乙型肝炎疫苗免疫接种现状及对无(弱)应答的探索

乙型肝炎疫苗接种在预防和控制乙型肝炎流行方面具有重要意义。本文综述乙型肝炎疫苗免疫接种现状,试从机体免疫、遗传等因素分析乙型肝炎疫苗接种后无(弱)应答发生的原因,并阐述目前对乙型肝炎疫苗无(弱)应答解决方法的研究。     

Correlation between humoral and cellular immune responses and the expression of the hepatitis A receptor HAVcr-1 on T cells after hepatitis A re-vaccination in high and low-responder vaccinees

Introduction

We recently published a study on the persistence of seroprotection 10 years after primary hepatitis A vaccination in an unselected study population of 1014 vaccinees. The majority of these vaccinees still exhibited sufficient protective antibody levels, while 2% displayed antibody concentrations below detection level. In order to investigate whether the low antibody levels were due to decline after primary vaccination or due to an intrinsic inability to sufficiently respond to hepatitis A antigen, we sought to recruit these low/no responder vaccinees to characterize their immune responses in more detail after booster vaccination in comparison to high responder vaccinees.

Materials and methods

Prior to and one week after booster vaccination with a hepatitis A vaccine, antibody levels, cytokine levels (IL-2, IFN-γ and IL-10) and CD surface marker expression on peripheral blood mononuclear cells were determined in a study population comprised of 52 individuals. Additionally, the hepatitis A HAV cellular receptor 1 (HAVcr-1) TIM-1, being also expressed on CD4+ T cells and associated with immunomodulatory properties, was measured by RT-PCR before and after hepatitis A booster.

Results

Our data indicate that there is indeed a small group of hepatitis A vaccinees that can be classified as low/no responders as their antibody levels remain below the seroprotection level of 20 mIU/ml after booster vaccination. We further describe a good correlation between antibody concentrations and cellular responses, showing that low antibody production is associated with low antigen specific cytokine levels (IL-2, IFN-γ, IL-10) and vice versa. While there was no significant difference in the expression of the most common surface markers on T and B cells before and after booster vaccination in low and high responder vaccinees, the expression of HAVcr-1 on CD4 T cells correlated significantly with the antibody responses and cytokine levels, suggesting this receptor as cellular prediction marker of immune responsiveness to hepatitis A.

Conclusion

Whether hepatitis A low/non-responders deserve particular attention as a risk group or might display certain resistance to hepatitis A infection due to a lack of the hepatitis A receptor needs further investigations. At this stage we suggest that persons at high exposure risk should be carefully observed.     

Prediction of non-responsiveness to standard high-dose gamma-globulin therapy in patients with acute Kawasaki disease before starting initial treatment

Clinical, laboratory, and echocardiographic data were retrospectively analyzed in 112 patients with acute Kawasaki disease who received high-dose (2 g/kg) intravenous gamma-globulin (IVIG) treatment within 2 days and were compared for those who were responsive and non-responsive to initial IVIG treatment. Coronary arteries adjusted for body surface area (BSA) were evaluated quantitatively by comparison with the mean dimensions for 85 normal control subjects. The incidence of coronary abnormalities was higher in IVIG-non-responsive patients as compared to IVIG-responsive patients (71% versus 5%, p<0.0001). Univariate analysis of pre-IVIG data showed that the neutrophil count and serum levels of C-reactive protein (CRP), total bilirubin (TB), aspartate aminotransferase (AST), alanine aminotransferase, and lactate dehydrogenase (LDH) were significantly higher in IVIG-non-responsive versus responsive patients. Multivariate analysis selected CRP (p=0.009), TB (p<0.001), and AST (p=0.002) as independent predictors of non-responsiveness to initial IVIG treatment. By defining predictive values, patients with at least two of three predictors (CRP≥7.0 mg, TB≥0.9 mg, or AST≥200 IU/L) are considered to be non-responsive to IVIG for acute Kawasaki disease. Alternatively, more intense initial therapy may be a promising therapeutic strategy for patients who are predicted to be IVIG-non-responsive.     

