Pretreatment with 6-Gingerol Ameliorates Sepsis-Induced Immune Dysfunction by Regulating the Cytokine Balance and Reducing Lymphocyte Apoptosis

Sepsis is characterized by an initial net hyperinflammatory response, followed by a period of immunosuppression, termed immunoparalysis. During this immunosuppressive phase, patients may have difficulty eradicating invading pathogens and are susceptible to life-threatening secondary hospital-acquired infections. Due to progress in antimicrobial treatment and supportive care, most patients survive early sepsis. Mortality is more frequently attributed to subsequent secondary nosocomial infections and multiorgan system failure. 6-Gingerol is the major pharmacologically active component of ginger. Although it is known to exhibit a variety of biological activities, including anti-inflammation and antioxidation, the role of 6-gingerol in sepsis-induced immune dysfunction remains elusive. Thus, we investigated whether 6-gingerol improves septic host response to infections during sepsis. 6-Gingerol-treated mice showed significantly lower mortality in polymicrobial sepsis induced by cecal ligation and puncture LPS via enhanced bacterial clearance in the peritoneum, blood, and organs (liver, spleen, and kidney) and inhibited the production of TNF-α and IL-6 in TLR2 and/or TLR4-stimulated macrophages. In addition, we demonstrated that survival improvement of secondary infection following septic insult was associated with an initial response of enhanced neutrophil numbers and function at the infection site, reduced apoptosis of immune cells, and a shift from a T helper cell type 2 (Th2) to a T helper cell type 1 (Th1) cytokine balance in the hypoinflammation phase. Our overall findings suggest that 6-gingerol potentially restores sepsis-induced immune dysfunction by shifting the balance of Th1/Th2 and by regulating apoptosis of immune cells.

Cytokine profiles in highly active antiretroviral treatment non-adherent, adherent and naive HIV-1 infected patients in Western Kenya

Background: Cytokines play an important role in signaling the immune system to build an adequate immune responseagainst HIV. HIV distorts the balance between pro and anti-inflammatory cytokines causing viral replication. Highly active antiretroviral treatment (HAART) acts by trying to restore pro and anti-inflammatory cytokine balance. It is not clear how HAART non-adherence influences circulating cytokine levels. This study therefore determined cytokine levels in HAART non-adherent individuals. Methods: This cross-sectional study recruited 163 participants (51 controls, 23 HIV-1+ HAART naive, 28 HAART-adherent6 months, 19 HAART-adherent 12 months and 42 HAART non-adherent). Cytokines were analyzed by ELISA while CD4 T cells determined in 3.0 μl of whole blood using BD FACSCaliburTM and viral load in 0.2ml plasma sample using Abbott Molecular m2000sp sample preparation and m2000rt real-time amplification and detection systems (Abbott MolecularInc., Illinois, USA) according to the manufacturer’s methods. Results: IL-4, IL-6, IL-10, TNF-α and TGF-β were significantly elevated in HIV-1 HAART non-adherent compared withHIV-1 HAART adherent and healthy controls P<0.01. IFN- γ was significantly decreased in HIV-1 HAART non-adherentcompared with HIV-1 HAART adherent and healthy controls P<0.01. TNF-α and TGF-β were significantly reduced in HIV-1 HAART adherent patients at 12 months compared to those at 6 months P<0.01. IL-4 and IL-10 correlated positively withviral load. IL-4, IL-6, IL-10, TNF-α and TGF- β associated inversely with CD4 T cell counts and body mass index (BMI). Conclusion: This study established that HAART adherence is immunologically beneficial to the pro and anti-inflammatory cytokine balance milieu while non-adherence appears to cause alterations in pro and anti-inflammatory cytokines warping the balance in this dichotomy. Keywords: Cytokines; non-adherence; HAART.

