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A new murine CD4+ T cell subset with an unrestricted cytokine profile.

CD4+ T cell clones were derived from mice immunized to keyhole limpet hemocyanin to characterize the cytokine profiles of newly isolated clones. Surprisingly, several of the clones had an unrestricted profile, producing IL-2, IL-3, IL-4, IFN-gamma, and TNF after either Con A or Ag stimulation. The coproduction of IL-2 and IL-4 was confirmed at the mRNA level. Subclones were derived which contained RNA transcripts for, as well as secreted, both IL-2 and IL-4 thus confirming the clonality of the original T cell clones. CD4+ T cell clones that expressed an unrestricted cytokine profile upon Con A stimulation were also isolated from mice immunized to other Ag (hen egg lysozyme, OVA, or type II collagen). These data indicate that CD4+ T cell clones newly isolated from immunized mice do not necessarily segregate into the Th1 and Th2 subsets. We propose this new murine CD4+ cell subset with an unrestricted pattern of cytokine production be called Th0.

Animals↗

Dysregulation of the Th1/Th2 cytokine profile is associated with immunosuppression induced by hypothalamic-pituitary-adrenal axis activation in mice.

Hypothalamic-pituitary-adrenal (HPA) axis activation-induced immunosuppression is associated with increased concentration of circulating corticosterone and impaired cellular immune responses. The purpose of this study was to investigate the effect of chronic HPA axis activation on the cellular immune response, Th1/Th2 cytokine profile, and concentration of corticosterone. Mice were divided into two groups: a control group comprised of healthy, untreated mice that received no stress, and an HPA axis-activated group that received stress through electric shock (ES). The delayed-type hypersensitivity reaction to dinitrofluorobenzene, splenocyte proliferative response to mitogens Concanavalin A and lipopolysaccharide, Th1 and Th2 profile, and TGF-beta1 production were measured in plasma and in culture supernatants. The corticosterone concentration was also measured in plasma. In the ES group, elevated plasma corticosterone concentration was associated with immunosuppression and a significant decrease in plasma concentrations of IL-2, IL-4, and TGF-beta1. In vitro IL-2 production in response to Con A was significantly lower in the ES group than in the control group. TGF-beta1 production in nonstimulated and stimulated cultures in response to either mitogen was significantly lower in the ES group than in the control group. Plasma concentrations of IFN-gamma and IL-10 did not differ significantly between groups. The concentrations of IFN-gamma, IL-4, and IL-10 in the supernatants of splenocytes stimulated with either mitogen and IL-4 production by nonstimulated cells were significantly higher in the ES group than in the control group. These results suggest that corticosterone mediates the immunosuppression induced by HPA axis activation, and induces dysregulation of the Th1/Th2 cytokine profile.

Animals↗

Effects of linear cationic alpha-helical antimicrobial peptides on immune-relevant genes in trout macrophages.

There are increasing evidence of the potential role of antimicrobial peptides in the regulation of immune responses in mammalian species. However, the effects of these peptides in fish have yet to be investigated. In this study, we examined the transcriptional expression profile of representative immune-relevant genes in a trout macrophage cell line, RTS11, in response to three linear cationic alpha-helical antimicrobial peptides (insect cecropin B, fish pleurocidin and a cecropin analogue CF17). The expression levels of two pro-inflammatory genes, interleukin-1 beta (IL-1 beta) and cyclo-oxygenase-2 (COX-2), increased in the peptide-treated RTS11 cells. The peptides did not appear to affect the expression levels of representative genes associated with antigen presentation, interferon response or JAK/STAT signal transduction. Furthermore, the induction of IL-1 beta and COX-2 in RTS11 by lipopolysaccharide was not adversely affected by these three antimicrobial peptides. Overall, the data indicate a pro-inflammatory effect of the three cationic antimicrobial peptides in the inflammatory response of salmonid species, suggesting a potential application of these peptides as immune adjuvant for fish vaccination.

Aeromonas↗

Submicroscopic profile of Isospora belli enteritis in a patient with acquired immune deficiency syndrome.