乙型肝炎疫苗接种无应答原因与机制

已正式批准使用的乙型肝炎（乙肝）疫苗包括早期的血源疫苗和目前普遍使用的重组疫苗，按标准方案实施一个免疫程序后确有一定比例的接种者乙肝表面抗体（抗HBs）滴度不能达到保护水平即10U／L，在健康人群中该比例波动在2％～15％之间。这些无应答者对乙肝病毒（HBV）仍然易感，一旦感染仍可发病或成为病毒携带者。对其原因及机制的研究已取得很多重要发现，为该问题的解决奠定了基础。     

乙型病毒性肝炎疫苗免疫无应答者进行再免疫的临床研究

目的探讨乙型病毒性肝炎疫苗(乙肝疫苗)全程接种后免疫无应答者获得免疫保护的方法。方法从接种过全程乙肝疫苗的成人中,筛选血清乙肝病毒标志物均阴性者为研究对象,并将其随机分为对照组、观察组1和观察组2。对照组单独应用乙肝疫苗10μg肌肉注射;观察组1加量应用乙肝疫苗20μg肌肉注射;观察组2联合使用卡介菌多糖核酸和乙肝疫苗。疗程结束30d检测保护性抗体(抗-HBs)。结果对照组、观察组1和观察组2的抗-HBs阳转率分别为10%、50%和90%。观察组2抗-HBs阳转率与对照组、观察组1比较,差别有统计学意义(P<0.01)。结论对乙肝疫苗全程免疫后无应答者联合使用卡介菌多糖核酸和乙肝疫苗可诱导或激发机体抗-HBs阳转,疗效满意。     

Association of HLA alleles with the responsiveness to hepatitis B virus vaccination in Korean infants

Hepatitis B virus (HBV) vaccination is the most effective means of countering HBV-related morbidity and mortality, and individuals who do not respond to vaccination (non-responders) are problematic. The aim of the present study was to investigate associations between HLA and responsiveness to HBV vaccine in Korean infants. A total of 944 healthy Korean infants 9–12 months old were enrolled, and HLA distribution was compared among subgroups in accordance with the response to HBV vaccination. The HLA distribution of the subjects was similar to known Korean population data and did not deviate from the HWE proportions. The alleles that showed positive associations with non-responsiveness (<10 mIU/mL) or low antibody titer (<100 mIU/mL) were HLA-A*33, B62, DRB1*04, and DRB1*07, while the alleles A*02 and DRB1*08 showed negative associations. Among these alleles, B62, DRB1*07 and DRB1*08(−) showed significant associations with a poor or decreased response to vaccination even after correction (OR = 1.83, 1.99, 5.63; p_c < 0.05) and also showed dose effects. After stratification by other associated alleles at different loci, B62 and DRB1*07 were independently associated with non-responsiveness, but A*02(−) and DRB1*08(–) lost their individual associations. The combined association of A*02(–)–DRB1*08(–) and B62–DRB1*08(–) was significant (OR = 25.2 and 24.5; p_c < 0.05). Although the hierarchy is not clear, we can assume the following: (i) B62 and DRB1*07 have independent effects, (ii) DRB1*08(–) has a very strong and synergic effect, and (iii) there is probability of a third factor controlling A*02(–) and DRB1*08(–) with an effect on non-responsiveness to HBV vaccination in Korean infants.     

静脉用丙种球蛋白联合糖皮质激素治疗丙种球蛋白无反应型川崎病临床研究

目的：探讨静脉用丙种球蛋白（IVIG）无反应型川崎病（KD）的进一步治疗方法及效果。方法：将我院住院确诊为IVIG无反应型川崎病的患儿45例根据进一步治疗方案分为观察组（同时追加IVIG和糖皮质激素）38例和对照组（单纯追加糖皮质激素）7例,观察治疗效果并进行随访。结果：两组患儿体温均在24 h内恢复正常。观察组和对照组治疗前血清C反应蛋白（CRP）水平分别为（115.8±55.9）mg/L、（130.1±59.4）mg/L（P>0.05）,治疗第3天分别为（22.0±8.5）mg/L、（27.7±16.3）mg/L（P>0.05）,两组患儿治疗后CRP水平均降低（P均<0.05）。观察组和对照组急性期（发病0~21 d）冠状动脉损害（CAL）发生率分别为15.8%、71.4%（P<0.05）,随访6个月内CAL发生率分别为5.3%、14.3%（P<0.05）。结论：同时追加IVIG和糖皮质激素进一步治疗IVIG无反应型KD与单纯追加糖皮质激素效果相当,但急性期及远期CAL发生率较低,可为临床治疗IVIG无反应型KD提供依据。     

Assessment of clopidogrel responsiveness: measurements of maximum platelet aggregation, final platelet aggregation and their correlation with vasodilator-stimulated phosphoprotein in resistant patients

