Sodium alginate coating simultaneously increases the biosafety and immunotherapeutic activity of the cationic mRNA nanovaccine

The extraordinary advantages associated with mRNA vaccines, including their high efficiency, relatively low severity of side effects, and ease of manufacture, have enabled them to be a promising immunotherapy approach against various infectious diseases and cancers. Nevertheless, most mRNA delivery carriers have many disadvantages, such as high toxicity, poor biocompatibility, and low efficiency in vivo, which have hindered the widespread use of mRNA vaccines. To further characterize and solve these problems and develop a new type of safe and efficient mRNA delivery carrier, a negatively charged SA@DOTAP-mRNA nanovaccine was prepared in this study by coating DOTAP-mRNA with the natural anionic polymer sodium alginate (SA). Intriguingly, the transfection efficiency of SA@DOTAP-mRNA was significantly higher than that of DOTAP-mRNA, which was not due to the increase in cellular uptake but was associated with changes in the endocytosis pathway and the strong lysosome escape ability of SA@DOTAP-mRNA. In addition, we found that SA significantly increased the expression of LUC-mRNA in mice and achieved certain spleen targeting. Finally, we confirmed that SA@DOTAP-mRNA had a stronger antigen-presenting ability in E. G7-OVA tumor-bearing mice, dramatically inducing the proliferation of OVA-specific CLTs and ameliorating the antitumor effect. Therefore, we firmly believe that the coating strategy applied to cationic liposome/mRNA complexes is of potential research value in the field of mRNA delivery and has promising clinical application prospects.

The latest progress on novel coronavirus vaccination in kidney transplant recipients / 器官移植

Due to long-term use of immunosuppressant, poor immune function and a higher risk of critical diseases after novel coronavirus pneumonia in kidney transplant recipients, it is of significance to deliver prophylactic vaccination for this high-risk population. Studies have shown that the immune reaction of kidney transplant recipients to novel coronavirus vaccine is significantly lower than that of healthy counterparts. Standard vaccination program in the United States, such as 2 doses of messenger RNA (mRNA) vaccine, fails to provide sufficient protection for kidney transplant recipients. Many studies have proven that increasing the frequency of vaccination for kidney transplant recipients may enhance the vaccine efficacy. Nevertheless, the role of adjusting immunosuppressive therapy in increasing vaccine efficacy remains to be elucidated. In this article, the importance, effectiveness and particularity of novel coronavirus vaccine for kidney transplant recipients and the effect of immunosuppressive therapy on the efficacy of novel coronavirus vaccine were reviewed, aiming to provide reference on the vaccination for kidney transplant recipients.

Preparation and immunogenicity evaluation of mRNA vaccine against porcine epidemic diarrhea / 生物工程学报

Porcine epidemic diarrhea (PED) is a highly contagious disease that causes high mortality in suckling piglets. Although several licensed inactivated and live attenuated vaccines were widely used, the infection rate remains high due to unsatisfactory protective efficacy. In this study, mRNA vaccine candidates against PED were prepared, and their immunogenicity was evaluated in mice and pregnant sows. The mRNA PED vaccine based on heterodimer of viral receptor binding region (RBD) showed good immunogenicity. It elicited robust humoral and cellular immune responses in mice, and the neutralizing antibody titer reached 1:300 after a single vaccination. Furthermore, it induced neutralizing antibody level similar to that of the inactivated vaccine in pregnant sows. This study developed a new design of PED vaccine based on the mRNA-RBD strategy and demonstrated the potential for clinical application.

Prevalence and factors associated with flares following COVID-19 mRNA vaccination in patients with rheumatoid arthritis, psoriatic arthritis and spondyloarthritis: a national cohort study

