Evaluation of Various IVC Systems According to Mouse Reproductive Performance and Husbandry and Environmental Parameters.

IVC systems are marketed for improving the health and management of mouse colonies. The current study compared mouse reproductive performance and husbandry and environmental parameters among 3 high-density (HD) IVC rack systems (RS1, RS2, and RS3), which were present in separate but comparable rooms. Three breeding trios each of Swiss Webster (CFW) and BALB/c mice were placed in each rack (n = 36 female, n = 18 male). Reproductive indices were measured for 3 breeding cycles over 2 generations; indices included time to parturition, litter size and pup weight, survivability, and interbirth interval. Over 18 wk, personnel used scoring systems to evaluate each RS daily to every other week according to cage dirtiness, need for spot changing, ease of cage changing, daily health checks, and cage wash processing. Macroenvironmental parameters (temperature, relative humidity, noise, total particulate matter) were measured weekly over 14 wks. Microenvironmental parameters (temperature, relative humidity, NH3, CO2, O2) of 2 cages each of male and female CFW mice (4 mice/cage) on each RS were measured at 6 time points over 2 wks. RS1 had significantly smaller mean litter sizes of CFW mice (mean ± 1 SD, 6.5 ± 2.9 pups) as compared with both RS2 (9.5 ± 1.7 pups) and RS3 (9.3 ± 3.8 pups). RS1 scored as being significantly easier to process through the cage wash. RS2 had significantly lower room noise levels (46.0 ± 5.0 dBA) but higher humidity (58.6% ± 8.9%) as compared with both RS1 (43.7% ± 9.9%) and RS3 (46.0% ± 12.0%) over the 2-wk cycle, particularly at 8 and 12 d after cage change. In conclusion, in terms of mouse reproductive performance and husbandry and environmental parameters, each system had at least 1 advantage over the other 2. Therefore, various factors should be considered when choosing an IVC system for mice.

Pharmacokinetics and biodistribution of extracellular vesicles administered intravenously and intranasally to Macaca nemestrina.

Extracellular vesicles (EVs) have potential in disease treatment since they can be loaded with therapeutic molecules and engineered for retention by specific tissues. However, questions remain on optimal dosing, administration, and pharmacokinetics. Previous studies have addressed biodistribution and pharmacokinetics in rodents, but little evidence is available for larger animals. Here, we investigated the pharmacokinetics and biodistribution of Expi293F-derived EVs labelled with a highly sensitive nanoluciferase reporter (palmGRET) in a non-human primate model (Macaca nemestrina), comparing intravenous (IV) and intranasal (IN) administration over a 125-fold dose range. We report that EVs administered IV had longer circulation times in plasma than previously reported in mice and were detectable in cerebrospinal fluid (CSF) after 30-60 minutes. EV association with PBMCs, especially B-cells, was observed as early as one minute post-administration. EVs were detected in liver and spleen within one hour of IV administration. However, IN delivery was minimal, suggesting that pretreatment approaches may be needed in large animals. Furthermore, EV circulation times strongly decreased after repeated IV administration, possibly due to immune responses and with clear implications for xenogeneic EV-based therapeutics. We hope that our findings from this baseline study in macaques will help to inform future research and therapeutic development of EVs.

Sex Differences in Lung Imaging and SARS-CoV-2 Antibody Responses in a COVID-19 Golden Syrian Hamster Model.

In the coronavirus disease 2019 (COVID-19) pandemic caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), more severe outcomes are reported in males than in females, including hospitalizations and deaths. Animal models can provide an opportunity to mechanistically interrogate causes of sex differences in the pathogenesis of SARS-CoV-2. Adult male and female golden Syrian hamsters (8 to 10 weeks of age) were inoculated intranasally with 105 50% tissue culture infective dose (TCID50) of SARS-CoV-2/USA-WA1/2020 and euthanized at several time points during the acute (i.e., virus actively replicating) and recovery (i.e., after the infectious virus has been cleared) phases of infection. There was no mortality, but infected male hamsters experienced greater morbidity, losing a greater percentage of body mass, developed more extensive pneumonia as noted on chest computed tomography, and recovered more slowly than females. Treatment of male hamsters with estradiol did not alter pulmonary damage. Virus titers in respiratory tissues, including nasal turbinates, trachea, and lungs, and pulmonary cytokine concentrations, including interferon-ß (IFN-ß) and tumor necrosis factor-α (TNF-α), were comparable between the sexes. However, during the recovery phase of infection, females mounted 2-fold greater IgM, IgG, and IgA responses against the receptor-binding domain of the spike protein (S-RBD) in both plasma and respiratory tissues. Female hamsters also had significantly greater IgG antibodies against whole-inactivated SARS-CoV-2 and mutant S-RBDs as well as virus-neutralizing antibodies in plasma. The development of an animal model to study COVID-19 sex differences will allow for a greater mechanistic understanding of the SARS-CoV-2-associated sex differences seen in the human population. IMPORTANCE Men experience more severe outcomes from coronavirus disease 2019 (COVID-19) than women. Golden Syrian hamsters were used to explore sex differences in the pathogenesis of a human isolate of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). After inoculation, male hamsters experienced greater sickness, developed more severe lung pathology, and recovered more slowly than females. Sex differences in disease could not be reversed by estradiol treatment in males and were not explained by either virus replication kinetics or the concentrations of inflammatory cytokines in the lungs. During the recovery period, antiviral antibody responses in the respiratory tract and plasma, including to newly emerging SARS-CoV-2 variants, were greater in female than in male hamsters. Greater lung pathology during the acute phase combined with lower antiviral antibody responses during the recovery phase of infection in males than in females illustrate the utility of golden Syrian hamsters as a model to explore sex differences in the pathogenesis of SARS-CoV-2 and vaccine-induced immunity and protection.