ABSTRACT
The aim of this study is to investigate the distribution and sequence conservation of P6 outer membrane protein (OMP6)-encoding gene of nontypeable Haemophilus influenzae isolates as well as screen and identify the predominant T-and B-cell (T-B)-combined antigenic epitopes in OMP6 sequences and their immunogenicity.The entire omp6 genes of NTHi isolates were amplified by PCR and the amplification products were sequenced after T-A cloning.By using bioinformatic softwares,the sequence conservation and membrane location of OMP6 were analyzed as well as the T-B-combined antigenic epitopes in OMP6 were predicted.The immunogenicity and immunoreactivity of T-B-combined antigenic epitope peptides displayed by recombinant phage PⅢ proteins (rPⅢ) were determined by Western Blot assay and ELISA.The PCR showed that all the 35 NTHi isolates tested were detectable for omp6 gene.The identities of nucleotide and amino acid sequences of omp6 genes from 28 strains in the NTHi isolates were 98.3％-100％ and 99.3 ％-100％,respectively.OMP6 of NTHi was predicted as an outer membrane superficial protein that contains OMP6-2-25,OMP6-61-86 and OMP6-98-126 predicted T-B-combined antigenic epitopes.The immunoblotting assay and ELISA confirmed that OMP6-2-25 presented stronger hybridization band with NTHi antisera while 96.9％ (59/62),69.4％ (43/62) and 74.2％ (46/62) of serum samples from NTHi-infected children were positive for OMP6-2-25,OMP6-61-86 and OMP6-98-126 T-B-combined antigenic epitope peptides,respectively.All the results lead to a conclusion thatomp6 is an extensive distribution and sequence conserved gene of NTHi,and OMP6-2-25 is the predominant T-B-combined antigenic epitopes which can be used as the candidates for developing multiple antigenic peptide vaccine against NTHi.
ABSTRACT
Objective To explore the effect of maternal history of allergy on the cytokine level in umbilical cord blood. Methods A prospective study was conducted among pregnant women in Yiwu City,China.136 participants were selected from the Yiwu maternal and child healthcare hospital in 2012.A questionnaire interview was conducted.The umbilical cord blood sample were collected to detect the level of IgE,eosinophile granulocyte,eotaxin,IL -9,IL -6,IL -4,IL -5, IFN -γ,IL -10.The association between allergy history and cytokine levels was analyzed.Results All subjects were recruited and divided into two groups(pregnant women with allergy history and without allergy history).Compared with the pregnant women with allergy history,these subjects without allergy history had higher IL -13 (5.52 pg/ml vs 2.84 pg/ml) and IL4 (43.33 pg/ml vs 8.23 pg/ml)(P <0.05).Besides,there is a correlation between IL -13 and IL -4 (rs =0.87,P <0.01).Partial correlation analysis showed the correlaiots among IL -4,IL -5,IL -9,IL -10,IFN -γand eotaxin.Conclusion The levels of IL -13 and IL -4 are significantly associated with the history of allergy in pregnant women.It could be considered the indexes are sensitive for early screening and surveillance of allergy for babies.
ABSTRACT
Objective To explore the influence of parental history of allergy on the cytokine level of umbilical cord blood of neonates.Methods A prospective study was conducted which selected the pregnant women attending Yiwu maternal and child healthcare hospital from July to December in 2012 as objects.The umbilical cord blood sample were collected to detect the level of IgE,eosinophile granulocyte,eotaxin,IL -9,IL -6,IL -4,IL -5,IFN -γand IL -10. Results A total of 136 subjects were recruited and divided into three groups according to the status of parental allergy histories.There was no significant difference in social and demographic characteristics among the three groups (P>0.05). Three groups have significantly different median concentration of IL-13 (1.82pg/mL vs.3.24pg/mL vs.6.12pg/mL), eotaxin (29.33pg/mL vs.50.71pg/mL vs.60.15pg/mL),IL-9 (43.75pg/mL vs.111.17pg/mL vs.183.19pg/mL), IL-6 (11.49pg/mL vs.19.35pg/mL vs.26.09pg/mL),IL-4 (3.67pg/mL vs.21.27pg/mL vs.49.51pg/mL),IL-5 (4.26pg/mL vs.7.69pg/mL vs.10.66pg/mL),IFN-γ(338.65pg/mL vs.649.17pg/mL vs.834.14pg/mL),IL-10 (7.13pg/mL vs.10.56pg/mL vs.14.64pg/mL),P <0.05.Conclusion Parental history of allergy could have influence on cytokine level of umbilical cord blood.Early screening and surveillance in neonates whose parents have history of allergy should be taken to guide early interventions.