CD56(+) T cells are abundant in liver and play an important role in defense against viral infections. However, the role of CD56(+) T cells in control of hepatitis C virus (HCV) infection remains to be determined. We investigated the noncytolytic anti-HCV activity of primary CD56(+) T cells in human hepatocytes. When HCV Japanese fulminant hepatitis-1 (JFH-1)-infected hepatocytes were co-cultured with CD56(+) T cells or incubated in media conditioned with CD56(+) T cell culture supernatants (SN), HCV infectivity and replication were significantly inhibited. The antibodies to <em>interferon</em> (IFN)-gamma or IFN-gamma receptor could largely block CD56(+) T cell-mediated anti-HCV activity. Investigation of mechanism(s) responsible for CD56(+) T cell-mediated noncytolytic anti-HCV activity showed that CD56(+) T SN activated the multiple elements of janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway and enhanced the expression of IFN regulatory factors (IRFs) 1, <em>3</em>, 7, 8, and 9, resulting in the induction of endogenous IFN-<em>alpha</em>/beta expression in hepatocytes. Moreover, CD56(+) T SN treatment inhibited the expression of HCV-supportive micro RNA (miRNA)-122 and enhanced the levels of anti-HCV miRNA-196a in human hepatocytes.

CONCLUSIONS

These findings provide direct in vitro evidence at cellular and molecular levels that CD56(+) T cells may have an essential role in innate immune cell-mediated defense against HCV infection. (HEPATOLOGY 2009.).

OBJECTIVE

Two types of interferons (IFNs), type I (IFN-alpha/beta) and type III (IFN-lambdas), utilize distinct receptor complexes to induce similar signalling and biological activities, including recently demonstrated for IFN-lambdas antitumour activity. However, ability of type III IFNs to regulate cell population growth remains largely uncharacterized.

METHODS

Intact and modified human colorectal adenocarcinoma HT29 cells were used to study regulation of apoptosis by IFN-lambdas.

CONCLUSIONS

We report that the IFN-lambdaR1 chain of the type III IFN receptor complex possesses an intrinsic ability to trigger apoptosis in cells. Signalling induced through the intracellular domain of IFN-lambdaR1 resulted in G(1)/G(0) phase cell cycle arrest, phosphatidylserine surfacing and chromosomal DNA fragmentation. Caspase-3, caspase-8 and caspase-9 were activated; however, pancaspase inhibitor Z-VAD-FMK did not prevent apoptosis. In addition, the extent of apoptosis correlated with the level of receptor expression and was associated with prolonged IFN-lambda signalling. We also demonstrated that the ability to trigger apoptosis is a unique intrinsic function of all IFN receptors. However, more robust apoptosis was induced by signalling through type III IFN receptor than through type I or type II (IFN-gamma) receptors, suggesting higher cytotoxic potential of type III IFNs. In addition, we observed that IFN-gamma treatment sensitized HT29 cells to IFN-lambda-mediated apoptosis. These results provide evidence that type III IFNs, alone or in combination with other stimuli, have the potential to induce apoptosis.

Type I <em>interferons</em> (IFNs) play critical roles in the host defense by modulating the expression of various genes via the IFN-dependent activation of signal transducers and activators of transcription and NF-kappaB (nuclear factor kappa B) transcription factors. Previous studies established that IFN<em>alpha</em>/beta activates NF-kappaB to promote cell survival through a phosphatidylinositol <em>3</em>-kinase (PI<em>3</em>K)/Akt pathway, which involves serine phosphorylation and degradation of IkappaB <em>alpha</em>. We now describe a second pathway by which IFNs activate NF-kappaB that is independent of IkappaB degradation. This pathway involves NF-kappaB-inducing kinase (NIK) and the tumor necrosis factor receptor-associated factor-2 (TRAF2) and results in IFN<em>alpha</em>/beta-induced processing of the p100/NF-kappaB2 precursor into p52. IFN<em>alpha</em>/beta stimulates NF-kappaB DNA binding and NF-kappaB-dependent transcription. Whereas expression of NIK and TRAF2 constructs causes NF-kappaB activation, expression of dominant negative NIK and TRAF2 constructs blocks IFN-promoted NF-kappaB activation and IFN-stimulated kappaB-dependent transcription and IFN<em>alpha</em>/beta-induced processing of the p100/NF-kappaB2 precursor into p52. In contrast, PI<em>3</em>K does not mediate IFN<em>alpha</em>/beta-induced p100 processing, although PI<em>3</em>K is involved in the pathway resulting in IkappaB <em>alpha</em> degradation. Moreover, whereas IFN promotes cell survival in lymphoblastoid cells, expression of dominant negative NIK and TRAF2 constructs enhances IFN-induced apoptosis. Our results for the first time place NIK and TRAF2, previously shown to function in TNF signaling, within the IFN signal transduction pathway. Thus, IFN induces NF-kappaB activation to mediate IFN-dependent cell survival signals through a "canonical" pathway of IkappaB <em>alpha</em> proteolysis mediated by PI<em>3</em>K/Akt and a "noncanonical" pathway of p100 processing mediated by NIK/TRAF.

