Investigation of serum IL-12, IL-16, and IL-17A as diagnostic biomarkers in children with cow’s milk protein allergy

Background Cow’s milk protein allergy (CMPA) is an abnormal immune response caused by milk proteins and is most common in infancy and early childhood. Statistics revealed up to 7.5% of children suffered from milk allergy. Its clinical symptoms were characterized by diversity, non-specificity, and can affect multiple systems, including the digestive tract, skin, and respiratory tract. In this study, we aimed to investigate the effects of IL-12, IL-16, and IL-17A on diagnosing and monitoring CMPA in children for clinical treatment.Methods A total of 158 infants with CMPA and 89 healthy babies were recruited and evaluated. Demographic and clinical information of all participants were recorded. An extensive analysis of inflammatory cytokine levels, including IL-12, IL-16, and IL-17A, was performed in blood samples from 247 infants younger than 9 months. Meanwhile, the serological specificity immunoglobulin E (sIgE) levels were evaluated. In addition, the area under the curve (AUC) values of IL-12, IL-16, and IL-17A in differentiating CMP from healthy babies were measured by receiver operating characteristic analysis. Finally, the correlation between sIgE and IL-12, IL-16, and IL-17A levels were detected using Spearman correlation analysis.Results Compared with healthy control, infants who developed CMPA had decreased IL-12, increased IL-16, and IL-17A. Moreover, a significant correlation between serum IL-12, IL-16, IL-17A and sIgE levels was observed in the CMPA group. In addition, AUC values of IL-12, IL-16, and IL-17A in discriminating CMPA from healthy infants were 0.8425, 0.9196, and 0.8813, respectively. Finally, IL-12 was increased while IL-16 and IL-17A levels were decreased in the CMPA group after three months of milk avoidance treatment.Conclusions We found that IL-12, IL-16, and IL-17A levels in children with CMPA were associated with SCORAD scores, sIgE levels (AU)

Generation of an alpaca derived nanobody recognizing human Her2 antigen / 军事医学

Objective To obtain alpaca single domain antibody targeting Her2.Methods An alpaca was immunized with human recombination Her2 protein mixed with Freund's adjuvant.Total RNA was extracted from the alpaca's blood and was used to synthesize first strand cDNA.Single domain antibody variable region (VHH) gene of the alpaca was amplified by PCR and cloned into pMES4 vector for library construction.After screening, E.coli BL21 (DE3) was transformed with selected clones and was induced with IPTG for the expression of recombinant proteins.The nanobody was purified by nickel ion affinity chromatography column.The affinity of the nanobodies to Her2 was tested.Results After the second round of screening, two antibody clones were selected, H3 and H5.The affinity of H5 was 8.106×10-10mol/L.Histochemistry results showed that H5 could recognize Her2 antigen in breast tumor tissue.Conclusion An Her2 specific nanobody derived from alpaca is obtained through phage display library screening, which can recognize human Her2 antibody in human breast tumor tissue.