IL-10- and TGFβ-mediated Th9 Responses in a Human Helminth Infection

Th9 cells are a subset of CD4+ T cells that express the protoypical cytokine, IL-9. Th9 cells are known to effect protective immunity in animal models of intestinal helminth infections. However, the role of Th9 cells in human intestinal helminth infections has never been examined.To examine the role of Th9 cells in Strongyloidis stercoralis (Ss), a common intestinal helminth infection, we compared the frequency of Th9 expressing IL-9 either singly (mono-functional) or co-expressing IL-4 or IL-10 (dual-functional) in Ss-infected individuals (INF) to frequencies in uninfected (UN) individuals.INF individuals exhibited a significant increase in the spontaneously expressed and/or antigen specific frequencies of both mono- and dual-functional Th9 cells as well as Th2 cells expressing IL-9 compared to UN. The differences in Th9 induction between INF and UN individuals was predominantly antigen-specific as the differences were no longer seen following control antigen or mitogen stimulation. In addition, the increased frequency of Th9 cells in response to parasite antigens was dependent on IL-10 and TGFx since neutralization of either of these cytokines resulted in diminished Th9 frequencies. Finally, following successful treatment of Ss infection, the frequencies of antigen-specific Th9 cells diminished in INF individuals, suggesting a role for the Th9 response in active Ss infection. Moreover, IL-9 levels in whole blood culture supernatants following Ss antigen stimulation were higher in INF compared to UN individuals.Thus, Ss infection is characterized by an IL-10- and TGFβ dependent expansion of Th9 cells, an expansion found to reversible by anti-helmintic treatment

Systemic Cytokine Profiles in Strongyloides stercoralis Infection and Alterations following Treatment

Strongyloides stercoralis is a soil-transmitted helminth organism that infects ∼50 to 100 million people worldwide. Despite its widespread prevalence, very little is known about the immune response that characterizes human S. stercoralis infection. To study the systemic cytokine profile characteristic of Strongyloides infection, we measured the circulating levels of a large panel of pro- and anti-inflammatory cytokines in asymptomatic, infected individuals (n = 32) and compared them to those in uninfected, controls (n = 24). Infected individuals exhibited significantly lower circulating levels of proinflammatory cytokines (gamma interferon [IFN-γ], tumor necrosis factor alpha [TNF-α], and interleukin-1β [IL-1β]) and significantly higher levels of anti-inflammatory cytokines (IL-4, IL-5, IL-9, IL-10, IL-13, IL-27, IL-37, and transforming growth factor β [TGF-β]). Moreover, treatment of Strongyloides infection resulted in a significant reversal of the cytokine profile, with increased levels of proinflammatory (IFN-γ, TNF-α, IL-2, IL-17A, IL-17F, IL-22, IL-23, and IL-1β) and decreased levels of anti-inflammatory (IL-4, IL-5, IL-9, IL-10, IL-13, IL-27, IL-37, and TGF-β) cytokines following treatment. Thus, S. stercoralis infection is characterized by alterations in the levels of systemic cytokines, reflecting major alterations in the underlying immune response to this chronic helminth infection

Hygrothermal effects on the mechanical properties of glass fibre-epoxy composite material

In this study, the hygrothermal effects on the mechanical properties of Glass Fiber Reinforced Plastic (GFRP) composite material was investigated. Unidirectional GFRP composite material was tested to determine tensile, flexure and Inter-laminar shear strength (ILSS) under both room temperature (RT) and hot-wet conditions. Prior to testing, hot-wet specimens were hygrothermally aged in an environmental chamber, maintained at 710C and 85 % relative humidity (RH) for 1%wt moisture absorption. Tests were performed in a computer controlled 50 kN servo-hydraulic test machine. For hot-wet tests, a split type environmental chamber was fixed to the machine which was also maintained at 710C and 85 % RH during the tests. The test data obtained was statistically analyzed to determine mean strength and B-basis design allowables. A comparative study was made to investigate the effect of moisture in the GFRP material at hot-wet and room temperature conditions, which showed a reduction in the strength. Presence of moisture in the epoxy matrix, in the fiber-matrix interface and the chemical attack of moisture on the glass fibers are the main reasons for reduced strength of GFRP material in hot-wet condition

Purification of heat labile toxin from Bordetella pertussis vaccine strain 134 employed indigenous technology

Aim and objective: The aim of the study was the evaluation of purified HLT from B. pertussis vaccine strain 134 by employing indigenous technology and examining the immuno-biochemical aspects of the purified protein. Materials and methods: Shaker cultivation of B. pertussis strain 134, sterility, opacity confirmation, TCA precipitation of cellular proteins, G50 purification subsequent DEAE purification, purity analysis, specific activity of HLT, and immune response analysis. Results: The shaker cultivated B. pertussis strain 134 passed its quality attributes such as sterility, opacity and purity. During TCA precipitation the B. pertussis desired proteins were precipitated and confirmed. The indigenous bed height was optimized and recovery was also calculated in G50 purification. The fractions were analyzed for OD, the total protein concentration and the results were 0.074–0.214, and the total protein content was found between 12.33 μg/ml and 35.67 μg/ml. Subsequent DEAE purification of selected G50 fractions was done and the fractions 9 and 14 had higher OD values of 0.675 and 0.397. Furthermore the DEAE purified samples were structurally analyzed through SDS PAGE and it was found that HLT in the single polypeptide band was around 140 kDa. The qualitative immune response analysis of DEAE purified selected fractions pool showed positive immune response in ODD assay, and in the case of guinea pig antisera it led to the development of diagnostic kits such as ELISA and other vital techniques. Here, in case of guinea pig experiment, the hemorrhagic necrosis analysis showed the necrosis on the skin at the injection site. Conclusion: The B. pertussis HLT could be purified through two phase with G50 and DEAE, cost effective techniques, the G50 purification has reduced the bioburden problems during DEAE purification and at the same time the quality of the product was high

