Journal: PLOS Pathogens

Article Title: B cells promote granulomatous inflammation during chronic Mycobacterium tuberculosis infection in mice

doi: 10.1371/journal.ppat.1011187

Figure Lengend Snippet: Mice were treated with the anti-CD20 mAb 5D2 beginning 2 days prior to a low-dose (100 CFU) aerogenic challenge with M . tuberculosis Erdman, as described in "Materials and Methods". B cell depletion was maintained throughout the duration of the experiment. The control mouse group (WT) received non-specific rat IgG. The lung tissues were examined at 5 months post-infection. The levels of granulomatous inflammation response were analyzed histologically by light microscopy on H&E-stained lung sections (A) and enumeration of total number of lung cells (B). The level of lung CD4 + T cell response was assessed by in vivo BrdU labeling to examine the proliferation capacity of this T cell subset (C), as well as by enumeration of IFN-γ-producing CD4 + T cells (D). Ex vivo evaluation of lung cells for the level of IL-10 production (E) was conducted as described in . Four to 5 mice per group were evaluated per group. Data depicted in B, C, D, and E are presented as means ± SEM. The data shown are representative of two experiments. The results demonstrated that the inflammation, Th1 response, and IL-10 phenotypes observed in the μMT mice are recapitulated in mice depleted for B cells.

Article Snippet: At day 90 post infection, 1 mg of anti-mouse IL-10 receptor (IL-10R; clone 1B1.3A; BioXcell) or isotype control Rat IgG1 antibody (BioXcell) was administered intraperitoneally (i.p.) per mouse.

Techniques: Control, Infection, Light Microscopy, Staining, In Vivo, Labeling, Ex Vivo

Journal: PLOS Pathogens

Article Title: B cells promote granulomatous inflammation during chronic Mycobacterium tuberculosis infection in mice

doi: 10.1371/journal.ppat.1011187

Figure Lengend Snippet: C57BL/6 mice were depleted for B cells via administration of 5D2 beginning 2 days prior to infection with a low dose (100 CFU) of M . tuberculosis Erdman delivered by aerosol. B cell depletion was maintained for the duration of the experiment. At 3 months after the infection, IL-10R blockade was initiated using the anti-mouse IL-10R antibody clone 1B1.3A. The control group received non-specific rat IgG. The IL-10R blockade was continued for two months. At five months post-infection (2 months after initiation of IL-10R blockade), mice were sacrificed and analyzed for the levels of inflammation in the lungs, as assessed by histological examination (A) and enumeration of total lung cells (B), CD4 + T cells proliferation via BrdU labeling (C), and Th1 response (D). Data shown are representation of two experiments. Three to four mice were analyzed per group. Data depicted in (B), (C), and (D) denote mean ± SEM.

Article Snippet: At day 90 post infection, 1 mg of anti-mouse IL-10 receptor (IL-10R; clone 1B1.3A; BioXcell) or isotype control Rat IgG1 antibody (BioXcell) was administered intraperitoneally (i.p.) per mouse.

Techniques: Infection, Aerosol, Control, Labeling

Journal: eLife

Article Title: Discovery of surrogate agonists for visceral fat Treg cells that modulate metabolic indices in vivo

doi: 10.7554/eLife.58463

Figure Lengend Snippet:

Article Snippet: 12-week-old B6.CD45.1 + mice were fed with HFD and injected intraperitoneally with 10 μg/g body weight anti-TNFα (XT3.11, BioXCell) or isotype control IgG (HRPN, BioXCell) twice a week for 6 weeks, transferred with 0.4 million Treg cells from pooled Spl/LNs of 6-8wk-old male CD45.2 + Tg + Foxp3-GFP KI/y mice.

