ABSTRACT
Objective To explore the impact of ketogenic diet (KD) on follicular helper T cells(TFH) in children with intractable epilepsy.Methods Thirty-three cases with intractable epilepsy were selected between Jul.2013 and Jan.2014 in Shenzhen Children's Hospital,19 boys and 14 girls; average age was 39.6 months,and seventeen age-matched healthy children who took a physical examination in the same hospital were assigned as the healthy control group.Blood samples were collected from the children with refractory epilepsy before and after 1 week of KD treatment.The proportions of the various stages of B cells and TFH cells were detected by flow cytometry.The plasma concentration of interleukin-21 (IL-21) was determined by enzyme-linked immunosorbent assay(ELISA),and realtime quantitative PCR(RT-PCR) was performed to detect the levels of peroxisome proliferator-activated receptor gamma (PPAR-γ),B-lymphocyte-induced maturation protein-1 (Blimp-1),B-cell lymphoma 6 (Bcl6) and IL-21 mRNA expression in CD4 + T cells.Results (1) The number of TFH cells in children with intractable epilepsy [(3.57 ± 0.58) ％] was remarkably decreased after KD treatment(P ＜ 0.01),while there were no difference between after KD treatment and healthy control group[(4.93 ±0.70)％ vs (5.03 ±0.63)％,P ＞0.05].(2) The levels of transcription factor Bcl6 expression after treatment were significantly decreased,while inhibitory factor Blimp-1 expression increased (P ＜ 0.05).(3)The plasma concentration of IL-21 had a trend to decrease (P ＞ 0.05),while there were no difference before and after KD treatment,and levels of IL-21 mRNA expressions in CD4 +T cells were significantly decreased after the treatment (8.28 × 10-3 ± 1.19 × 10-3 vs 1.72 × 10-2 ± 0.81 × 10-2,t =3.08,P ＜ 0.05).(4) There was no significant difference in CD27-IgD + B cells before and after KD treatment (P ＞ 0.05),CD27 + IgD + B cell and CD27-IgD-B cells had a trend to decrease after KD treatment(P ＞0.05),and CD27 + IgD-B cells and CD27 + IgD-CD38 high plasma cells were significantly decreased after KD treatment (P ＜ 0.05).(5) The number of TFH cells were correlated positively with the number of CD27 + IgD-B cells and CD27 + IgD-CD3g high plasma cells (r =0.785,0.745,P ＜ 0.05).(6) The levels of PPAR-γmRNA in CD4 + T cells expression were significantly up-regulated after KD treatment (3.49 × 10-3 ± 1.10 × 10-3 vs 2.28 ± 10-3 ± 1.30 × 10-3,t =3.41,P ＜0.05),and the number of TFH cells and PPAR-γgene expression was correlated negatively (r =-0.619,P ＜ 0.05).Conclusions KD might down regulate TFH cell number and function through inducing PPAR-γexpression and could inhibit B cell differentiation,which might be one of the factors for hypogammaglobuinemia by KD treatment.
ABSTRACT
Objective To investigate the possible factors for differentiation affecting of neonatal regulatory T cells(Treg). Methods Umbilical cord blood was collected from 200 newborns. Treg number was detected by DNA demethylation in the Foxp3 of Treg - cell - specific demethylatedregion(TSDR)based on high resolution melting anal-ysis(HRMA),concentrations of 7,8 - dihydroxy - 9,10 - epoxy - benzo(a)pyrene(BPDE - DNA)adducts and interleukin - 4( IL - 4)in the supernatants of cord blood by enzyme - linked immunosorbent assay( ELISA),and follow - up questionnaires were carried out till 1. 0 - 1. 5 years,for recurrent wheezing or stubborn eczema in infants and related information on parental history of atopic diseases. Results (1)In wheezing group[(0. 48 ± 0. 05)% ]and ec-zema group[(0. 76 ± 0. 05)% ],the number of Tregs was significantly lower compared with that of the asymptomatic group[(1. 14 ± 0. 08)% ](t = 2. 62,2. 83,all P ﹤ 0. 05);the number of Treg in parental history of atopic group was significantly lower than that of the non - atopic group(P ﹤ 0. 05);but the Treg numbers in the non - atopic group was still lower than that of the asymptomatic group(P ﹤ 0. 05).(2)The concentrations of BPDE - DNA adducts in the wheezing group[(236. 30 ± 6. 59)ng/ L]and the eczema group[(173. 40 ± 7. 38)ng/ L]were higher than those of the asymptomatic group[(111. 01 ± 3. 36)ng/ L](t = 10. 35,6. 53,all P ﹤ 0. 05),while BPDE - DNA adduct concen-trations in the atopic group with parental history of wheezing or eczema in infants were lower than those of the non -atopic group(P ﹤ 0. 05).(3)The concentrations of IL - 4 in the wheezing or eczema group in the supernatants of cord blood was higher than the asymptomatic group(P ﹤ 0. 05). Conclusions Neonatal genetic factors and BPDE - DNA adducts could affect Treg differentiation,which are probably the reasons for the formation of allergic diseases.
ABSTRACT
Objective To investigate the possible mechanism of hypogammaglobuinemia caused by ketongenic diet (KD).Methods Thirty-six children with intractable epilepsy (IP) and seventeen age-matched healthy children were recruited in this study.The percentages of B cells at various stages of devel-opment and follicular helper T ( Tfh) cells were detected by flow cytometry.The plasma concentrations of IL-21 were determined by ELISA.Real-time quantitative PCR was performed to detect the expression of mam-malian target of rapamycin ( mTOR) , Blimp-1, Bcl-6 and IL-21 at mRNA level in CD4+T cells.Results mTOR at mRNA level was significantly down-regulated after KD treatment (P<0.05).The numbers of Tfh cells were positively correlated with the transcriptional level of mTOR (r=-0.691, P<0.05).Conclusion KD treatment might down-regulate Tfh and B cells through suppressing the expression of mTOR at mRNA level, suggesting a possible mechanism of hypogammaglobuinemia induced by KD treatment.