Robust Antibody and Cytokine Response to Hepatitis B Vaccine Among Not-in-Treatment Patients With Chronic Hepatitis C: An Open-Label Control Study in China.

Background: Hepatitis B vaccine is an effective measure to prevent hepatitis B virus infection. Whether chronic hepatitis C virus (HCV) infection decreases humoral and cell-mediated immunity responses to hepatitis B vaccination is still controversial. Methods: Patients with chronic HCV infection who were not in treatment and healthy controls, matched at a 1:2 ratio for community, sex, and age (within 5 years), were identified from a community-based screening. All participants received 3 doses of hepatitis B vaccine. Antibody to hepatitis B surface antigen was tested 1 month after the third vaccine dose and was compared between 2 groups. Spot-forming cells (SFCs) of interferon γ and interleukin 2, 4, 5, and 6 were counted by means of enzyme-linked immunospot, and SFC counts were compared between the 2 groups. Results: The rates of nonresponse and low, normal, and high response were 3.80%, 10.13%, 45.57%, and 40.50% respectively, in the HCV group, and the corresponding rates in the healthy control group were 1.26%, 10.13%, 39.24%, and 49.37% (all P > .05). There were no significant differences in SFC counts between the 2 groups for interferon γ or interleukin 2, 4, or 5 (all P > .05). Conclusions: This study provided preliminary evidence of the good immunogenicity and safety of hepatitis B vaccination among patients in China with chronic hepatitis C who are not in treatment. Clinical Trials Registration: NCT 02898922.

Extended planar defects of oxygen vacancies in ferroelectric [Formula: see text] and impact on ferroelectricity.

Extended defects of vacancies in ferroelectrics (FE), where vacancies spread over an extended space, are of critical importance in terms of understanding the long-standing problems such as polarization fatigue and aging. However, extended defects in FEs are poorly understood. Here we investigate the extended planar oxygen vacancies in ferroelectric [Formula: see text] using density functional theory and the modern theory of polarization. Oxygen vacancies of different charge states, namely [Formula: see text], [Formula: see text], and [Formula: see text], are studied. We obtain interesting results such as: (i) The formation energy of planar [Formula: see text] vacancies can be very small (merely 0.54 eV) even under the oxygen-rich condition, which is considerably smaller than the formation energy (4.0 eV) of planar [Formula: see text] vacancies; (ii) Planar [Formula: see text] vacancies drastically reduce the ferroelectric polarization in [Formula: see text] by more than one order of magnitude, which provides a pivotal (theoretical) evidence that the planar oxygen vacancies could be the origin of polarization fatigue and imprinting. The polarization dead layer caused by planar oxygen vacancies is shown to be around 72 Å. Microscopic origin and insight, based on the local Ti-O relative displacements, are provided to understand these interesting phenomena.

Significant transcriptome and cytokine changes in hepatitis B vaccine non-responders revealed by genome-wide comparative analysis.

Individuals fail to elicit protective antibody after hepatitis B vaccination remain at risk for hepatitis B virus infection. Analysis of the transcriptome of peripheral blood mononuclear cells (PBMCs) is essential to elucidate the characteristics of gene expression in non-responders. In this study, we enrolled seven responders who had received three injections and seven non-responders who had six injections of hepatitis B vaccine before. All the participants were then vaccinated with a three-dose boost regimen. Microarray analysis and Luminex assay were applied to examine mRNA expression and Th1/Th2/Th9/Th17/Th22/Treg cytokine and chemokine profiles in non-responders and responders. Differentially expressed genes in PBMCs of non-responders at 5 time points, i.e. pre-vaccination, 3rd, 7th, 28th day post the first dose vaccination and 7th day post the second dose vaccination indicated a dense network trend. Compared with responders, nine coding genes (BPI, DEFA1B, DEFA4, CEACAM8, MMP8, FOLR3, LTF, TCN1 and TKTL1) were significantly up-regulated in non-responders at all 5 time points, which could probably be the characteristic genes in hepatitis B vaccine non-responsiveness. Gene ontology analysis revealed that most of the DEGs were related with immune responses. Validation results of these 9 genes using quantitative real-time polymerase chain reaction were mostly consistent with the results of microarray. Cytokine analysis demonstrated that IL-27 and CXCL12 concentrations in responders were significantly higher than non-responders on the 3rd day after the first dose and 7th day after the second dose of vaccination, respectively. No significant difference was observed in other cytokine and chemokine signatures between the two groups. In conclusion, our results revealed characteristic transcriptome and cytokine changes in hepatitis B vaccine non-responders after boost immunization.

Immune responses to vaccines involving a combined antigen-nanoparticle mixture and nanoparticle-encapsulated antigen formulation.

