Results in immunology最新文献

HLA/peptide tetramers are frequently used for ex vivo monitoring of disease- or vaccine-induced T cell immune responses and for T cell epitope identification. However, when low-levels HLA/peptide tetramer-positive T cell populations are encountered, it is difficult to ascertain whether this represents a true T cell receptor (TCR)-mediated interaction or background signal. To address this issue, we have developed a method for both HLA class I and class II tetramer assays to confirm tetramer-binding to the TCR/CD3 complex. Preincubation of T cells with anti-CD3 mAb SPV-T3b and subsequent crosslinking interferes with the binding of HLA/peptide tetramers to the TCR/CD3 complex and thereby indicates to what extent HLA/peptide tetramer binds through interaction with TCR/CD3 complex. SPV-T3b pretreatment results in a 2- to 10-fold decrease in tetramer-binding intensity to antigen-specific CD8+ or CD4+ T cells, whereas background reactivity of HLA/peptide tetramers containing HIV-derived peptide in HIV-negative donors remained unchanged. SPV-T3b pretreatment forms a valuable tool to verify tetramer-based detection of antigen-specific T cells during the monitoring of immune responses in clinical studies.

Crystal toxins from Bacillus thuringiensis bind to glycolipids and glycoproteins using two different lectin domains in the toxin protein. Our previous observations suggested that the sequestration of crystal toxin depends on the functional interaction of a toxin lectin with glycolipids. Given the finding that competition of a galectin LEC-8 with Cry5B for binding to glycolipids resulting in reduced Bt toxicity in nematode, it is interesting to explore the role of LEC-8 in insects. Here, we reported that the LEC-8 can also be exploited by insect for their survival when they were fed with Bt toxin food. Bioassay with LEC-8 showed that pre-feeding of Helicoverpa armigera larvae reduced the Cry1Ac susceptibility. Both LEC-8 and Cry1Ac bind to the midgut glycolipid in a similar way. Further ELISA indicated that LEC-8 interacts with glycolipid from insect midgut, thus reduce Cry1Ac binding to glycolipid. This in turn enhances insect tolerance to Cry1Ac toxin. The sugar determinants of LEC-8 were studied by using haemagglutination (HA) and haemagglutination inhibition (HAI) assay. It was suggested that the terminal sugar of LEC-8 has multiple sugar binding property.

To examine possible determinants of autoantibody levels at type 1 diabetes mellitus (T1DM) onset.

We assessed levels of glutamic acid decarboxylase 65 islet cell antigen (GADA) and anti-insulin antibodies (IAA) in 247 incident T1DM cases presenting <15 years of age in Melbourne from 1st March 2008 to 30th June 2010.

58.9% (142/241) of cases were GADA seropositive and 42.3% (94/222) were IAA seropositive. Factors associated with elevated IAA antibodies included younger age and red hair phenotype. Factors associated with elevated GAD antibodies included lower birthweight and recent eczema. Intriguingly, low recent or past sun exposure was only associated with elevated GADA levels among children presenting at age <5 years, not older (difference in effect, p<0.05 for 4 of 5 associations).

These findings show that environmental and phenotypic factors are associated with autoantibody levels at time of presentation for T1DM. We recommend such environmental and phenoytypic factors should be examined in further detail.

Aplastic anemia (AA) is a marrow failure syndrome mediated by aberrant T-cell subsets. Mesenchymal stem cells (MSCs) play an important role in maintaining immune homeostasis through modulating a variety of immune cells. However, little is known about the immunomodulation potential of bone marrow MSCs (BM-MSCs) in AA. Here, we reported that BM-MSCs from AA patients were reduced in suppressing the proliferation and clonogenic potential of CD4⁺ T cells and the production of tumor necrosis factor-α (TNF-α) and interferon-γ (IFN-γ), which was associated with decreased prostaglandin E₂ (PGE₂). Meanwhile, BM-MSCs from AA patients were defective to promote CD4⁺CD25⁺FOXP3⁺ regulatory T cells expansion through reduced transforming growth factor-β (TGF-β). No significant difference between AA and normal BM-MSCs was observed in affecting the production of interleukins (IL)-4, IL-10 and IL-17. Our data indicate that BM-MSCs were impaired in maintaining the immune homeostasis associated with CD4⁺ T cells, which might aggravate the marrow failure in AA.