We investigated the primary cellular immune responses to human immunodeficiency virus type 1 (HIV-1) Env and Gag proteins elicited by recombinant vesicular stomatitis viruses (rVSVs). The primary response to Env peaked 5 to 7 days after intraperitoneal vaccination, at which time 40% of CD8(+) cells were Env tetramer positive and activated (CD62L(Lo)). These freshly isolated cells actively lysed target cells pulsed with the p18-I10 peptide and secreted gamma <em>interferon</em> and tumor necrosis factor <em>alpha</em> after stimulation with the Env p18-I10 peptide. The primary response to Env elicited by rVSVs was sixfold higher than that elicited by recombinant vaccinia viruses (rVVs) at 5 days postvaccination. An intranasal route of vaccination with VSV-Env also elicited a strong primary response to Env. The primary immune response to Gag elicited by rVSV peaked 7 days after vaccination, at which time <em>3</em>% of CD8(+) cells were Gag tetramer positive and CD62L(Lo) and functional by intracellular cytokine staining. This response was eightfold higher than that elicited by rVV expressing Gag. VSV-GagEnv, which expresses both Gag and Env from a single recombinant, also induced strong cytotoxic T-lymphocyte (CTL) responses to both Env and Gag. Our quantitative analyses illustrate the potency of the VSV vector system in CTL induction.

OBJECTIVE

To determine the circulating levels of Th1 and Th2 cytokines in patients with systemic lupus erythematosus (SLE) and to elucidate their association with disease activity and autoimmune response.

METHODS

We included 52 patients and 25 healthy controls. Serum levels of tumor necrosis factor (TNF) <em>alpha</em>, <em>interferon</em> (IFN) gamma, interleukin (IL)-12p70, IL-10, and IL-4, as well as anti-DNA, -Ro, -La, -RNP, and -Sm antibodies were determined by enzyme-linked immunosorbent assay. Disease activity was recorded according to the Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) and classified as very active (SLEDAI>> or = 1<em>3</em>), moderately active (SLEDAI: <em>3</em>-12), or inactive (SLEDAI < or = 2).

RESULTS

The mean age of the patients was <em>3</em>4.2 +/- 12.6 years, and the mean duration of disease was 4.9 +/- 7.6 years. Twelve patients (2<em>3</em>%), 20 patients (<em>3</em>4.5%), and 20 patients (<em>3</em>4.5%) had highly, moderately, and inactive SLE, respectively. Levels of IFN-gamma, TNF-<em>alpha</em>, and IL-12 were significantly higher in patients than in healthy controls (P <.0<em>3</em>), as well as the IL-12/IL-10, IL-12/IL-4, IFN/IL-10, IFN/IL-4, TNF/IL-10, and TNF/IL-4 ratios (P <.01), suggesting a major participation of Th1 over Th2 cytokines. Nevertheless, a direct correlation between Th1 (IFN-gamma and TNF-<em>alpha</em>) and Th2 (IL-4 and IL-10) cytokines was observed in patients (r>>.5, P <.01), indicating a mutual Th1-Th2 participation. TNF-<em>alpha</em> levels and the TNF/IL-10 ratio were higher in patients with inactive disease compared with patients with very active disease and controls (P <.04). IL-12 levels and IL-12/IL-4, as well as IL-12/IL-10, ratios were higher in patients with very active disease than in those with inactive SLE and controls (P <.01). IL-10 levels were associated with anti-DNA, anti-Ro, and anti-La response (P <.01).

CONCLUSIONS

Our results suggest that TNF-alpha could be a protective factor in SLE patients, whereas IL-12p70 participates in disease activity and IL-10 influences the autoimmune response (autoantibody production).

Infection by hepatitis C virus (HCV) usually results into chronic hepatitis that can ultimately lead to cirrhosis and hepatocellular carcinoma. Type 1 <em>interferons</em> (IFN-<em>alpha</em>/beta) constitute the primary cellular defense against viral infection including HCV. IFN binding to their receptors activates associated Jak1 and Tyk2 kinases, which ultimately leads to phosphorylation and assembly of a signal transducer and activator of transcription protein (STAT)1-STAT2-<em>interferon</em> regulatory factor (IRF)9 trimetric complex called <em>interferon</em>-stimulated gene factor <em>3</em> that translocates into the nucleus and binds to the <em>interferon</em>- stimulated response elements (ISRE), leading to transcriptional induction of several antiviral genes, including double-stranded RNA-activated protein kinase (PKR), 2',5'- oligoadenylate synthetase (OAS), and myxovirus resistance protein A (MxA). Understanding the mechanisms of how the virus evades this cellular innate defense and establishes a chronic infection is the key for the development of better therapeutics against HCV infection. Here, we demonstrate that p5<em>3</em> could have a crucial role in the cellular innate defense against HCV. We observed significantly higher levels of HCV RNA replication and viral protein expression in the Huh7 cells when their p5<em>3</em> expressions were knocked down. Moreover, IFN treatment was less effective in inhibiting the HCV RNA replication in the p5<em>3</em>-knocked-down (p5<em>3</em>kd) Huh7 cells. In fact, the activation of the ISRE and the induction of ISGs were significantly attenuated in the p5<em>3</em>kd Huh7 cells and p5<em>3</em> was found to directly interact with IRF9.

CONCLUSIONS

These observations underscore the potential contributions of the tumor suppressor p5<em>3</em> in cellular antiviral immunity against HCV with possible therapeutic implications.

BACKGROUND

Based on a 2-year survival of 43%, the Gastrointestinal Tumor Study Group (GITSG) recommended adjuvant 5-FU-based chemoradiation for resected patients with adenocarcinoma of the pancreatic head. Here we report improved survival over the GITSG protocol with a novel adjuvant chemoradiotherapy based on interferon-alpha (IFNalpha).

