Use of Nanomaterials-based Enzymes in Vaccine Production and Immunization

Immunologic characterization of HIV-specific DNA vaccine.

The history of successful vaccination against severe viral diseases such as smallpox poliomyelitis or measles led to the initial hope that a vaccine against AIDS would be developed quickly. However an effective vaccine against HIV needs to overcome substantial obstacles that emerged as research progressed. Due to life cycle HIV can effectively hide from the host immune response integrating itself as proviral DNA in the host cell genome. A strategy pursued to deal with this difficulty is to include early viral regulatory proteins such as Tat Rev or Nef as vaccine antigens for induction of immune responses that can recognize and destroy HIV-infected cells as soon as the virus life cycle is activated [12]. The virus preferentially targets and destroys host immune cells such as T-helper lymphocytes macrophages and dendritic cells that are probably essential to maintain an effective antiviral immune response. This would imply that vaccine-elicited immunity unlikely to be able to prevent infection itself must be able to quickly control virus replication to prevent harm to the immune system. The high antigenic variability of HIV can be considered as an extremely effective immune-evasion strategy. Because of the low fidelity of the viral RNA polymerase virus progeny always represents a collection of RNA genomes (quasi-species) with random mutations. In vivo selection of immunodeficiency virus variants that can evade the recognition of neutralizing antibodies is common and strong virus-specific cytotoxic T-cell responses can select for escape variants already during resolution of primary viremia [3]. Most HIV infections are acquired sexually via the genital or rectal mucosae; however at these entry sites it appears difficult to induce strong antiviral immunity by vaccination. Finally HIV infection is a poverty-related disease that is particularly threatening health in societies of the developing world. Therefore vaccine candidates must be safe and feasible in production and administration to be eligible for use where most needed. (excerpt)

Chronic Ethanol Consumption Impairs Cellular Immune Responses Against HCV NS5 Protein Due to Dendritic Cell Dysfunction

Longevity and Clinical Effectiveness of the Humoral and Cellular Responses to SARS-CoV-2 Vaccination in Hemodialysis Patients