Abstract Objective To determine prevalence and factors associated with flares post Coronavirus disease 2019 (COVID-19) mRNA vaccination in patients with rheumatoid arthritis (RA), psoriatic arthritis (PsA) and spondyloarthritis (SpA). Methods A retrospective multi-centre study was conducted (January 2021 to February 2022). Data were collected during index visit, defined as first post-vaccine visit in which the patient had a physician-defined flare, or if at least 3 months had elapsed since first vaccine dose, whichever came first. Factors associated with flares were identified using mixed effects Cox regression and expressed as hazard ratio (HR) and 95% confidence interval (CI). Results Total of 2377 patients were included (1563 RA, 415 PsA and 399 SpA). Among patients with RA, PsA and SpA, 21.3%, 24.1% and 21.8% experienced a flare respectively. Of those who experienced a flare, only 10.2%, 11.0% and 14.9% were severe in patients with RA, PsA and SpA respectively. Patients with low or moderate/high disease were more likely to flare compared to those in remission in patients with RA only (HR: 1.68, 95% CI 1.22-2.31; HR: 2.28, 95% CI 1.50-3.48, respectively). Receiving the Moderna vaccine was associated with a higher HR of flare compared to the Pfizer vaccine in patients with PsA only (HR: 2.21, 95% CI 1.20-4.08). Patients who had two vaccine doses were found to be less likely to flare (HR: 0.08, 95% CI 0.06-0.10). HRs of flares were not significantly different among RA, PsA and SpA. Conclusion About one-fifth of patients experienced a disease flare post COVID-19 mRNA vaccination, but most flares were non-severe. Patients with active disease prior to vaccination should be monitored closely for disease flares, especially in patients with RA.

Comparison of methods for detection of antibody level in serum immunized with SARS-CoV-2 mRNA vaccine / 中国生物制品学杂志

@#Objective To compare three methods for detection of antibody level in serum immunized with SARS-CoV-2mRNA vaccine. Methods Enzyme-linked immunosorbent assay(ELISA),pseudo virus-based neutralization assay(PBNA)and micro-cytopathic effect neutralization test(MCPENT)were used to detect the antibody levels of a total of 120 serum samples(40 before immunization and 80 after immunization)before and after 2 doses of mRNA vaccine immunization,and the consistency and correlation of the three methods were analyzed. Results The consistency rates of the three methods detecting 120 serum samples were all over 90%,the Kappa coefficients were all more than 0. 7,and each P was less than0. 01. The correlation coefficient(r)between the antibody potency results of positive serum samples detected by the three methods was 0. 825～0. 902,and each P was less than 0. 01. Conclusion The three methods have good consistency and correlation in detecting antibody level of serum immunized with SARS-CoV-2 mRNA vaccine.

Effects of poly(A) tails with different lengths on mRNA expression in vitro and stability of transcription template in Escherichia coli / 中华微生物学和免疫学杂志

Objective:To investigate the effects of poly(A) tails with different lengths on mRNA expression in vitro and the passage stability of transcription template with poly (A) tail in Escherichia coli ( E. coli). Methods:Plasmids with poly(A) tails of 38, 60, 103, 125 and 126 (60 nt+ 6 nt spacer+ 60 nt) nt were designed and constructed. Then the plasmids were linearized by single enzyme digestion and used as transcription template for preparing enhanced green fluorescent protein (EGFP)-mRNA. EGFP-mRNA containing poly(A) tails of different lengths were transfected into 293T cells and the expression of EGFP was detected by flow cytometry. As to stability test, the template plasmids with poly (A) tail of 125 and 126 nt were transformed into E. coli TransStbl3 and Top10 competent cells. Seven clones were selected for culture and plasmid extraction, and then the plasmids were digested by restriction enzyme and detected by capillary electrophoresis. For passage stability, three correctly sequenced clones of each group were selected for continuous passage at 37℃, and the plasmids were extracted and digested every two generations for capillary electrophoresis. At the same time, the correctly sequenced clones of 125 nt group were also passaged at 30℃, and the plasmids were also extracted and digested every two generations for capillary electrophoresis. Results:The transcription templates with poly(A) tail of different lengths were successfully constructed. Flow cytometry showed that the fluorescence expression of the template plasmids with poly (A) tail of 103 and 125 nt were significantly higher than that of 38 and 60 nt. The fluorescence expression of the plasmid with poly (A) tail of 126 nt was significantly higher than that of all other groups. The percentages of stable sequences of the template plasmid with poly(A) tail of 125 nt in TransStbl3 and Top10 competent cells were 76% and 91%, respectively. The results of continuous passage showed that poly(A) tail of 125 nt could be stable to the 4th generation at 37℃ in both TransStbl3 and Top10 competent cells, and stable to the 16th and 10th generations at 30℃. The percentages of stable sequences of the template plasmid with poly(A) tail of 126 nt in TransStbl3 and Top10 competent cells were 95% and 48%, respectively. The results of continuous passage showed that poly(A) tail of 126 nt could be stable to the 12th generation at 37℃ in both TransStbl3 and Top10 competent cells.Conclusions:The length and composition of poly(A) tail in mRNA affected the expression of target protein. Adding a spacer with a length of 6 nt to poly(A) tail and low temperature culture were both helpful to improve the stability of the template plasmid, which provided a reference for the design and preparation of in vitro transcription template of mRNA vaccine.

