Immune changes and increased susceptibility to infection are often reported in elite athletes. Infectious episodes can often impair training and performance with consequences for health and sporting success. This study monitored the occurrence of episodes of upper respiratory symptoms (URS) and the variation in circulating NK cells, CD56(bright) and CD56(dim) NK cells subpopulations, over a winter swimming season. Nineteen national elite swimmers and 11 non-athlete controls participated in this study. URS episodes were monitored using daily log books. Blood samples were taken at rest at four time points during the season: before the start of the season (t1--middle September), after 7 weeks of an initial period of gradually increasing training load (t2--early November), after 6 weeks of an intense training cycle (t3--late February) and 48 h after the main competition (t4--early April) and from the controls at three similar time points (t1--early November; t2--late February; t3--early April). In the swimmers, the occurrence of URS clustered around the periods of elevated training load (67 %). No URS were reported at equivalent time points in the non-athletes. Athletes showed a decrease in the percentage (t2 = 21 %; t3 = 27 %; t4 = 17 %) and absolute counts of circulating NK cells (t2 = 35 %; t3 = 22 %; t4 = 22 %), coinciding with the periods of increased training load, never recovering to the initial values observed at the start of the season. The reduction in the CD56(dim) and an increase in the CD56(bright) NK cell subpopulations were significant at t2 and t3 (p < 0.05). Concomitant with the fall in values of NK cells, in athletes that shown more than three URS episodes, a moderate correlation (r = 0.493; p = 0.036) was found between CD56(bright)/CD56(dim) ratio and the number of URS episodes after the more demanding training phase (t3). At t3, a lower value of CD56 cell counts was found in the group who reported three or more URS episodes (t = 2.239; p = 0.032). A progressive significant decrease in the expression of CD119, the receptor for IFN-γ, on the CD56(dim) cells was found over the season and an elevation in Granzyme B expression was coincident with the more demanding training phases. Periods of highly demanding training seem to have a negative impact on innate immunity mediated by NK cell subsets, which could partially explain the higher frequency of URS observed during these training phases.

[Cytokine regulation in patients with brain tumours].

The aim of the study was to investigate serum levels, as well as the ability to produce in vitro, some pro-inflammatory and anti-inflammatory cytokines in patients with brain tumours (BT). Serum concentrations of IFN-gamma, IFN-alpha, TNF-alpha, IL-4, IL-6, IL-8, and RA IL-1 were determined in 17 patients with brain tumours, and in 21 healthy donors. The results showed that baseline levels of cytokines studied was above normal in most patients (78%) had the highest rates of IFN-gamma. During the whole observation period, slightly higher in patients compared with healthy individuals (t=1.8) was the content of IFN-alpha. Serum concentrations of IL-4 in patients before treatment were significantly higher than in control group (p<0.01), remaining at about the same level, and after treatment. Levels of Rail-1, IL-6 and IL-8 in patients before treatment were within normal limits, while remaining unchanged at the end of the survey. Indicators induced in vitro production of cytokines studied to some extent correlated with their serum levels. Hyperproduction in vitro IFN-gamma was observed in 61% of patients, low production - 28% of patients, and only in 11% of cases, these figures were within normal limits. In patients with malignant brain tumors were identified violations of serum levels and production of cytokines studied. The most significant was the increase of PHA-induced production of IFN-gamma, which was not reduced by the end of treatment. High serum and, in particular, PHA-induced levels of IFN-gamma at the end of treatment may be recommended as an additional criterion for immunodiagnosis in a dynamic laboratory monitoring.

Detection of HIV gp120 in plasma during early HIV infection is associated with increased proinflammatory and immunoregulatory cytokines.

