Neutralizing Antibody Responses against Five SARS-CoV-2 Variants and T Lymphocyte Change after Vaccine Breakthrough Infections from the SARS-CoV-2 Omicron BA.1 Variant in Tianjin, China: A Prospective Study / 生物医学与环境科学（英文）

OBJECTIVE@#To investigate whether Omicron BA.1 breakthrough infection after receiving the SARS-CoV-2 vaccine could create a strong immunity barrier.@*METHODS@#Blood samples were collected at two different time points from 124 Omicron BA.1 breakthrough infected patients and 124 controls matched for age, gender, and vaccination profile. Live virus-neutralizing antibodies against five SARS-CoV-2 variants, including WT, Gamma, Beta, Delta, and Omicron BA.1, and T-lymphocyte lymphocyte counts in both groups were measured and statistically analyzed.@*RESULTS@#The neutralizing antibody titers against five different variants of SARS-CoV-2 were significantly increased in the vaccinated population infected with the Omicron BA.1 variant at 3 months after infection, but mainly increased the antibody level against the WT strain, and the antibody against the Omicron strain was the lowest. The neutralizing antibody level decreased rapidly 6 months after infection. The T-lymphocyte cell counts of patients with mild and moderate disease recovered at 3 months and completely returned to the normal state at 6 months.@*CONCLUSION@#Omicron BA.1 breakthrough infection mainly evoked humoral immune memory in the original strain after vaccination and hardly produced neutralizing antibodies specific to Omicron BA.1. Neutralizing antibodies against the different strains declined rapidly and showed features similar to those of influenza. Thus, T-lymphocytes may play an important role in recovery.

Research progress of broadly neutralizing antibodies against human immunodeficiency virus / 中国生物制品学杂志

@#Acquired immune deficiency syndrome(AIDS)is a global public health issue,which has a major impact on human life and health.As the main pathogen of AIDS,human immunodeficiency virus type 1(HIV-1)has high variability and latent characteristics,there are no available vaccines and drugs to prevent and cure AIDS.The broadly neutralizing antibodies(bNAbs)against HIV-1 can induce effective immune response to HIV-1 infection in vivo,and neutralize different types of strains at the same time,which have great application potential in preventing HIV-1 infection and controlling the process of AIDS.In this paper,the production and acquisition,classification and characteristics as well as application of HIV-1bNAbs are reviewed so as to provide a reference for the subsequent research on HIV-1 vaccine and development of antibody drugs.

Comparison of immunization strategies for human rabies pre-exposure prophylaxis in laboratory animals / 中华微生物学和免疫学杂志

Objective:To compare the differences in the safety, efficacy and protective effects of rabies vaccine using the current pre-exposure prophylaxis schedule in China (0-7-21 or 28) and the newly recommended immunization program of WHO (0-7), aiming to provide data support for modifying the related content of Technical Guideline for Human Rabies Prevention and Control. Methods:The mice were randomly divided into five groups, namely 0-7-21 group (3-injection regimen), 0-7 group (2-injection regimen), 0-14 group, 0-21 group and control group, according to the current 3-injection regimen (0-7-21) in China and the 2-injection regimen (0-7) recommended by WHO. The survival status of the mice was observed. The mice were weighed every five days to compare the safety of different immunization procedures. Rabies virus neutralizing antibodies (RVNA) were detected 7, 14, 21, 28 and 35 d after the initial immunization. On day 35, the mice in each group were challenged with lethal dose of CVS-11 rabies virus to evaluate the protective effects of different pre-exposure immunization procedures.Results:There was no significant difference in weight gain of mice after vaccination. The positive rate of RVNA was 100% in all immunized groups from day 14, which could provide complete protection to mice. There was a significant difference in RVNA levels between 0-7-21 and 0-7 groups at 35 d( P<0.05), but there was no statistical difference at other time points ( P>0.05). RVNA level had a significant difference between 0-7 and 0-21 groups at 21 d and 35 d ( P<0.05). There was no statistical difference in RVNA level between other groups at each time point ( P>0.05). In the protective test, the survival rates of mice in all immune groups were 100%. Conclusions:The current 3-injection pre-exposure immunization procedure for rabies vaccine (0-7-21) and the newly recommended 2-injection immunization procedure (0-7) had similar efficacy and protective effects in animal tests. In view of the cost saving and better compliance of the 2-injection immunization procedure, it was recommended that relevant departments should conduct clinical trials as soon as possible to promote the implementation of this program.