COVID-19 mRNA vaccine triggering dermatomyositis: A case report

The battle against SARS-COV-2 is rising and the hope in the safety and effectiveness of immunization against this virus is growing up, even though serious and severe adverse events are scarcely observed. In this article, we report a case of mRNA vaccine induced an autoimmune dermatomyositis with features of severity that are managed by immunosuppressants medications and still in regular follow-up. Inflammatory dermatomyositis can be triggered after vaccination with COVID vaccine in the same mechanism that COVID-19 infection-induced myositis.

Drug-induced liver injury after covid-19 mRNA vaccine: case report

Case description: A 22-year-old female patient received the first dose of Pfizer-BioNTech vaccine (RNAm) against COVID-19; 6 days later, she presented abdominal pain located in the right hypochondrium and epigastrium, associated with emetic episodes. Re-consultation 21 days later due to the same symptoms; three days after the second dose of the vaccine was administered. Clinical findings: Pain on palpation in the right hypochondrium. Laboratories reported hepatocellular lesion and cholestasis, with negative amylase, hepatotropic virus and autoimmune hepatitis tests. Liver and biliary tract ultrasound and cholangioresonance were normal. Treatment and Results: Hyoscine and intravenous fluids as support therapy. She presented improvement in abdominal pain and progressive decrease of transaminases and bilirubin levels until normalization, and was discharged on the fifth day of hospitalization. A drug-associated hepatotoxicity (DILI) diagnosis was considered probable, in this case, secondary to vaccination against COVID-19. Clinical Relevance: The current SARS CoV-2 pandemic has spurred the development of new vaccines, the safety of which remains a concern. There is a likely causal relationship between vaccination and liver involvement in this clinical case, rather than simply a sporadic occurrence.

Descripción del caso: Paciente femenina de 22 años, quien recibió primera dosis de vacuna Pfizer-BioNTech (RNAm) contra COVID-19; presenta 6 días después, dolor abdominal localizado en hipocondrio derecho y epigastrio, asociado a episodios eméticos. Reconsulta a los 21 días por la misma sintomatología; tres días posteriores a la aplicación de la segunda dosis de la vacuna. Hallazgos clínicos: dolor a la palpación en hipocondrio derecho. Los laboratorios reportaron lesión hepatocelular y colestasis, con amilasa, estudios para virus hepatotrópos y hepatitis autoinmune negativos. La ecografía de hígado, vías biliares y colangioresonancia fueron normales. Tratamiento y Resultados: hioscina 20 mg vía oral cada 8 horas y líquidos endovenosos como terapia de soporte. Presentó mejoría del dolor abdominal y descenso progresivo de transaminasas y bilirrubinas, hasta su normalización y se dio egreso al quinto día de hospitalización. Se consideró probable diagnóstico de hepatotoxicidad asociada a medicamentos (DILI), en este caso, secundario a la vacunación contra COVID-19. Relevancia Clínica: La pandemia actual por el virus SARS CoV-2 ha impulsado el desarrollo de nuevas vacunas, cuya seguridad sigue siendo un motivo de preocupación. En este caso clínico, hay una probable relación causal entre la vacunación y el compromiso hepático, en lugar de una simple aparición esporádica.

Particle size and size distribution of mRNA vaccines under different testing conditions / 中华微生物学和免疫学杂志

Objective:To investigate the factors that could influence the particle size and size distribution of mRNA vaccines.Methods:The influences of several factors including the ionic strength and pH values of buffers, solutions, dilution folds and testing equipments on the particle size and size distribution of three batches of mRNA vaccines were analyzed by dynamic light scattering.Results:The particle size increased with increasing ionic strength, but no significant change in size distribution was observed. The particle size also increased with increasing pH values and the size distribution showed significant change when the buffer solution was weakly alkaline. Solution types could affect the particle size, but had no influence on size distribution. There was no significant change in the particle size or size distribution when the dilution was limited to 100 folds. Moreover, the particle size and size distribution detected by different equipments showed no significance difference.Conclusions:The particle size and size distribution of mRNA vaccines could be affected by solution, dilution fold and testing equipment, which should be concerned during the vaccine production and quality control.