METHODS

From July 1993 to September 1998, 33 patients with adenocarcinoma of the pancreatic head underwent pancreaticoduodenectomy (PD) and subsequently went on to adjuvant therapy (GITSG-type, n = 16) or IFNalpha-based (n = 17) typically given between 6 and 8 weeks after surgery. The latter protocol consisted of external-beam irradiation at a dose of 4,500 to 5,400 cGy (25 fractions per 5 weeks) and simultaneous three-drug chemotherapy consisting of (1) continuous infusion 5-FU (200 mg/m2 per day); (2) weekly intravenous bolus cisplatin (30 mg/m2 per day); and (3) IFNalpha (3 million units subcutaneously every other day) during the 5 weeks of radiation. This was then followed by two 6-week courses of continuous infusion 5-FU (200 mg/m2 per day, given weeks 9 to 14 and 17 to 22). Risk factors for recurrence and survival were compared for the two groups.

RESULTS

A more advanced tumor stage was observed in the IFNalpha-treated patients (positive nodes and American Joint Committee on Cancer [AJCC] stage III = 76%) than the GITSG group (positive nodes and stage III = 44%, P = 0.052). The 2-year overall survival was superior in the IFNalpha cohort (84%) versus the GITSG group (54%). With a mean follow-up of 26 months in both cohorts, actuarial survival curves significantly favored the IFNalpha group (P = 0.04).

CONCLUSIONS

With a limited number of patients, this phase II type trial suggests better survival in the interferon group as compared with the GITSG group even though the interferon group was associated with a more extensive tumor stage. The 2-year survival rate in the interferon group is the best published to date for resected pancreatic cancer. The interferon/cisplatin/5-FU-based adjuvant chemoradiation protocol appears to be a promising treatment for patients who have undergone PD for adenocarcinoma of the pancreatic head.

OBJECTIVE

The aim of this study was to clarify the correlation between cytokine profile in colonic mucosa with disease activity and response to granulocytapheresis (GCAP) in patients with ulcerative colitis (UC), using a reliable, reproducible quantitative method.

METHODS

Colonoscopic biopsies of inflamed colonic mucosa (16 patients, 21 cases) and uninflamed colonic mucosa (25 patients, 33 cases) were obtained from UC patients. Messenger (m)RNA was extracted and subjected to realtime polymerase chain reaction for quantitative measurement of interleukin (IL)-12, interferon-gamma, tumor necrosis factor-alpha, IL-4, IL-8, and IL-18 mRNAs. In seven patients with high disease activity despite prednisolone (PSL) treatment >> or = 20 mg/day), one course of GCAP was conducted, and pre- and post-GCAP cytokine profiles were determined.

RESULTS

In inflamed colonic mucosa of UC patients, three cytokine profiles were observed: 1) high expression of interferon-gamma, tumor necrosis factor-alpha, and IL-4 mRNAs but low expression of IL-8 mRNA; 2) high expression of IL-8 mRNA and low expression of others; and 3) low expression of all cytokines examined. Inflamed colonic mucosa of patients with high disease activity showed the second pattern. Inflamed colonic mucosa of patients who were not treated with PSL and who had low disease activity showed the first pattern, whereas those on high-dose PSL exhibited the second pattern. IL-8 mRNA was significantly higher in inflamed UC samples than in uninflamed samples. GCAP was effective in five of seven PSL-resistant patients (71.4%). IL-8 was the only cytokine that correlated with effectiveness of GCAP. Compared with GCAP nonresponders, responders had significantly higher IL-8 mRNA before GCAP and showed marked reduction of IL-8 mRNA after GCAP.

CONCLUSIONS

IL-8 mRNA was significantly increased in inflamed mucosa of UC. Patients with high IL-8 mRNA expression in colonic mucosa despite PSL treatment were responsive to GCAP. Therefore, quantitative measurement of mucosal IL-8 mRNA may be useful in predicting the response to GCAP.

OBJECTIVE

There is evidence to suggest that hepatitis C virus (HCV) infection is a high-risk condition for developing type 2 diabetes. However, there are no interventional studies that confirm that HCV infection causes diabetes. The main aim of this study was to compare the incidence of glucose abnormalities (diabetes plus impaired fasting glucose) between HCV-infected patients with or without sustained virological response (SVR) after antiviral therapy.

METHODS

Patients with normal fasting glucose (<100 mg/dl) with biopsy-proven chronic hepatitis C without cirrhosis and with at least <em>3</em> years of follow-up after finishing antiviral therapy were included in the study (n = 2<em>3</em>4). Patients received <em>interferon</em> <em>alpha</em>-2b (alone or with ribavirin) for 6 or 12 months according to genotype. Cumulative incidence of glucose abnormalities was evaluated by using the Kaplan-Meier method comparing subjects with and without a SVR to antiviral treatment. A multivariate Cox proportional hazards analysis was performed to explore the variables independently associated with the development of glucose abnormalities.

RESULTS

During follow-up, 14 of 96 (14.6%) patients with SVR and 47 of 1<em>3</em>8 (<em>3</em>4.1%) nonsustained responders developed glucose abnormalities (P = 0.001). Patients with SVR did not develop diabetes during follow-up, whereas nine cases of diabetes were detected in nonsustained responders (P = 0.007). After adjustment for the recognized predictors of type 2 diabetes, the hazard ratio for glucose abnormalities in patients with SVR was 0.48 (95% CI [0.24-0.98], P = 0.04).

CONCLUSIONS

Our results provide evidence that eradication of HCV infection significantly reduces the incidence of glucose abnormalities in chronic hepatitis C patients. In addition, this study supports the concept that HCV infection causes type 2 diabetes.