Serosurvey for SARS-CoV-2 among blood donors in Wuhan, China from September to December 2019

The emerging of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused COVID-19 pandemic. The first case of COVID-19 was reported at early December in 2019 in Wuhan City, China. To examine specific antibodies against SARS-CoV-2 in biological samples before December 2019 would give clues when the epidemic of SARS-CoV-2 might start to circulate in populations. We obtained all 88,517 plasmas from 76,844 blood donors in Wuhan between 1 September and 31 December 2019. We first evaluated the pan-immunoglobin (pan-Ig) against SARS-CoV-2 in 43,850 samples from 32,484 blood donors with suitable sample quality and enough volume. Two hundred and sixty-four samples from 213 donors were pan-Ig reactive, then further tested IgG and IgM, and validated by neutralizing antibodies against SARS-CoV-2. Two hundred and thirteen samples (from 175 donors) were only pan-Ig reactive, 8 (from 4 donors) were pan-Ig and IgG reactive, and 43 (from 34 donors) were pan-Ig and IgM reactive. Microneutralization assay showed all negative results. In addition, 213 screened reactive donors were analyzed and did not show obviously temporal or regional tendency, but the distribution of age showed a difference compared with all tested donors. Then we reviewed SARS-CoV-2 antibody results from these donors who donated several times from September 2019 to June 2020, partly tested in a previous published study, no one was found a significant increase in S/CO of antibodies against SARS-CoV-2. Our findings showed no SARS-CoV-2-specific antibodies existing among blood donors in Wuhan, China before 2020, indicating no evidence of transmission of COVID-19 before December 2019 in Wuhan, China.

Neutralizing antibody response to COVID vaccination among the young and the elderly

Objective: The objective of the study was to assess the levels of neutralizing antibody after COVID vaccination in the elderly and compare it with that of the younger persons. The study also aimed at determining the association between the age, sex, and comorbidities and levels of neutralizing antibodies in the young and the old. Subjects and Methods: This was a single-center, cross-sectional, analytical study, conducted in the General Medicine Unit of ACS Medical College, Chennai, from August 2021 to October 2021. Forty?five elderly persons aged 60 years and above and 103 young adults aged 18 years and above and <60 years who were vaccinated with either COVISHIELD or COVAXIN were randomly selected to participate in this study. A detailed history regarding vaccination status, vaccination type, comorbidities, and breakthrough infection was obtained. Blood samples were collected from the participants to analyze the levels of neutralizing antibodies developed after COVID vaccination. Results: The mean age of the older participants was 66.13 ± 5.3 years and the mean age of the younger participants was 36.48 ± 10.9 years. The median level of neutralizing antibody in the younger participants was 97.4% (interquartile range [IQR]: 96.4%–98.0%) and in the older participants was 97.1% (IQR: 93.1%–97.6%). There was a significant difference in the neutralizing antibody level between the younger and the older participants (P = 0.033). There was no significant difference in the neutralizing antibody levels after two doses of either of the two vaccines among both the groups of participants. There was no significant association between the neutralizing antibody titer and sex and comorbidities in both the groups of participants. Five young and two old participants had breakthrough infections after vaccination. The antibody level was higher in persons with breakthrough infection than in those with no breakthrough infection in both the study groups. Conclusion: There was a significant difference in the neutralizing antibody level between the younger and the older participants after COVID vaccination. No significant difference existed in neutralizing antibody response with respect to the type of vaccine in both the study groups. There was no significant association between sex, comorbid status, and neutralizing antibody levels in both the groups.

Respuesta humoral inducida por la vacuna BBIBP-CorV/Sinopharm COVID-19 en trabajadores de primera línea / BBIBP-CorV/Sinopharm COVID-19 vaccine-induced humoral response in frontline workers

Con la llegada de la vacuna contra el COVID-19 se evidenció la disminución de casos de Infección por SARS-CoV-2. El objetivo del estudio fue cuantificar la producción de anticuerpos neutralizantes (An) e inmunoglobulina G (S-IgG) en trabajadores de primera línea inmunizados con dos dosis de la vacuna BBIBP-CorV/Sinopharma. Se realizó un estudio observacional analítico transversal en personal de salud inmunizado con la vacuna inactivada (BBIBP-CorV). Sus muestras sanguíneas se recogieron tres meses después de la segunda dosis, para medir las respuestas de anticuerpos (An, S-IgG). De un total de 172 personas evaluadas, 110 (64%) personas ya habían tenido COVID-19 antes de ingresar al estudio, los títulos de An fueron superiores a 10 AU/mL en el 60,5% de los vacunados y el 89,3% mostró S-IgG superior a 50 UA/mL. Los trabajadores mayores de 60 años fueron el grupo que no desarrolló suficientes anticuerpos. La correlación de An y S-IgG fue positiva (r=0,84) (p<0,001). La administración de dos dosis de la vacuna inactivada BBIBP-CorV/Sinopharma provocó una notable respuesta An y S-IgG, excepto en mayores de 60 años(AU)

With the arrival of the vaccine against COVID-19, the decrease in cases of SARS-CoV-2 infection was evidenced. The objective of the study was to quantify the production of neutralizing antibodies (An) and immunoglobulin G (S-IgG) in frontline workers immunized with two doses of the BBIBP-CorV/Sinopharma vaccine. A cross-sectional analytical observational study was carried out in health personnel immunized with the inactivated vaccine (BBIBP-CorV). Their blood samples were collected three months after the second dose, to measure antibody responses (An, S-IgG). Of a total of 172 people evaluated, 110 (64%) people already had COVID-19 before entering the study, An titers were greater than 10 AU/mL in 60.5% of those vaccinated and 89, 3% showed S-IgG greater than 50 AU/mL. Workers older than 60 years were the group that did not develop enough antibodies. The correlation of An and S-IgG was positive (r=0.84) (p<0.001). The administration of two doses of the inactivated BBIBP-CorV/Sinopharma vaccine caused a notable An and S-IgG response, except in those over 60 years of age(AU)

