GLT-1-Dependent Disruption of CNS Glutamate Homeostasis and Neuronal Function by the Protozoan Parasite Toxoplasma gondii
Fig 4
Ceftriaxone does not alter the immune response to Toxoplasma infection.
Treatment of infected mice with ceftriaxone was conducted for one or three weeks starting at 5 weeks post-infection and compared to naïve and untreated infected mice. A) Parasite burden was quantified by RT-PCR of forebrain DNA extracted at one week post treatment (6 weeks post infection) from infected (n = 4) and ceftriaxone (n = 5) treated animals (Student’s t-test: infected vs. ceftriaxone p = 0.2730) and at three weeks post treatment (8 weeks post infection)(Student’s t-test: infected (n = 5) vs. ceftriaxone (n = 4) p = 0.2902). B) Brain mononuclear cells (BMNC) were extracted following one week of treatment from infected (n = 4) and ceftriaxone (n = 5) treated brains and counted (Student’s t-test: infected vs. ceftriaxone p = 0.0826) and after 3 weeks of ceftriaxone treatment (Student’s t-test: infected (n = 5) vs. ceftriaxone (n = 4) p = 0.7794). C) Hematoxylin and eosin staining was performed on 3 week treated mice and images of the frontal cortex (10X) and blood vessels (25X) within the frontal cortex were taken. D) Flow cytometry of BMNC reveals the lymphocyte (CD45+CD11b-), macrophage (CD45+CD11bhi) and microglial (CD45intCD11bint) populations following 3 weeks of treatment with ceftriaxone. E) Quantification of the proportions of BMNC between infected (n = 5) and ceftriaxone (n = 4) treated mice for microglia (p = 0.7804), macrophages (p = 0.5013) and lymphocytes (p = 0.4164). F) Flow cytometry of BMNC for CD4 and CD8 (Student’s t-test: CD4 infected vs. ceftriaxone p = 0.2075; CD8 infected vs. ceftriaxone p = 0.4122). G) Immunohistochemistry for the microglial marker Iba-1 and the astrocytic marker GFAP was performed on 12μm frozen sections from the frontal cortex (scale bar: 80μm); inserts, high magnification images of cell morphology (scale bar: 5μm).