Small bowel mucosal fragments from a human immunodeficiency virus-positive female patient with chronic diarrhea were investigated by transmission electron microscopy, and Isospora belli enteritis was documented. The submicroscopic profile was characterized by a moderate abnormality of mucosal architecture with reduction in height of villi and hypertrophy of crypts. Stages of both asexual (trophozoite, schizont and merozoite) and sexual (macrogametocyte) phases of the life cycle of the parasite were identified in the epithelium, always enclosed within a parasitophorous vacuole. Moreover, the presence of occasional extracellular merozoites in the intestinal lumen and in the lamina propria near or within lymphatic vessels was documented. These findings expand the current knowledge of this parasite regarding its capacity to survive in an extracellular environment and document a possible mechanism by which extraintestinal infection can take place.

AIDS-Related Opportunistic Infections↗

Long-term effect of primary immunization on subsequent immune responsiveness.

Specific antigen/adjuvant combinations preferentially induce type 1 or type 2 cytokine responses. For example, BALB/c mice primed with TNP-ovalbumin in complete Freund's adjuvant (TNP-OVA/CFA) produce a type 2-dominated response characterized by the activation of IL-4-secreting cells and the production of IgG1 and IgE anti-TNP antibodies. In contrast, mice primed with TNP conjugated to Brucella abortus (TNP-BA) produce a type 1 response dominated by the secretion of IFN-gamma and IgG2a anti-TNP antibodies. We examined whether treating young mice with these antigen/adjuvant combinations altered the cytokine profile of their subsequent immune responses. Mice immunized with TNP-BA and boosted several months later with TNP-OVA/CFA developed a cytokine and antibody profile similar to the priming rather than boosting antigen. This was also observed in mice immunized with TNP-OVA/CFA and boosted with TNP-BA. Both the ratio of IL-4:IFN-gamma-secreting cells and the isotype of antibodies produced by these mice were altered by primary immunization. Analysis of Con A-responsive cells from these animals showed that long-lived changes in the frequency of T lymphocytes available to secrete type 1 versus type 2 cytokines were induced by strong primary immunogens.

Animals↗

[Frequency of immunologic disorders in acute schizophrenia].

Immunological examinations in schizophrenic patients have shown that there are many alterations in both arms of the immune system, i.e. cellular and humoral activities. The results are quite heterogeneous, as not even all schizophrenics show these pathological changes. Immunological findings are assumed to be etiopathogenetically related to the disease process or to be an epiphenomenon. The present study supposes that immunological alterations as they can be found during the course of schizophrenia may be an indicator for somatic vulnerability or an epiphenomenon. 60 male inpatients, fulfilling DSM-IV criteria of schizophrenia where examined during their acute phases of psychosis and during their phases of clinical improvement, by means of a serological profile including cellular and humoral parameters. The control group consisted of 42 healthy male volunteers. It was the aim of this study to find out if there were (a) overall differences in the immune profiles between patients and control group and (b) differences between different categories of schizophrenic disorder. During the acute phase nearly half of the schizophrenic patients showed pathologic immunological parameters, whereas none of the controls did. During the phase of clinical improvement the number of patients with normal immunological findings predominated. Furthermore there was a difference between the Paranoid and the Disorganized Subtype, the latter showing more immunological abnormalities. The results of this study give further support to the hypothesis that immunological aberrations should not be seen as closely etiopathogenetically related to schizophrenic disorders, but rather as an epiphenomenon (e.g. as a stress marker) and/or as indicators for somatic vulnerability.

Acute Disease↗

Evaluation of the immunocontraceptive potential of Escherichia coli expressed recombinant non-human primate zona pellucida glycoproteins in homologous animal model.