Absence of neutralizing antibodies against the Omicron SARS-CoV-2 variant in convalescent sera from individuals infected with the ancestral SARS-CoV-2 virus or its Gamma variant

Abstract Objectives: The aim of the present study was to evaluate if neutralizing antibody responses induced by infection with the SARS-CoV-2 strain that was dominant at the beginning of the pandemic or by the Gamma variant was effective against the Omicron variant. Methods: Convalescent sera from 109 individuals, never exposed to a SARS-CoV-2 vaccine, who had mild or moderate symptoms not requiring hospitalization following either a documented SARS-CoV-2 ancestral strain infection or a Gamma variant infection, were assayed for in vitro neutralizing antibody activity against their original strains and the Omicron variant. Results: Following an infection with the ancestral strain, 56 (93.3%), 45 (77.6%) and 1 (1.7%) serum sample were positive for neutralizing antibodies against the ancestral, Gamma variant, and Omicron variant, respectively. After infection with the Gamma variant, 43 (87.8%) and 2 (4.1%) sera were positive for neutralizing antibodies against the Gamma and Omicron variants, respectively. Conclusions: Neutralizing antibodies generated following mild or moderate infection with the SARS-CoV-2 ancestral strain or the Gamma variant are not protective against the Omicron variant. HIGHLIGHTS Individuals previously infected with SARS-CoV-2 ancestral strain do not develop neutralizing antibodies against the Omicron variant. Omicron variant escapes immune response after SARS CoV-2 previous infection with the SARS CoV-2 Gamma variant. Individuals previously infected with SARS-CoV-2 ancestral strain or with SARS-CoV-2 Gamma variant will likely have little protection if subsequently exposed to the Omicron variant.

Elevacion de anticuerpos neutralizantes contra sars cov-2 post vacunacion con sinopharm y astrazeneca hasta la tercera dosis, en el personal del banco de sangre materno infantil cns la paz - bolivia / Elevation of neutralizing antibodies against SARS CoV-2 after vaccination with Sinopharm and AstraZeneca up to the third dose, in the staff of the Maternal and Child Blood Bank CNS La Paz - Bolivia

Introducción: Después de la COVID-19, surgieron luego de muchas investigaciones un suministro de vacunas aprobadas a nivel mundial, dichas vacunas deben conferir una protección eficaz por un tiempo prolongado, poseer un buen perfil de seguridad, ser asequible y ser fácilmente accesible para todos, un reto difícil de conseguir por el tiempo y las características del virus. Objetivo: determinar la concentración de anticuerpos neutralizantes (A. N.) post vacunación en una población de trabajadores del Banco de Sangre Material y métodos: Se realizó un estudio prospectivo, descriptivo, transversal, tomando como población de estudio a todo el personal del Banco de Sangre, varones y mujeres con un rango de edad entre 26 y 72 años, se evaluó el aumento de A. N. después de inoculada la segunda dosis de Sinopharm en fecha 19/05/21, luego se midió la cantidad de anticuerpos generados en fecha 20/10/21 previo a la tercera dosis de refuerzo de Astrazeneca, evaluando nuevamente a los 35 días luego de la tercera dosis 02/12/2021 para finalmente evaluar estos niveles en fecha 26/01/2022. La técnica utilizada fue (ELISA) de la marca EUROIMMUN Anti-SARS- CoV-2 S1 del tipo IgG. Resultados y conclusiones: Se puede verificar que la concentración de A. N. producidos por la vacunación desde Sinopharm y el refuerzo con Astrazeneca favoreció a que dichos anticuerpos se mantengan altos en el tiempo (322 días luego de la primera dosis) llegando a un 80% de la concentración máxima en la lectura final. Concluimos que con cada refuerzo de vacuna anti SARS-CoV-2 el título de A.N. sube de manera significativa, motivo por el cual consideramos importante en nuestro pais una cuarta dosis como método preventivo y de inmunidad.

Introduction: After COVID-19, emerged after much research a supply of vaccines approved worldwide, these vaccines must confer effective protection for a prolonged time, possess a good safety profile, be affordable and be easily accessible to all, a challenge difficult to achieve due to the time and characteristics of the virus. Objective: to determine the concentration of neutralizing antibodies post vaccination in a population of Blood Bank workers; Material and methods: A prospective, descriptive, observational, cross-sectional study was carried out, taking as study population all the Blood Bank personnel, among men and women with an age range between 26 and 72 years old. the increase of Neutralizing Antibodies was evaluated after inoculation of the second dose of Sinopharm on 05/19/21, then the amount of antibodies generated was measured on 10/20/21 prior to the third booster dose of Astrazeneca, evaluating again 35 days after the third dose on 12/02/2021 and finally evaluating these levels on 01/26/2022. The technique used was the EUROIMMUN Anti-SARS-CoV-2 S1 ELISA. Results and conclusions: It can be verified that the concentration of N.A. produced by vaccination from Sinopharm and the booster with Astrazeneca favored that these antibodies remained high over time (322 days after the first dose) reaching 80% of the maximum concentration in the final reading. We conclude that with each booster of anti SARS CoV 2 vaccine, the titer of N.A. rises significantly, which is why we consider important in our country a fourth dose as a preventive and immunity method.