This study was performed to detect and characterise the possible occurrence of natural and inducible lectins in human serum by hemagglutination method, wherein, the serum was treated using exogenous elicitors, namely, proteases and detergents.

Natural and inducible lectins were detected and characterised in human serum. Untreated serum agglutinated buffalo and rabbit RBC, while serum treated with pronase, trypsin, α-chymotrypsin or SDS for the very first time, agglutinated hen/hen and sheep RBC within 15 min in a dosimetric manner. Cross adsorption test revealed that both trypsin and α-chymotrypsin-treated serum showed similar RBC adsorption pattern. The lectin activity in untreated, pronase-treated serum was cation independent and moderately sensitive/insensitive to calcium chelator EDTA, whereas, trypsin-treated serum was cation dependent as well as EDTA sensitive (sheep RBC), cation independent and EDTA insensitive (hen RBC). Hemagglutination of untreated serum was inhibited by certain glycosides and di-, oligo-saccharides, whereas, activity in pronase-treated serum was inhibited by hexosamines. By contrast, hemagglutination of trypsin-treated serum showed specificity for acetylated mannosamine as well as sialic acid for sheep RBC and certain glycoproteins for hen RBC.

Thus, we have detected inducible lectins with distinct ligand binding specificity, upon treatment of human serum with proteases, namely, pronase and trypsin. Nevertheless, lectin activity was found in untreated human serum too with different ligand specificity.

The aim of this study is to investigate the long-term immunogenicity of inactivated split-virion 2009 pandemic influenza A H1N1 vaccine after a single immunization. We recruited 480 adults, aged 18–60 years, for a placebo-controlled, observer-masked, single-center clinical study. We randomly assigned subjects into four groups: 15 μg, 30 μg and 45 μg of hemagglutinin (HA) dosage groups, and a placebo control group. Finally, 259 subjects completed the entire study. The rates of seroconversion and seroprotection and the geometric mean increase (GMI) fulfilled the criteria of the European Medicines Agency (EMEA) for influenza vaccine for 180 days after vaccination in all three dosage groups. However, the seroprotection rates of all dosage groups were below 70% at day 360 post vaccination, while the seroconversion rates and the GMI continued to meet the licensure criteria at this time point. In conclusion, a single dose of 15 μg HA vaccine could induce a protective immune response persisting for at least six months in adults. This study could be beneficial for the future development of influenza vaccines conferring long-term immunity.

Infectious bursal disease (IBD) is a highly contagious disease of chickens which leads to immunosuppression. In our previous study it was demonstrated that, possibly, CD4⁺ and CD8⁺ T cells may employ perforin and granzyme-A pathway for the clearance of IBDV-infected bursal cells. In this study, we evaluated the cytotoxic T cell responses involving two independently functioning but complementary mechanisms: Fas–Fas ligand and perforin–granzyme pathways in IBDV-infected chickens. As demonstrated previously, infection of chickens with IBDV was accompanied by influx of CD8⁺ T cells in the bursa and spleen. There was an upregulation in the gene expression of cytolytic molecules: Fas and Fas ligand (FasL), perforin (PFN) and granzyme-A (Gzm-A) in bursal and in the splenic tissues of IBDV inoculated chickens. Additionally, for the first time, we detected Fas, Fas ligand, Caspase-3 and PFN producing CD8⁺ T cells in the bursa and spleen of IBDV-infected chickens. The infiltration and activation of CD8⁺ T cells was substantiated by the detection of Th1 cytokine, IFN-γ. These data suggest that T cells may be involved in the clearance of virus from the target organ bursa and peripheral tissues such as spleen. The findings of these studies provide new insights into the pathogenesis of IBD and provide mechanistic evidence that the cytotoxic T cells may act through both Fas–FasL and perforin–granzyme pathways in mediating the clearance of virus-infected cells.