Events that occur during acute HIV infection likely contribute to the immune dysfunction common in HIV-infected individuals. During this early stage, there is high-level viral replication, loss in CD4(+) T cell number and function, and an up-regulation of proinflammatory and immunoregulatory cytokines. The mechanisms responsible for this are not completely understood. We hypothesize that the HIV envelope glycoprotein, gp120, contributes to immune dysfunction during early HIV infection. Using a cohort of subjects enrolled during acute and early HIV infection, we determined the amount of gp120, TNF-α, IL-6, IL-10, IFN-α, and IFN-γ in plasma at baseline and 6 months. At matched time points, we also measured CD4(+) T cell proliferation, T cell activation, and apoptosis. Plasma from 109 subjects was screened for gp120. Thirty-six subjects (33%) had detectable gp120 (0.5-15.6 ng/ml). Subjects with greater than 1 ng/ml of gp120 at baseline had similar levels at all time points tested, even when viral replication was undetectable due to therapy. Subjects with detectable gp120 had higher levels of plasma IL-6, IL-10, and TNF-α. There was no difference in the level of T cell activation, proliferation, or apoptosis in subjects with gp120 compared to those without. We conclude that persistent expression of gp120 occurs in a subset of individuals. Furthermore, the presence of gp120 is associated with higher levels of plasma IL-6, IL-10, and TNF-α, which may contribute to immune dysfunction during early HIV infection.

Comparison of cytokine levels in depressed, manic and euthymic patients with bipolar disorder.

BACKGROUND: The neurobiology of bipolar disorder is not completely understood. Cytokines have received increasing attention as potential mediators of the interaction with immune, neuroendocrine system and specific pathways involved in mood, energy, and activity control. Previous reports have suggested the association of mania and bipolar depression with a proinflammatory state. However, they did not compare cytokine levels in all phases of bipolar disorder. METHODS: Sixty-one bipolar patients were recruited for assessment of serum cytokine levels. Of these, 14 were in euthymic state, 23 and 24 were in manic and depressive episodes, respectively. A healthy comparison group included 25 healthy volunteers. Cytokines involved in Th1/Th2 balance, such as TNF-alpha, IL-2, IL-4, IL-6, IL-10, IFN-gamma, were examined by flow cytometry. RESULTS: During mania, proinflammatory cytokines, IL-2, IL-4 and IL-6, were increased in comparison with healthy subjects. Patients in depressive episode showed only increased IL-6 levels. There were no significant differences in cytokine levels between patients in remission and healthy subjects, except for IL-4. Mood symptoms showed a positive correlation with IL-6 and IL-2. DISCUSSION: These findings suggest that mania, and to a less extent, depression are associated with a proinflammatory state. These changes seem to be related to mood state, as changes in cytokine profile were more pronounced during acute episodes than in euthymia. This study provides further support to investigate the immune system as a target for future treatment development.

Age-dependent decrease in serum soluble interferon-gamma receptor (sIFN-gammaR) in healthy Japanese individuals; population study of serum sIFN-gammaR level in Japanese.

We planned to investigate the clinical significance of serum soluble interferon-gamma receptor (sIFN-gammaR) level in pediatric patients. The diagnostic application of the measurement of serum sIFN-gammaR level depends critically on the control value. However, there is no information of the control value of serum sIFN-gammaR for children. In the present study, we determined the serum sIFN-gammaR level of healthy Japanese children using an ELISA. The serum sIFN-gammaR level of children (0-14 years old) was significantly higher than that of adults (over 15 years old) (p < 0.01, n = 104). Thus, it is recommended that, when the serum sIFN-gammaR level of patients is evaluated, it should be compared against age-matched controls. We also preliminarily applied this assay as a diagnostic parameter for the patients with diarrhea positive (D+) hemolytic uremic syndrome (HUS).

Interferon inhibitory activity in patients with multiple sclerosis.

BACKGROUND: Interferon inhibitory activity (IIA) is a logical candidate for explaining neutralizing antibody-negative partial responsiveness to interferon beta in multiple sclerosis (MS), but its role has not been evaluated. OBJECTIVE: To investigate the role of IIA and soluble interferon-alpha/beta receptor (sIFNR) in determining response of patients with MS to interferon beta therapy. DESIGN: Parallel-group, open-label study. SETTING: Baird Multiple Sclerosis Center, Buffalo, NY. Patients Blood was obtained before and 24 hours after injection of interferon beta-1a from 38 anti-interferon beta neutralizing antibody-negative patients with relapsing-remitting MS and 16 untreated healthy controls. On the basis of clinical parameters of response to interferon beta therapy, the patients were divided into stable or good-responder (n = 20) and active or partial-responder (n = 18) groups. MAIN OUTCOME MEASURES: Quantitative analyses of magnetic resonance imaging were obtained; the IIA and sIFNR levels were measured using bioassay and enzyme-linked immunosorbent assay, respectively. RESULTS: The IIA and sIFNR levels were elevated in MS patients compared with controls (P<.001). The IIA levels were higher in active or partial responders compared with stable or good responders (P<.001); the sIFNR levels were not different between groups. The Extended Disability Status Score and T2 lesion volumes were higher in the active or partial-responder group compared with the stable or good-responder group. Interferon beta-1a did not have short-term effects on the IIA and sIFNR levels. In univariate general linear model and stepwise regression analyses, IIA levels were associated with T2 lesion volume. CONCLUSION: The levels of IIA are associated with increased MS disease activity and with responsiveness to interferon beta therapy in anti-interferon beta neutralizing antibody-negative MS patients.