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Event Abstract Back to Event DNA vaccine encoding Hantavirus glycoprotein N-terminal, targeted to the major histocompatibility complex II compartment by lysosome-associated membrane protein, significantly elicits both specific humoral and cellular immune responses and induces immune protection against Hantavirus challenge in Balb/c mice Dongbo Jiang1, Yuanjie Sun1, Linfeng Cheng1, Kun Yang1* and Fanglin Zhang1* 1 The Forth Military Medical University, China Endogenous antigen encoded by conventional naked DNA was processed by antigen-presenting cells (APCs) and only presented to CD8+ T cell through forming peptide/MHCI (major histocompatibility complex type I molecules) complex. Whereas, antigen presentation by MHCII molecules and activation of CD4+ helper T cells are critical for the generation of effective immue response and long-term immune memory. Our previous studies showed DNA vaccine encoding SARS Coronavirus nucleocapsid (CoV-N), even epitope-based SARS CoV-N70-122 DNA vaccine with the lysosome-associated membrane protein (LAMP) as a chimera can not only elicit both humoral and cellular immune responses, but also promote a long-lasting T-cell memory response. Here, LAMP was as a key molecule trafficking DNA vaccine-encoding target protein to the MHC II compartment. Hantaan virus (HTNV), an old world Hantavirus, is mainly distributed in Asia. Haemorrhage fever with renal sydrome (HFRS) caused by HTNV is one of three Hantavirus-causing diseases (EN, HFRS and HPS). The fatality is among 3%-15%. In the past two years, sporatic cases of HPS obviously increased with a high fatality in the North America. There is no effective treatment to Hantavirus infection yet. Hence, vaccination prophylaxis is considered preferable solution against HV. Currently, inacitve HTNV vaccines are only applied in Asia, mainly China, Japan and South Korea. A continued reduction in HFRS was obseved while these vaccines distributed widely. However, short term of immunogenicity and low efficacy are disadvantages of inactive vaccine. development of a more effective vaccine of HTNV is of great need. Hantavirus glycoprotein N-terminal, named Gn, was reported that could induce neutralizing antibody production although with a low serum titer as natural infection. No study yet evaluates immune efficiency elicited by Gn as a vaccine. Therefore, we constructed DNA vaccine with HTNV Gn targeted to the MIIC by trafficking molecule LAMP. In this study, we have investigated the cellular and humoral immune responses in ex vivo and immune protection in vivo of immunized mice by DNA vaccine encoding HTNV glycoprotein N-terminal (Gn). The cDNA encoded HTNV Gn was acquired by PCR from a full-length HTNV M segment. First of all, effective expression of the proteins encoded by the DNA plasmid constructs in transfected 293 cells was confirmed. And then, Four groups of Balb/c mice were immunized with four different plasmids: ① pVAX1 vector with the Food and Drug Administration (FDA) document as a negative control. ② DNA encoding unmodified Gn as an endogenous antigen (pVAX-Gn); ③ DNA encoding Gn as a LAMP chimera targeted to the MIIC (pVAX-LAMP/Gn); ④ DNA encoding LAMP alone as a control (pVAX-LAMP) of pVAX-LAMP/Gn group. We tested specific antibody production in serum two weeks after each immunization by ELISA and neutralizing activity of antibody against HTNV by the cell microculture neutralization assay for humoral response evaluation. At the same time, we measured the peptide-specific IFN-γ secretion by ELISpot and specific CTL cytotoxicity against P815 cells which were loaded with Gn overlaping peptide-pool by LDH assay to evaluate cellular immune response. After Balb/c mice were immunized three times, animal protection experiment in vivo was also performed with Hantavirus challenging, and virus load in different tissues was detected by Sandwich ELISA and qRT-PCR. The results demonstrated that the immune responses differed markedly. Twice immunizations with pVAX-LAMP/Gn were sufficient to generate strong antibody response while no response in other groups. ELISpot analyses indicated that pVAX-LAMP/Gn elicited much greater IFN-γ response than pVAX-Gn. Whereas, pVAX-LAMP and pVAX induced very low levels, respectively. The cell microculture neutralization assay suggested that neutralizing antibodies against HTNV were producted in immunized mice with pVAX-LAMP/Gn and neutralizing titer in serum was the highest comparing with other three groups (p<0.05). The low level of neutralizing antibodies was detected in serum of immunized mice with pVAX-Gn, but the antibody neutralizing activity was undetectable in serum from pVAX or pVAX-LAMP group. For nonradioactive cytotoxicity assay, P815 cells were loaded with 83 overlaping peptides-pool of Gn as target cells. Among the experimental groups, splenocytes from mice immunized with the pVAX-LAMP/Gn showed higher specific cytotoxicity than that from mice immunized with the pPVAX-Gn at E/T ratios of 20:1, 10:1 and 5:1 (p<0.05). In contrast, no cytotoxicity against peptides loaded P815 cells in control mice immunized with pPVAX-LAMP or pVAX was detectable at each E/T ratios. Moreover, we investigated the protection efficacy in vivo after three times administration with naked DNA plasmids and carried out HTNV challenge. As no available animal model existed for HTNV challenge, which reflects the disease manifestations of severe HFRS, we made an observation of the Balb/c mice infected with HTNV 76-118 strain and established an evaluation system about animals infected with HTNV 76-118 before the animal protection experiment. We ran a set of programs (post-immunization viral intramuscular injection method) to infected the Balb/c mice with HTNV 76-118. Four groups of Balb/c mice were exposured to HTNV 76-118 at day 14 after third immunizations. And then, experimental mice were sacrificed and brain, heart, liver, spleen, lung and kidney were exteriorized after three days. Evaluation of viral load was performed by sandwich ELISA and qRT-PCR, and significant differences among the groups were observed. That is, the HTNV specific antigen could be detected in the brains and livers of the Balb/c mice immunized with pVAX-LAMP or pVAX, but undetected in that of all mice immunized with pVAX-LAMP/Gn or pVAX-Gn (p<0.01) . In summary, HTNV Gn usually cannot induce a strong immune response. However, Gn showed a strong immunogenicity to elicit both humoral and cellular responses with LAMP as a chimera. And results of protection assay in vivo indicated that the immune response established was HTNV specific and protective. These findings not only demonstrate that the LAMP as a trafficking molecule can introduce Gn to MHCII presenting pathway and significantly enhance HTNV specific immune response, but also suggest that the pVAX-LAMP/Gn as a DNA vaccine has potential application on clinic for HFRS immunoprophylaxis. Acknowledgements This work was supported by the National High Technology Research and Development Program 863 (No. 2006AA02A237) and the Military Key Scientific and Technological Project of 531 PR China (08G112) Keywords: Gene Vaccine, Hantaan Virus Gn, lysosome associated membrane protein, major histocompatibility complex II compartment, immune protection Conference: 15th International Congress of Immunology (ICI), Milan, Italy, 22 Aug - 27 Aug, 2013. Presentation Type: Abstract Topic: Translational immunology and immune intervention Citation: Jiang D, Sun Y, Cheng L, Yang K and Zhang F (2013). DNA vaccine encoding Hantavirus glycoprotein N-terminal, targeted to the major histocompatibility complex II compartment by lysosome-associated membrane protein, significantly elicits both specific humoral and cellular immune responses and induces immune protection against Hantavirus challenge in Balb/c mice. Front. Immunol. Conference Abstract: 15th International Congress of Immunology (ICI). doi: 10.3389/conf.fimmu.2013.02.01047 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 10 Jul 2013; Published Online: 22 Aug 2013. * Correspondence: Prof. Kun Yang, The Forth Military Medical University, Xi'an, China, yangkunkun@fmmu.edu.cn Prof. Fanglin Zhang, The Forth Military Medical University, Xi'an, China, flzhang@fmmu.edu.cn Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Dongbo Jiang Yuanjie Sun Linfeng Cheng Kun Yang Fanglin Zhang Google Dongbo Jiang Yuanjie Sun Linfeng Cheng Kun Yang Fanglin Zhang Google Scholar Dongbo Jiang Yuanjie Sun Linfeng Cheng Kun Yang Fanglin Zhang PubMed Dongbo Jiang Yuanjie Sun Linfeng Cheng Kun Yang Fanglin Zhang Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

