9–10-week-old female BALB/cAnNHsd mice were purchased from Envigo. Eight mice per group were intranasally challenged with 105 TCID50 of SARS-CoV-2 (MA10 strain) and 24 hours p.i. administered through the intraperitoneal route with 100 mg/kg of the 54043-5 antibodies (wt or LALA-PG version) or with 200 mg/kg of an isotype control (DENV-2D22) or 10 mg/kg of a SARS-CoV-2 human neutralizing antibody (COV2–2381). Weight, clinical signs, and survival were monitored for 5 days. Four mice per group were sacrificed at day 2 and 5 p.i. and lungs collected for determining viral titers, cytokine analysis and histopathology.
Balb cannhsd mice
Manufactured by Inotiv
BALB/cAnNHsd mice are a commonly used inbred mouse strain. They exhibit a white coat color and are known for their susceptibility to certain diseases and immune responses. This strain is widely used in biomedical research, but a detailed description of their core function is not available while maintaining an unbiased and factual approach.
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Lab products found in correlation
Low ph igg elution buffer, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
A20 cells, by American Type Culture Collection (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
L glutamine, by Thermo Fisher Scientific (1 mentions)
C57bl 6j mice, by Jackson ImmunoResearch (1 mentions)
B6 cg tg k18 ace2 2prlmn j, by Jackson ImmunoResearch (1 mentions)
Balb c, by Inotiv (1 mentions)
9 protocols using balb cannhsd mice
1
SARS-CoV-2 Challenge and Antibody Treatment
2
Murine Model Characterization for Immunology
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Male 10-12-week-old mice were obtained from The Jackson Laboratory. Rag2−/− mice on a C57BL/6 background have a disruption of the recombination activating gene 2 (Rag2) and fail to produce mature T or B lymphocytes [19 (link)]. Mice were housed in the animal facility at Tulane University School of Medicine. The Institutional Animal Care and Use Committee of Tulane University reviewed and approved all procedures for sample handling, inactivation, and removal from a BSL3 containment (permit number 4267). One-year-old female BALB/cAnNHsd mice (strain 047) (herein referred to as “BALB/c” female mice throughout the manuscript) were obtained from Envigo and were housed at the University of North Carolina at Chapel Hill.
3
Mammary Gland Development in BALB/c Mice
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BALB/cAnNHsd mice were purchased from Envigo at either 5- (pubertal) or 10-(adult) weeks old for mammary gland development studies or 6–8 weeks old for tumor transplantations. All experiments were performed using female mice housed in specific pathogen-free facilities. All animal care and procedures were approved by the Institutional Animal Care and Use Committee of the University of Minnesota and were in accordance with the procedures detailed in the Guide for the Care and Use of Laboratory Animals [38 ].
4
H7N9 Influenza Protection in Mice
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BALB/cAnNHsd mice (Envigo) were given a 0.1 mg of FI6v3 (~5 mg kg−1) or 0.2 mg of purified H7-specific or control naïve Igs (~10 mg kg−1) intraperitoneally (N = 10). Twenty-four hours later, the mice were infected intranasally with 10 × LD50 of H7N9 A/Anhui/01/2013 (H7N9) at Bioqual. The animals were monitored twice daily for development of clinical signs and weighed daily for 14 days. Any animals that had lost 20% or more of their initial body weight were euthanized.
5
Passive Transfer of Hyper-immune Ig Against Influenza
6
Passive Transfer of Hyper-immune Ig Against Influenza
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To generate hyper-immune Ig for passive transfer, the immune serum samples from each NHP were diluted 1:50 with PBS, added to protein A columns, and incubated overnight at 4°C. After washing the columns briefly, captured antibodies were eluted with low-pH IgG elution buffer (ThermoFisher Scientific) and the eluates were immediately neutralized by adding 1 M Tris-HCl (pH 8.0) to a final concentration of 100 mM. Purified polyclonal antibodies were dialyzed two times against PBS, concentrated to ~20 mg ml−1 and stored at −80°C until use. BALB/cAnNHsd mice (Envigo) were given intraperitoneally 0.2 mg of FI6v3 (approximately 10 mg kg−1) or 10 mg of purified polyclonal Ig from individual NHPs. Twenty-four hours later, the mice were infected intranasally with 25× or 10× LD50 of H5N1 or H7N9 viruses (Supplementary Table 4 ) at Bioqual. The animals were monitored twice daily for development of clinical signs of infection and weighed daily for 14 days. Any animals that lost 20% or more of their initial body weight were euthanized.
7
In Vivo Tumor Microinjection Profiling
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A20 cells (ATCC) were cultured in RPMI 1640 with l -Glutamine (Thermo Fisher Scientific), 10% fetal bovine serum (Thermo Fisher Scientific), and 50 nM β-Mercaptoethanol at 37 °C and 5% CO2. All experiments in mice were approved by IACUC Board of Presage Biosciences, Seattle, WA (Protocol number PR-001) and were performed at Presage in accordance with relevant guidelines and regulations. For generating A20 allografts, female BALB/cAnNHsd mice (Envigo) were inoculated with 1 × 106 A20 cells. CIVO IT microinjections were performed as described previously28 (link). Briefly, mice (n = 6 per time point, 4 and 24 h) were enrolled in microinjection studies when implanted tumors reached the following approximate dimensions: 14 mm (length), 10 mm (width) and 7 mm (depth). The CIVO device was configured with 6 thirty-gauge injection needles with a total volume delivery of 2 μL. Presage’s fluorescent tracking marker (FTM, 5% by volume) was added to the injection contents for spatial orientation. At 4 and 24 h following CIVO microinjections, mice were euthanized using CO2 inhalation for biomarker analyses.
8
SARS-CoV-2 Infection Model in Mice
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Twelve-week-old male B6.Cg-Tg(K18-ACE2)2Prlmn/J (strain number 034860, K18-hACE2; n = 28) and 14-week-old male C57BL/6J mice (strain no. 000664; n = 12) were obtained from The Jackson Laboratory (JAX, Bar Harbor, ME). Additionally, 10-week-old male BALB/cAnNHsd mice (strain number 047, BALB/c; n = 16) were obtained from Envigo (Indianapolis, IN). Mice were housed in groups of 3 to 4 per cage under a 12-h light/12-h dark cycle with ad libitum access to water and a standard chow diet.
9
Balb/c Mouse Husbandry and Necropsy
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Seven-to-eight-week-old female BALB/cAnNHsd mice (Envigo; Indianapolis, IN) were housed in clean rooms controlled to 70 ± 2°F and 30–70% humidity. Animals were fed irradiated Teklad 2918.15 Rodent Diet and water ad libitum.
Clinical observations were performed at least once daily, and body weights were recorded three times weekly. Animals found in distress or moribund condition were euthanized. Necropsies were performed and potential cause of death was assessed; target organs for toxicity were evaluated and presence/absence of metastases was noted.
Clinical observations were performed at least once daily, and body weights were recorded three times weekly. Animals found in distress or moribund condition were euthanized. Necropsies were performed and potential cause of death was assessed; target organs for toxicity were evaluated and presence/absence of metastases was noted.
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