The acute phase response (APR) is a systemic first-line defense against challenges including infection, trauma, stress, and neoplasia. Alteration of acute phase protein (APP) levels in plasma is the most important change during acute phase response. C-reactive protein (CRP), which increases dramatically during inflammation onset, is an indicator of inflammation. To monitor the process of APR, we generated human CRP promoter-driven luciferase transgenic (hCRP-Luc) mice to quantify the hCRP promoter activation in vivo. The naïve female hCRP-Luc mice express low basal levels of liver bioluminescence, but the naïve male hCRP-Luc mice do not. Thus, female hCRP-Luc mice are suitable for monitoring the process of APR. The liver bioluminescence of female hCRP-Luc mice can be induced by several toll-like receptor (TLR) ligands. The expression of liver bioluminescence was highly sensitive to endotoxin stimulation in a dose-dependent manner. On-off-on bioluminescence response was noted in female hCRP-Luc mice upon two endotoxin stimulations one month apart. The LPS-induced bioluminescence of the female hCRP-Luc mice was IL-6-mediated and associated with APP alpha-1-acid glycoprotein expression. In conclusion, the female hCRP-Luc mouse is a non-invasive, sensitive and reusable reporter tool for APR.

Determining the onset of diabetes based on blood glucose (BG) levels can be challenging in mouse models, as thresholds can vary from 200 to 400 mg/dl in one or two consecutive tests. Urine glucose (UG) levels can be detected non-invasively as another criterion of diabetic condition, but it is considered a lagging indicator due to physiological downstream from BG. In this study, we demonstrate that the lagging period is practically unnoticeable in spontaneously model of non-obese diabetic (NOD) mice which develop autoimmune diabetes randomly from 12 to 32 weeks of age. After comprehensive measurements across entire onset window in 60 female NOD mice, we concluded that BG measurements before UG reaches 250 mg/l contribute nearly nothing to diabetic identification. Refined protocol encompasses UG survey twice-weekly to select positive candidates for further intensive BG measurements is recommended and tested in another batch of 60 mice. This protocol precisely identified every newly onset individual with average BG of 350 mg/dl which is lower than conventional once-weekly BG survey alone around 400 mg/dl. Moreover, intensive measurements near onset indicate two BG+ within four days can serve as a refined onset criterion, allowing for dynamic arrangement of sampling time to make the process even more efficient in practice. From 3R perspectives, this protocol potentially saves dozens of bleeding procedures in one individual mouse and hundreds of lancets, BG strips, labors, and unnecessary animal suffering during batch screening that serve as a convenient alternative for newly onset identification of diabetes.

Triple-negative breast cancer (TNBC) is one of the most fatal subtype of breast cancer. Tumor immune microenvironments (TIMEs) have critical influences on TNBC therapies. However, syngeneic mouse tumor models are insufficient to evaluate immunotherapeutic efficacy, because the different immune responses between mice and human. Therefore, human xenograft tumor mouse models with humanized immune systems (HIS) are more suitable for evaluation of immunotherapeutic efficacy. In these study, two immune humanized mouse models transferred with human hematopoietic stem cells (hHSC-HIS) or human peripheral mononuclear cells (hPBMC-HIS) in our advanced severe immune deficiency (ASID) mice were generated. Both systemic immune profiling and TIMEs of TNBC MDA-MB-231 tumors from hHSC- and hPBMC-HIS ASID models were compared. Generation of the comprehensive human immune system and large amounts of human macrophages/bone marrow-derived stromal cells around the TIMEs were noted in hHSC-HIS ASID mice, but the growth of TNBC was not affected. In contrast, T cells dominated the systemic immune system and TIMEs in hPBMC-HIS ASID mice, and inhibited the tumor growth. In summary, the TIMEs of hHSC-HIS ASID model is suitable for validations of immunotherapies reversing immune suppression. The TIMEs of hPBMC-HIS ASID can be applied for cytotoxicity test of tumor specific T cells.