Many physicochemical characteristics significantly influence the adjuvant effect of micro/nanoparticles; one critical factor is the kinetics of antigen exposure to the immune system by particle-adjuvanted vaccines. Here, we investigated how various antigen-nanoparticle formulations impacted antigen exposure to the immune system and the resultant antigen-specific immune responses. We formulated antigen with poly(lactic-co-glycolic acid) (PLGA) nanoparticles by encapsulating antigen within nanoparticles or by simply mixing soluble antigen with the nanoparticles. Our results indicated that the combined formulation (composed of antigen encapsulated in nanoparticles and antigen mixed with nanoparticles) induced more powerful antigen-specific immune responses than each single-component formulation. Mice immunized with the combined vaccine formulation displayed enhanced induction of antigen-specific IgG antibodies with high avidity, increased cytokine secretion by splenocytes, and improved generation of memory T cell. Enhanced immune responses elicited by the combined vaccine formulation might be attributed to the antigen-depot effect at the injection site, effective provision of both adequate initial antigen exposure and long-term antigen persistence, and efficient induction of dendritic cell (DC) activation and follicular helper T cell differentiation in draining lymph nodes. Understanding the effect of antigen-nanoparticle formulations on the resultant immune responses might have significant implications for rational vaccine design.

Biodegradable polylactide microspheres enhance specific immune response induced by Hepatitis B surface antigen.

Hepatitis B (HB) infection caused by Hepatitis B virus (HBV) is the most common liver disease in the world. HB vaccine, when administered in conjunction with alum adjuvants, induces Th2 immunity that confers protection against HBV. However, currently available vaccine formulations and adjuvants do not elicit adequate Th1 and CTL responses that are important for prevention of maternal transmission of the virus. Microspheres synthesized from poly (D, L-lactide-co-glycolide) (PLGA) or poly (D, L-lactide) (PLA) polymers have been considered as promising tools for in vivo delivery of antigens and drugs. Here we describe PLA microspheres synthesized by premix membrane emulsification method and their application in formulating a new microsphere based HB vaccine. To evaluate the immunogenicity of this microsphere vaccine, BALB/c mice were immunized with microsphere vaccine and a series of immunological assays were conducted. Results of Enzyme-linked ImmunoSpot (ELISPOT) assays revealed that the number of interferon-gamma (IFN-γ)-producing splenocytes and CD8(+) T cells increased significantly in the microsphere vaccine group. Microsphere vaccine group showed enhanced specific cell lysis when compared with HB surface antigen (HBsAg) only group in (51)Cr cytotoxicity assays. Moreover, microsphere vaccine elicited a comparable level of antibody production as that of HB vaccine administered with alum adjuvant. We show that phagocytosis of HBsAg by dendritic cells is more pronounced in microsphere vaccine group when compared with other control groups. These results clearly demonstrate the potential of using PLA microspheres as effective HB vaccine adjuvants for an enhanced Th1 immune response.

[Comparison of the kinesis of immune responses in mice vaccinated by different kinds of recombinant hepatitis B vaccines].

OBJECTIVE: To evaluate the kinesis of cellular and humoral immune responses to different kinds of recombinant hepatitis B(rHB) vaccines in the immunized mice. METHODS: At serial time points, the levels of IFN-gamma and IL-2 secreted by spleens mononuclear cells (MNC) of the vaccinated mice were detected by enzyme-linked immunospot methods (ELISPOT) after stimulation in vitro with HBsAg MHC class I peptide S28-39 or HBsAg. The lymphocytotoxicity of the immunized mice were also detected (CTL) by a specific lysis assay and the levels of anti-HBs were measured by the Abbott IMX kit. RESULTS: The peak values of IFN-gamma and IL-2 in vaccinated mice were detected by ELISPOT, 10 - 14 days after immunization. The CTL and the level of IFN-gamma induced by rHB vaccine derived from yeast cells (Hansenula polymorpha) (rHP vaccine) were significantly higher than the other two vaccines (P < 0.05). The maximum lysis of CTL appeared in the vaccinated mice on day 10 after immunization, with the percentage of 39.8%. The levels of IL-2 induced by rHP vaccine were significantly higher than the other two vaccines (P < 0.05). However, the IL-2 levels in the rSC (saccharomyces cerevisiae) vaccine group were higher as compared with the rCHO vaccine group at day 7 and day 14 (7 d t = 4.595, P = 0.001 < 0.05; 14 d t = 5.721, P = 0.000 < 0.05) after immunization. The cellular immune response to the rHP vaccine was the strongest while it was the lowest to the rCHO vaccine at day 7 after immunization. The sero-positive rates and the titers of anti-HBs in the vaccinated mice increased with time after vaccination. The titers of anti-HBs in the rCHO vaccine group at day 7 were similar to the rSC vaccine group, but significantly higher than that of the rHP vaccine group (P = 0.044 < 0.05). The anti-HBs titers of the rCHO vaccine group at day 14 were significantly higher as compared to the rSC (P = 0.012 < 0.05) and rHP (P = 0.009 < 0.05) vaccine groups. CONCLUSION: The immune responses induced by the three kinds of rHB vaccines were different in their patterns and levels. According to the intensity of early cellular immune response, the two yeast HB vaccines were superior to the rCHO vaccine, especially to the rHP vaccine. In contrast, the rCHO vaccine induced early seroconversion and high levels of anti-HBs.