Evaluación de la respuesta humoral inducida por la vacuna BBIBP-CorV mediante la determinación de anticuerpos neutralizantes en personal sanitario peruano / Evaluation of the humoral response induced by BBIBP-CorV vaccine by determining neutralizing antibodies in peruvian healthcare personnel

RESUMEN Objetivo. Determinar el título de anticuerpos frente al dominio de unión al receptor (RBD) de la proteína espiga (S) en personal de salud entre la 4.ª y 12.ª semana luego de haber recibido la vacuna BBIBP-CorV (Sinopharm). Materiales y métodos. Se incluyeron 168 trabajadores de salud de dos hospitales de la región, quienes cumplían el esquema completo con vacuna de Sinopharm, y se realizó la medición de anticuerpos en suero mediante la prueba Elecsys®Anti-SARS-CoV-2. Resultados. Todos los participantes desarrollaron anticuerpos frente al dominio RBD. El valor mínimo fue de 1,78 U/mL. En 70 (41,7%) participantes se encontraron niveles iguales o por encima de 250. La media geométrica fue de 82,6 (IC 95% 67,8-100,6). Las mujeres presentaron un mayor nivel de anticuerpos. El grupo de participantes en los que se midieron anticuerpos entre las semanas 4 y 7 posvacunación mostró niveles de anticuerpos significativamente mayores que los pacientes cuyas determinaciones fueron realizadas entre las 10 y 12 semanas posvacunación. Entre los pacientes con antecedente de COVID-19, los niveles de anticuerpos se encontraron en cifras iguales o por encima de 250 U/mL en el 88% de casos, en comparación con 6% entre aquellos sin antecedente de COVID-19, (p<0,001). Conclusión. Todos los participantes inmunizados con la vacuna BBIBP-CorV presentaron positividad a anticuerpos frente al RBD de la proteína S del SARS-CoV-2. Es necesario evaluar la correlación entre la magnitud de los títulos y la protección frente a COVID-19 y el tiempo de protección conferido por la vacuna.

ABSTRACT Objective. To determine the titer of antibodies against the receptor binding domain (RBD) of the spike protein (S) in health personnel between the 4th and 12th week after receiving the BBIBP-CorV vaccine (Sinopharm). Materials and methods. We included a total of 168 healthcare workers from two hospitals in the region, who complied with the complete Sinopharm vaccine schedule; serum antibodies were measured using the Elecsys® Anti-SARS-CoV-2 test. Results. All participants developed antibodies to the RBD domain. The lowest antibody titer level was 1.78 U/mL. Levels equal to or above 250 were found in 70 (41.7%) participants. The geometric mean was 82.6 (95% CI: 67.8-100.6). Women had higher antibody levels. Participants whose antibodies were measured between 4- and 7-weeks post-vaccination showed significantly higher antibody levels than patients whose antibody levels were measured between 10- and 12-weeks post-vaccination. Among patients with a history of COVID-19, antibody levels were found to be at or above 250 U/mL in 88% of cases, compared to 6% among those without a history of COVID-19, (p<0.001). Conclusion. All participants immunized with BBIBPCorV vaccine were positive for antibodies against the SARS-CoV-2 spike protein RBD. The correlation between the titer level and protection against COVID-19, as well as the length of the protection provided by the vaccine, needs to be evaluated.

Efectividad de inducción de tolerancia inmune en niños con hemofilia A y aloanticuerpos neutralizantes / Immunotolerance induction effectivity in hemophilia A children and neutralizing alloantibodies

Resumen: Introducción: El desarrollo de aloanticuerpos neutralizantes anti-factor VIII en hemofilia A es la complicación más seria relacionada al tratamiento. La inducción de tolerancia inmune (ITI) o inmunotolerancia es el único tratamiento que erradica inhibidores, permitiendo utilizar nuevamente factor VIII para el tratamiento o profilaxis de eventos hemorrágicos. Objetivo: reportar la experiencia en niños sometidos a inmunotolerancia en la red pública del país. Pacientes y Método: Análisis retrospectivo y descriptivo de 13 niños con Hemofilia A severa e inhibidores persistentes de alto título, que recibieron ITI y seguimiento completo. Se utilizó concentrado de FVIII plasmático en dosis de 70-180 UI/Kg/diarias, definiendo éxito como la negativización del inhibidor y recu peración de la vida media del FVIII. Resultados expresados en media (rango). Resultados: En 13 pacientes se identificó el inhibidor, a una edad de 17,6 meses (2-48), tras 35,2 días (9-112) de exposición a FVIII. Once pacientes (84,6%) recuperaron la vida media del FVIII, tras 49,6 meses (26-70) de tratamiento. En los pacientes que respondieron, el título del inhibidor se negativizó en 7,3 meses (1-20). Conclusiones: En niños con hemofilia A e inhibidores persistentes de alto título, la ITI tiene un elevado éxito. Dado que el tiempo de respuesta es variable, la inmunotolerancia debe ser personalizada.