B19-VLPs as an effective delivery system for tumour antigens to induce humoral and cellular immune responses against triple negative breast cancer

In the last 2 years, several studies investigated the immunological responses to SARS-CoV-2 infections and/or COVID-19 vaccinations,1-6 but long-term immunological effects in response to different vaccination combinations are poorly defined. As recently demonstrated, humoral and cellular immune responses to SARS-CoV-2 vaccines wane with time.7-9 Correspondingly, epidemiological data show a reduction in protection against (symptomatic) COVID-19 with increasing time following vaccination.10, 11 In the COV-ADAPT cohort, we recently studied the humoral and cellular immune responses and their interdependencies following different vaccine combinations before (T1) and up to 3 months after the second immunization (T2).12 This follow-up investigated the stability of long-term immune responses and aimed to identify predictive markers. Thus, we assessed humoral (anti-spike-RBD-IgG, neutralization capacity and avidity) and cellular (spike-induced T-cell interferon-γ release) immune responses 3–7 months after the second immunization (T3) in blood samples of 320 healthcare workers of the COV-ADAPT cohort with previous homologous ChAdOx1 nCoV-19 (ChAdOx1, n = 26), homologous BNT162b2 (n = 49), heterologous ChAdOx1/BNT162b2 (n = 243) or heterologous ChAdOx1 /mRNA-1273 (n = 2) vaccinations (Figure S1; all participants provided written informed consent). The last group was not analyzed separately due to the low n-number (for characterization of study participants see Table S1). The study was approved by the local ethics committee (21/5/21) and registered with the German Clinical Trials Register (DRKS00026029). We detected no nucleocapsid (NCP)-IgG seroconversion between T2 and T3 indicating the absence of breakthrough infections (Figure S2). At T3, homologous ChAdOx1 vaccination resulted in significantly lower anti-spike-RBD-IgG vs. heterologous ChAdOx1/BNT162b2 and homologous BNT162b2 (Figure 1A, Table S1). ChAdOx1/BNT162b2 and BNT162b2/BNT162b2 did not significantly differ. Despite the decrease from T2 to T3 in all groups, anti-spike-RBD-IgG was still significantly higher at T3 vs. T1 (Figure 1A). T-cell interferon-γ release (i.e., the cellular response) also decreased from T2 to T3 in all regimes. Only the heterologous ChAdOx1/BNT162b2 group still showed significantly higher T-cell responses at T3 vs. T1 (Figure 1B), and no differences were observed between the regimes at T3 (Figure 1B, Table S1). For the groups with BNT162b2 as a second vaccination, anti-spike-RBD-IgG was negatively associated with the days elapsed since the second vaccination (Figure 1C, left panel) suggesting higher antibody dynamics for vaccination regimes including BNT162b2. Such an association was not found for the cellular response (Figure 1C, right panel). Similar to our previous findings at T1/T2, humoral and cellular immune responses showed significantly positive associations at T3 for the study population as a whole and the ChAdOx1/BNTb162b2 group (Figure S3). Between T2 and T3, strong associations with high predictive power were observed for cellular and humoral immune responses for all groups (Figure S4). The early cellular response (at T1) emerged as a predictor of long-term immune responses as it was significantly associated with late (T3) humoral (ChAdOx1/BNT162b2 and BNT162b2/BNT162b2) and cellular responses (all groups) (Figure 2). Antibody neutralization and avidity indices were significantly higher at T3 vs. T1 in all groups (Figure S5) indicating durable antibody quality. Neutralization capacity was higher in the groups with a second BNT162b2 vaccination vs. homologous ChAdOx1 at T3. Interestingly, subjects with a negative neutralization index (as per the manufacturer's instructions) did not present generally lower anti-spike-RBD-IgG levels (Table S2). In conclusion, we identified important long-term interactions between the humoral and the cellular immune systems and observed distinct long-term dynamics following different SARS-CoV-2 vaccination regimes. In this regard, vaccination regimes including BNT162b2 elicit strong immune responses with a more rapid decline, whereas vector-based vaccinations yield lower and comparably stable immunological effects. The immunological drawbacks of either homologous vaccination regime appear to be somewhat mitigated by the combination of both vaccination principles in the form of a heterologous vaccination. We additionally identified the early T-cell response to predict long-term immune responses in different vaccination regimes. It will be of utmost importance to determine how the observed interdependencies and long-term dynamics of immune response react to booster vaccinations and breakthrough infections. This work was supported by the German Research Foundation (Deutsche Forschungsgemeinschaft [DFG]), grant number ER723/3-1 (to LE) and grant number ZA697/6-1 (to AEZ). Open Access funding enabled and organized by Projekt DEAL. The authors have no conflicts of interest to declare. Appendix S1 Please note: The publisher is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.