The effects of arachidonic acid ethanolamide (anandamide), palmitoylethanolamide and delta9-tetrahydrocannabinol on the production of tumor necrosis factor-<em>alpha</em> (TNF-<em>alpha</em>), interleukin-4, interleukin-6, interleukin-8, interleukin-10, <em>interferon</em>-gamma, p55 and p75 TNF-<em>alpha</em> soluble receptors by stimulated human peripheral blood mononuclear cells as well as [<em>3</em>H]arachidonic acid release by non-stimulated and N-formyl-Met-Leu-Phe (fMLP)-stimulated human monocytes were investigated. Anandamide was shown to diminish interleukin-6 and interleukin-8 production at low nanomolar concentrations (<em>3</em>-<em>3</em>0 nM) but inhibited the production of TNF-<em>alpha</em>, <em>interferon</em>-gamma, interleukin-4 and p75 TNF-<em>alpha</em> soluble receptors at higher concentrations (0.<em>3</em>-<em>3</em> microM). Palmitoylethanolamide inhibited interleukin-4, interleukin-6, interleukin-8 synthesis and the production of p75 TNF-<em>alpha</em> soluble receptors at concentrations similar to those of anandamide but failed to influence TNF-<em>alpha</em> and <em>interferon</em>-gamma production. The effect of both compounds on interleukin-6 and interleukin-8 production disappeared with an increase in the concentration used. Neither anandamide nor palmitoylethanolamide influenced interleukin-10 synthesis. delta9-Tetrahydrocannabinol exerted a biphasic action on pro-inflammatory cytokine production. TNF-<em>alpha</em>, interleukin-6 and interleukin-8 synthesis was maximally inhibited by <em>3</em> nM delta9-tetrahydrocannabinol but stimulated by <em>3</em> microM delta9-tetrahydrocannabinol, as was interleukin-8 and <em>interferon</em>-gamma synthesis. The level of interleukin-4, interleukin-10 and p75 TNF-<em>alpha</em> soluble receptors was diminished by <em>3</em> microM delta9-tetrahydrocannabinol. [<em>3</em>H]Arachidonate release was stimulated only by high delta9-tetrahydrocannabinol and anandamide concentrations (<em>3</em>0 microM). These results suggest that the inhibitory properties of anandamide, palmitoylethanolamide and delta9-tetrahydrocannabinol are determined by the activation of the peripheral-type cannabinoid receptors, and that various endogenous fatty acid ethanolamides may participate in the regulation of the immune response.

Little is known about the regulation and coordinated expression of genes involved in the innate host response to Candida albicans. We therefore examined the kinetic profile of gene expression of innate host defense molecules in normal human monocytes infected with C. albicans using microarray technology. Freshly isolated peripheral blood monocytes from five healthy donors were incubated with C. albicans for 0 to 18 h in parallel with time-matched uninfected control cells. RNA from monocytes was extracted and amplified for microarray analysis, using a 42,421-gene cDNA chip. Expression of genes encoding proinflammatory cytokines, including tumor necrosis factor <em>alpha</em>, interleukin 1 (IL-1), IL-6, and leukemia inhibitory factor, was markedly enhanced during the first 6 h and coincided with an increase in phagocytosis. Expression of these genes returned to near baseline by 18 h. Genes encoding chemokines, including IL-8; macrophage inflammatory proteins 1, <em>3</em>, and 4; and monocyte chemoattractant protein 1, also were strongly up-regulated, with peak expression at 4 to 6 h, as were genes encoding chemokine receptors CCR1, CCR5, CCR7, and CXCR5. Expression of genes whose products may protect monocyte viability, such as BCL2-related protein, metallothioneins, CD71, and SOCS<em>3</em>, was up-regulated at 4 to 6 h and remained elevated throughout the 18-h time course. On the other hand, expression of genes encoding T-cell-regulatory molecules (e.g., IL-12, gamma <em>interferon</em>, and transforming growth factor beta) was not significantly affected during the 18-h incubation. Moreover, genes encoding IL-15, the IL-1<em>3</em> receptor (IL-1<em>3</em>Ra1), and CD14 were suppressed during the 18-h exposure to C. albicans. Thus, C. albicans is a potent inducer of a dynamic cascade of expression of genes whose products are related to the recruitment, activation, and protection of neutrophils and monocytes.

BACKGROUND

Hepatocellular carcinoma (HCC) patients with major vascular involvement or extrahepatic metastasis are not good candidates for surgery or transarterial chemoembolization (TACE). In this study, the authors evaluated the efficacy of combined therapy with intraarterial cisplatin infusion and systemic administration of interferon-alpha (IFN-alpha) as a palliative treatment for these patients.

METHODS

Sixty-eight HCC patients with major portal vein thrombosis (n = 47) or distant metastasis (n = 27) were randomly allocated to 1 of 3 groups. Group I (n = 19) received combined therapy consisting of intraarterial cisplatin infusion and systemic IFN-alpha, Group II (n = 23) received intraarterial cisplatin infusion, and Group III (n = 26) was managed with only supportive care. Cisplatin 2 mg/kg was infused through the proper hepatic artery every 8 weeks, and IFN-alpha 3 million IU/m2 was administered subcutaneously 3 times a week.

RESULTS

The partial response (defined as a 50% or greater reduction in the product of the 2 longest perpendicular tumor measurements) rate of Group I was significantly higher than that of Group II (33% vs. 14%; P < 0.05). Also, the 1-year survival rate of Group I (27%) was higher than that of Group II (9%) or Group III (0%) (P < 0.05 and P < 0.01, respectively). The median survival period of Group I was 19 weeks, which was significantly longer than that of Group II (11 weeks) or Group III (5 weeks) (P < 0.05 and P < 0.01, respectively).

