Trend analysis of hemagglutinin and ovalbumin content for influenza virus split vaccines in 2021 / 中国生物制品学杂志

@#Objective To analyze the trend of the hemagglutinin(HA) and ovalbumin contents in the lot release of influenza virus split vaccines in 2021,and evaluate the quality and quality control level of the vaccines.Methods The HA and ovalbumin content data of influenza virus split vaccines from two domestic enterprises in 2021 were collected and collated. The mean value and standard deviation were calculated according to the first 40 batches of data of the enterprise in the year,and the warning limit and action limit were established. The trend analysis of the above indexes was carried out to evaluate the stability and consistency of the product quality of the enterprise. Statistical data comparison and consistency analysis were made between the test results of the batch inspected by the lot release institution and the results of the enterprise.Results Through the retrospective data analysis of quadrivalent influenza virus split vaccines from two vaccine enterprises A and B,it was found that the content of H1N1 subtype HA and ovalbumin in the two enterprises and the content of Bv HA in the B enterprise had out of trend(OOT)situations,while the trend of other items was stable. The results of paired student's t test or Wilcoxon signed-rank test of the samples inspected by the lot release institution showed that except Bv subtype HA(t = 1. 094 and 0. 742 respectively)and ovalbumin(w =-64 and 36 respectively)contents showed no statistically significant difference(P > 0. 05),the HA contents of H1N1(t = 3. 862,w = 232),H3N2(t = 8. 225 and3. 473 respectively)and By(t = 5. 616 and 4. 934 respectively)of the two enterprises had significant differences(P <0. 05). The results of enterprises were generally higher than the lot release institution. Bland-Altman test analysis found that the consistency between the test data of enterprise A's HA content and the data of the lot release institution was better than that of enterprise B.Conclusion The stability and consistency of data trends of active ingredients and main impurity ingredients of quadrivalent influenza virus split vaccine batches in 2021 were generally good. The trend analysis can identify potential problems in vaccine production,and enterprises should carefully implement trend analysis and effectively monitor the product quality of vaccines.

Immunogenicity of MDCK cell-based quadrivalent influenza split vaccine with different adjuvants in mice / 中华微生物学和免疫学杂志

Objective:To evaluate the immunogenicity of Madin-Darby canine kidney (MDCK) cell-based quadrivalent influenza split vaccine (MDCK-Va) combined with different adjuvants.Methods:Different doses of MDCK-Va and chicken embryo-based quadrivalent influenza split vaccine (egg-Va) were intramuscularly immunized BALB/c mice twice with an interval of three weeks. Serum samples were collected to detect antibody titers using hemagglutination inhibition (HI) assay. BALB/c mice were immunized with different doses of MDCK-Va combined with QS21, AddVax, PolyI∶C, CpG ODN 1826 and AddVax/PolyI∶C (Add/Poly), respectively. HI and microneutralization assays were used to detect antibody titers 21 d after the first and booster immunization. Spleen tissues were collected from the mice immunized with 10 μg MDCK-Va combined with the above adjuvants 5 d after the booster immunization to analyze spleen index and the types of spleen cells.Results:The immunoprotective effect of MDCK-Va was not inferior to that of egg-Va. MDCK-Va combined with each of the above adjuvants could induce higher HI antibody titer than MDCK-Va alone, especially the QS21/Va and Add/Poly/Va groups, and the differences were statistically significant. For H1N1 vaccine, the Pearson′s correlation coefficient ( r) between HI antibody and neutralizing antibody was 0.737-0.910, and for H3N2 subtype vaccine, the value of r was 0.839-0.947. Compared with the MDCK-Va group, the QS21/Va group showed significantly increased spleen index and decreased proportion of single lymphocytes. QS21 and Add/Poly were much better than other adjuvants in stimulating mouse splenic neutrophils and CD4/CD8 cells. Conclusions:Add/Poly had a stronger immune enhancement effect on MDCK-Va, suggesting that it was a potential adjuvant for MDCK-Va. The antibody titer detected by HI and MN assays had a strong positive correlation.

Intranasal Immunization Using CTA1-DD as a Mucosal Adjuvant for an Inactivated Influenza Vaccine / 生物医学与环境科学（英文）

OBJECTIVE@#To evaluate the effect of intranasal immunization with CTA1-DD as mucosal adjuvant combined with H3N2 split vaccine.@*METHODS@#Mice were immunized intranasally with PBS (negative control), or H3N2 split vaccine (3 μg/mouse) alone, or CTA1-DD (5 μg/mouse) alone, or H3N2 split vaccine (3 μg/mouse) plus CTA1-DD (5 μg/mouse). Positive control mice were immunized intramuscularly with H3N2 split vaccine (3 μg/mouse) and alum adjuvant. All the mice were immunized twice, two weeks apart. Then sera and mucosal lavages were collected. The specific HI titers, IgM, IgG, IgA, and IgG subtypes were examined by ELISA. IFN-γ and IL-4 were test by ELISpot. In addition, two weeks after the last immunization, surivival after H3N2 virus lethal challenge was measured.@*RESULTS@#H3N2 split vaccine formulated with CTA1-DD could elicit higher IgM, IgG and hemagglutination inhibition titers in sera. Furthermore, using CTA1-DD as adjuvant significantly improved mucosal secretory IgA titers in bronchoalveolar lavages and vaginal lavages. Meanwhile this mucosal adjuvant could enhance Th-1-type responses and induce protective hemagglutination inhibition titers. Notably, the addition of CTA1-DD to split vaccine provided 100% protection against lethal infection by the H3N2 virus.@*CONCLUSION@#CTA1-DD could promote mucosal, humoral and cell-mediated immune responses, which supports the further development of CTA1-DD as a mucosal adjuvant for mucosal vaccines.

