ABSTRACT
@#Abstract: Objective To observe the expression of peripheral blood stimulating molecules CD80 and CD86 in children with severe hand, foot, and mouth disease (HFMD), and to analyze the relationship between them and the therapeutic effects of children. Methods The clinical data of 252 children with severe HFMD treated in Wuhan Hospital of Traditional Chinese Medicine from March 2017 to March 2021 were collected retrospectively. All children were treated with standardized treatment and the therapeutic effects was evaluated. The baseline data and laboratory test results of children were recorded, and the positive rates of CD80 and CD86 cells in peripheral blood were detected by flow cytometry. Logistic regression was used to analyze the relationship between the above indexes and the therapeutic effects of children. The receiver operating curve (ROC) was drawn to evaluate the value of the above indicators in predicting the therapeutic effects of children. Results After standardized treatment, 48 children were ineffective, and 204 children were effective; the levels of serum CD80 [(2.28±0.84)% vs (2.12±0.33 )%] and CD86 [(3.35±0.96)% vs (2.23±0.41)%] in children were significantly lower than those at admission (t=2.851, 16.991; P<0.05). The levels of blood lactic acid, serum C-reactive protein (CRP), matrix metalloproteinase-9 (MMP-9), CD80 and CD86 at admission in the ineffective group were significantly higher than those of the effective group (P<0.05). Logistic regression analysis showed that the overexpression of serum CRP (OR=10.929), MMP-9 (OR=1.926), CD80 (OR=3.943) and CD86 (OR=1.947) at admission might be the risk factors of ineffective (all P<0.05). The results of the goodness of fit test for the model showed that, the goodness of fit was high (χ2=6.245, P=0.620); the model collinearity results showed that the variance inflation factors (VIF) values of each variable were <2, and there was no collinearity among the main indicators; the results of the individual independence test for the model showed that Durbin-Watson statistics (D-W)=0.279 and there was poor mutual independence among main indicators. ROC curve analysis showed that the area under the curve(AUC) of serum CD80 at admission in predicting the therapeutic effects of children was 0.762, the cut-off value was 2.390%, and the specificity, sensitivity and Youden index were 0.598, 0792 and 0.390 respectively; the AUC predicted by CD86 was 0.739, the cut-off value was 3.280%, and the specificity, sensitivity and Youden index were 0.510, 0.896 and 0.406 respectively; the AUC by combined prediction was 0.823, and the specificity, sensitivity and Youden index were 0.696, 0.833 and 0.529 respectively. Conclusions Peripheral blood stimulating molecules CD80 and CD86 are involved in the progression of HFMD. Their overexpression may suggest a high risk of treatment ineffectiveness in children with severe HFMD. Early dynamic monitoring of the expression of serum CD80 and CD86 has a certain predictive value for the therapeutic effect of children.
ABSTRACT
Nitric oxide (NO) mediated mechanisms have been implicated in killing of some life-stages of Brugia malayi/Wuchereria bancrofti and protect the host through type 1 responses and IFN-γ stimulated toxic mediators' release. However, the identity of NO stimulating molecules of the parasites is not known. Three predominantly NO-stimulating SDS-PAGE resolved fractions F8 (45.24-48.64 kDa), F11 (33.44-38.44 kDa) and F12 (28.44-33.44 kDa) from B. malayi were identified and their proteins were analyzed by 2-DE and MALDI-TOF/TOF. Tropomyosin, calponin and de novo peptides were identified by 2-DE and MALDI-TOF/TOF in F8 and immunization with F8 conferred most significant protection against L3-initiated infection in Mastomys coucha. Immunized animals showed upregulated F8-induced NO, IFN-γ, TNF-α, IL-1ß, IL-10, TGF-ß release, cellular proliferative responses and specific IgG and IgG1. Anti-IFN-γ, anti-TNF-α, and anti-IL-1ß significantly reduced F8-mediated NO generation and iNOS induction at protein levels. Anti-IFN-γ treated cells showed maximum reduction (>74%) in NO generation suggesting a predominant role of IFN-γ in iNOS induction. In conclusion, the findings suggest that F8 which contains tropomyosin, calponin and de novo peptides protects the host via IFN-γ mediated iNOS induction and may hold promise as vaccine candidate(s). This is also the first report of identification of tropomyosin and calponin in B. malayi.