The survival rate, weight loss, immune parameters, resistance against Vibrio alginolyticus and white-spot syndrome virus (WSSV), and expressions of lipopolysaccharide- and ß-glucan-binding protein (LGBP), peroxinectin (PX), prophenoloxidase-activating enzyme (ppA), prophenoloxidase (proPO) I, proPO II, α2-macroglobulin (α2-M), integrin ß, heat shock protein 70 (HSP70), cytosolic manganese superoxide dismutase (cytMnSOD), mitochondrial manganese superoxide dismutase (mtMnSOD), and extracellular copper and zinc superoxide dismutase (ecCuZnSOD) were examined in the white shrimp Litopenaeus vannamei (8.18 ± 0.86 g body weight) which had been denied food (starved) for up to 14–28 days. Among shrimp which had been starved for 7, 14, 21, and 28 days, 100%, 90%, 71%, and 59% survived, and they lost 3.2%, 7.3%, 9.2%, and 10.4% of their body weight, respectively. Hyaline cells (HCs), granular cells (GCs, including semi-granular cells), the total haemocyte count (THC), phenoloxidase (PO) activity, respiratory bursts (RBs), and SOD activity significantly decreased in shrimp which had been starved for 1, 1, 1, 5, 14, and 3 days, respectively. The expression of integrin ß significantly decreased after 0.5–5 days of starvation, whereas the expressions of LGBP, PX, proPO I, proPO II, ppA, and α2-M increased after 0.5–1 days. Transcripts of all genes except ecCuZnSOD decreased to the lowest level after 5 days, and tended to background values after 7 and 14 days. Cumulative mortality rates of 7-day-starved shrimp challenged with V. alginolyticus and WSSV were significantly higher than those of challenged control-shrimp for 1–7 and 1–4 days, respectively. In another experiment, immune parameters of shrimp which had been starved for 7 and 14 days and then received normal feeding (at 5% of their body weight daily) were examined after 3, 6, and 12 h, and 1, 3, and 5 days. All immune parameters of 7-day-starved shrimp were able to return to their baseline values after 5 days of re-feeding except for GCs, whereas all parameters of 14-day-starved shrimp failed to return to the baseline values even with 5 days of re-feeding. It was concluded that shrimp starved for 14 days exhibited three stages of modulation of gene expression, together with reductions in immune parameters, and decreased resistance against pathogens.

Histone H2A participates in host defense responses by producing antimicrobial peptides (AMPs). The present study deals with identification of a putative antimicrobial sequence, Himanturin from the histone H2A of Round Whip Ray, Himantura pastinacoides. A 204 bp fragment encoding 68 amino acid residues was amplified from cDNA of Round Whip Ray, H. pastinacoides. Himanturin exhibited high similarity to previously reported histone H2A derived AMPs indicating the presence of an antimicrobial sequence motif. Physicochemical properties of Himanturin suggest it to be a potential antimicrobial candidate.

Penaeidins are members of a special family of antimicrobial peptide existing in penaeid shrimp and play an important role in the immunological defense of shrimp. Here, we report a penaeidin sequence cloned from the Indian white shrimp Fenneropenaus indicus (Fein-Penaeidin). The Fein-Penaeidin open reading frame encodes a 77 amino acid peptide including a 19 amino acid signal peptide. The deduced amino acid sequences of Fein-Penaeidin include a proline rich N-terminal domain and a carboxyl-domain that contains six cysteine residues. Structural analysis revealed an alpha-helix in its secondary structure and the predicted 3D structure indicated two-disulphide bridges in the alpha-helix. Phylogenetic analysis and sequence comparison with other known peaneidin suggest the gene shows high similarity to that of penaeidin from Peneaus monodon (95%), F. indicus (80%) and Fenneropenaeus chinensis (74%). Fein-Penaeidin was examined in normal and microbial challenged shrimp and was found to be constitutively expressed in haemocytes, Heart, gills, muscles, intestine, hepatopancreas and eyestalk. Bacterial challenge resulted in mRNA up-regulation, inducing expression at 6 h post injection indicating the penaeidin involved in the innate immunity.