Research progress of nano delivery system in mRNA tumor vaccines / 药学学报

Tumor vaccine is one of the most promising therapeutic strategies in tumor immunotherapy. It promotes the antigen presentation process by delivering tumor antigen and then activates the anti-tumor immune response. As a new class of vaccines, messenger RNA (mRNA) vaccines can activate the immune system to achieve the purpose of immunotherapy by delivering the mRNA sequence of a specific antigen into the body and expressing the corresponding antigen protein. Compared with traditional vaccines, mRNA vaccines have the advantages of a short production cycle, high effectiveness, and strong immunogenicity. In recent years, the application of mRNA vaccines in tumor immunotherapy has attracted widespread attention, but the instability and low delivery efficiency of mRNA limit its application. Nano delivery system can effectively solve the problem of mRNA vaccine delivery, greatly promote the research process and clinical application of mRNA tumor vaccines, and has become a hot spot in the research of mRNA vaccines. In this review, we introduced the mRNA tumor vaccines, focusing on the application of nano delivery system in mRNA tumor vaccines, in order to provide new ideas and new methods for the efficient delivery of mRNA tumor vaccines and tumor immunotherapy.

SARS-CoV-2 infection rates after different vaccination schemes: An online survey in Turkey

Objective: To identify effects of various nationwide vaccination protocols on the evolution of new SARS-CoV-2 infections among adult population and to evaluate the safety of mRNA (BioNTech/ Pfizer) vaccine. Methods: Totally 10 735 adult volunteers that received at least one dose of BioNTech/Pfizer or triple doses of CoronaVac participated in this cross-sectional-online survey between 1 and 10 September 2021. The information was collected covering a 5-month period from April 2021 to September 2021. Information about people who were vaccinated with only single and double dose CoronaVac were not included in this study. Results: At least one side effect after single and double dose of BioNTech/Pfizer and triple doses of CoronaVac were observed in 42.1%, 42.5% and 10.9%, respectively. The most common side effects were shoulder/arm pain, weakness/fatigue, muscle/joint pain and headache. The side effects were the most frequent in single BioNTech/Pfizer, while it was the least in triple CoronaVac. The rate of positive PCR tests before vaccination was 17.6%, and decreased to 3.0% after vaccination. The rates of positive SARS CoV-2-PCR were 18.8%, 3.5%, 3.1%, 0.5% and 4.6% in single BioNTech/Pfizer, double BioNTech/Pfizer, double CoronaVac+single BioNTech/Pfizer, double CoronaVac+double BioNTech/Pfizer and triple CoronaVac, respectively. While 1.8% of PCR positive COVID-19 cases needed intensive unit care in the pre-vaccination period, intensive care unit was required in 0%, 1.5%, 2.4%, 0% and 4.2% after single BioNTech/ Pfizer, double BioNTech/Pfizer, double CoronaVac+single BioNTech/ Pfizer, double CoronaVac+double BioNTech/Pfizer and triple CoronaVac, respectively. Reinfection rate after vaccination was 0.4%. Conclusions: The rarity of COVID-19 infection after vaccination suggests that efficacy of vaccines is maintained. On the other hand, the data underscore the critical importance of continued public health mitigation.

Immune responses elicited by influenza A mRNA vaccine based on lipopolyplex-encapsulated virus nucleoprotein and matrix protein 2 extracellular domain fusion in mice / 中华微生物学和免疫学杂志

