Is Serum TGF-ß1 and TGF-ß2 levels Correlated to Children with Autism Intensity?

Objective: Transforming growth factor-beta (TGF-ß), a group of multifunctional growth factors, plays an important role in the neuron survival and neurodevelopmental functions. Some studies have evaluated the correlation between TGF-ß1 and TGF-ß2 abnormalities and autism spectrum disorders. In this study, we compared the TGF-ß1 and TGF-ß2 levels between autistic and intellectually normal individuals. Materials & Methods: The study population consisted of 39 autistic and 30 age-matched intellectually normal individuals (control group). Blood samples were taken from all individuals, and all patients were divided into 2 groups (mild-to-moderate and severe) according to the childhood autism rating scale. The cytokines levels were measured by Enzyme Linked Immunosorbent Assay (ELISA). Results: The mean concentration of TGF-ß1 was significantly lower (P < 0.0001) in children with autism compared to the control group (25.3 ± 6.5 versus 35.1 ± 9.4 ng/mL, respectively). Also, the mean concentration of TGF-ß2 in children with autism (32.35± 7.75 ng/ mL) was higher compared to those in the control group (30.47± 4.36 ng/mL); however, this difference did not reach statistical significance (P = 0.21). A positive correlation was observed between TGF-ß1 concentration and autism severity (r = 0.41; P = 0.02), whereas a negative correlation was found between TGF-ß2 concentration and autism severity (r = -0.41; P = 0.02). severity (r = 0.41; P = 0.02), whereas a negative correlation was found between TGF-ß2 concentration and autism severity (r = -0.41; P = 0.02). Conclusion: The results of the present investigation suggest that there is a decrease in the levels of TGF-ß1 in the serum of patients with autism and this cytokine may be effective in the treatment of the pathophysiological aspects of autism.

Determining the Effect of Amino Acids on the Allergenic Activity of Pollen Extracts.

BACKGROUND: The diagnosis and treatment of allergic diseases require high quality pollen allergen extracts for reliable test results and effective treatments. The quality of the pollen allergen extracts is influenced by pharmacologically inert ingredients, such as stabilizers which are added to prevent the degradation of the allergenic activity. This study was conducted to develop a stabilizer formulation in order to protect the allergenic activity of the pollen's extracts. METHODS: Pine and orchard grass pollen allergen extracts were incubated for 40 days at 37 °C. The effects of chemicals were examined via inhibition ELISA on days 7, 14, 21, 28, and 40 to evaluate the ability of the pollen allergen extracts to inhibit specific IgE in the sera of sensitized patients. RESULTS: Our findings showed that the pine pollen and orchard grass allergen extracts treated with Lys/Glu had the best stabilizing effect resulting in a 97% IgE inhibition following the 40 days of incubation. In the non-treatment group, the IgE inhibition decreased to 23% at the end of the 40 days. The orchard grass pollen allergen extracts receiving no treatment decreased to 12% IgE inhibition following the 40-day incubation. CONCLUSION: Amino acids are able to act as an effective stabilizer for pollen allergen extracts and prevent the degradation of their activity over time. Particularly applying Lys/ Glu in pollen allergenic extracts can protect allergenic activity and potency of the pollen extracts to inhibit specific IgE in human sera.

Production and Characterization of Monoclonal Antibody against Vit v1: A Grape Allergen Belonging to Lipid Transfer Protein Family.