In order to evaluate the immunocontraceptive potential of zona pellucida (ZP) glycoproteins, recombinant bonnet monkey (Macaca radiata) zona pellucida glycoprotein-1 (r-bmZP1) and -2 (r-bmZP2) were expressed as polyhistidine fusion proteins in Escherichia coli. Female bonnet monkeys were immunized with the purified r-bmZP1 (n=5) and r-bmZP2 (n=4) conjugated to diphtheria toxoid (DT). Immunization led to generation of antibodies against r-bmZP1, r-bmZP2 and DT as determined by enzyme linked immunosorbent assay. The immunized animals exhibited normal menstrual cyclicity and progesterone profile, except during the summer amenorrhoea. Immunized animals, when mated with males of proven fertility, showed protection from conceiving for cumulative 45 ovulatory cycles in r-bmZP1-DT immunized group and 32 ovulatory cycles in r-bmZP2-DT immunized group. Ovarian histopathology of both the immunized groups revealed the presence of atretic follicles with degenerated oocytes, which may have been the principle cause for the failure of immunized animals to conceive in spite of the decline in either anti-r-bmZP1 or anti-r-bmZP2 antibody titres to background levels. These studies demonstrate, for the first time, that the block of fertility subsequent to immunization with r-bmZP1 and r-bmZP2, in a homologous non-human primate model, may be mediated due to ovarian dysfunction.

Animals↗

Hepatitis C virus-like particles induce virus-specific humoral and cellular immune responses in mice.

We have recently described the production of hepatitis C virus-like particles (HCV-LPs) in insect cells that resemble the putative virions. Here we evaluate the humoral and cellular immunogenicity of the virus-like particles with or without viral p7 protein, a small viral polypeptide that resides between the structural and nonstructural regions of the HCV polyprotein and whose function has not been defined. Immunized BALB/c mice developed high titers of anti-E2 antibodies and virus-specific cellular immune responses including cytotoxic T lymphocytes and T helper responses with gamma interferon production. The virus-like particles without p7 generated a higher cellular immune response with a more T(H)1 profile than the particles with p7. Immunization of heat-denatured particles resulted in substantially lower humoral and cellular responses, suggesting that the immunogenicity is strongly dependent on particle formation. Administration of CpG oligonucleotide or cationic lipid 3beta-[N-(N',N'-dimethylaminoethane)carbamoyl]cholesterol (DC-Chol), two potent adjuvants, did not significantly enhance the immunogenicity of HCV-LPs. Our results indicate that HCV-LPs can induce humoral and cellular immune responses and offer a promising approach to vaccine development.

Adjuvants, Immunologic↗

Effect of active immunization against inhibin on hormonal concentrations and semen characteristics in Shiba bucks.

Active immunization against inhibin increased ovulation rate in females; in males, the effects of active immunization against inhibin on hormonal concentrations and sperm production need more investigation. To test the hypothesis that active immunization against inhibin increases FSH secretion and sperm output, the present study was undertaken to determine the effects of active immunization against inhibin on hormonal profile and sperm production in Shiba bucks. The bucks were actively immunized against inhibin alpha-subunit (immunized group, n=6) or Freund adjuvant (control group, n=5) four times, at 5-weeks intervals. Blood samples were collected twice-weekly and two successive ejaculates of semen were collected (with an artificial vagina) once-weekly. Plasma concentrations of FSH, LH and testosterone were measured by radioimmunoassay (RIA) and sperm motility characteristics were measured by computer-assisted sperm analysis (CASA). All inhibin-immunized bucks produced antibodies against inhibin. Relative to control bucks, in immunized bucks there were significant increases in plasma FSH concentrations and in sperm concentrations from 5 to 9 weeks and from 8 to 11 weeks, respectively, after primary immunization. However, plasma concentrations of LH and testosterone, semen volume, percentage of motile spermatozoa and motility parameters (straight-line velocity, curvilinear velocity and linearity index) were similar in both groups. In conclusion, active immunization against inhibin alpha-subunit increased FSH secretions and enhanced sperm production in bucks, whereas LH and testosterone concentrations, semen volume and sperm motility parameters were unaffected. Active immunization against inhibin could be used to improve fertility in Shiba bucks.

Animals↗

Hydrogen peroxide increases interleukin-12 p40/p70 molecular ratio and induces Th2-predominant responses in mice.