Blood and urinary cytokine balance and renal outcomes at cardiac surgery

Background Increased perioperative pro-inflammatory biomarkers, renal hypoperfusion and ischemia reperfusion injury (IRI) heighten cardiac surgery acute kidney injury (CS-AKI) risk. Increased urinary anti-inflammatory cytokines attenuate risk. We evaluated whether blood and urinary anti-inflammatory biomarkers, when expressed as ratios with biomarkers of inflammation, hypoperfusion and IRI are increased in CS-AKI patients. Methods Preoperative and 24-h postoperative blood and urinary pro-inflammatory and anti-inflammatory cytokines, blood VEGF and H-FABP (hypoperfusion biomarkers), and MK, a biomarker for IRI, were measured in 401 cardiac surgery patients. Pre- and postoperative concentrations of biomarkers and selected ratios thereof, were compared between non-CS-AKI and CS-AKI patients. Results Compared with non-CS-AKI, blood pro-inflammatory (pre- and post-op TNFα, IP-10, IL-12p40, MIP-1α, NGAL; pre-op IL-6; post-op IL-8, MK) and anti-inflammatory (pre- and post-op sTNFsr1, sTNFsr2, IL-1RA) biomarkers together with urinary pro-inflammatory (pre- and post-op uIL-12p40; post-op uIP-10, uNGAL) and anti-inflammatory (pre- and post-op usTNFsr1, usTNFsr2, uIL-1RA) biomarkers, were significantly higher in CS-AKI patients. Urinary anti-inflammatory biomarkers, when expressed as ratios with biomarkers of inflammation (blood and urine), hypoperfusion (blood H-FABP and VEGF) and IRI (blood MK) were decreased in CS-AKI. In contrast, blood anti-inflammatory biomarkers expressed as similar ratios with blood biomarkers were increased in CS-AKI. Conclusions The urinary anti-inflammatory response may protect against the injurious effects of perioperative inflammation, hypoperfusion and IRI. These finding may have clinical utility in bioprediction and earlier diagnosis of CS-AKI and informing future therapeutic strategies for CS-AKI patients.

Acupoint Autohemotherapy Attenuates DNCB-induced Atopic Dermatitis Lesions by Regulating Th1/Th2 Cytokine Balance in BALB/c Mice

Objective Acupoint autohemotherapy (A-AHT) is considered an effective therapy for atopic dermatitis (AD) with few side-effects. Previous experiments showed the treatment had the potential to regulate T helper (Th) 1 and Th2 cytokines, like interferon (IFN)- gamma and interleukin (IL)- 4. This study focuses on the effects of A-AHT on the AD-like skin lesions through regulating Th1/Th2 immune responses. Methods The treatments of A-AHT, sham acupoint autohemotherapy and acupoint injection of normal saline were administered in the AD mice once every other day for 4 weeks. The total immunoglobulin (Ig) E, IL-4 and IFN-γ cytokine levels in the serum were examined after animal sacrifice. Th1/Th2 expression was analyzed in murine spleen cells via flow cytometry and immunohistochemical analysis of GATA-3 and T-bet in skin lesions were further assessed. Results Either type of repeated autologous whole blood (AWB) injection (into acupoint or sham acupoint) reduced the severity of AD-like symptoms and level of serum IgE. All of the three treatments had the similar inhibitory effect on levels of IL-4 and upregulation on the ratio of IFN-γ/IL-4, while differed on Th1/Th2 ratio as A-AHT regulates the body’s Th1/Th2 shift. This treatment also increased the related transcription factors T-bet expression, and upregulated T-bet/GATA3 ratio compared with the DNCB group. These differences were significant only in A-AHT group. Conclusion A-AHT effectively reduces AD symptoms and serum IgE levels in a mouse model and may act by regulating Th1/Th2 immune responses.

