Egg-Derived Anti-SARS-CoV-2 Immunoglobulin Y (IgY) With Broad Variant Activity as Intranasal Prophylaxis Against COVID-19.

COVID-19 emergency use authorizations and approvals for vaccines were achieved in record time. However, there remains a need to develop additional safe, effective, easy-to-produce, and inexpensive prevention to reduce the risk of acquiring SARS-CoV-2 infection. This need is due to difficulties in vaccine manufacturing and distribution, vaccine hesitancy, and, critically, the increased prevalence of SARS-CoV-2 variants with greater contagiousness or reduced sensitivity to immunity. Antibodies from eggs of hens (immunoglobulin Y; IgY) that were administered the receptor-binding domain (RBD) of the SARS-CoV-2 spike protein were developed for use as nasal drops to capture the virus on the nasal mucosa. Although initially raised against the 2019 novel coronavirus index strain (2019-nCoV), these anti-SARS-CoV-2 RBD IgY surprisingly had indistinguishable enzyme-linked immunosorbent assay binding against variants of concern that have emerged, including Alpha (B.1.1.7), Beta (B.1.351), Delta (B.1.617.2), and Omicron (B.1.1.529). This is different from sera of immunized or convalescent patients. Culture neutralization titers against available Alpha, Beta, and Delta were also indistinguishable from the index SARS-CoV-2 strain. Efforts to develop these IgY for clinical use demonstrated that the intranasal anti-SARS-CoV-2 RBD IgY preparation showed no binding (cross-reactivity) to a variety of human tissues and had an excellent safety profile in rats following 28-day intranasal delivery of the formulated IgY. A double-blind, randomized, placebo-controlled phase 1 study evaluating single-ascending and multiple doses of anti-SARS-CoV-2 RBD IgY administered intranasally for 14 days in 48 healthy adults also demonstrated an excellent safety and tolerability profile, and no evidence of systemic absorption. As these antiviral IgY have broad selectivity against many variants of concern, are fast to produce, and are a low-cost product, their use as prophylaxis to reduce SARS-CoV-2 viral transmission warrants further evaluation. Clinical Trial Registration: https://www.clinicaltrials.gov/ct2/show/NCT04567810, identifier NCT04567810.

Affordable IgY-based antiviral prophylaxis for resource-limited settings to address epidemic and pandemic risks.

Background: The COVID-19 pandemic caused by SARS-CoV-2 exposed a global problem, as highly effective vaccines are challenging to produce and distribute, particularly in regions with limited resources and funding. As an alternative, immunoglobulins produced in eggs of immunized hens (IgY) can be a simple and inexpensive source for a topical and temporary prophylaxis. Here, we developed a method to extract and purify IgY antibodies from egg yolks of hens immunized against viral pathogen-derived proteins using low-cost, readily available materials, for use in resource-limited settings. Methods: Existing protocols for IgY purification and equipment were modified, including extraction from yolks and separation of water-soluble IgY using common household reagents and tools. A replacement for a commercial centrifuge was developed, using a home food processor equipped with a 3D printed adapter to enable IgY precipitation. IgY purification was verified using standard gel electrophoresis and Western blot analyses. Results: We developed a step-by-step protocol for IgY purification for two settings in low- and middle-income countries (LMIC): a local laboratory, where commercial centrifuges are available, or a more rural setting, where an alternative for expensive centrifuges can be used. Gel electrophoresis and Western blot analyses confirmed that the method produced highly enriched IgY preparation; each commercial egg produced ~ 90 mg of IgY. We also designed a kit for IgY production in these two settings and provided a cost estimate of the kit. Conclusion: IgY purified from eggs of immunized local hens can offer a fast and affordable prophylaxis, provided that purification can be performed in a resource-limited setting. Here, we created a low-cost method that can be used anywhere where electricity is available using inexpensive, readily available materials in place of costly, specialized laboratory equipment and chemicals. This procedure can readily be used now to make an anti-SARS-CoV-2 prophylaxis in areas where vaccines are unavailable, and can be modified to combat future threats from viral epidemics and pandemics.

Plasma glutamine changes after high-intensity exercise in elite male swimmers.

The aim of this study was to establish the pattern and time course of plasma glutamine recovery after acute, high-intensity exercise in well-trained swimmers. In Study 1, elite male swimmers (n=8) performed 15 x 100 m swimming intervals (ITS) at 70% and 95% of maximal 100m freestyle time. Resting plasma glutaminle levels were determined on a nonexercise control day (0% ITS). Venous blood samples were obtained prior to, immediately afte;, and 30, 60, 120, and 150 mini postexercise. In Study 2, the 95% ITS was repeated in elite male swuimmers (n=8), while control subjects (n=8) did not exercise, to test for any diurnal variation in plasma glutamine levels. Venous blood samples were obtained prior to and 2, 4, 6, and 8 h postexercise. In Study 1, no change was observed in plasma glutamine following the 0% (control) and 70% ITS, but following the 95% ITS glutamine decreased significantly (p < 0.01) over the recovery period. In Study 2, plasma glutamine again decreased over the recovery period in the swimmers, but no changes were observed in the controls. It was concluded that intensive swim traininlg results in postexercise decreases in plasma glutamine levels. Because glutamine has been suggested as a marker of overtraining, a need to measure glutaminle at standard times within training programs is indicated.

Plasma glutamine responses to high-intensity exercise before and after endurance training.

Glutamine responses to strenuous interval exercise were examined before and after 6 weeks of endurance training. Glutamine measures were obtained before and after the interval exercise sessions and training in untrained males assigned to training (T; n = 10) or control (C; n = 10) groups. Before training, C and T group glutamine progressively decreased (p < 0.05) by 18% and 16%, respectively, by 150-min postinterval exercise. Over the training period C group glutamine did not change, while T group values increased (p < 0.05) by 14%. After training, glutamine again decreased (p < 0.05) by similar percentages (C = 16% and T = 15%) by 150-min postinterval exercise, but the T group recorded higher (p < 0.05) resting and postexercise glutamine concentrations than the C group. Training induced increases in glutamine may prevent the decline in glutamine levels following strenuous exercise falling below a threshold where immune function might be acutely compromised.