To investigate the influence of oxidative stress on the immune response, mice were injected with H(2)O(2), and peritoneal macrophages were isolated and stimulated in vitro with lipopolysaccharide (LPS). H(2)O(2) significantly augmented both interleukin (IL)-12p40 and IL-12p70 production and increased the p40/p70 molecular ratio. This was confirmed by mRNA analysis, which showed that H(2)O(2) increased LPS-induced mRNA expression of both IL-12p40 and IL-12p35 subunits with an increased p40/p35 ratio. Analysis of anti-ovalbumin (OVA) antibodies revealed that H(2)O(2) injection significantly increased the production of type 2 helper T cell (Th2)-associated antibody classes [immunoglobulin (Ig)E and IgG1] but not a Th1-associated antibody class (IgG2a). To confirm the Th2-predominant immune response, we analyzed the profile of cytokine production by spleen T cells of OVA-immunized and H(2)O(2)-injected mice. H(2)O(2) significantly increased the production of IL-4 but not that of interferon-gamma. Together, these results suggest that H(2)O(2)-induced overproduction of IL-12p40 promotes the Th2-predominant response through increased production of IL-12p40-homodimers, which could serve as an antagonist of the Th1-inducing cytokine IL-12p70.

Animals↗

Immune suppression and Th1/Th2 balance in pregnancy revisited: a (very) personal tribute to Tom Wegmann.

PROBLEM: The paradigm of local suppression necessary to understand the survival of the fetal allograft is often compared with the host-tumor relationship. METHODS: We investigated two components of local immune suppression: placenta-induced immunosuppression, which is mediated at least in part by a soluble factor of low molecular weight that can induce anergy in lymphocytes, and interleukin-10 (IL-10). RESULTS: We show that enhancement of IL-10 production in the decidua and placenta after alloimmunization requires the presence of Asialo GM1+ cells. Placenta-induced immunosuppression is linked with defects in phosphorylation of some components of the T cell receptor. CONCLUSION: NK cells could be in fact regulatory cells pushing maternal immune response toward a Th2 profile, beneficial for fetal survival, or toward a Th1 type of immune response, which acts in synergy. Modulation of TcR may represent a new mechanism for maternal-fetal tolerance.

Animals↗

Effects of passive immunization of goats against inhibin on follicular development, hormone profile and ovulation rate.

This study was conducted to investigate the effect of immunoneutralization against endogenous inhibin on FSH secretion and ovulation rate, with the aim of developing a new superovulation method using inhibin antiserum in goats. Two groups of goats received an i.v. injection of either 10 ml normal goat serum (control; n = 6) or 10 ml inhibin antiserum developed against [Tyr(30)]-inhibin alpha (1-30) (n = 6) 48 h before treatment with prostaglandin F(2alpha) (PGF(2alpha)). Blood samples were collected at 6 h intervals and ovaries were examined each day using a B-mode ultrasound scanner equipped with a 7.5 MHz transducer during the experimental period. Immunization against inhibin resulted in a four- to fivefold increase (P < 0.01) in plasma concentrations of FSH. After luteolysis, plasma concentrations of oestradiol increased markedly to reach a preovulatory peak, which was about two times higher (P < 0.01) than that of the controls. The treatment was accompanied by a significant increase in the total number of follicles of > or = 3 mm in diameter at 24 (8.2 +/- 0.4 in inhibin antiserum group versus 4.8 +/- 0.3 in control group) and 96 h later (13.5 +/- 1.0 in inhibin antiserum group versus 5.3 +/- 0.6 in control group). The ovulation rate was significantly (P < 0.01) higher in goats treated with inhibin antiserum (4.2 +/- 0.5; n = 6) than in control goats (1.8 +/- 0.3; n = 6). These results indicate that inhibin is an important factor in the regulation of FSH secretion in goats and demonstrate that passive immunization against inhibin at 48 h before treatment with PGF(2alpha) induces the development of more follicles and increases ovulation rate. Thus, inhibin antiserum treatment may be an alternative to the use of exogenous gonadotrophins for induction of superovulation in goats.

Animals↗

An overview of real-time quantitative PCR: applications to quantify cytokine gene expression.

