Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Complement C1q and C3 are critical for the innate immune response to Streptococcus pneumoniae in the central nervous system.

doi: 10.4049/jimmunol.178.3.1861

Figure Lengend Snippet: FIGURE 3. Effect of C1q and C3 deficiency on cytokine mRNA ex- pression within the brain following intracisternal pneumococcal infection. A mouse inflammatory cytokine and receptor oligonucleotide array was used to screen for differences in the mRNA expression of cytokines, che- mokines, and other inflammatory factors from PBS-injected controls (a and b), C3-deficient controls (c), pneumococci-infected wt mice (d and e), and infected C3-deficient mice (f and g). Equal amounts of total RNA were pooled from eight mice in each experimental group and analyzed twice (a plus b, d plus e, f plus g) (except for the C3-deficient control group which had four mice and was analyzed once; c). For analysis, the expression signal of each gene probe was normalized to the signal of the internal GADPH standard on the same membrane and expressed as percentage expression of GADPH. Gene probes were excluded from analysis if their relative expression levels (a) differed by more than 2-fold within a group (see E for representative examples) or (b) was 5% of the internal GAPDH standard. Fourteen gene probes were discarded from further anal- ysis due to 2-fold differences in expression levels within a group. Among the remaining 98 gene probes, no differences were found in the relative expression levels between wt and C3-deficient control brains. Pneumococ- cal infection was associated with an induction or up-regulation of 15 in- flammatory cytokines and receptors, including the cytokines IL-1 and IL-6, the chemokines CXCL1, CXCL2, CXCL10, and CCL2, the TLR receptor TLR7, as well as the complement component C3 (see rectangles). The expression level of all these meningitis-induced immune factors was blunted in brains from infected C3-deficient mice.

Article Snippet: Mice brains were screened for 62 cytokines/chemokines using a mousespecific Cytokine Ab Array (Array 3.1; Ray Biotech).

Techniques: Infection, Expressing, Injection, Control, Membrane

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Complement C1q and C3 are critical for the innate immune response to Streptococcus pneumoniae in the central nervous system.

doi: 10.4049/jimmunol.178.3.1861

Figure Lengend Snippet: FIGURE 4. Effect of C1q and C3 deficiency on the cytokine protein expression profile within the brain following intracisternal pneumococcal infection. Mouse cytokine Ab arrays were used to determine the differences in the cytokine protein expression in brains from (a) PBS-injected controls, (b) pneumococci-infected wt mice, (c) infected C1q-deficient mice, (d) C3-deficient mice, and (e) C3-deficient control mice. The brain cytokine expression pattern did not differ between wt and C3-deficient controls (a; e). Compared with the cytokine protein expression in PBS-injected control brains (a), 14 of 62 cytokines were induced or up-regulated in brains of infected wt mice (b) (see rectangles). Among them were the cytokines IL-1, IL-1, IL-6, IL-12p40/p70, and IL-12p70, the cytokine receptor TNFRI, the CXC chemokines KC, MIP-2, and CXCL16, the CC chemokines CCL2, CCL12, and MIP-1, the hemopoietin G-CSF, and the adhesion molecule L-selectin. With the exceptions of KC, CCL12, and G-CSF, the expression level of all these cytokines was 2-fold lower in brain homogenates from infected C3-deficient mice than in brain homogenates of infected wt mice (d). The differences in cytokine expression between infected C1q-deficient mice (c) and infected wt mice (b) were all below the 2-fold difference level. The four lower bar charts present the results of the densitometric analysis of the brain protein expression of the immune factors IL-1 (f), IL-12p79 (g), L-selectin (h), and MIP-1 (i) which were also measured by ELISA (see Fig. 5).

Article Snippet: Mice brains were screened for 62 cytokines/chemokines using a mousespecific Cytokine Ab Array (Array 3.1; Ray Biotech).

Techniques: Expressing, Infection, Injection, Control, Enzyme-linked Immunosorbent Assay

Journal: Journal of immunology (Baltimore, Md. : 1950)

Article Title: Complement C1q and C3 are critical for the innate immune response to Streptococcus pneumoniae in the central nervous system.

doi: 10.4049/jimmunol.178.3.1861

Figure Lengend Snippet: FIGURE 6. Effect of C1q and C3 deficiency on the cytokine protein expression profile in blood. Mouse cytokine Ab arrays were used to determine the differences in the cytokine protein expression in blood samples from (a) PBS-injected controls, (b) pneumococci-infected wt mice, (c) infected C1q-deficient mice, and (d) C3-deficient mice. Twenty-four hours after intracisternal pneumococcal infection, 7 of 62 cytokines were induced or up-regulated in blood of infected wt mice (b), compared with that of PBS-injected control mice (a). These factors include IL-6, IL-12p40/p70, IL-12p40, CXCL9, CCL2, CCL12, and G-CSF (see rectangles). The expression levels of IL-6, IFN-, CXCL9, CXCL13, and CCL12 were 2-fold higher in blood from infected C3-deficient mice (d) than wt mice (b), whereas the other cytokines were equally expressed in both mice strains. Infected C1q-deficient mice (c) also showed a substantial increase in blood levels of IL-6, IFN-, and CXCL13. The lower bar charts present the results of the densitometric analysis of the blood protein expression of four representative immune factors, namely G-CSF (e), IFN- (f), IL-6 (g), and CCL12 (h).

Article Snippet: Mice brains were screened for 62 cytokines/chemokines using a mousespecific Cytokine Ab Array (Array 3.1; Ray Biotech).

Techniques: Expressing, Injection, Infection, Control