[Cellular immune responses of recombinant hepatitis B (rHB) vaccine and HBsAG derived from Hansenula polymorpha cells].

OBJECTIVE: To study the kinetics of immune response in mice and human immunized with rHB vaccine or rHBsAg derived from yeast cells (Hansenula polymorpha). METHODS: With different doses,the level of IFN-gamma secreted by spleen mononuclear cells (MNC) including CD8+ T cells by MACs of mice were detected by enzyme-linked immunospot (ELISPOT) methods after stimulation in vitro with HBsAg MHC class I peptide S28-39, respectively. At serial time points, the immunized mice were detected for IFN-gamma by ELISPOT as above and for the lymphocytotoxicity test (CTL) by specific lysis assay. The levels of IFN-gamma, IL-2, IL-5 and anti-HBs in mice induced by rHB vaccine were detected after single or three doses. Four adults were vaccinated with rHB vaccine according to 0, 1 and 2 month schedule. The peripheral blood mononuclear cells (PBMCs) were collected at the 3, 8, 21, 34 and 65 days after the first dose. The CD8+ T cells with high purity obtained by sorting from PBMCs were stimulated with rHBsAg or HBsAg peptides. The SFC of IFN-gamma, IL-2 and IL-4 of CD4+ and CD8+ T cells were determined by ELISPOT. RESULTS: The cytokine of IFN-gamma became detectable on day 7 and its peak value appeared on day 14 by ELISPOT. The CTL was detected on day 7 and the maximum lysis of CTL appeared on day 28. The cellular immune response of IFN-gamma of MNCs were significantly correlated with the doses vaccinated from 1 microg to 8 microg (r(positive rates) = 0.951, P(positive rates) = 0.049 < 0.05; r(SFC) = 0.996, P(SFC) = 0.000 < 0.05). IFN-gamma SFC of CD+ T cells were significantly associated with the doses from 1 microg to 4 microg (r = 0.999, P = 0.025 < 0.05). The HBsAg specific cellular immune an d humoral responses of mice immunized with three doses were significantly higher than that with a single dose (P < 0.05). The characteristics of IFN-gamma, IL-2 and IL-4 of CD4+ and CD8+ T cells were variable between individuals immunized with the same rHB vaccine. The level of IL-2 and IL-4 of responders were significantly related to the titer of anti-HBs. CONCLUSION: Data from this study showed the kinesis of cellular immunity in mice and adults vaccinated with rHBsAg or rHB vaccine respectively, and the characteristics of cellular immune response in adults induced by the vaccine. Our data provided the basis of standardizing the analysis of cellular immune response to rHB vaccine.

[Study on the kinesis of cellular immunity in adults vaccinated with recombinant hepatitis B vaccine].

OBJECTIVE: To evaluate the kinesis of cellular immunity in adults who were vaccinated with yeast recombinant hepatitis B(rHB) vaccine and the correlation between cellular and humoral immune responses induced by the vaccine. METHODS: Eight adults were vaccinated with rHB vaccine according to 0, 1,2 month schedule. The peripheral blood mononuclear cells(PBMCs) were collected at the 3, 8, 21, 34 and 65 days after the first dose. The high purity of CD4+ and CD8+ T cells obtained by sorting from PBMCs were restimulated with recombinant hepatitis B surface antigens (rHBsAg) or peptides. The spot forming cell (SFC) of IFN-gamma, IL-2 and IL-4 of CD4+ and CD8+ T cells were detected by enzyme-linked immunospot (ELISPOT). RESULTS: The characteristics of IFN-gamma, IL-2 and IL-4 of CD4+ and CD8+ T cells appeared different after immunization with rHB vaccine. IFN-gamma of CD8+ and CD4+ T cells could be detected early with stable SFC, while the IL-2 and IL-4 of CD4+ T cells appeared late but increased after the second and third dose of vaccination. The positive rate of IL-4 of CD4+ T cells were significantly correlated with the positive rate of anti-HBs, while the SFCs of IL-4 and IL-2 of CD4+ T cells were also significantly related to the titers of anti-FIBs. CONCLUSION: IFN-gamma could be detected early after rHB vaccination in adults, and the positive rates of IL-4 and IL-2 were correlated with that of anti-HBs.