CONCLUSIONS

These results suggest that combined therapy consisting of intraarterial cisplatin infusion and systemic IFN-alpha may be useful as a palliative treatment for HCC patients with major vascular involvement or extrahepatic metastasis.

BACKGROUND

The acute-phase response and anaemia of chronic disease are characterized by hypoferraemia associated with an increased ferritin synthesis, which might be mediated by the activated cytokine cascade.

METHODS

We examined the prolonged effects of isolated limb perfusion (ILP) with recombinant human tumour necrosis factor alpha (rTNF), recombinant human interferon gamma (rIFN-gamma) and melphalan on interleukin (IL) 6 and acute-phase protein levels, iron status and serum transferrin receptor (sTfR) levels in 12 patients with melanoma or sarcoma. Patients were treated with ILP during 90 min after pretreatment with rIFN-gamma during 2 days.

RESULTS

After ILP, leakage of TNF resulted in systemic peak levels at 3 min followed by an increase in IL-6 with maximum levels at 4h. C-reactive protein (CRP) rose at 4 h to peak levels at day 2, whereas alpha 1-antitrypsin and alpha 1-acid glycoprotein increased to maximum levels at day 3. Albumin and transferrin levels decreased after ILP and recovered after day 2. Serum iron and sTfR levels decreased during pretreatment and after ILP to minimum levels at 8 h and day 1 respectively. This was associated with an increase in serum ferritin levels, which paralleled CRP values.

CONCLUSIONS

Our data point to a central role for the cytokine network in the modulation of iron metabolism in the acute-phase response and anaemia of chronic disease. TNF, possibly via induction of IL-6, and IFN-gamma induce hypoferraemia, which may in part result from a decrease in tissue iron release based on a primary stimulation of ferritin synthesis. The fall in sTfR levels may reflect an impaired erythroid growth and/or TfR expression mediated by TNF and IFN-gamma.

Certain cytokines such as <em>interferon</em>-<em>alpha</em> and interleukin-2 are often used in the treatment certain cancers and chronic diseases such as melanoma, hepatitis C infection and multiple sclerosis. Several neuropsychiatric side effects such as depression, anxiety, psychosis, suicidal ideation, hypomanic mood and cognitive impairment were reported in those patients who received those medications. In certain patients with those neuropsychiatric side effects, the symptoms ceased when the medication was stopped. However, in some cases, the cognitive impairment persisted even for years after cessation of the medication. In animal studies, those cytokines could induce sickness behaviour, anxiety behaviour and social anhedonia. The increased in pro-inflammatory cytokines in certain neuropsychiatric disorders was widely reported. In addition, in animal studies, the treatment with <em>interferon</em>-<em>alpha</em> or interleukin-1 could induce depressive like behaviour. Recently, the role of certain pro-inflammatory cytokines that could enhance the activity of the enzyme, indoleamine 2-<em>3</em>, dioxygenase (IDO) which in turn would increase tryptophan degradation into kynurenine and decrease tryptophan availability of tryptophan in the brain to synthesize serotonin, a neurotransmitter which is necessary for the normal mood state became of interest in pathophysiology of psychiatric disorders. Furthermore, the imbalance in the further downward catabolic kynurenine pathway and their interactions with other neurotransmitters has been proposed to play an important role. The presence of such an imbalance in patients being treated with cytokines and in patients with psychiatric disorders and the possible consequence of those changes on the neuroprotective function in the brain are discussed in this review.

Modified vaccinia virus Ankara (MVA) is an attenuated poxvirus that has been engineered as a vaccine against infectious agents and cancers. Our goal is to understand how MVA modulates innate immunity in dendritic cells (DCs), which can provide insights to vaccine design. In this study, using murine bone marrow-derived dendritic cells, we assessed type I <em>interferon</em> (IFN) gene induction and protein secretion in response to MVA infection. We report that MVA infection elicits the production of type I IFN in murine conventional dendritic cells (cDCs), but not in plasmacytoid dendritic cells (pDCs). Transcription factors IRF<em>3</em> (IFN regulatory factor <em>3</em>) and IRF7, and the positive feedback loop mediated by IFNAR1 (IFN <em>alpha</em>/beta receptor 1), are required for the induction. MVA induction of type I IFN is fully dependent on STING (stimulator of IFN genes) and the newly discovered cytosolic DNA sensor cGAS (cyclic guanosine monophosphate-adenosine monophosphate synthase). MVA infection of cDCs triggers phosphorylation of TBK1 (Tank-binding kinase 1) and IRF<em>3</em>, which is abolished in the absence of cGAS and STING. Furthermore, intravenous delivery of MVA induces type I IFN in wild-type mice, but not in mice lacking STING or IRF<em>3</em>. Treatment of cDCs with inhibitors of endosomal and lysosomal acidification or the lysosomal enzyme Cathepsin B attenuated MVA-induced type I IFN production, indicating that lysosomal enzymatic processing of virions is important for MVA sensing. Taken together, our results demonstrate a critical role of the cGAS/STING-mediated cytosolic DNA-sensing pathway for type I IFN induction in cDCs by MVA. We present evidence that vaccinia virulence factors E<em>3</em> and N1 inhibit the activation of IRF<em>3</em> and the induction of IFNB gene in MVA-infected cDCs.