Abstract: Introduction: The development of anti-factor VIII neutralizing antibodies in hemophilia A is the most severe com plication related to treatment. Immune tolerance induction (ITI) is the only known treatment for eradicating inhibitors. A successful ITI allows using factor VIII (FVIII) again for the treatment or prophylaxis of hemorrhagic events. Objective: To report the experience of pediatric patients who underwent ITI in the country's public health care network. Patients and Method: Retrospective and descriptive analysis of 13 pediatric patients with severe Hemophilia A and high-titer inhibitors persis tence who underwent ITI and complete follow-up. Plasma-derived FVIII concentrate was used at 70 180 IU/kg/day doses. The success of the treatment is defined by achieving a negative titer and a half life recovery of the FVIII. The results were expressed in median (range). Results: In 13 patients, the inhibitor was identified at an average age of 17.6 months, after 35.2 days of exposure to the FVIII. 11 patients (84.6%) recovered the half-life of FVIII after 49.6 months of treatment. In the patients who responded to treatment, the inhibitor titer was negative at 6 months on average. Conclusions: ITI is the treatment of choice for patients with hemophilia A and inhibitors persistence. ITI must be perso nalized since the time response is variable in each patient.

Immunogenicity in sheep of Uruguayan commercial vaccines against bovine alphaherpesvirus 1, 5 and bovine pestiviruses / Imunogenicidade em ovelhas de vacinas comerciais Uruguaias para alfaherpesvírus bovino 1, 5 e pestivírus de bovinos

ABSTRACT: The serological responses induced by four commercial inactivated Uruguayan vaccines against bovine alphaherpesviruses (BoHV)-1 and -5 and bovine pestiviruses (BVDV-1, BVDV-2, and HoBiPeV) were evaluated in sheep. Thirty-seven sheep were immunized twice (day 0 and 25) and their serum samples were tested at different intervals (days 0, 25, 40, 60, and 90) post-vaccination (PV). Among the four vaccines tested, only one (G4) could induce the production of moderate neutralizing antibody titers against BoHV-1 and -5 and BVDV-1 and -2. The G3 vaccine showed a neutralizing serological response against the bovine alphaherpesviruses only. The G1 and G2 vaccines produced extremely low levels of antibodies in a few vaccinated animals only (geometric mean titers (GMT) 2.2). Similar levels of immunological responses were induced by the G4 vaccine against BoHV-1 and -5, and titers of neutralizing antibodies induced in approximately 70% of the animals are known to confer protection (GMT > 8). For bovine pestiviruses, the vaccine stimulated response of G4 against BVDV-2 was higher compared to that against BVDV-1, and extremely low for HoBiPeV. The peak of neutralizing antibodies to BoHV-1 and BVDV-1 was observed on days 40 and 60 PV, respectively. Thereafter, a remarkably decrease in neutralizing antibody response was observed at day 90 PV. These results demonstrated that tested commercial Uruguayan vaccines did not induce a serological response of adequate magnitude and duration. Thus, it is important to periodically review formulations and compositions of commercial vaccines against bovine alphaherpesviruses and pestiviruses.

RESUMO: A resposta sorológica induzida por quatro vacinas comerciais uruguaias inativadas contra os alfaherpesvírus bovinos (BoHV-1 e -5) e pestivírus de bovinos (BVDV-1, BVDV-2 e HoBiPeV) foi avaliada em ovinos. Os animais foram imunizados duas vezes (dia 0 e dia 25) e o soro testado em diferentes intervalos (dias 0, 25, 40, 60 e 90) após a vacinação (PV). Dentre as quatro vacinas testadas, apenas uma (G4) apresentou títulos de anticorpos neutralizantes moderados para os BoHV-1 e -5, BVDV-1 e 2. A vacina G3 apresentou resposta somente para alfaherpesvírus bovinos. As vacinas G1 e G2 estimularam resposta somente em alguns animais vacinados. Para a vacina G4, observou-se que a resposta imunológica frente ao BoHV-1 e 5 foi semelhante e pelo menos 70% dos animais apresentaram níveis protetivos de anticorpos neutralizantes. Para os pestivírus bovinos, a vacina G4 estimulou resposta para o BVDV-2 mais elevada quando comparada com o BVDV-1, e quase que indetectável para HoBiPeV. O pico de anticorpos neutralizantes para o BoHV-1 foi observado no dia 40 PV e no dia 60 PV para o BVDV-1. Após isso, observou-se um decréscimo considerável na resposta de anticorpos neutralizantes. Os resultados demonstraram que vacinas comerciais uruguaias testadas não induziram resposta sorológica de magnitude e duração adequadas. Assim, ressalva-se a importância de rever periodicamente a formulação e composição das vacinas comerciais para alfaherpesvírus e pestivírus bovinos.