Allergy to non-specific lipidtransfer protein (nsLTP), the major allergen of grape (Vit v1), is considered as one of the most common fruit allergies in Iran. Therefore, a specific monoclonal antibody (mAb) can be used for the characterization and assessment of. Accordingly, this study aimed to generate and characterize a mAb against Vit v1 with a diagnostic purpose. To this end, Vit v1 allergen (9 kDa) was extracted using a modified Bjorksten extraction method. Natural Vit v1-immunized mouse splenocytes were fused with SP2/0Ag-14 myeloma cells for generating hybridoma cells. Specific antibody-secreting Hybridoma cells were selected using ELISA. Finally, anti-Vit v1 mAb was characterized by western blotting, ELISA, and isotyping methods. In the current study, a 9 kDa (Vit v1) protein was attained fromcrude and fresh juice of grape extracts and the isotype of desired anti-Vit v1 mAb was determined as IgM with k light chain. In addition, The ELISA results demonstrated that anti-Vit v1 mAb was specified against natural Vit v1 in the grape cultivar and related LTP allergens, such as Pla or 3 (p<0.0001). In the present study, a specific mAb was produced for detecting the LTP allergen. This mAb with a confirmed specificity can be utilized for evaluating the LTP allergens and their allergenicity in different grape cultivars.

The novel immunogenic chimeric peptide vaccine to elicit potent cellular and mucosal immune responses against HTLV-1.

This study reports on the immunogenicity assessment of a novel chimeric peptide vaccine including Tax, gp21, gp46, and gag immunodominant epitopes of human T-cell lymphotropic virus type 1 (HTLV-1) to induce immunity against HTLV-1 after subcutaneous (SC) or intranasal administration in a mice model. Additionally, to elevate the efficacy of the HTLV-1 vaccine, the chimera was physically mixed with monophosphoryl lipid A (MPLA) or ISCOMATRIX (IMX) adjuvants. For this purpose, the ISCOMATRIX with a size range of 40-60 nm were prepared using lipid film hydration method. Our investigation revealed that the mixture of IMX and chimera could significantly increase antibody titers containing IgG2a, and mucosal IgA, as well as IFN-γ and IL-10 cytokines and decrease the level of TGF-ß1, compared to other vaccine formulations. The intranasal delivery of chimera vaccine in the absence or presence adjuvants stimulated potent mucosal sIgA titer relative to subcutaneous immunization. Furthermore, the SC or nasal delivery of various vaccine formulations could shift the immunity toward cell-mediated responses, as evident by higher IgG2a and IFN-γ, as well as suppressed TGF-ß1 level. Our findings suggest that proper design, construction, and immunization of multi-epitope vaccine are essential for developing an effective HTLV-1 vaccine.

Lung-derived innate cytokines: new epigenetic targets of allergen-specific sublingual immunotherapy.

OBJECTIVES: Sublingual allergen-specific immunotherapy is a safe and effective method for treatment of IgE-mediated respiratory allergies; however, the underlying mechanisms are not fully understood. This study was planned to test whether sublingual immunotherapy (SLIT) can exert epigenetic mechanisms through which the airway allergic responses can be extinguished. MATERIALS AND METHODS: BALB/c mice were sensitized intraperitoneally and challenged intranasally. Then, they received sublingual treatment with recombinant Che a 2 (rChe a 2), a major allergen of Chenopodium album. After SLIT, allergen-specific antibodies in sera, cytokine profiles of spleen cell cultures, mRNA and protein expression of lung-derived IL-33, IL-25, and TSLP (thymic stromal lymphopoietin), and histone modifications of these three genes were assessed. RESULTS: Following Immunotherapy, systemic immune responses shifted from Th2 to Th1 profile as demonstrated by significant decrease in IgE and IL-4 and substantial increase in IgG2a and IFN-γ. At local site, mRNA and protein levels of lung-derived pro-inflammatory cytokines IL-33 and TSLP were markedly down-regulated following SLIT that was associated with marked enrichment of trimethylated lysine 27 of histone H3 at promoter regions of these two cytokines. CONCLUSION: In our study, sublingual immunotherapy with recombinant allergen effectively attenuated allergic immune responses, at least partly, by induction of distinct histone modifications at specific loci. Additionally, the lung-derived pro-allergic cytokines IL-33 and TSLP could be promising mucosal candidates for either monitoring allergic conditions or therapeutic approaches.