The analysis of cytokine profiles helps to clarify functional properties of immune cells, both for research and for clinical diagnosis. The real-time reverse transcription polymerase chain reaction (RT-PCR) is becoming widely used to quantify cytokines from cells, body fluids, tissues, or tissue biopsies. Being a very powerful and sensitive method it can be used to quantify mRNA expression levels of cytokines, which are often very low in the tissues under investigation. The method allows for the direct detection of PCR product during the exponential phase of the reaction, combining amplification and detection in one single step. In this review we discuss the principle of real-time RT-PCR, the different methodologies and chemistries available, the assets, and some of the pitfalls. With the TaqMan chemistry and the 7700 Sequence Detection System (Applied Biosystems), validation for a large panel of murine and human cytokines and other factors playing a role in the immune system is discussed in detail. In summary, the real-time RT-PCR technique is very accurate and sensitive, allows a high throughput, and can be performed on very small samples; therefore it is the method of choice for quantification of cytokine profiles in immune cells or inflamed tissues.

Animals↗

[Expression map of immune-related genes on PBMCs in familial clustering patients with chronic virus hepatitis B].

AIM: To investigate peripheral blood mononuclear cells (PBMCs) immune-related gene expression profile in patients with chronic virus hepatitis B(CHB) by oligonucleotide gene array technique. METHODS: PBMCs were collected from the members of a family of clustering hepatitis B virus (HBV) infection including 5 CHB patients and 4 healthy spouses and RNA prepared from PBMCs was hybridized to high-density oligonucleotide arrays(HG-U133A 2.0 Human Gene Chips, Affymetrix), covering the expression of 22 000 human ESTs. Primary scanned image was analyzed with DNT software package. RESULTS: Out of the 22 000 ESTs, 24 different immune-related genes were identified. Among the 24 genes, 7 genes showed increased expression and 17 genes showed decreased expression in CHB compared with those in healthy spouses. The up-regulated genes were mainly associated with adaptive immunity, while the down-regulated genes were associated with innate immunity. CONCLUSION: Our findings suggest that HBV infection alters a broad range of immunity genes expression and innate immunity-associated genes are important in the defense against HBV chronic infection.

Down-Regulation↗

A distinct psoriasis-atopic dermatitis overlapping phenotype in adults with dual type 2 and type 3 immune features and favorable response to Janus kinase 1 inhibition.

BACKGROUND: Patients exhibiting overlapping histopathologic features of psoriasis (Pso) and atopic dermatitis (AD) present a diagnostic and therapeutic challenge. OBJECTIVES: To delineate the clinicopathological and immunologic portrait of psoriasis-atopic dermatitis overlapping (Pso-Ec) for integrated diagnosis and therapeutic decision-making. METHODS: We conducted a 2-center prospective study comparing 30 Pso-Ec patients with typical Pso and AD cohorts. Clinicopathological characteristics and immunologic profiling of lesional skin and peripheral blood were analyzed, and treatment courses were assessed. RESULTS: Pso-Ec patients (aged 13-72 years; mean age: 49.7 years) presented with ill-defined erythematous plaques with thin scales, excoriation, intense itching, and mixed psoriatic-eczematous histology. Immune profiling showed co-existence of helper T cell 2/cytotoxic T cell 2 and helper T cell 17/cytotoxic T cell 17 in skin and blood, with Janus kinase (JAK) 1 signal transducer and activator of transcription 2/6 signaling involvement. Responses to prior Pso-targeted (n = 18) and AD-targeted (n = 15) biologics were often inadequate. In contrast, JAK1 inhibitors achieved minimal disease activity (body surface area &#x2264; 2, numerical rating scale &#x2264;1) during a median follow-up of 17 months. No progression to classic Pso or AD phenotypes was observed. LIMITATIONS: Sample size was limited and immunologic analyses were performed in a representative subset of patients. CONCLUSIONS: Pso-Ec represents a distinct, predominantly adult-onset phenotype driven by dual type 2 and type 3 inflammation, and JAK1 inhibitors appear as an effective option.

Humans↗

Immune activation of peripheral blood and mucosal CD3+ lymphocyte cytokine profiles in children with autism and gastrointestinal symptoms.

