A reporter mouse for non-invasive detection of toll-like receptor ligands induced acute phase responses.

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.

Latent tuberculosis among injection drug users in a methadone maintenance treatment program, Taipei, Taiwan: TSPOT.TB versus tuberculin skin test.

BACKGROUND: Taiwan has a growing HIV/AIDS epidemic that has recently shifted to an increase among injection drug users (IDUs). IDUs co-infected with HIV and tuberculosis (TB) have a high risk of progression from latent tuberculosis infection (LTBI) to active TB. METHODS: This study aimed to determine the prevalence and correlates of LTBI among IDUs by TSPOT.TB and tuberculin skin test (TST), in a large methadone program in Taipei, Taiwan. Consenting participants were interviewed by a trained worker regarding sociodemographics, substance use history, and health factors. RESULTS: Multivariate analysis was used to determine risks associated with each test outcome. Of 287 participants, 165 (58.7%) tested TSPOT.TB-positive and 244 (85.0%) tested TST-positive. The mean age was 44 y, and 7.3% were HIV-infected. Kappa statistics indicated slight concordance between TSPOT.TB and TST. In multivariate analysis, after controlling for potential confounders, TSPOT.TB positivity was significantly associated with age ≥ 50 y (reference, 20-34 y). A history of ever having had contact with a TB-infected person was associated with TST positivity, whereas HIV infection was inversely associated with TSPOT.TB positivity and TST positivity. CONCLUSIONS: This study shows a high prevalence of LTBI in individuals at risk for HIV infection in Taipei, Taiwan. Future TB prevention programs should particularly focus on IDUs.

Coat color-tagged green mouse with EGFP expressed from the RNA polymerase II promoter.

Laborious molecular genotyping and variegated gene expression are two widely encountered issues for transgenic mouse studies. To facilitate genotyping in the FVB/N albino background and to reduce variegated expression, we successfully generated double-tagged transgenic mice for direct visual genotyping with the coat color phenotype derived from tyrosinase cDNA driven by the tyrosinase promoter and with simultaneous high enhanced green fluorescent protein (EGFP) expression driven by the promoter of RNA polymerase II large subunit gene. Incorporation of insulator into a transgene construct achieved high efficiency of transgene expression in more than 90% of the founders. EGFP was detected as early as the one-cell fertilized egg and lasted for the whole embryo development, as well as in all of the adult tissues examined. The coat color-tagged green mice offer opportunities in applications such as tissue transplantation, lineage tracing, chimera biology, RNA interference, and other transgenic studies.