Techniques: Recombinant, In Vivo, Control, Staining, Enzyme-linked Immunosorbent Assay, Adjuvant, Software

Journal: NPJ Vaccines

Article Title: A candidate subunit vaccine induces protective immunity against Mycobacterium avium subspecies paratuberculosis in mice

doi: 10.1038/s41541-023-00675-1

Figure Lengend Snippet: a Schematic diagram of the vaccination timeline. C57BL/6 mice were subcutaneously injected two times (three weeks apart) with 50 μg 66NC fusion protein formulated in Montanide ISA 61 VG adjuvant. Control groups include mice immunized with Montanide ISA 61 VG adjuvant alone as a negative control or 74 F mixed MPL adjuvant as subunit vaccine control. Three weeks after the booster immunization, the mice were bled and sacrificed. b Antigen-specific IgG1 and IgG2a isotypes were measured by ELISA. c Pictures of the IFN-γ and IL-4 ELISpot assay (representative of one experiment with at least five independent replicates). d , e Quantitation of the IFN-γ and IL-4 ELISpot assay. f The percentage of cytokine-positive (%Cyt + ) antigen-specific T-lymphocytes was quantified by intracellular cytokine staining (ICS) in CD4 + and CD8 + following stimulation of splenic T lymphocytes with the indicated antigen. g Serum cytokines levels of IFN-γ, TNF-α, and IL-17A were measured by ELISA. d – g Error bars indicate the Standard Error of Mean (SEM). One-way ANOVA was used to analyze the statistical significance, followed by Tukey’s multiple comparisons tests. ** P < 0.01, *** P < 0.001.

Article Snippet: Antibody PE/Cy5 Rat IgG1 (1:50, RG1, NBP1-43076, Novus Biologicals, Littleton, Co, USA), Alexa Fluor 647 Rat IgG1 (1:50, KLH/G1-2-2, 0116-31, SouthernBiotech, Birmingham, AL, USA), and eFluor 450 Rat IgG1 (1:50, eBRG1, 48-4301-82, ThermoFisher) were used according to the manufacturer’s instructions for isotype control.

Techniques: Injection, Adjuvant, Control, Negative Control, Enzyme-linked Immunosorbent Assay, Enzyme-linked Immunospot, Quantitation Assay, Staining

Journal: NPJ Vaccines

Article Title: A candidate subunit vaccine induces protective immunity against Mycobacterium avium subspecies paratuberculosis in mice

doi: 10.1038/s41541-023-00675-1

Figure Lengend Snippet: a Schematic diagram of the vaccination and MAP challenge timeline. C57BL/6 mice were subcutaneously injected two times (three weeks apart) with 50 μg 66NC fusion protein formulated in Montanide ISA 61 VG (61 VG) adjuvant. Control groups include mice immunized with Montanide ISA 61 VG adjuvant alone as a negative control or 74 F mixed MPL adjuvant as subunit vaccine control. Three weeks after the booster immunization, the mice were challenged with MAP K-10 by intraperitoneal injection. b Body weight was measured for 2, 4, 8, and 12 weeks after the MAP challenge ( n = 6 per group). Monitor IgG ( c ) and IgM ( d ) antibody levels. Sera were collected and measured for specific anti-66NC or 74 F antibody response of both IgG and IgM at 2, 4, 6, 8, 10, 12, 14, 16, and 18 weeks after prime vaccination by ELISA. Bacterial load quantification was evaluated in the liver ( e ) and intestine ( f ) from mice challenged by intraperitoneal injection with MAP K-10 for 2, 4, and 8 weeks after vaccination. g Representative images of Ziehl-Neelsen acid-fast stain in the liver of vaccinated mice, two weeks after being challenged with MAP, are presented (scale bars, 100 μm (top) and 10 μm (bottom)). The bottom images are enlarged from the outlined areas of the top images. b , e , f Data indicate cumulative results from at least three to six independent replicates. Mean ± SEM and Two-way ANOVA were used to analyze the statistical significance, followed by Tukey’s multiple comparisons tests. ns, non-significant; * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Antibody PE/Cy5 Rat IgG1 (1:50, RG1, NBP1-43076, Novus Biologicals, Littleton, Co, USA), Alexa Fluor 647 Rat IgG1 (1:50, KLH/G1-2-2, 0116-31, SouthernBiotech, Birmingham, AL, USA), and eFluor 450 Rat IgG1 (1:50, eBRG1, 48-4301-82, ThermoFisher) were used according to the manufacturer’s instructions for isotype control.

Techniques: Injection, Adjuvant, Control, Negative Control, Enzyme-linked Immunosorbent Assay, Ziehl-Neelsen Stain