Immunological aspects of bacterial keratites in patients with diabetes mellitus

Background. The purpose was to analyze the cytokine balance of lacrimal fluid in patients with bacterial keratitis and diabetes mellitus (DM) at the first visit and to identify the immunological aspects of the disease. Materials and methods. The analysis of pro- and anti-inflammatory cytokine concentration in the lacrimal fluid was performed in 17 patients with type 1 DM and bacterial keratitis and 15 nondiabetic patients with bacterial keratitis at the first visit. Data from 14 healthy individuals were used for comparison. In addition to standard ones, ophthalmic examination methods included bacteriological examination, fluorescein test, anterior segment optical coherence tomography, non-contact corneal aesthesiometry. The levels of interleukin (IL) 1β, IL-6 and IL-10 in the lacrimal fluid of the sick and the contralateral eye were determined by a quantitative colorimetric enzyme-linked immunosorbent assay using ELISA kits. Results. In DM patients with bacterial keratitis, the concentration of IL-1β and IL-6 in the lacrimal fluid of the sick eye exceeded that in healthy individuals (p < 0.05) and did not differ significantly from nondiabetic patients with bacterial keratitis (p > 0.05). In the lacrimal fluid of the contralateral eye of DM patients with bacterial keratitis, the level of IL-1β and IL-6 exceeded the corresponding indicators of nondiabetic patients with bacterial keratitis and healthy individuals (p < 0.05). The concentration of IL-10 in the lacrimal fluid of the contralateral eye in DM patients with bacterial keratitis exceeded that in healthy individuals (p < 0.05) and did not significantly differ from those in nondiabetic patients with bacterial keratitis (p > 0.05). Conclusions. DM patients with bacterial keratitis have immunological features of the disease.

The State of Cellular Immunity and Cytokine Balance in Pregnant Women at Intrauterine Infection

The question of studying changes in the immune responses of mother and foetus against the background of infections acquired via various routes remains relevant. This study aimed to assess the state of T-cell immunity and cytokine balance in pregnant women with various pathways of intrauterine infection and the condition of newborns. The study involved 205 pregnant women at high risk for intrauterine infection. In the 1st trimester, bacteriological and DNA analysis of the lower urogenital tract and cytological examination of the cervical canal were performed, blood levels of immunoglobulins M and G for herpes simplex virus type 1 and 2, cytomegalovirus infection, toxoplasmosis, and rubella were determined. In the whole blood of the women, lymphocyte immunophenotyping was performed using CYTO-STAT® triCHROMETM CD8-FITC/CD4-RD1/CD3-FITC monoclonal antibodies; the content of cytokines (IL-6, IL-10) was analysed by means of enzyme-linked immunosorbent assay. After delivery, a pathomorphological examination of the placenta was performed in line with the generally accepted method. According to the results of the study, the following groups of women were identified: 1) without infectious or inflammatory changes (n = 59); 2) with confirmed ascending infection (n = 69); 3) with haematogenous infection (n = 33); 4) with mixed infection (n = 44). The condition of newborns was assessed with the help of laboratory and instrumental methods, using the INTERGROWTH-21st charts and the Apgar score. We found that the functioning of the immune system of pregnant women is affected by viral infections acquired via the haematogenous route, resulting in a relative increase in suppressor T cells and a decrease in helper T cells, as well as ina growing absolute number of lymphocytes in the blood. The identified inhibition of IL-6 and IL-10 production in the groups with signs of placental lesions due to infection at 16–18 weeks can indicate a strain on the immune processes and development of placental insufficiency. Newborns with morphological signs of haematogenous infection are characterized by changes in the cytological parameters of residual cord blood, signs of placental insufficiency, low birth weight, and hypoxic-ischemic damage to the central nervous system.

Analysis of the Cytokine Balance in Tear Fluid of Diabetic Patients with Bacterial Keratitis Depending on the Severity of the Disease