Efficacy analysis of severe acute respiratory syndrome coronavirus 2 DNA vaccine and recombinant subunit vaccine inducing neutralizing antibodies in mice / 第二军医大学学报

Objective To investigate the efficacy of neutralizing antibodies induced by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) receptor-binding domain (RBD) and spike (S) protein S1 subunit. Methods The SARS-CoV-2 RBD and mouse immunoglobulin G1 (IgG1) Fc fragment (mFc) fusion protein expression plasmid pVRCRBD- mFc was constructed and transfected into human embryonic kidney 293T cells. The RBD-mFc fusion protein in the cell supernatants was detected by Western blotting. The effect of RBD-mFc in cell supernatants and CHO recombinant S1-human IgG1 Fc (S1-hFc) fusion protein on SARS-CoV-2 infection was detected by microneutralization test. BALB/c mice were immunized with plasmid pVRC-RBD-mFc and S1-hFc fusion protein via intramuscular injection. Anti-S1 IgG antibodies in mouse sera were detected by enzyme-linked immunosorbent assay (ELISA), and the virus neutralization activity of mouse sera was detected by microneutralization test. Results The RBD-mFc fusion protein could be detected in the culture supernatants of 293T cells transfected with the plasmid pVRC-RBD-mFc, the concentrated supernatants and the S1- hFc fusion protein could inhibit SARS-CoV-2 infection on Vero E6 cells in a concentration-dependent manner. Anti-S1 IgG antibodies could be detected in the sera of mice immunized with plasmid pVRC-RBD-mFc and S1-hFc fusion protein, and the sera of both groups could neutralize SARS-CoV-2 infection. The serum antibody titers and virus neutralization activity of S1- hFc fusion protein immunized mice were significantly higher than those of plasmid pVRC-RBD-mFc immunized mice (both P<0.01). Conclusion Both SARS-CoV-2 RBD and S1 subunit may be used as effective vaccine antigens. Compared with DNA vaccine, recombinant subunit vaccine can induce neutralizing antibody more effectively..

Durability of neutralizing antibodies and T-cell response post SARS-CoV-2 infection / 医学前沿

The ongoing pandemic of Coronavirus disease 19 (COVID-19) is caused by a newly discovered β Coronavirus named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). How long the adaptive immunity triggered by SARS-CoV-2 can last is of critical clinical relevance in assessing the probability of second infection and efficacy of vaccination. Here we examined, using ELISA, the IgG antibodies in serum specimens collected from 17 COVID-19 patients at 6-7 months after diagnosis and the results were compared to those from cases investigated 2 weeks to 2 months post-infection. All samples were positive for IgGs against the S- and N-proteins of SARS-CoV-2. Notably, 14 samples available at 6-7 months post-infection all showed significant neutralizing activities in a pseudovirus assay, with no difference in blocking the cell-entry of the 614D and 614G variants of SARS-CoV-2. Furthermore, in 10 blood samples from cases at 6-7 months post-infection used for memory T-cell tests, we found that interferon γ-producing CD4

Respuesta inmune y terapia gó©nica con vectores virales adenoasociados / Immune response and gene therapy with adenoassociated viral vectors

En los ó;ºltimos aó;±os la terapia gó;©nica se ha posicionado como una opció;n real y segura en el desarrollo de alternativas terapó;©uticas para la cura y la prevenció;n de diferentes enfermedades. Consiste en la inserció;n de material genó;©tico en un tejido o có;©lula defectuosa, mediante el uso de un vector. Existen varias consideraciones para seleccionar el vector más apropiado, incluyendo el potencial de unió;n y entrada a la có;©lula diana, la capacidad de transferencia del material genó;©tico al nó;ºcleo, la habilidad de expresió;n del inserto y la ausencia de toxicidad. En el panorama actual, los vectores virales más utilizados son los derivados de los virus adenoasociados (AAV). Características como su bioseguridad, baja toxicidad y tropismo selectivo, han posibilitado su evaluació;n como opció;n terapó;©utica en un amplio nó;ºmero de enfermedades monogó;©nicas o complejas. A pesar de sus ventajas, los vectores AAV presentan inconvenientes, siendo el más importante la respuesta inmune del paciente al vector, especialmente la respuesta mediada por anticuerpos neutralizantes (NAb). Los NAb disminuyen la transducció;n del vector e impiden la expresió;n del gen que transporta, limitando su aplicació;n clínica. Por lo tanto, identificar y cuantificar la presencia y actividad de los NAbs, es el primer paso en cualquier protocolo de terapia gó;©nica con vectores AAV. La presencia de NAb depende principalmente de la exposició;n al virus en la naturaleza y varía drásticamente segó;ºn edad, localizació;n geográfica y estado de salud de la persona evaluada.

In recent years, gene therapy has been positioned as a real and safe option in the development of therapeutic alternatives for the cure and prevention of different diseases. It consists in the insertion of genetic material in a defective tissue or cell, through the use of a vector. There are several considerations for selecting the most appropriate vector, including the potential for binding and entry to the target cell, the ability of the genetic material to transfer to the nucleus, the ability to express the insert, and the absence of toxicity. In the current scenario, the most commonly used viral vectors are those derived from adeno-associated viruses (AAV). Characteristics such as biosafety, low toxicity and selective tropism have enabled its evaluation as a therapeutic option in many monogenic or complex diseases. Despite their advantages, AAV vectors have drawbacks, the most important being the patient’s immune response to the vector, especially the response mediated by neutralizing antibodies (NAb). NAbs decrease the transduction of the vector and prevent the expression of the gene it transports, limiting its clinical application. Therefore, identifying and quantifying the presence and activity of NAbs is the first step in any gene therapy protocol with AAV vectors. The presence of NAbs depends mainly on exposure to the virus in nature and varies drastically according to age, geographic location and health status of the person evaluated.

