Journal: International Journal of Molecular Sciences

Article Title: H-Ferritin Produced by Myeloid Cells Is Released to the Circulation and Plays a Major Role in Liver Iron Distribution during Infection

doi: 10.3390/ijms23010269

Figure Lengend Snippet: Evolution of infection by Mycobacterium avium in mice deficient in H-ferritin in myeloid cells. Fth1 +/+ (blue) and Fth1 −/− (red) mice were intravenously infected with 10 6 CFU of M. avium 25291. ( A ) Animals’ body weight throughout the experiment. Data are expressed as percentage of the initial weight, presented as the mean ± SD of 5 to 8 animals per group. Empty circles: noninfected; filled circles: infected. ( B ) Bacterial burden 60 days post-infection in the liver, spleen, and bone marrow of 8 Fth1 +/+ and 7 Fth1 −/− mice. Bars represent the mean, and circles represent each animal. ( C ) Kaplan–Meier survival curve corresponding to 12 Fth1 +/+ and 16 Fth1 −/− animals. Mice were euthanized when they lost 20% of the initial weight (between day 43 and day 152 post infection). ( D ) Bacterial burden in the liver, spleen, and bone marrow, at the time of euthanasia in the survival curve, of 12 Fth1 +/+ and 16 Fth1 −/− mice. Bars represent the mean and circles represent each animal. Statistics: multiple t -test in ( A , B , D ); log-rank (Mantel–Cox) test in ( C ). ** p < 0.01 and *** p < 0.001 when comparing Fth1 −/− with Fth1 +/+ mice.

Article Snippet: FTH1 and FTL were quantified by ELISA in the mice serum samples, using rabbit polyclonal anti-mouse FTH1 or FTL (DIBIT-IRCCS San Raffaele Scientific Institute, Milan, Italy) as primary antibodies, and biotinylated rabbit polyclonal anti-mouse FTH1 or FTL as the secondary antibodies.

Techniques: Infection

Journal: International Journal of Molecular Sciences

Article Title: H-Ferritin Produced by Myeloid Cells Is Released to the Circulation and Plays a Major Role in Liver Iron Distribution during Infection

doi: 10.3390/ijms23010269

Figure Lengend Snippet: Inflammatory response in mice infected with M. avium . Fth1 +/+ (blue) and Fth1 −/− (red) mice were intravenously infected with 10 6 CFU of M. avium 25291 and euthanized 60 days later. ( A ) TNF-alpha, ( B ) IFN-gamma, and ( C ) IL-6 were measured in the serum, using a cytometric bead array kit. Bars represent the mean, and the circles represent individual values of 4 to 8 animals per group. Statistics: two-way ANOVA followed by Sidak multiple-comparison post hoc test. * p < 0.05, ** p < 0.01, *** p < 0.001 when comparing Fth1 +/+ to Fth1 −/− mice. ## p < 0.01, ### p < 0.001 when comparing infected to noninfected mice of the same genotype. ( D – G ) Representative images of H&E-stained liver sections from noninfected (NI) and infected (60 dpi) Fth1 +/+ and Fth1 −/− mice. Representative granulomas are outlined. Scale bar: 200 µm.

Article Snippet: FTH1 and FTL were quantified by ELISA in the mice serum samples, using rabbit polyclonal anti-mouse FTH1 or FTL (DIBIT-IRCCS San Raffaele Scientific Institute, Milan, Italy) as primary antibodies, and biotinylated rabbit polyclonal anti-mouse FTH1 or FTL as the secondary antibodies.

Techniques: Infection, Comparison, Staining

Journal: International Journal of Molecular Sciences

Article Title: H-Ferritin Produced by Myeloid Cells Is Released to the Circulation and Plays a Major Role in Liver Iron Distribution during Infection

doi: 10.3390/ijms23010269

Figure Lengend Snippet: Impact of infection on iron distribution. Sera from Fth1 +/+ (blue) and Fth1 −/− (red) mice infected for 60 days with M. avium 25291 (60 dpi) or from noninfected mice (NI), were used to measure ( A ) iron, ( B ) transferrin saturation, and ( C ) total ferritin ( n = 5 to 8). ( D , E ) Serum FTL and FTH1 subunits were measured by ELISA ( n = 2 to 4). ( F ) Liver sections were collected, and the iron concentration was determined by atomic absorption spectrometry ( n = 5 to 8). Bars represent the mean, and circles represent each animal. Statistics: two-way ANOVA followed by Sidak multiple-comparison post hoc test. ** p < 0.01, *** p < 0.001 when comparing Fth1 −/− with Fth1 +/+ mice in the same experimental condition. # p < 0.05, ## p < 0.01, ### p < 0.001 when comparing infected to noninfected mice of the same genotype. ( G – J ) Perls’ staining of liver sections was used to evaluate iron distribution. Arrows indicate regions of iron accumulation (blue staining). Scale bar = 200 µm.