Interferon-alpha receptor 1 mRNA expression in peripheral blood mononuclear cells is associated with response to interferon-alpha therapy of patients with chronic hepatitis C.

Interferon (IFN)-alpha receptor mRNA expression in liver of patients with chronic hepatitis C has been shown to be a response to IFN-alpha therapy. The objective of the present study was to determine whether the expression of mRNA for subunit 1 of the IFN-alpha receptor (IFNAR1) in peripheral blood mononuclear cells (PBMC) is associated with the response to IFN-alpha in patients with chronic hepatitis C. Thirty patients with positive anti-HCV and HCV-RNA, and abnormal levels of alanine aminotransferase in serum were selected and treated with IFN-alpha 2b for one year. Those with HBV or HIV infection, or using alcohol were not included. Thirteen discontinued the treatment and were not evaluated. The IFN-alpha response was monitored on the basis of alanine aminotransferase level and positivity for HCV-RNA in serum. IFNAR1-mRNA expression in PBMC was measured by reverse transcription-polymerase chain reaction before and during the first three months of therapy. The results are reported as IFNAR1-mRNA/beta-actin-mRNA ratio (mean +/- SD). Before treatment, responder patients had significantly higher IFNAR1-mRNA expression in PBMC (0.67 +/- 0.15; N = 5; P < 0.05) compared to non-responders (0.35 +/- 0.17; N = 12) and controls (0.30 +/- 0.16; N = 9). Moreover, IFNAR1-mRNA levels were significantly reduced after 3 months of treatment in responders, whereas there were no differences in IFNAR1 expression in non-responders during IFN-alpha therapy. Basal IFNAR1-mRNA expression was not correlated with the serum level of alanine and aspartate aminotransferases or the presence of cirrhosis. The present results suggest that IFNAR1-mRNA expression in PBMC is associated with IFN-alpha response to hepatitis C and may be useful for monitoring therapy in patients with chronic hepatitis C.

Expression of IFN-gamma and its receptor alpha in the peripheral blood of patients with chronic hepatitis C.

BACKGROUND: It has been known that intra-cellular immunity is important for defense against viral infections and this function lies with interferon gamma (INF-gamma). Here we evaluated the role of IFN-gamma system in the pathogenesis of chronic hepatitis C (CHC). METHODS: The levels of interferon gamma receptor alpha (IFNGR alpha) on the peripheral lymphocyte membrane were assayed with flow cytometry. The plasma concentrations of the cytokines IFN-gamma and IL-10 in CHC patients and normal controls were assayed by enzyme-linked-immunosorbent assay (ELISA). The samples were collected randomly from Xinjiang Autonomous Region, Zhejiang and the northern regions of Jiangsu Province in China. RESULTS: The levels of IFNGR alpha in CHC patients were significantly lower than that of normal controls (NC), especially among patients during the stable stage (P < 0.001), whereas there were no significant differences between CHC in active and stable stages. Among the patients of the three regions, there were no significant differences between patients from Xinjiang and Zhejiang provinces, but both had statistically significant difference compared with the patients from Jiangsu Province (P < 0.001). Plasma IFN-gamma and IL-10 concentrations in CHC patients decreased significantly, IFN-gamma in particular, but there were no significant differences in these levels between various stages of the disease. The IFN-gamma/IL-10 (Th1/Th2) ratio in patients was reversed. CONCLUSION: There may be defects in the IFN-gamma system in chronic HCV infected subjects and a low immune response, which may play an important role in the persistence of HCV infection.

