Immune Responses and Protective Immunity in Pangasius Pangasius (Hamilton, 1822) as Induced by Outer Membrane Proteins of Edwardsiella Tarda and Aluminium Hydroxide Adjuvant Complex

Edwardsiella tarda is considered one of the important bacterial fish pathogens. The outer membrane proteins (OMPs) of E. tarda are structurally and functionally conserved, and immunogenic. This study assessed the effects of the OMPs of E. tarda CGH9 as a vaccine without aluminium hydroxide [AH] (T1) and with AH adjuvant (T2) on the respiratory burst (ROB) activity, lymphocyte proliferation of head kidney (HK) leukocytes, and serum antibody production in pangas catfish Pangasius pangasius. The ROB activity and lymphocyte proliferation of HK leukocytes increased in both vaccinated groups compared to control. Nonetheless, the T2 group showed a gradual increase in ROB activity and lymphocyte proliferation of HK leukocytes up to 3-weeks post-vaccination (wpv). The serum antibody production in the T1 group decreased initially for up to 2-wpv and increased from 3-wpv; whereas, in the T2 group, the serum-specific antibody levels were significantly high from 1-wpv compared to control. Simultaneously, the protective efficacy in terms of relative percentage survival (RPS) in the T2 group after injecting with a lethal dose of E. tarda CGH9 was high (89.00±15.56) compared to the T1 group (78.00±0.00). Furthermore, the catfish administered with a booster dose of E. tarda OMPs with or without AH adjuvant showed no additional increase in immune response or protective immunity. These results suggested that E. tarda OMPs and AH adjuvant complex has a higher potential to induce protective immunity, which may be a good choice as a vaccine to combat E. tarda infection in catfish.

A subset of follicular helper-like MAIT cells can provide B cell help and support antibody production in the mucosa

We identify a MAIT cell subset expressing T follicular helper markers and show the ability of MAIT cells to support B cell responses in the mucosa.

Nanodiamonds in Oil Emulsions as Effective Vaccine Adjuvants and Antitumor Therapeutic Agents

Background Composed of mineral oil and mycobacteria pathogens, complete Freund’s adjuvant (CFA) is one of the most commonly used adjuvants for antibody production and scientific research due to its high efficiency. However, the dead mycobacteria in CFA can cause many allergic reactions. We propose here a new formulation based on the use of nanodiamonds (NDs) as biocompatible non-allergic additives in incomplete Freund’s adjuvant (IFA) to avoid these adverse effects. Methods Chicken egg ovalbumin (OVA) was used as the antigens and 100-nm NDs after purification by air oxidation and strong oxidative acid washes were used as the additives. Levels of OVA-specific IgG antibody in mouse sera were measured by using enzyme-linked immunosorbent assays (ELISA) after the second and third immunizations of healthy mice with OVA and OVA/ND in IFA or CFA. Abilities of the OVA/ND/IFA vaccination to inhibit the tumor growth of mice inoculated with EL4 cells or OVA-expressing E.G7 cells were examined over 1 month. Results The new formulation worked well as a potent vaccine adjuvant, which could boost the immune responses and reduce the consumption of antigens in producing antibodies of interest in model animals like mice. Additionally, the composites showed distinct therapeutic activities, as proven by the OVA/ND/IFA treatment that effectively inhibited the tumor progression of E.G7-inoculated mice, allowing the animals to survive over 35 days post tumor-cell challenges. About 0.2% of the injected ND particles were found in mouse spleens on day 24 after vaccination of the E.G7-inoculated mice with OVA/ND/IFA. Conclusions The multiple functionality of ND makes it useful as an active and trackable component of a vaccine adjuvant not only to enhance antibody production but also to suppress tumor growth in vivo. The ND-based new formulation can be developed into single-dose vaccines with promising potential for real-world applications.

Augmenting recombinant antibody production in HEK293E cells: Optimising transfection and culture parameters

Background Optimising recombinant antibody production is important for cost-effective therapeutics and diagnostics. With impact on commercialisation, higher productivity beyond laboratory scales is highly sought, where efficient production can also accelerate antibody characterisations and investigations. Methods Investigating HEK293E cells for mammalian antibody production, various transfection and culture parameters were systematically analysed for antibody light chain production before evaluating them for whole antibody production. Transfection parameters investigated include seeding cell density, the concentration of the transfection reagent and DNA, complexation time, temperature, and volume, as well as culture parameters such as medium replacement, serum deprivation, use of cell maintenance antibiotic, incubation temperature, medium volume, post-transfection harvest day and common nutrient supplements. Results Using 2 mL adherent HEK293E cell culture transfections with 25 kDa linear Polyethylenimine in the most optimised parameters, we demonstrated a ~ 2-fold production increase for light chain alone and for whole antibody production reaching 536 and 49 μg respectively in a cost-effective manner. With the addition of peptone, κ light chain increased by ~ 4-fold to 1032 μg while whole antibody increased to a lesser extent by ~ 2.5-fold to 51 μg, with benefits potentially for antibodies limited by their light chains in production. Conclusions Our optimised findings show promise for a more efficient and convenient antibody production method through transfection and culture optimisations that can be incorporated to scale up processes and with potential transferability to other mammalian-based recombinant protein production using HEK293E cells. Statement of Significance Recombinant antibody production is crucial for antibody research and development. Systematically investigating transfection and culture parameters such as PEI/DNA concentrations, complexation time, volume, and temperature, supplements, etc., we demonstrated a ~ 4-fold light chain alone production increase to 1032 μg and a 2.5-fold whole antibody production increase to 51 μg from 2 mL transfections.