Absence of neutralizing activity in serum 1 year after successful treatment with antivirals and recovery from MERS in South Korea

We evaluated the neutralizing activity in serum from three patients >1 year after recovery from Middle East respiratory syndrome (MERS) associated with mild pneumonia treated with antivirals during the MERS outbreak in South Korea at 2015. The neutralizing activity in serum was measured by pseudovirus inhibition assays. Three-fold diluted serum of subjects showed only 9.7%, 10.3%, and 2.2% reductions in relative light units. So, significant neutralizing activity was not demonstrated in any sera of three patients with mild pneumonia >1 year after being successfully treated with antiviral agents and recovering from MERS coronavirus infection.

Investigation on neutralizing antibody level of Japanese encephalitis virus in adults from Guizhou Province / 中华疾病控制杂志

@# Objective To investigate the level of neutralizing antibodies against Japanese encephalitis (JE) virus in adults from Guizhou Province. Methods A multi-stage random sampling method was used to collect 360 serum of healthy adults in 6 age groups of 3 cities (states) from May to June 2017. Neutralizing antibodies against Japanese encephalitis virus of healthy adults were detected by plaque reduction neutralization test. Results The positive rate of neutralizing antibodies against Japanese encephalitis virus was 55.28%, and the geometric mean titer (GMT) was 1 :17.52 in 360 subjects. The difference of the positive rates of neutralizing antibodies against Japanese encephalitis virus of adults in different genders(2=10.798, P=0.001) and in different incidence regions(2=6.090, P=0.048)was statistically significant(both P<0.05). The positive rate of different age groups was 45.00%-60.00%(2=4.236, P=0.516). The positive rate of Buyi nationality was the highest (79.17%). Conclusions The level of neutralizing antibodies against Japanese encephalitis virus of adults in Guizhou province is low, and there is a risk of epidemic encephalitis in adults.

The development of HIV vaccines targeting gp41 membrane-proximal external region (MPER): challenges and prospects

A human immunodeficiency virus type-1 (HIV-1) vaccine which is able to effectively prevent infection would be the most powerful method of extinguishing pandemic of the acquired immunodeficiency syndrome (AIDS). Yet, achieving such vaccine remains great challenges. The membrane-proximal external region (MPER) is a highly conserved region of the envelope glycoprotein (Env) gp41 subunit near the viral envelope surface, and it plays a key role in membrane fusion. It is also the target of some reported broadly neutralizing antibodies (bNAbs). Thus, MPER is deemed to be one of the most attractive vaccine targets. However, no one can induce these bNAbs by immunization with immunogens containing the MPER sequence(s). The few attempts at developing a vaccine have only resulted in the induction of neutralizing antibodies with quite low potency and limited breadth. Thus far, vaccine failure can be attributed to various characteristics of MPER, such as those involving structure and immunology; therefore, we will focus on these and review the recent progress in the field from the following perspectives: (1) MPER structure and its role in membrane fusion, (2) the epitopes and neutralization mechanisms of MPER-specific bNAbs, as well as the limitations in eliciting neutralizing antibodies, and (3) different strategies for MPER vaccine design and current harvests.

Preliminary assessment of correlation between T-lymphocyte responses and control of porcine reproductive and respiratory syndrome virus (PRRSV) in piglets born after in-utero infection of a type 2 PRRSV / 대한수의학회지

A preliminary study into the protective mechanisms of adaptive immunity against porcine reproductive and respiratory syndrome virus (PRRSV) in piglets (n = 9) born to a gilt challenged intranasally with a type-2 PRRSV. Immune parameters (neutralizing antibodies, CD3⁺CD4⁺, CD3⁺CD8⁺, CD3⁺CD4⁺CD8⁺ T-lymphocytes, and PRRSV-specific interferon (IFN)-γ secreting T-lymphocytes) were compared with infection parameters (macro- and microscopic lung lesion, and PRRSV-infected porcine alveolar macrophages (CD172α⁺PRRSV-N⁺ PAM) as well as with plasma and lymphoid tissue viral loads. Percentages of three T-lymphocyte phenotypes in 14-days post-birth (dpb) peripheral blood mononuclear cell (PBMC) had significant negative correlations with percentages of CD172α⁺PRRSV-N⁺ PAM (p 0.1) with infection parameters. The results indicate that T-lymphocytes contribute to controlling PRRSV replication in young piglets born after in-utero infection.