[The clinical significance of interferon-gamma and its receptor in patients with schistosomiasis].

OBJECTIVE: To study the expression of interferon-gamma (IFN-gamma) and type I interferon-y receptor (IFN-gammaRI) in the chronic and advanced patients of schistosomiasis japonica, and to discuss its clinical relationship with hepatic fibrosis. Methods IFN-yR1 in the peripheral blood lymphocytes was detected by ELISA, and the hyaluronic acid (HA) , collagen type IV (C-IV), procollagen type III (PC III), laminin (LN) were detected by radio-immunoassay (RIA) in schistosomiasis patients. The level of IFN-y, IFN-gammaR1 and serum markers of hepatic fibrosis were observed, and the relationship with each other was analyzed by statistical method. RESULTS: There was no difference in the expression of IFN-gamma and IFN-gammaR1 between the patients with chronic schistosomiasis and the normal group (P > 0. 05 ), the IFN-gammaR1 in advanced cases without splenectomy was low (P <0. 05) , but IFN-y was high (P < 0. 01). The two indicators in the advanced schistosomiasis patients with splenectomy returned to normal. There was no corresponding relationship between the two indicators and HA, C-IV, PC III, LN with a r value of 0. 19, 0.20, 0. 14, and 0.21 respectively. CONCLUSION: There is a corresponding relationship between IFN-gamma and IFN-gammaR1; the expression of IFN-gammaR1 is related to the course of schistosomiasis, and the relationship with hepatic fibrosis needs further study.

Free interferon-alpha/beta receptors in the circulation of patients with adenocarcinoma.

BACKGROUND: Many viral and neoplastic diseases are resistant to interferon-alpha/beta (IFN-alpha/beta) therapy or develop resistance during the course of IFN treatment. In patients with viral diseases, the authors identified four IFN inhibitors, of which the most important, most likely is a free IFN receptor of type 1 appearing in the circulation that captures and neutralizes IFN-alpha/beta. METHODS: Ninety-one cancer patients and 25 healthy individuals were studied. Free circulating IFN receptor-alpha/beta type 1 was studied. The patients were ages 35-75 years. The diagnoses were 24 cases of colon carcinoma, 7 cases of prostate carcinoma, 16 cases of breast carcinoma, 8 cases of ovarian carcinoma, 9 cases of uterine carcinoma, 5 cases of lung carcinoma, 3 cases of astrocytoma, 4 cases of transitional cell carcinoma of the bladder, 1 case of osteosarcoma, 3 cases of multiple myeloma, 4 cases of Hodgkin disease, 2 cases of non-Hodgkin lymphoma, 3 cases of myelodysplastic syndrome, and 2 disseminated tumors of unknown origin. RESULTS: All patients were found to have increased free IFN receptor-alpha/beta type 1 in the circulation, with the highest levels reported in patients with adenocarcinoma. CONCLUSIONS: High IFN inhibitory activity in patients with cancer may be a significant factor in their increased susceptibility to progressive disease, infectious complications, and resistance to IFN therapy. Ongoing studies are being performed with the objective of overcoming this inhibitory activity.

Integrated measurements by flow cytometry of the cytokines IL-2, IFN-gamma, IL-12, TNF-alpha and functional evaluation of their receptors in human blood.

Immunodeficiencies might be caused not only by the lack of cytokine production, but also by defective expression and/or function of the cytokine receptors. We have measured by flow cytometry, within 2 days, not only the production of IL-2, IFN-gamma, IL-12 and TNF-alpha, but also the functional expression of the receptors for these cytokines in blood samples obtained from 15 healthy donors and 13 patients suffering from tuberculosis. Cytoplasmic and surface staining with monoclonal antibodies (mAbs) was used to assess the production of cytokines and their receptors, respectively, after polyclonal stimulation. To evaluate receptor activity, peripheral blood mononuclear cells (PBMC) were first incubated with the corresponding recombinant human (rh) cytokine. CD69 was detected on lymphocytes after incubation with rhIL-2; IFN-gamma was detected in lymphocytes after co-stimulation with rhIL-12 plus PHA; iNOS induction and upregulation of major histocompatibility complex (MHC) II and MHC I was detected on monocytes after recombinant human interferon-gamma (rhIFN-gamma) stimulation; finally, COX-2 expression and MHC II upregulation were detected on monocytes after rhTNF-alpha stimulation. The assay that was developed can be used clinically to assess the activities of components of the cytokine signaling pathways of patients with immunodeficiencies or those with chronic intracellular infections such as tuberculosis.

