75 research outputs found
The Effect of IFN-γ, Alum and Complete Freund Adjuvant on TNP-KLH Induced Ig.G1, IgE and IgG2a Responses in Mice
Adjuvants are considered to play an important role in directing the
isotype and amount of antibodies produced upon immunization by
conducting the development of either Th-1 or Th-2 cells upon T-cell
stimulation. This is based on the different cytokine production
patterns that were observed after in vitro resttmulation of T cells
isolated from mice immunized with antigen either adsorbed on alum or
emulsified in complete Freund adjuvant (CFA). However, other studies
suggest that primarily the type of antigen determines which isotypes
are produced and to what extent. In these studies, however, IgE was
not determined. Therefore, this study examined whether alum and CFA
influenced the amount and/or ratio of IgG1, IgE and IgG2a produced
after TNP-KLH immunization. Similar levels of IgG1, IgE and IgG2a
antibodies were found upon immunization with TNP-KLH either adsorbed
on alum or emulsified in CFA. Moreover, administration of IFN-γ in
combination with TNP-KLH adsorbed on alum did not increase the
amount of IgG2a produced. IFN-γ treatment resulted in an increased
IL-6 and decreased IFN-γ production by spleen cells upon Con A
stimulation, whereas it did not change the IL-4 production in
similar conditions. The presented results suggest that upon
immunization with TNP-KLH high IL-4 levels are produced, resulting
in an antibody response that is dominated by IgG1, independent of the
adjuvant employed. The IL-4 inducing property of TNP-KLH is
substantiated by the finding that repeated immunization of mice with
TNP-KI, without adjuvant, increases the serum total IgE level. The
presented data suggest that the carrier part of TNP-KLH
preferentially results in Th-2 cell activity after which the
adjuvant merely enhances the antibody responses generated
Modulation of systemic cytokine levels by implantation of alginate encapsulated cells
The availability of cell lines that are transfected with IL-4, IL-5 and IFN-γ cytokine genes permits the prolonged in vivo delivery of functional cytokines in relatively large doses for the modulation of specific immune responses. Oft
Cytokine Detection and Modulation in Acute Graft vs. Host Disease in Mice
A murine model for acute lethal graft vs. host disease (GVHD) was
used to study the role that a number of cytokines play in the development of lethal GVHD. In this study we focused on the role of IL-1, IL-2, IL-4, IL-6, IFN-γ and TNF-α. Lethally irradiated (C57BL × CBA)F1 mice were reconstituted either with 107 allogeneic BALB/c spleen cells or with a similar number of syngeneic cells, as a control. A significant rise in serum levels of IL-6, TNF-α and IFN-γ levels was found in allogeneically reconstituted mice. This is in contrast to the
syngeneic control group in which no rise was seen. Serum IL-2 and IL-4 levels were below the detection limit. In the supernatant of Con A stimulated spleen cells from allogeneically reconstituted mice
IL-6, IFN-γ and TNF-α concentrations were increased. The
expression of mRNA for cytokines as detected by reverse
transcription PCR was studied in spleen cells. In the allogeneic
reconstituted mice the mRNA expression of IL-1α, IL-2, IL-6,
IFN-γ and TNF-α displayed faster kinetics compared with
that in syngeneic reconstituted mice. The effect of treatment with
recombinant cytokines, antibodies to cytokines and to cytokine
receptors on the development of GVHD was investigated.
Administration of recombinant IL-2 to allogeneically reconstituted
mice strongly increased the morbidity and mortality whereas
injection of IL-1α and TNF-α did not influence survival.
Administration of antibodies against IL-2 or the IL-2 receptor
decreased the morbidity and mortality. Anti-IL-6, anti-IFN-γ,
and anti-TNF-α mAB, on the other hand, did not affect the
morbidity and mortality of GVHD. The results of this study suggest
successive waves of cytokine-secreting cell populations consistent
with the induction of an inflammatory response in the development of
acute GVH disease
Type 2 diabetes monocyte microrna and mrna expression
There is increasing evidence that inflammatory macrophages in adipose tissue are involved in insulin resistance of type 2 diabetes (T2D). Due to a relative paucity of data on circulating monocytes in T2D, it is unclear whether the inflammatory changes of adipose tissue macrophages are reflected in these easily accessible cells. Objective To study the expression pattern of microRNAs and mRNAs related to inflammation in T2D monocytes. Design A microRNA finding study on monocytes of T2D patients and controls using array profiling was followed by a quantitative Real Time PCR (qPCR) study on monocytes of an Ecuadorian validation cohort testing the top over/under-expressed microRNAs. In addition, monocytes of the validation cohort were tested for 24 inflammation-related mRNAs and 2 microRNAs previously found deregulated in (auto)-inflammatory monocytes. Results In the finding study, 142 significantly differentially expressed microRNAs were identified, 15 having the strongest power to discriminate T2D patients from controls (sensitivity 66%, specificity 90%). However, differences in expression of these microRNAs between patients and controls were small. On the basis of >1.4 or <0.6-fold change expression 5 microRNAs were selected for further validation. One microRNA (miR-34c-5p) was validated as significantly over-expressed in T2D monocytes. In addition, we found over expression of 3 mRNAs (CD9, DHRS3 and PTPN7) in the validation cohort. These mRNAs are important for cell morphology, adhesion, shape change, and cell differentiation. Classical inflammatory genes (e.g. TNFAIP3) were only over-expressed in monocytes of patients with normal serum lipids. Remarkably, in dyslipidemia, there was a reduction in the expression of inflammatory genes (e.g. ATF3, DUSP2 and PTGS2). Conclusions The expression profile of microRNAs/mRNAs in monocytes of T2D patients indicates an altered adhesion, differentiation, and shape change potential. Monocyte inflammatory activation was only found in patients with normal serum lipids. Abnormal lipid values coincided with a reduced monocyte inflammatory state. Copyright
Study on inflammation-related genes and microRNAs, with special emphasis on the vascular repair factor HGF and miR-574-3p, in monocytes and serum of patients with T2D
Background: Recently, we reported signs of inflammation (raised IL-8, reduced miR-146a) and signs of vascular repair (raised HGF) in the serum of Ecuadorian patients with type 2 diabetes (T2
Antibody formation in mouse bone marrow: VI. The regulating influence of the spleen on the bone marrow plaque-forming cell response to Escherichia coli lipopolysaccharide
Mouse bone marrow is capable of a distinct plaque-forming cell (PFC) response after i.v. immunization with the thymus-independent antigen E. coli lipopolysaccharide (LPS). Both during the primary and secondary response to i.v. administered LPS the spleen contained the majority of PFC until about 5 days after immunization. In the course of the reaction the number of PFC in the bone marrow rose to a level which surpassed the level in the spleen. This paper deals with the regulating influence of the spleen on the primary and secondary anti-LPS PFC response in the bone marrow. Splenectomy prior to the first injection of 5 μg LPS i.v. initially did not affect the bone marrow PFC response. However, at the 7th day after immunization the PFC response in the bone marrow fell to only 10 per cent of the bone marrow PFC activity in sham-splenectomized mice. In contrast to the primary response no regulating influence of the spleen on the bone marrow PFC activity could be demonstrated during the secondary response. The influence of splenectomy on the appearance of B-memory cells in the bone marrow depended on the priming dose. The appearance of LPS-specific B-memory cells in the bone marrow was not affected by splenectomy at priming doses of LPS as high as 1 and 0.1 μg. On the other hand splenectomy before 0.01 μg LPS i.v. reduced, and splenectomy prior to 0.001 μg LPS i.v. completely prevented the appearance of B-memory cells in the bone marrow
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