Do immunoglobulin G and immunoglobulin E anti-l-asparaginase antibodies have distinct implications in children with acute lymphoblastic leukemia? A cross-sectional study

Abstract Background l-Asparaginase is essential in the treatment of childhood acute lymphoblastic leukemia. If immunoglobulin G anti-l-asparaginase antibodies develop, they can lead to faster plasma clearance and reduced efficiency as well as to hypersensitivity reactions, in which immunoglobulin E can also participate. This study investigated the presence of immunoglobulin G and immunoglobulin E anti-l-asparaginase antibodies and their clinical associations. Methods Under 16-year-old patients at diagnosis of B-cell acute lymphoblastic leukemia confirmed by flow cytometry and treated with a uniform l-asparaginase and chemotherapy protocol were studied. Immunoglobulin G anti-l-asparaginase antibodies were measured using an enzyme-linked immunosorbent assay. Intradermal and prick skin testing was performed to establish the presence of specific immunoglobulin E anti-l-asparaginase antibodies in vivo. Statistical analysis was used to investigate associations of these antibodies with relevant clinical events and outcomes. Results Fifty-one children were studied with 42 (82.35%) having anti-l-asparaginase antibodies. In this group immunoglobulin G antibodies alone were documented in 10 (23.8%) compared to immunoglobulin E alone in 18 (42.8%) patients. Immunoglobulin G together with immunoglobulin E were simultaneously present in 14 patients. Children who produced exclusively immunoglobulin G or no antibodies had a lower event-free survival (p-value = 0.024). Eighteen children (35.3%) relapsed with five of nine of this group who had negative skin tests suffering additional relapses (range: 2-4), compared to none of the nine children who relapsed who had positive skin tests (p-value < 0.001). Conclusion Children with acute lymphoblastic leukemia and isolated immunoglobulin G anti-l-asparaginase antibodies had a higher relapse rate, whereas no additional relapses developed in children with immunoglobulin E anti-l-asparaginase antibodies after the first relapse.

Serological response to rabies virus induced by commercial vaccines in cattle / Resposta sorológica ao vírus da raiva induzida por vacinas comerciais em bovinos

ABSTRACT: The antibody response to rabies virus (RABV) induced by commercial vaccines in heifers was investigated. For this, 84 heifers were vaccinated twice (30 days interval) with each of four vaccines (G1 = 14 animals; G2 = 24; G3 = 22 and G4 = 24) and received a booster vaccination 360 days later. Serum samples collected at different intervals after vaccination and 30 days after booster were submitted to a virus neutralizing (VN) assay for RABV antibodies. Thirty days after the second vaccine dose, 92% of the immunized animals presented VN titers ≥0.5UI/mL (geometric medium titers [GMT] 1.7 to 3.8UI/mL). At the day of the booster (360 days post-vaccination); however, the percentage of animals harboring antibody titers ≥0.5UI/mL had dropped to 31% (0-80% of the animals, depending on the vaccine), resulting in lower GMT (0.1 to 0.6UI/mL). Booster vaccination at day 360 resulted in a detectable anamnestic response in all groups, resulting in 83% of animals (65 to 100%) harboring VN titers ≥0.5UI/mL thirty days later (GMT 0.6 to 4.3UI/mL). These results indicated that these vaccines were able to induce an adequate anti-RABV response in all animals after prime vaccination (and after booster as well). However, the titers decreased, reaching titers <0.5UI/mL in approximately 70% of animals within the interval before the recommended booster. Thus, booster vaccination for rabies in cattle using the current vaccines should be performed before the recommended one-year interval, as to maintain neutralizing antibodies levels in most vaccinated animals.

RESUMO: A resposta sorológica contra o vírus da raiva (RABV) induzida por vacinas comerciais foi investigada em bovinos. Para isso, 84 novilhas foram vacinadas duas vezes (30 dias de intervalo) com cada vacina (G1 = 14 animais; G2 = 24; G3 = 22 e G4 = 24) e receberam uma vacinação de reforço 360 dias depois. Amostras de soro coletadas em diferentes momentos após a vacinação e após o reforço vacinal foram submetidas ao teste de vírus neutralização (VN) para detecção de anticorpos contra o RABV. Trinta dias após a segunda dose vacinal, 92% dos animais apresentaram títulos neutralizantes ≥0,5UI/mL (GMT 1,7 a 3,8UI/mL). Porém, no dia do reforço (360 dias pós-vacinação), a porcentagem de animais que ainda apresentava títulos ≥0,5UI/mL havia se reduzido a 31% dos animais (0 a 80%, dependendo da vacina), resultando em baixos TMGs (0,1 a 0,6UI/mL). A vacinação de reforço no dia 360 resultou em resposta anamnéstica em todos os grupos, resultando em 83% (65 a 100%) de animais com títulos VN ≥0,5UI/mL trinta dias após (GMT 0,6 a 4,3UI mL-1). Esses resultados indicam que as vacinas avaliadas induzem uma resposta adequada de anticorpos anti-RABV após a vacinação (e também após o reforço). No entanto, os títulos reduzem-se, atingindo níveis <0,5UI/mL em 70% dos animais durante o intervalo antes do reforço. Assim, vacinação de reforço contra a raiva em bovinos, utilizando-se as vacinas atuais, deve ser realizada em intervalo inferior a um ano, de forma a manter os níveis de anticorpos neutralizantes na maioria dos animais.