Determination of Bacterial Endotoxin in Influenza Split Vaccine by a Dynamic Turbidity Method / 中国药师

Objective:To investigate the feasibility of dynamic turbidity method for the detection of bacterial endotoxin content in influenza split vaccine. Methods: According to the bacterial endotoxin detection method described in general rule 1143 in Chinese Pharmacopoeia (2015 edition), the reliability test for standard curves of influenza split vaccine, the interference initial screening test, the interference verification test and the endotoxin content were performed or determined, and the results were compared with those by the gel method for the same batches of vaccine. Results:The results of reliability test for standard curves were accordance with the reg-ulations. In the interference initial screening test, vaccine was diluted by 160 times, 320 times and 640 times, and the recovery was between 50% and 200%, which showed no interference. The results of the interference verification test further proved that vaccine with 640 times dilution had no interference effect on test. The bacterial endotoxin contents of 10 batches of influenza split vaccine deter-mined by the turbidity method were less than the limit value of 20 EU·ml-1 , and the results were the same as those determined by the gel method. Conclusion:It is feasible to detect the content of bacterial endotoxin in influenza split vaccine by the dynamic turbidity method, which is worthy of promoted application.

Immunogenicity and Safety of Trivalent Split Influenza Vaccine in Healthy Korean Adults with Low Pre-Existing Antibody Levels: An Open Phase I Trial

PURPOSE: A phase I clinical trial was conducted to evaluate the immunogenicity and safety of newly developed egg-cultivated trivalent inactivated split influenza vaccine (TIV) in Korea. MATERIALS AND METHODS: The TIV was administered to 43 healthy male adults. Subjects with high pre-existing titers were excluded in a screening step. Immune response was measured by a hemagglutination inhibition (HI) assay. RESULTS: The seroprotection rates against A/California/7/2009 (H1N1), A/Perth/16/2009 (H3N2) and B/Brisbane/60/2009 were 74.42% [95% confidence interval (CI): 61.38–87.46], 72.09% (95% CI: 58.69–85.50), and 86.05% (95% CI: 75.69–96.40), respectively. Calculated seroconversion rates were 74.42% (95% CI: 61.38–87.46), 74.42% (95% CI: 61.38–87.46), and 79.07% (95% CI: 66.91–91.23), respectively. There were 25 episodes of solicited local adverse events in 21 subjects (47.73%), 21 episodes of solicited general adverse events in 16 subjects (36.36%) and 5 episodes of unsolicited adverse events in 5 subjects (11.36%). All adverse events were grade 1 or 2 and disappeared within three days. CONCLUSION: The immunogenicity and safety of TIV established in this phase I trial are sufficient to plan a larger scale clinical trial.

Adjuvant effect of Epimedium polysaccharide for H1N1 influenza viruses split vaccine / 国际药学研究杂志

Objective To study the adjuvant effect of Epimedium polysaccharide co-immunized with H1N1 influenza virus split vaccine. Methods BALB/c mice were intramuscularly injected H1N1 virus lysate premixed with Epimedium polysaccharide G1, G4 and saline, respectively. The titer of serum antibody of mice was determined by ELISA two weeks after the initial immunization. The proliferation of T lymphocytes was measured by MTT assay and the levels of lymphocyte cytokines IFN-γ and IL-4 were determined by ELISA. CD4 +, CD8 +, CD3 + and CD19+ lymphocyte populations were determined by flow cytometry. Results Epimedium polysaccharides, G1 and G4, could rapidly induce the production of high levels of serum antibody against H1N1 in mice. The polysaccharides could significantly promote lymphocytes proliferation and IFN-γ secretion. The contents of CD4 +, CD8 + and CD3 +, the ratio of CD3 + and CD19+ (CD3+/CD19+) were promoted in the polysaccharides groups. Conclusion These polysaccharides could induce significant immune response against H1N1 in mice without observable toxicity. They should be further evaluated as a useful adduvant candidate for H1N1 influenza virus split vaccine.

Safety and immunogenicity on three lots of influenza split vaccines among adults / 中华流行病学杂志

Objective To evaluate the irnmunogenicity, safety and stability of the manufacture process regarding three consecutive lots of influenza split vaccines (Anflu ). Methods A double-blind, randomized and controlled clinical trial was conducted in healthy volunteers. A total of 566 subjects aged 18 to 60 years were recruited and stratified into four age groups before randomly assigned into four groups. Each group would receive one dose of influenza vaccine from either one of the three lots ofAnflu or one lot of the licensed control vaccine. Each dose of the vaccines contained 15 μg of each of the H1N1, H3N2 and B type antigen. Safety was assessed through 30-minute observation for immediate allergic reaction and three-day observation after vaccination. HI antibody titers were determined before vaccination and on day 21, after vaccination. Results Mild adverse reaction was reported and the overall incidence rates on fever of the four groups were from 1.4% to 2.8% but no significant difference was observed between groups. Seroconversion rates of the three viral strains in four groups were 80.3% and above with fold increase as≥11.1 and protection rate was≥93.4%. For the three lots of investigated vaccines, all of the indexes of the three viral strains in four groups exceeded the standards on EMEA and FDA for influenza vaccine. Conclusion The three consecutive lots of Anflu appeared to be good, with both consistent immunogenieity and safety, indicating the stability of manufacture process.