Article Snippet: FTH1 and FTL were quantified by ELISA in the mice serum samples, using rabbit polyclonal anti-mouse FTH1 or FTL (DIBIT-IRCCS San Raffaele Scientific Institute, Milan, Italy) as primary antibodies, and biotinylated rabbit polyclonal anti-mouse FTH1 or FTL as the secondary antibodies.

Techniques: Infection, Enzyme-linked Immunosorbent Assay, Concentration Assay, Comparison, Staining

Journal: International Journal of Molecular Sciences

Article Title: H-Ferritin Produced by Myeloid Cells Is Released to the Circulation and Plays a Major Role in Liver Iron Distribution during Infection

doi: 10.3390/ijms23010269

Figure Lengend Snippet: Effect of iron overload on the growth of M. avium in the liver. Groups of Fth1 +/+ (blue) and Fth1 −/− (red) mice were intraperitoneally injected with 10 mg of iron, in the form of iron–dextran, 10 days before intravenous infection with 10 6 CFU of M. avium 25291. Sixty days after infection, noninfected (NI) and infected (60 dpi) mice were euthanized. ( A ) Liver iron was quantified by atomic absorption spectrometry ( n = 4 to 8 per group). ( B ) Bacterial burden in the livers, 60 days after infection ( n = 6 to 8 per group). The bars represent the mean of the group, and each circle corresponds to one animal. Statistics: two-way ANOVA followed by Sidak multiple-comparison post hoc test. *** p < 0.001 when comparing Fth1 −/− to Fth1 +/+ mice in the same experimental condition. @ p < 0.001 when comparing iron-overloaded with non-iron-overloaded mice of the same genotype and infection status.

Article Snippet: FTH1 and FTL were quantified by ELISA in the mice serum samples, using rabbit polyclonal anti-mouse FTH1 or FTL (DIBIT-IRCCS San Raffaele Scientific Institute, Milan, Italy) as primary antibodies, and biotinylated rabbit polyclonal anti-mouse FTH1 or FTL as the secondary antibodies.

Techniques: Injection, Infection, Comparison

Journal: International Journal of Molecular Sciences

Article Title: H-Ferritin Produced by Myeloid Cells Is Released to the Circulation and Plays a Major Role in Liver Iron Distribution during Infection

doi: 10.3390/ijms23010269

Figure Lengend Snippet: Iron-related gene expression induced by mycobacterial infection.

Article Snippet: FTH1 and FTL were quantified by ELISA in the mice serum samples, using rabbit polyclonal anti-mouse FTH1 or FTL (DIBIT-IRCCS San Raffaele Scientific Institute, Milan, Italy) as primary antibodies, and biotinylated rabbit polyclonal anti-mouse FTH1 or FTL as the secondary antibodies.

Techniques: Gene Expression, Infection

Journal: International Journal of Molecular Sciences

Article Title: H-Ferritin Produced by Myeloid Cells Is Released to the Circulation and Plays a Major Role in Liver Iron Distribution during Infection

doi: 10.3390/ijms23010269

Figure Lengend Snippet: Effects of infection and myeloid Fth1 deficiency in iron and ferroportin distribution in the liver. Liver sections were obtained from ( A , B ) noninfected mice or ( C – H ) mice infected with M. avium for 60 days. Sections were stained for ( A – D , G , H ) ferroportin, using immunohistochemistry (brown staining) or for ( E , F ) iron, using the Perls’ method (blue staining). ( E – H ) Mice were intraperitoneally injected with 10 mg of iron, as iron–dextran, 10 days before infection. Representative images of at least three animals per experimental condition. Representative granulomas are outlined. Scale bar: 200 µm.

Article Snippet: FTH1 and FTL were quantified by ELISA in the mice serum samples, using rabbit polyclonal anti-mouse FTH1 or FTL (DIBIT-IRCCS San Raffaele Scientific Institute, Milan, Italy) as primary antibodies, and biotinylated rabbit polyclonal anti-mouse FTH1 or FTL as the secondary antibodies.

Techniques: Infection, Staining, Immunohistochemistry, Injection

Journal: International Journal of Molecular Sciences

Article Title: H-Ferritin Produced by Myeloid Cells Is Released to the Circulation and Plays a Major Role in Liver Iron Distribution during Infection

doi: 10.3390/ijms23010269

Figure Lengend Snippet: Primers sequences.

Article Snippet: FTH1 and FTL were quantified by ELISA in the mice serum samples, using rabbit polyclonal anti-mouse FTH1 or FTL (DIBIT-IRCCS San Raffaele Scientific Institute, Milan, Italy) as primary antibodies, and biotinylated rabbit polyclonal anti-mouse FTH1 or FTL as the secondary antibodies.

Techniques: Sequencing