Objective:To evaluate the immunogenicity of a novel influenza virus mRNA vaccine based on conserved antigens delivered by lipopolyplex (LPP) platform in a mouse model.Methods:Four copies of genes coding for extracellular domain of matrix 2 protein (M2e) and nucleoprotein (NP) of influenza A virus were synthetized after codon optimization. The fusion antigens were transcribed in vitro and delivered by LPP platform, named as LPP-4M2eNP. Expression of M2e and NP in eukaryotic cells was detected by immunofluorescence assay (IFA). BALB/c mice were inoculated intramuscularly twice with 10 μg or 30 μg LPP-4M2eNP vaccine at an interval of four weeks. Antibody response was detected by ELISA and cellular-mediated immunity (CMI) was detected by enzyme-linked immunospot assay (ELISPOT). Results:IFA showed that NP and M2e were expressed correctly in eukaryotic cells. Single dose immunization could induce significant antigen (NP, M2e)-specific CMI and antigen (NP, M2e)-specific antibody response was induced in mice with Th1 type bias after boost immunization. Moreover, NP-specific CMI was increased significantly after the second immunization, while no significant change in M2e-specific CMI was observed.Conclusions:Stronger CMI was triggered in mice by single dose of LPP-4M2eNP vaccine. Furthermore, robust humoral and cellular immune responses were induced after boost immunization. This study suggested that LPP-4M2eNP vaccine, which based on conserved antigen of influenza A and delivered by LPP platform, had great potential for development and application.

Development of mRNA vaccine and its research in influenza / 中华微生物学和免疫学杂志

Influenza is an acute infectious disease of the respiratory tract and has posed a serious threat to human health. Vaccination is the most effective way to prevent influenza. The traditional inactivated or attenuated live vaccine based on chicken embryo has a long production cycle. Additionally, the surface antigens of vaccine strains and epidemic strains often do not match due to the continuous antigen drift or shift of influenza virus. As a result, the effectiveness of influenza vaccine is significantly reduced, so there is an urgent need to find a rapid and efficient influenza vaccine production strategy to overcome this limitation. In recent years, with the development of genetic engineering technology and various delivery systems, nucleic acid vaccine has been well developed, especially mRNA vaccine has attracted wide attention because of its simple production process and good safety. Therefore, mRNA-based vaccine immunization strategy provides a new direction for the prevention of influenza. This article reviews the molecular design, delivery system and research in influenza in the development of mRNA vaccine, in order to provide a reference basis for the research and development of influenza mRNA vaccine.

Innate and adaptive immune responses toward nanomedicines

Since the commercialization of the first liposomes used for drug delivery, Doxil/Caelyx® and Myocet®, tremendous progress has been made in understanding interactions between nanomedicines and biological systems. Fundamental work at the interface of engineering and medicine has allowed nanomedicines to deliver therapeutic small molecules and nucleic acids more efficiently. While nanomedicines are used in oncology for immunotherapy or to deliver combinations of cytotoxics, the clinical successes of gene silencing approaches like patisiran lipid complexes (Onpattro®) have paved the way for a variety of therapies beyond cancer. In parallel, the global severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) pandemic has highlighted the potential of mRNA vaccines to develop immunization strategies at unprecedented speed. To rationally design therapeutic and vaccines, chemists, materials scientists, and drug delivery experts need to better understand how nanotechnologies interact with the immune system. This review presents a comprehensive overview of the innate and adaptative immune systems and emphasizes the intricate mechanisms through which nanomedicines interact with these biological functions.

Covid-19 vaccine management (Comirnaty and mrna-1273 Moderna) in a teaching hospital in Italy: a short report on the vaccination campaign

OBJECTIVES@#In this article, we aim to share our experience in the hospital reorganization made to conduct the SARS-CoV-2 vaccination campaign, based on the principles of flexibility and adaptability.@*STUDY DESIGN@#A descriptive study.@*METHODS@#The data concerning the organization of the vaccination campaign were taken from the operative protocol developed by the hospital dedicated task force, composed by experts in hygiene, public health, occupational medicine, pharmacists, nurses, hospital quality, and disaster managers. Data about the numbers of vaccine administered daily were collected by the Innovation and Development Operative Unit database.@*RESULTS@#Vaccinations against COVID-19 started across the EU on the 27th of December 2020. The first phase of the vaccination campaign carried out in our hospital was directed to healthcare workers immunization including medical residents, social care operators, administrative staff and technicians, students of medicine, and health professions trainees. The second phase was enlarged to the coverage of extremely fragile subjects. Thanks to the massive employment of healthcare workers and the establishment of dynamic pathways, it was possible to achieve short turnaround times and a large number of doses administered daily, with peaks of 870 vaccines per day. From the 27th of December up to the 14th of March a total of 26,341 doses of Pfizer have been administered. 13,584 were first doses and 12,757 were second doses. From the 4th to the 14th of March, 296 first doses of Moderna were dispensed. It was necessary to implement adequate spaces and areas adopting anti-contagion safety measures: waiting area for subjects to be vaccinated, working rooms for the dilution of the vaccine and the storage of the material, vaccination rooms, post-vaccination observation areas, room for observation, and treatment of any adverse reactions, with an emergency cart available in each working area.@*CONCLUSIONS@#The teaching hospital of Pisa faced the beginning of the immunization campaign readjusting its spaces, planning an adequate hospital vaccination area and providing an organization plan to ensure the achievement of the targets of the campaign. This represented a challenge due to limited vaccine doses supplied and the multisectoral teams of professionals to coordinate in the shortest time and the safest way possible. The organizational model adopted proved to be adequate and therefore exploited also for the second phase aimed to extremely fragile subjects.