T cell responses to SARS-CoV-2 in healthy controls and primary immunodeficiency patients

Understanding the T cell response to SARS-CoV-2 is key in patients who lack antibody production. We demonstrate the applicability of a functional assay to measure the T cell response in a cohort of patients with immunodeficiency.

Polyphenols in Matily Herbal & Spices Boosts Antibodies in an Elderly Following COVID 19 Infections – A Case Study

An elderly patient had COVID-19 infection in August 2020 and started the home remedies treatment for the first 18 hours, followed by Azithromycin 250 mg for 6 days. The patient revered well, and the infection was confirmed by antibodies in the patient's serum. Later on, the patient was on Matily Herbal Drink and Matily Herbal & Spices Mix to avoid complications connected with COVID-19 re-infection. After completing 12 months of post COVID-19 infection, the antibodies were assessed to find the status. It was found to be increased in qualitative assessment. The quantitative assessment of antibodies showed a much higher value compared with individuals who had both the dose of vaccines and tested the blood after two weeks since the second dose of vaccine. We suggest that the increased antibodies could be because of the bioavailability of polyphenols present in the Matily Herbal Drink and Matily Herbal & Spices Mix. Polyphenols inactivate COVID-19 virus and this may have helped the body to increase its antibody production. The bioavailability of polyphenols depends on various factors, including acidulants in the diet. Based on the present studies, we suggest India should have her own strategies to increase antibodies in the population instead of just following the norms laid by International Agencies for the booster dose of vaccine Keywords: COVID-19, Matily Herbal Drink, Matily Herbal, Spices Mix

NFATc1/αA and Blimp-1 Support the Follicular and Effector Phenotype of Tregs

CD4+CXCR5+Foxp3+ T-follicular regulatory (TFR) cells control the germinal center responses. Like T-follicular helper cells, they express high levels of Nuclear Factor of Activated T-cells c1, predominantly its short isoform NFATc1/αA. Ablation of NFATc1 in Tregs prevents upregulation of CXCR5 and migration of TFR cells into B-cell follicles. By contrast, constitutive active NFATc1/αA defines the surface density of CXCR5, whose level determines how deep a TFR migrates into the GC and how effectively it controls antibody production. As one type of effector Treg, TFR cells express B lymphocyte-induced maturation protein-1 (Blimp-1). Blimp-1 can directly repress Cxcr5 and NFATc1/αA is necessary to overcome this Blimp-1-mediated repression. Interestingly, Blimp-1 even reinforces the recruitment of NFATc1 to Cxcr5 by protein-protein interaction and by those means cooperates with NFATc1 for Cxcr5 transactivation. On the contrary, Blimp-1 is necessary to counterbalance NFATc1/αA and preserve the Treg identity. This is because although NFATc1/αA strengthens the follicular development of Tregs, it bears the inherent risk of causing an ex-Treg phenotype.

The influence of the gut microbiota on influenza vaccine-induced immunity

<p>Currently, annual vaccination is widely considered the most effective method for preventing and controlling influenza virus infection. However, many individuals mount suboptimal immune responses to vaccination and the factors leading to poor immune responses are yet to be elucidated. Interestingly, it has been proposed that microorganisms that inhabit the intestinal tract, the gut microbiota, can profoundly influence many facets of the host immune system, including the strength of the immune response to influenza vaccination. In line with these observations, we observed that short-term administration of antibiotics drastically reduced influenza vaccine-specific antibody production. In particular, antibiotic treatment diminished the frequency and activation status of multiple myeloid cell subsets in the draining lymph nodes at steady-state and following vaccination, with associated impairments in B and TFH cell responses. Composition and function of gut microbiota communities can be rapidly altered through dietary changes. Therefore, the impact of potential prebiotic and probiotic nutritional interventions on the immune response to influenza vaccination and subsequent infection was assessed. No improvement in antibody responses to influenza vaccination was observed following the nutritional interventions studies. However, oral administration of a propolis formulation led to some improvement in viral control following infection. Collectively, this investigation indicates that alterations in microbial-associated signals leads to severe impairments in cellular responses crucial to humoral immunity and subsequent vaccine-induced antibody production. Furthermore, by altering the gut microbiota through dietary interventions, there is potential to improve immune responses to vaccination.</p>