Gastrointestinal pathology, characterized by lymphoid nodular hyperplasia and entero-colitis, has been demonstrated in a cohort of children with autistic spectrum disorder (ASD). Systemic and intestinal mucosal immune dysregulation was assessed in ASD children with gastrointestinal (GI) symptoms (n = 18), and typically developing controls (n = 27), including non-inflamed controls (NIC) and inflamed GI control children with Crohn's disease (CD), by analysis of intracellular cytokines in CD3+ lymphocytes. In both peripheral blood and mucosa, CD3+ TNFalpha+ and CD3+ IFNgamma+ were increased in ASD children compared with NIC (p < 0.004) and reached levels similar to CD. In contrast, peripheral and mucosal CD3+ IL-10+ were markedly lower in ASD children with GI symptoms compared with both NIC and CD controls (p < 0.02). In addition, mucosal CD3+ IL-4+ cells were increased (p < 0.007) in ASD compared with NIC. There is a unique pattern of peripheral blood and mucosal CD3+ lymphocytes intracellular cytokines, which is consistent with significant immune dysregulation, in this ASD cohort.

Autistic Disorder↗

Expression profiling of B cell chronic lymphocytic leukemia suggests deficient CD1-mediated immunity, polarized cytokine response, altered adhesion and increased intracellular protein transport and processing of leukemic cells.

We used oligonucleotide microarrays to profile the expression of chronic lymphocytic leukemia (CLL) B cells from eight patients compared with CD5-expressing normal B cells from four donors and with pooled normal circulating B cells. Of 6790 genes examined, we identified 87 genes that were differentially expressed at least two-fold between CLL and the normal B cells. CLL cells significantly down-regulated transcripts from CD1c and CD1d genes, which encode proteins known to present lipid antigen and mediate innate and adaptive immunity. The expression pattern was also consistent with reduced signaling by interferon gamma but increased response to interleukin 4 in leukemic cells. CLL cells increased the expression of several collagen-associated extracellular matrix and adhesion molecules, up-regulated many genes involved in intracellular protein transport and processing, while downregulating genes involved in proliferation and metabolism. Based on the expression pattern, we propose that CLL-B cells prolong their survival through increased interaction with survival factors such as IL-4, and through various mechanisms of evading the immune response, such as turning off the expression of CD1c and CD1d, reducing immunogenic response to interferon gamma, inactivating T cell in B-T interaction and increasing the expression of immunoglobulin receptors which neutralize antibody-dependent cell-mediated cytotoxicity.

Adult↗

SYT8 Drives Colorectal Cancer Progression and Immune Evasion via the SETD1A-H3K4me3 Axis.

By integrating transcriptomic data from The Cancer Genome Atlas, Gene Expression Omnibus, and a self-established colorectal cancer (CRC) cohort, it was identified that synaptotagmin 8 (SYT8) is significantly up-regulated in tumors and is predictive of poor prognosis. Single-cell RNA sequencing, immunohistochemistry, and immunofluorescence experiments demonstrate that SYT8 expression is largely confined to tumor cells, predominantly in the nucleus. Functional assays reveal that depletion of SYT8 impairs, whereas its overexpression enhances, CRC cell proliferation and invasion. Transcriptomic profiling indicates an enrichment of cell cycle and epithelial-mesenchymal transition signatures. Mechanistically, co-immunoprecipitation/mass spectrometry identifies SET domain containing 1A (SETD1A) as a direct SYT8-interacting partner. The SYT8-SETD1A axis forms a positive-feedback loop that increases histone H3 lysine 4 trimethylation (H3K4me3) levels and drives the transcription of protumorigenic genes. Immune profiling further indicates that high SYT8 expression correlates with increased regulatory T-cell infiltration, suggesting an immunosuppressive microenvironment and potential resistance to immunotherapy. Collectively, SYT8 promotes CRC progression through the SETD1A/H3K4me3-mediated activation of the cell cycle, induction of epithelial-mesenchymal transition, and remodeling of the immune microenvironment. Therefore, SYT8 is established as a prognostic biomarker and serves as a therapeutic target in colorectal cancer.

Humans↗