Pineal melatonin secretion, but not ocular melatonin secretion, is sufficient to maintain normal immune responses in Japanese quail ( Coturnix coturnix japonica)

Recombinant African swine fever virus p30-flagellin fusion protein promotes p30-specific humoral and cellular immune responses in mice.

Diurnal variation in the cellular and humoral immune responses of Japanese quail: Role of melatonin

Cellular and humoral immune response to SARS-CoV-2 vaccination and booster dose in immunosuppressed patients: An observational cohort study.

Continuous evaluation of the coronavirus disease 2019 (COVID-19) vaccine effectiveness in hemodialysis (HD) patients is critical in this immunocompromised patient group with higher mortality rates due to severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection. The response towards vaccination in HD patients has been studied weeks after their first and second SARS-CoV-2 vaccination dose administration, but no further studies have been developed in a long-term manner, especially including both the humoral and cellular immune response. Longitudinal studies that monitor the immune response to COVID-19 vaccination in individuals undergoing HD are therefore necessary to prioritize vaccination strategies and minimize the pathogenic effects of SARS-CoV-2 in this high-risk group of patients. We followed up HD patients and healthy volunteers (HV) and monitored their humoral and cellular immune response three months after the second (V2+3M) and after the third vaccination dose (V3+3M), taking into consideration previous COVID-19 infections. Our cellular immunity results show that, while HD patients and HV individuals secrete comparable levels of IFN-γ and IL-2 in ex vivo stimulated whole blood at V2+3M in both naïve and COVID-19-recovered individuals, HD patients secrete higher levels of IFN-γ and IL-2 than HV at V3+3M. This is mainly due to a decay in the cellular immune response in HV individuals after the third dose. In contrast, our humoral immunity results show similar IgG binding antibody units (BAU) between HD patients and HV individuals at V3+3M, independently of their previous infection status. Overall, our results indicate that HD patients maintain strong cellular and humoral immune responses after repeated 1273-mRNA SARS-CoV-2 vaccinations over time. The data also highlights significant differences between cellular and humoral immunity after SARS-CoV-2 vaccination, which emphasizes the importance of monitoring both arms of the immune response in the immunocompromised population.

Immune responses to recombinant Mycobacterium smegmatis expressing fused core protein and preS1 peptide of hepatitis B virus in mice

<b>Background:</b> SARS-CoV-2 vaccines are expected to induce both cellular and humoral immune responses, however, the dynamics and correlation between the two types of immunity are not precisely understood. <b>Aims:</b> Assessing IgG levels and T-cell response induced by SARS-CoV-2 vaccines and investigating the correlation between cellular and humoral immune responses. <b>Methods:</b> Blood samples were taken from 166 respiratory healthcare professionals at four time-points: first before administering the booster vaccine, then on day 28, 56 and 120 post-vaccination. For the assessment of humoral immune response anti-Spike protein IgG ELISA was used, while T-cell response was tested by interferon-gamma-release-assay. <b>Results:</b> Out of 166 patients, 120 individuals presented positive result for interferon-gamma, while 100 had positive results for IgG. The positivity rate of cellular immune response was found to be significantly higher than humoral between the second and third doses of anti-COVID vaccines (P&lt;0,05). Participants, who have had SARS-CoV-2 infection before the first two shots, the immune response was detectable at a statistically higher rates than in the COVID naϊve group. The third dose triggered different dynamics regarding the IgG titers and T-cell response. Four months after the administration of the booster shot both humoral and cellular immune response were detectable simultaneously. <b>Conclusions:</b> After the first two doses, the cellular immune response was found to last longer than humoral, especially in previously infected individuals. Measuring the T-cell responses to SARS-CoV-2 vaccines may complement the antibody tests currently used in clinical practice.