Systemic Cytokine Profiles in Strongyloides stercoralis Infection and Alterations following Treatment

Strongyloides stercoralis is a soil-transmitted helminth organism that infects ∼50 to 100 million people worldwide. Despite its widespread prevalence, very little is known about the immune response that characterizes human S. stercoralis infection. To study the systemic cytokine profile characteristic of Strongyloides infection, we measured the circulating levels of a large panel of pro- and anti-inflammatory cytokines in asymptomatic, infected individuals (n = 32) and compared them to those in uninfected, controls (n = 24). Infected individuals exhibited significantly lower circulating levels of proinflammatory cytokines (gamma interferon [IFN-γ], tumor necrosis factor alpha [TNF-α], and interleukin-1β [IL-1β]) and significantly higher levels of anti-inflammatory cytokines (IL-4, IL-5, IL-9, IL-10, IL-13, IL-27, IL-37, and transforming growth factor β [TGF-β]). Moreover, treatment of Strongyloides infection resulted in a significant reversal of the cytokine profile, with increased levels of proinflammatory (IFN-γ, TNF-α, IL-2, IL-17A, IL-17F, IL-22, IL-23, and IL-1β) and decreased levels of anti-inflammatory (IL-4, IL-5, IL-9, IL-10, IL-13, IL-27, IL-37, and TGF-β) cytokines following treatment. Thus, S. stercoralis infection is characterized by alterations in the levels of systemic cytokines, reflecting major alterations in the underlying immune response to this chronic helminth infection

Strongyloides infection is associated with increased spontaneously expressed and antigen-induced frequencies of CD4⁺ mono- and dual-functional Th9 cells.

<p>Whole blood was cultured with media alone or with antigens for 6 h and the baseline and antigen-stimulated frequencies of Th9 cells determined. The baseline (A) as well as SsAg (B), NIE (C), PPD (D) and P/I (E) stimulated frequencies of mono- and dual-functional CD4⁺ Th9 cells in INF (n = 28) and UN (n = 23) individuals are shown. The MFI of IL-9 expression on CD4⁺ T cells in INF and UN individuals at baseline and following stimulation are shown (F). Each circle represents a single individual and the bars represent the geometric mean values. Net frequencies were calculated by subtracting baseline frequencies from the antigen-induced frequencies for each individual. <i>P</i> values were calculated using the Mann-Whitney test.</p

Strongyloides infection is associated with increased spontaneously produced and antigen-induced production of IL-9.

<p>Whole blood was cultured with media alone or with Ss Ag or P/I for 18 h and the baseline and antigen-stimulated levels of IL-9 determined by ELISA. (A) The baseline as well as SsAg and P/I stimulated levels of IL-9 in INF (n = 28) and UN (n = 23) individuals are shown. Each circle represents a single individual and the bars represent the geometric mean values. <i>P</i> values were calculated using the Mann-Whitney test. (B) The IL-9 levels following stimulation with media alone, SsAg and P/I before and after treatment with a standard dose of ivermectin and albendazole in a subset of INF individuals (n = 15). Each line represents a single individual. P values were calculated using the Wilcoxon signed rank test.</p

Strongyloides infection is associated with expansion of mono- and dual-functional Th9 cells.

<p>Whole blood was cultured with media alone for 6 h and the baseline and antigen-specific frequencies of Th9 cells determined. A representative whole-blood intracellular cytokine assay flow data from a INF individual showing expression of IL-9, IL-4 and IL-10 at baseline (A) and following stimulation with SsAg (B) or P/I (C). The plots shown are gated on CD3⁺CD4⁺ T cells.</p

IL-10 and TGFβ mediate the expansion of mono- and dual-functional Th9 cells in Strongyloides infections.

<p>(A) IL-10 neutralization (with anti-IL-10 antibody) significantly diminishes the frequencies of mono- functional or dual-functional CD4⁺ Th9 cells following stimulation with NIE in a subset of INF individuals (n = 15). (B) TGFβ neutralization (with anti-TGFβ antibody) significantly diminishes the frequencies of mono- functional or dual-functional CD4⁺ Th9 cells following stimulation with NIE in a subset of INF individuals (n = 15). (C) Both IL-10 and TGFβ neutralization significantly reduce the MFI of IL-9 expression on CD4⁺ T cells following stimulation with NIE in a subset of INF individuals (n = 15). Antigen-stimulated frequencies are shown as net frequencies with the baseline levels subtracted. Each line represents a single individual. P values were calculated using the Wilcoxon signed rank test.</p