Safety of BNT162b2 COVID-19 vaccine in adolescent patients of UP-PGH

Background@#In response to the pandemic brought about by COVID-19, vaccines were developed immediately. Together with adhering to safety protocols, vaccines are needed to help decrease the mortality and morbidity. As with any other, COVID-19 vaccines are evaluated based on efficacy and safety. Real world data is important in the recommendation of vaccines. @*Objectives@#This study aims to assess the short-term safety of BNT162b2 COVID-19 vaccines administered to Filipino adolescents from October 15, 2021 to December 15, 2021 at the Philippine General Hospital. The number and type of local and systemic reaction within 7 days of vaccination were determined. @*Methods@#This is a retrospective cohort study. The review of the recorded events was done through an electronic diary that was accessed from the official Electronic Medical Records of University of the Philippines-Philippine General Hospital (UP-PGH). This included solicited and prespecified local and systemic reactions that occurred within 7 days of receipt of vaccine dose. Descriptive statistics was used to present the data. @*Results@#Out of the 1,756 BNT162b2 vaccines administered (Dose 1- 890; Dose 2- 866), 13% (N=221) indicated having adverse reaction. Injection site pain was the overall most common reaction with majority (81%) experiencing it within 7 days of vaccination. Systemic reactions made up 60% of the reactions after Dose 1 and 85% of the reactions after Dose 2. This includes tiredness, headache and fever. None of the reactions required hospitalization or further workup. @*Conclusion@#BNT162b2 vaccine has a good safety profile among adolescents vaccinated at UP-PGH, since most of the reported adverse events within 7 days of vaccination were local and systemic reactogenic reactions that did not necessitate hospitalization or work-up. No serious adverse events were reported. Further follow-up is suggested to assess longer term safety.

Progress in mRNA vaccines against influenza viruses / 中华微生物学和免疫学杂志

Influenza poses a serious threat to global public health and causes serious economic los-ses. Vaccination is the most effective measure to prevent influenza. However, the mismatch between the vac-cine strain and the epidemic strain, which results from antigenic drift and antigen conversion of influenza vi-rus, often invalidates the conventional vaccine stockpiles. mRNA vaccine is a relatively safe platform com-posed of nucleic acid. Improvements in genetic engineering technology and novel delivery systems have great-ly promoted the research and development of mRNA vaccines against influenza. mRNA vaccines are promis-ing candidates for the rapid and efficient prevention of seasonal influenza epidemics and influenza pandemics.

mRNA vaccine delivery system based on liposome: research advances / 国际药学研究杂志

mRNA vaccines are a new class of nucleic acid vaccines. Compared with DNA vaccines, mRNA vaccines do not need to enter the nucleus, and there is no risk of integration into the genome, which makes them relatively safer. However, the poor stability of mRNA vaccines and the facile degradation by nuclease in vitro and in vivo are the bottlenecks restricting its development. Therefore, mRNA vaccines need to have a suitable delivery vehicle to deliver them to the body in order to have a better immune effect. Currently, mRNA delivery vectors include viral vectors and non-viral vectors. In this paper, a brief overview of non-viral vector liposome is presented, mainly focusing on the mRNA vaccines, liposome, endosomal escape and enhancing liposome delivery, and the research prospects of mRNA and liposome delivery vectors are also commented.