We investigated cytokine profiles in interleukin (IL)-4 transgenic (Tg) mice with a skin inflammatory disease resembling human atopic dermatitis. cDNA microarray revealed that the mRNAs encoding IL-1beta, IL-2, IL-<em>3</em>, IL-4, IL-5, IL-6, IL-10, IL-12p40, IL-1<em>3</em>, tumour necrosis factor (TNF)-<em>alpha</em>, TNF-beta and <em>interferon</em> (IFN)-gamma were up-regulated in the skin of late lesion Tg mice and to a lesser degree in non-lesion Tg mice when compared to those of non-Tg mice. Real time reverse transcription-polymerase chain reaction (RT-PCR) analyses indicated that the cDNA copy numbers of IL-1beta, IL-4, IL-6, IL-10, TNF-<em>alpha</em> and IFN-gamma from the skin of late, early and non-lesions increased significantly compared to non-Tg mice. IL-2 and IL-12p40 cDNA copy numbers were increased significantly in early, but not late, lesions. Interestingly, IL-1beta, IL-<em>3</em>, IL-4, IL-5, IL-6, IL-10, IL-1<em>3</em>, TNF-<em>alpha</em>, and IFN-gamma cDNAs were increased significantly the skin of before-onset and/or non-lesion mice. Flow cytometry analyses demonstrated an increased percentage of keratinocytes producing IL-4 as the disease progressed. The percentage of IL-2, IL-4, IL-10 and IFN-gamma-producing T cells and IL-12-producing antigen-presenting cells in skin-draining lymph nodes and inflammatory skin also increased, particularly in mice with late lesion. These results suggest that disease induction is primarily triggered by Th2 cytokines and that Th1, Th2 and non-Th proinflammatory cytokines are all involved in the disease process.

We describe the clinical characteristics, the patterns of association, and the role of antiviral therapies in patients with sarcoidosis associated with chronic hepatitis C virus (HCV) infection. Sixty-eight patients were included in the current study, 56 cases identified in the literature search plus 12 unpublished cases from our department. In 50 HCV patients, sarcoidosis appeared after starting antiviral therapy. Antiviral therapy associated with triggered sarcoidosis consisted of <em>alpha</em>-<em>interferon</em> monotherapy in 20 cases and combined therapy with <em>alpha</em>-<em>interferon</em> and ribavirin in <em>3</em>0. Sarcoidosis appeared during the first 6 months after starting therapy in 66% of patients. The clinical picture of sarcoidosis included predominantly pulmonary disease in <em>3</em>8 (76%) patients and cutaneous sarcoidosis in <em>3</em>0 (60%). Antiviral therapy was discontinued in 60% of patients and continued or adjusted in 14%, while sarcoidosis appeared after completed therapy in the remaining cases. Specific therapy for sarcoidosis was started in only 21 patients, mainly with oral corticosteroids. The outcome of patients was detailed in 46 cases: remission or improvement was observed in <em>3</em>8/46 (8<em>3</em>%) patients, stabilization of sarcoidosis in 5/46 (11%), and reactivation of sarcoidosis after an initial improvement in <em>3</em>/46 (6%). Finally, 18 treatment-naive HCV patients presented sarcoidosis, with 14/18 (87%) patients presenting with pulmonary involvement and 8/18 (44%) with cutaneous involvement. In summary, sarcoidosis may be observed in HCV patients in 2 different situations: triggered by antiviral therapy (in 75% of cases) and unrelated to treatment. Sarcoidosis during antiviral therapy may present mainly as cutaneous or pulmonary disease, with a benign, uncomplicated evolution in more than 85% of cases. However, more complicated cases are observed, especially in HCV patients with preexisting sarcoidosis and/or with previous antiviral treatment. Clinicians should be aware of the possibility that sarcoidosis may initially manifest or be reactivated during or shortly after treatment with antiviral therapy in patients with chronic HCV infection.

Many randomized trials have evaluated <em>alpha</em>-<em>interferon</em> as myeloma therapy, some suggesting a benefit and others not. Most were too small to give reliable answers, so a systematic overview has been performed to provide a more reliable estimate of the effect of <em>interferon</em>. The main end-points were response rates (induction trials), progression-free survival (PFS) and overall survival (OS). Individual patient data were supplied for 24 trials involving 4012 patients, 12 induction trials (2469 patients) and 12 maintenance trials (154<em>3</em> patients). In induction, response rates were slightly better with <em>interferon</em> (57.5% versus 5<em>3</em>.1%, P = 0.01). PFS was better with <em>interferon</em> (<em>3</em><em>3</em>% versus 24% at <em>3</em> years, P < 0.00001), an effect seen in both induction (P = 0.000<em>3</em>) and maintenance (P < 0.00001) trials. Median time to progression was increased by about 6 months in both settings. OS was somewhat better with <em>interferon</em> (5<em>3</em>% versus 49% at <em>3</em> years, P = 0.01) with median survival increased by about 4 months. This benefit was restricted to the smaller trials. The effect of <em>interferon</em> was not significantly related to the dose or duration of <em>interferon</em> or to patients' characteristics. PFS was improved with <em>interferon</em>, but the survival benefit, if any, was small and needs balancing against cost and toxicity.