[Modulation and implications in pathogenesis of interferon gamma receptor 1 in patients with chronic hepatitis B].

OBJECTIVE: To study the possible differences in the interferon gamma receptor (IFN-gamma R1) response among a variety of clinical types in patients with chronic hepatitis B (CHB) and implications in pathogenesis. METHODS: The serum level of IFN-gamma and the expression level of IFN-gamma R1 in peripheral leucocytes, from 53 CHB patients, were examined by ELISA and flow cytometry respectively, which were compared with the baseline levels of 15 healthy controls and were performed correlation analysis with alanine aminotransferase (ALT), total bilirubin (TBil) in serum and morphological change in hepatic tissues. RESULTS: The results showed that the level of IFN-gamma R1 (28.89% 11.77%) expressed on the membranes of lymphocytes in CHB patients, which correlated with liver inflammation (r=0.621, P<0.01) and serum TBil level (r=0.575, P<0.01), was much higher than that (9.23% 1.30%) of the healthy controls (Z=3.988, P<0.05), and no obvious difference on the membranes of leucocytes. The serum levels of IFN-gamma in patients with cirrhosis and severe hepatitis were higher than those of healthy controls. And the two was no difference from each other, but the standard deviation in each group was relatively large. CONCLUSION: These findings suggest that the IFN-gamma signal transduction pathway is modulated through up-regulating the expression of IFN-gamma R1 on the membranes of lymphocytes, which takes part in the immuno-pathogenesis in CHB patients.

High levels of interferon gamma in the plasma of children with complete interferon gamma receptor deficiency.

We have found that children with complete interferon gamma (IFNgamma) receptor deficiency, unlike patients with other genetic defects predisposing them to mycobacterial diseases, have very high levels of IFNgamma in their plasma. This unexpected observation provides a simple and accurate diagnostic method for complete IFNgamma receptor deficiency in children with clinical disease caused by bacille Calmette-Guérin vaccines or environmental nontuberculous mycobacteria.

Interleukin-18 binding protein in acute graft versus host disease and engraftment following allogeneic peripheral blood stem cell transplants.

Dysregulation of the cytokine network plays an important role in graft-versus-host disease (GVHD). Interleukin-18 (IL-18) is an obligatory cytokine for interferon-gamma (IFN-gamma) production and IFN-gamma and sIFN-gammaR are elevated in patients with GVHD. Because IL-18 binding protein (IL-18BP) is an inhibitor of IL-18-mediated IFN-gamma production, we evaluated IL-18BP levels in patients undergoing allogeneic peripheral blood stem cell transplantation (PBSCT). IL-18BP levels were assessed in 14 patients on day -10 (before conditioning), on the day of transplant, on the day of engraftment, and during transplant-related complications. A comparison of the kinetics of IL-18BP and soluble(s) IL-6R, sIFN-gammaR, IL-18 serum levels was performed. IL-18BP levels were assessed by specific monoclonal antibodies in a double-sandwich enzyme-linked immunosorbent assay (ELISA). In all patients IL-18BP levels decreased during conditioning and increased in parallel with engraftment (p < 0.05). Accordingly, during rejection, IL-18BP serum levels remained low and similar to pretransplant levels. The mean elevation of IL-18BP detected in association to acute GVHD was significantly higher in comparison to normal engraftment (p < 0.05). A correlation between IL-18BP, sIFNgammaR, and sIL-6R serum levels was found in all patients. No correlation between IL-18 and IL-18BP serum levels was found in patients undergoing uneventful PBSCT and rejection, whereas a marked increase in both IL-18 and IL-18BP levels was detected during acute GVHD (p < 0.01). Our data suggest that the dysregulation of IL-18 and IL-18BP may be important in the pathophysiology of transplant-related complications. Furthermore, because preliminary data from our group show that IL-18 blockage ameliorates GVHD in murine models, it is inferred that these cytokines may represent potential targets in the development of new therapeutic strategies in acute GVHD.