BACKGROUND: Controversy surrounds the optimal platelet aggregation measurement to assess clopidogrel non-responsiveness. The P2Y12 reactivity ratio (PRR) determined by vasodilator-stimulated phosphoprotein phosphorylation levels has been used to indicate the extent of P2Y(12) blockade. OBJECTIVES: We sought to compare the prevalence of non-responsiveness measured by maximum (MA) and 6 min aggregation (FA) and correlate these measurements with PRR in patients with non-responsiveness. METHODS: MA and FA were measured in stented patients (n=100) before and after clopidogrel treatment. The PRR was determined in 22 non-responsive patients. Responsiveness was defined as pre-treatment minus post-treatment aggregation; and non-responsiveness was defined as <10% change in platelet aggregation. RESULTS: Responsiveness was greater as determined by FA, p=0.006 (5 microM ADP) and p=0.003 (20 microM ADP)). There was a strong correlation between MA and FA stimulated by 5 microM (r=0.84, p<0.0001) and 20 microM ADP (r=0.90, p<0.001). The prevalence of non-responsiveness rose with agonist concentration but did not differ significantly between methods: 5 microM ADP [22% (MA) vs. 17% (FA), p=0.186] and 20 microM ADP [33% (MA) vs. 29% (FA), p=0.270]. PRR correlated with both MA (r=0.66, p<0.001) and FA (r=0.74, p<0.001) in non-responsive patients indicating incomplete receptor blockade. CONCLUSION: Clopidogrel responsiveness is higher when measured by FA as compared to MA. However, these measurements are equivalent in determining the prevalence of non-responsiveness: FA and MA are affected to the same degree in patients with non-responsiveness. These findings are relevant to ongoing studies assessing platelet inhibition by P2Y(12) inhibitors and support previous studies that employed MA to assess non-responsiveness.     

静脉丙种球蛋白无反应性川崎病的危险因素分析

目的:探讨首次静脉注射丙种球蛋白(IVIG)无反应性川崎病(KD)的发病率及相关危险因素。方法:总结2005年1月至2010年12月温州医学院附属第二医院KD患儿的临床资料,IVIG无反应性KD定义为首次IVIG治疗无效,36 h后体温仍超过38.5℃,根据对首次大剂量IVIG有无反应分成IVIG敏感组和IVIG无反应组,比较两组的临床特点。结果:515例KD患儿纳入研究对象,其中IVIG敏感者476例,IVIG无反应者39例,发生率为7.57%(39/515)。Logistic回归分析发现血白细胞、血红蛋白、C反应蛋白(CRP)、血沉、ALT、白蛋白及IVIG用药方案是IVIG无反应性的独立危险因素(P<0.05)。结论:7.57%KD患儿对初次IVIG治疗无反应。血白细胞、血红蛋白、CRP、血沉、ALT、白蛋白及IVIG用药方案是IVIG无反应的独立危险因素。     

Association of HLA-DP variants with the responsiveness to Hepatitis B virus vaccination in Korean Infants

Recently, HLA-DP single nucleotide polymorphisms (SNPs) have been reported to be related to responsiveness to hepatitis B virus (HBV) vaccination. The aim of this study was to investigate associations between HLA-DP SNPs and responsiveness to HBV vaccine in Korean infants. A total of 290 healthy Korean infants who were registered to Seoul Metropolitan Public Cord Blood Bank during the period of February 2007 to December 2011 were enrolled. Anti-HBs antibody level was analyzed after three doses of HBV vaccination. Genotyping of HLA-DPA1 SNPs (rs3077 and rs3830066) and HLA-DPB1 SNPs (rs7770370, rs7770501, rs3128961, and rs9277535) were performed by PCR-sequencing. HLA-A, -B, and –DRB1 genotyping was also performed by PCR-sequence-specific oligonucleotide probe kits. HLA-DPB1 SNPs (rs7770370, rs7770501, rs3128961, and rs9277535) were associated with HBV vaccine response. Allele frequencies of rs7770370 A, rs7770501 C, rs3128961 G, and rs9277535 A were significantly higher in responders than in non-responders (all p < 0.01). Anti-HBs antibody levels were different according to genotypes of DPB1 rs7770370, rs7770501, rs3128961, and rs9277535 (all p < 0.01). In multivariate analysis, HLA-DPB1 rs7770370 AA genotype was significantly associated with HBV vaccine response (relative risk, RR = 2.5, p = 0.033) and high-titer vaccine response (RR = 2.7, p < 0.001). In conclusion, HLA-DPB1 SNPs were significantly associated with responses to HBV vaccination in Korean infants.