We assessed the efficacy of <em>interferon</em> (IFN) <em>alpha</em>-2b plus ribavirin therapy in patients with hepatitis C virus (HCV)-related cirrhosis, and elucidated the risk factors for the development of hepatocellular carcinoma (HCC) to determine whether these therapies might reduce the incidence of HCC. One hundred and thirty-two HCV-cirrhotic patients receiving IFN <em>alpha</em>-2b (<em>3</em> or 5 MU thrice weekly) and oral ribavirin (1,000-1,200 mg/day) for 24 or 48 weeks were analysed. Cumulative incidence of HCC was estimated by the Kaplan-Meier method. The prognostic relevance of clinical variables and HCC occurrence was evaluated by univariate analysis with the log-rank test and by multivariate Cox's regression analysis. A total of 116 patients completed the treatment and 7<em>3</em> (55%) achieved a sustained virological response (SVR). Stepwise logistic regression analysis showed that nongenotype 1b (P < 0.001) and low viral load (P = 0.018) were independent variables of SVR. During a median follow-up period of <em>3</em>7 (12-6<em>3</em>) months, HCC developed in 11 patients with non-SVR and five with SVR (P = 0.0178), whereas there was no difference between those with transient biochemical response and nonresponse (P = 0.5970). The Kaplan-Meier method also showed that old age >>or=60 years) (P = 0.00<em>3</em>4) and genotype 1b (P = 0.0104) were associated with HCC occurrence. Using Cox's regression analysis, non-SVR (odds ratio = <em>3</em>.521, P = 0.0<em>3</em>6), male (odds ratio = 6.269, P = 0.011) and old age (odds ratio = <em>3</em>.076, P = 0.049) were independent significant risk factors contributing to HCC development. Our results suggest that achieving SVR by IFN <em>alpha</em>-2b plus ribavirin therapy may decrease the incidence of HCC in patients with HCV-related cirrhosis.

Treating acute hepatitis C with <em>interferon</em> <em>alpha</em> prevents chronicity in nearly all cases when therapy is initiated within <em>3</em> months after infection. However, 15-50% of untreated patients may clear the hepatitis C virus (HCV) spontaneously. Therefore, factors are needed to identify patients prior to therapy who have a higher or lower risk for developing a chronic course to avoid unnecessary treatment. The role of the HCV genotype for spontaneous recovery from acute hepatitis C has been discussed controversially. In the year 2002, all 1,176 new incoming prisoners in a Northern German prison for young men (age 16-24) were screened for anti-HCV antibodies and 92 tested positive. Ninety eight percent of these reported i.v.-drug abuse for a median of <em>3</em>2 months prior to imprisonment. HCV-RNA negative individuals (21%) were serotyped and HCV-RNA positive patients were genotyped. The prevalence of HCV genotype <em>3</em> was significantly higher among individuals who had cleared HCV spontaneously as compared to chronically infected patients (86% vs. <em>3</em>8%; P = 0.002). Ninety three percent of individuals exposed to HCV genotype 1 but only 6<em>3</em>% of individuals exposed to genotype <em>3</em> experienced a chronic course of the infection (P = 0.006). Thus, acute infection in young Caucasian men with HCV genotype <em>3</em> leads more often to spontaneous clearance than infection with HCV genotype-1. Considering also the high chance of successful treatment of chronic HCV genotype <em>3</em> infection with pegylated-<em>interferon</em> in combination with ribavirin, we suggest not to treat acute hepatitis C genotype <em>3</em> infection early but rather to wait at least <em>3</em> months after the onset of symptoms when chronicity becomes likely.

Recently, a collection of surface markers was exploited to isolate viable Lin(-) TdT(+) cells from murine bone marrow. These early pro-B cells were enriched for B-lineage lymphocyte precursor activity measured by short-term culture and had little responsiveness to myeloid growth factors. Early precursors can be propagated with remarkably high cloning frequencies in stromal cell-free, serum-free cultures, permitting this analysis of direct regulatory factors. Expression of the interleukin-7 receptor (IL-7R<em>alpha</em>) chain marks functional precursors and IL-7 is necessary for progression beyond the CD45RA(+) CD19(-) stage. Efficient survival and differentiation were only observed when stem cell factor and Flt-<em>3</em> ligand were also present. IL-7-responsive CD19(+) precursors are estrogen resistant. However, B-lineage differentiation was selectively abrogated when highly purified Lin(-) precursors were treated with hormone in the absence of stromal cells. In addition, early stages of B lymphopoiesis were arrested by limitin, a new <em>interferon</em> (IFN)-like cytokine as well as IFN-<em>alpha</em>, IFN-gamma, or transforming growth factor beta (TGF-beta), but not by epidermal growth factor (EGF). Lin(-) TdT(+) early pro-B cells are shown here to be CD27(+) AA4.1(+/-)Ki-67(+) Ly-6C(-) Ly-6A/Sca-1(Lo/-)Thy-1(-)CD4<em>3</em>(+) CD4(+/-)CD16/<em>3</em>2(Lo/-)CD44(Hi) and similar in some respects to the "common lymphoid progenitors" (CLP) identified by others. Although early pro-B cells have lost myeloid differentiation potential, transplantation experiments described here reveal that at least some can generate T lymphocytes. Of particular importance is the demonstration that a pivotal early stage of lymphopoiesis is directly sensitive to negative regulation by hormones and cytokines.