The aim. To analyze the cytokine balance of tear fluid in patients with bacterial keratitis at presentation depending on the severity of the disease and the presence of diabetes mellitus (DM). Materials and methods. The analysis was performed through the comparison of the level of pro- and anti-inflammatory cytokines in the tear fluid of 17 patients with type 1 DM and bacterial keratitis and 15 patients with bacterial keratitis without DM at presentation. Data from 14 healthy individuals of the appropriate age were also used for comparison. The patients with bacterial keratitis were divided into subgroups according to the severity of bacterial keratitis. The levels of IL-1β, IL-6 and IL-10 in the tear fluid of the sick and the contralateral eye were determined by quantitative colorimetric enzyme-linked immunosorbent assay. Results and discussion. At presentation, patients with bacterial keratitis, both with and without DM, showed increased levels of proinflammatory cytokines, namely IL-1β and IL-6, in the tear fluid of the sick eye, which correlated with the severity of the disease, and also increased level of anti-inflammatory cytokine IL-10 in the tear fluid of the contralateral eye. In addition, the concentrations of proinflammatory cytokines IL-1β, IL-6 in the tear fluid of the contralateral eye in DM patients were increased at all degrees of severity of bacterial keratitis. Conclusions. In patients with bacterial keratitis, the cytokine balance of the tear fluid of the sick and the contralateral eye depends on the severity of the disease and the presence of DM. Keywords: diabetes mellitus, bacterial keratitis, severity of keratitis, cytokines, interleukins.

Effect of biotherapics made from serum or rabbit serum in Trypanosoma cruzi infection

The use of biotherapics as an intervention in experimental models of infection is a possible means to understand the effects of these medications [1-3]. This study evaluated the immunological and parasitological effects of biotherapics that were prepared from mouse or rabbit serum uninfected (MUI or RUI groups) or chronically infected with T. cruzi (MI or RI groups), dynamization 13cH. Male Swiss mice, 28 days of age were infected with T. cruzi-Y strain and treated with the different biotherapics diluted in water (1mL/100mL). The study was approved by the Ethics Committee for Experiments in Animals-UEM (protocol no. 080/2012). Using biotherapics made with mouse serum, MUI group exhibited good outcome with a pronounced Th1 response that was attributable to a reduction of IL-4 concentrations and decrease in IL-17 concentrations compared with the control group. However, this cytokine balance was not sufficient to promote decreased parasitemia in treated animals, likely because of a decrease in IFN-gama, thus hindering a more effective beneficial Th1 response. In contrast, the MI group presented a pronounced Th2 response that was attributable to increase in IL-4 and decrease in IFN-gama concentration compared to control group. MI group exhibited the worst outcome where the cytokine balance suppressed the immune response to T. cruzi in murine infection, resulting in a significant increased parasitemia and decreased survival time. Using biotherapics made with rabbit serum, RUI group exhibited the best outcome, including decreased parasitemia, with pronounced Th1 response that was attributable to decrease in IL-4 concentrations, with no changes in TNF-alpha and IFN-gama, associated to decrease in IL-17 compared to control group. In contrast, RI group did not exhibit alterations in parasitemia but a pronounced Th2 response that was attributable to increase in IL-4 concentration, with no changes in TNF-alpha and IFN-gama, associated to decrease in IL-17 compared to control group. Results show that biotherapics that were prepared from mouse or rabbit serum uninfected or chronically infected with T. cruzi differentially modulate the immune system in mice infected with this protozoan. Also, results provide evidence that biotherapics prepared with serum from healthy animal performed better than one made with serum from infected animal. In the same way biotherapics prepared with serum from resistant specie performed better than one made with serum from susceptible specie.

Tildrakizumab and omalizumab: An interlocking therapy for autoimmune conditions

In Western countries the number of individuals suffering from an autoimmune condition is constantly growing and often patients suffering from autoimmune disease are susceptible to developing a second autoimmune disorder. We report a case of an adult female patient affected by psoriasis vulgaris and treated with tildrakizumab, a humanized monoclonal antibody targeting interleukin-23, who later developed chronic spontaneous urticaria and started omalizumab, a humanized antibody to IgE, showing a favorable outcome. We speculate that the two combined therapies have restored the cytokine balance bringing it towards tolerance and remission of the two pathologies. It is conceivable that tildrakizumab may have a synergic action with omalizumab in the treatment of urticaria in patients affected by both psoriasis and urticaria. Our case and the study of the mechanisms of action of the two drugs suggest how the two therapies can act with an interlocking mechanism in achieving the final therapeutic effect. Keywords: Chronic spontaneous urticarial; dual biologic therapy; omalizumab; psoriasis; tildrakizumab.