Animal and human studies have shown that greatly increasing the amounts of flax seed oil [rich in the (n-<em>3</em>) polyunsaturated fatty acid (PUFA) <em>alpha</em>-linolenic acid (ALNA)] or fish oil [FO; rich in the long chain (n-<em>3</em>) PUFA eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA)] in the diet can decrease mitogen-stimulated lymphocyte proliferation. The objective of this study was to determine the effect of dietary supplementation with moderate levels of ALNA, gamma-linolenic acid (GLA), arachidonic acid (ARA), DHA or FO on the proliferation of mitogen-stimulated human peripheral blood mononuclear cells (PBMC) and on the production of cytokines by those cells. The study was randomized, placebo-controlled, double-blinded and parallel. Healthy subjects ages 55-75 y consumed nine capsules/d for 12 wk; the capsules contained placebo oil (an 80:20 mix of palm and sunflower seed oils) or blends of placebo oil with oils rich in ALNA, GLA, ARA or DHA or FO. Subjects in these groups consumed 2 g of ALNA or 770 mg of GLA or 680 mg of ARA or 720 mg of DHA or 1 g of EPA plus DHA (720 mg of EPA + 280 mg of DHA) daily from the capsules. Total fat intake from the capsules was 4 g/d. The fatty acid composition of PBMC phospholipids was significantly changed in the GLA, ARA, DHA and FO groups. Lymphocyte proliferation was not significantly affected by the placebo, ALNA, ARA or DHA treatments. GLA and FO caused a significant decrease (up to 65%) in lymphocyte proliferation. This decrease was partly reversed by 4 wk after stopping the supplementation. None of the treatments affected the production of interleukin-2 or <em>interferon</em>-gamma by PBMC and none of the treatments affected the number or proportion of T or B lymphocytes, helper or cytotoxic T lymphocytes or memory helper T lymphocytes in the circulation. We conclude that a moderate level GLA or EPA but not of other (n-6) or (n-<em>3</em>) PUFA can decrease lymphocyte proliferation but not production of interleukin-2 or <em>interferon</em>-gamma.

Type I <em>interferon</em> (IFN) receptor consists of two chains (Hu-IFN-<em>alpha</em>R1 and Hu-IFN-<em>alpha</em>R2), and Hu-IFN-<em>alpha</em>R2 takes a soluble (Hu-IFN-<em>alpha</em>R2a), short (Hu-IFN-<em>alpha</em>R2b), or long (Hu-IFN-<em>alpha</em>R2c) form. We examined the expression of type I IFN receptor, the growth-suppression effect of IFN-<em>alpha</em>, and their relationship in 1<em>3</em> liver cancer cell lines. With reverse-transcription polymerase chain reaction (RT-PCR) analysis, the expressions of Hu-IFN-<em>alpha</em>R1, Hu-IFN-<em>alpha</em>R2a, and Hu-IFN-<em>alpha</em>R2c were confirmed in all cell lines, and that of Hu-IFN-<em>alpha</em>R2b in 12 cell lines. All cell lines expressed mRNAs of a transcriptional activator, <em>interferon</em> regulatory factor (IRF)-1, and its antagonistic repressor (IRF-2). Flow cytometry revealed weak expression of Hu-IFN-<em>alpha</em>R2 on the cell surface in 12 cell lines. The soluble-form protein of Hu-IFN-<em>alpha</em>R2 was detected at varying levels in culture supernatants of all cell lines with enzyme-linked immunosorbent assay (ELISA). Cell proliferation was suppressed in proportion to the dose of human natural IFN-<em>alpha</em> at 96 hours of culture, but it was not clearly related to the expression of Hu-IFN-<em>alpha</em>R2 protein on the cell surface. Investigations on the morphology, DNA, and cell cycle presented four growth suppression patterns as a result of IFN-<em>alpha</em>: 1) induction of apoptosis and blockage of cell cycle at the S phase (9 cell lines); 2) blockage at the S phase (2 cell lines); <em>3</em>) induction of apoptosis and blockage at the G2/M phase (1 cell line); and 4) blockage at the G1 phase (1 cell line). There was no evidence showing that changes in the expressions of Bcl-2, Bcl-xL, Bak, and Bax lead directly to IFN-<em>alpha</em>-mediated apoptosis. Our findings demonstrated that IFN-<em>alpha</em> would express growth-suppression effects at varying degrees by inducing inhibition of cell-cycle progression with or without apoptosis, regardless of the expression level of Hu-IFN-<em>alpha</em>R2 protein on the cell surface.

Poliovirus (PV) and enterovirus 71 (EV71) cause severe neurological symptoms in their infections of the central nervous system. To identify compounds with anti-PV and anti-EV71 activities that would not allow the emergence of resistant mutants, we performed drug screening by utilizing a pharmacologically active compound library targeting cellular factors with PV and EV71 pseudoviruses that encapsidated luciferase-encoding replicons. We have found that metrifudil (N-[2-methylphenyl]methyl)-adenosine) (an A2 adenosine receptor agonist), N(6)-benzyladenosine (an A1 adenosine receptor agonist) and NF449 (4,4',4'',4'''-[carbonylbis[imino-5,1,<em>3</em>-benzenetriyl bis(carbonyl-imino)]] tetrakis (benzene-1,<em>3</em>-disulfonic acid) octasodium salt) (a Gs-<em>alpha</em> inhibitor) have anti-EV71 activity, and that GW5074 (<em>3</em>-(<em>3</em>, 5-dibromo-4-hydroxybenzylidine-5-iodo-1,<em>3</em>-dihydro-indol-2-one)) (a Raf-1 inhibitor) has both anti-PV and anti-EV71 activities. EV71 mutants resistant to metrifudil, N(6)-benzyladenosine and NF449 were isolated after passages in the presence of these compounds, but mutants resistant to GW5074 were not isolated for both PV and EV71. The inhibitory effect of GW5074 was not observed in Sendai virus infection and the treatment did not induce the expression of OAS1 and STAT1 mRNA. Small interfering RNA treatment against putative cellular targets of GW5074, including Raf-1, B-Raf, Pim-1, -2, and -<em>3</em>, HIPK2, GAK, MST2 and ATF-<em>3</em>, did not consistently suppress PV replication. Moreover, downregulation of Raf-1 and B-Raf did not affect the sensitivity of RD cells to the inhibitory effect of GW5074. These results suggest that GW5074 has strong and selective inhibitory effect against the replication of PV and EV71 by inhibiting conserved targets in the infection independently of the <em>interferon</em> response.