mhcii deficient  (Charles River Laboratories)

Journal: Immunity

Article Title: The persistence of interleukin-6 is regulated by a blood buffer system derived from dendritic cells

doi: 10.1016/j.immuni.2020.12.001

Figure Lengend Snippet: cDCs set the circulating concentration of sIL-6R, and this regulates antibody responses (A) Histograms reporting surface IL-6R in immune cells from WT, cDC- Irf5 −/− , and cDC- IL6ra −/− mice (B cells = B220 + /CD3 − , T cells = CD3 + /B220 − , cDCs = CD3 − /B220 − /MHCII + /CD11c hi , tissue resident macrophages = CD3 − /B220 − /CD11b + /F4-80 + ). Quantification of these data is presented as the proportion of cells expressing surface IL-6R (mean ± SD, n = 3 per group, ∗∗∗ p < 0.001, ANOVA with Tukey’s test). (B) Total IRF5 was measured in cDCs (B220 − /CD3 − /CD11c hi /MHCII + ) isolated by fluorescence-activated cell sorting (FACS) on splenocytes of WT and cDC- Irf5 −/− animals. The sorted cDCs (200,000 cells) were lysed, separated by SDS-PAGE, and immunoblotted for either IRF5 or β-actin. (C) Serum IgM versus IgG responses against HA (with lipid A adjuvant) in transgenic mice (mean ± SD, n = 4 animals per group). The dilution curves were quantified by endpoint dilution; ∗∗ p < 0.01, ANOVA with Tukey’s test. (D) Serum sIL-6R as measured in WT, cDC- Irf5 −/− , and cDC- IL6ra −/− (mean ± SD, n = 4 animals per group); ∗∗∗∗ p < 0.0001, ANOVA with Tukey’s test. (E) Co-injection of recombinant murine sIL-6R rescued the IgM and IgG responses to HA (with lipid A adjuvant) in cDC- IL6ra −/− animals (mean ± SD, n = 4 animals per group). The dilution curves were quantified by endpoint dilution; ∗∗ p < 0.01, ANOVA with Tukey’s test. See also and for immune cell subsets in cDC- IL6ra −/− and cDC- IL6ra −/− , sIL6R concentration in cDC-DTR, and IL6R expression by cDC1 and cDC2 in cDC- IL6ra −/− and Batf3 −/− .

Article Snippet: The following mice were purchased from Jackson Laboratories: C57BL/6, Tcra −/− , Zbtb46 -DTR +/+ , MHCII deficient (homozygous H2 dlAb1-Ea ), Il6 −/− , Il12p40 −/− , Zbtb46 - cre +/+ , Il6ra flox/flox , Irf5 flox/flox , Adam17 flox/flox , Batf3 −/− , Tlr4 −/− , Tlr7 −/− and Ifnar1 −/− .

Techniques: Concentration Assay, Expressing, Isolation, Fluorescence, FACS, SDS Page, Adjuvant, Transgenic Assay, Injection, Recombinant

Journal: Immunity

Article Title: The persistence of interleukin-6 is regulated by a blood buffer system derived from dendritic cells

doi: 10.1016/j.immuni.2020.12.001

Figure Lengend Snippet: cDC-derived sIL-6R gates antibody responses by controlling plasma cell differentiation (A) Surface expression of gp130 on B cells (B220 + /CD3 − , gate I) versus plasmablasts (CD3 − /B220 lo /CD138 + , gate II) isolated from spleen of WT mice. The splenocytes were either left unstimulated or exposed to sIL-6Rα or IL6-sIL-6Rα (hyper-IL-6). (B) Same as in (A), except that animals were first immunized with lipid A adjuvant and spleen was harvested 3 days later. (C) Quantification of the data in (A). Signaling induced within gate I and gate II was measured as the percentage of STAT3-activated cells (sIL-6Rα-stimulated minus unstimulated and IL-6-sIL-6Rα-stimulated minus unstimulated; mean ± SD, n = 4 animals per group, ∗∗∗ p < 0.001, Student’s t test). (D) Quantification of the data in (B) (as in C). (E) Differentiation from plasmablasts (CD3 − /B220 lo /CD138 + /MHCII + ) to plasma cells (CD3 − /B220 lo /CD138 + /MHC class II lo /CD93 + ) was monitored within each genotype 1 week post-immunization with lipid A adjuvant, and cDC- IL6ra −/− mice were also co-injected with recombinant sIL-6R (as in Figure 2 E). The proportion of plasmablasts differentiating into plasma cells (indicated by a box in flow plots) was then quantified below (mean ± SD, n = 3 animals per group; ∗∗ p < 0.005, ANOVA with Tukey’s test). (F) Model for T cell-independent, cDC-dependent control over antibody responses.

Article Snippet: The following mice were purchased from Jackson Laboratories: C57BL/6, Tcra −/− , Zbtb46 -DTR +/+ , MHCII deficient (homozygous H2 dlAb1-Ea ), Il6 −/− , Il12p40 −/− , Zbtb46 - cre +/+ , Il6ra flox/flox , Irf5 flox/flox , Adam17 flox/flox , Batf3 −/− , Tlr4 −/− , Tlr7 −/− and Ifnar1 −/− .

Techniques: Derivative Assay, Clinical Proteomics, Cell Differentiation, Expressing, Isolation, Adjuvant, Injection, Recombinant, Control

Journal: Immunity

Article Title: The persistence of interleukin-6 is regulated by a blood buffer system derived from dendritic cells

doi: 10.1016/j.immuni.2020.12.001

Figure Lengend Snippet:

Article Snippet: The following mice were purchased from Jackson Laboratories: C57BL/6, Tcra −/− , Zbtb46 -DTR +/+ , MHCII deficient (homozygous H2 dlAb1-Ea ), Il6 −/− , Il12p40 −/− , Zbtb46 - cre +/+ , Il6ra flox/flox , Irf5 flox/flox , Adam17 flox/flox , Batf3 −/− , Tlr4 −/− , Tlr7 −/− and Ifnar1 −/− .

Techniques: Virus, Recombinant, Staining, Adjuvant, Lysis, Blocking Assay, Produced, Enzyme-linked Immunosorbent Assay, Software

Journal: Immunity

Article Title: The persistence of interleukin-6 is regulated by a blood buffer system derived from dendritic cells

doi: 10.1016/j.immuni.2020.12.001

Figure Lengend Snippet:

Article Snippet: Mouse: MHCII deficient ( H2 dlAb1-Ea ) , Jackson laboratory , Stock no. 003584.

Techniques: Virus, Recombinant, Staining, Adjuvant, Lysis, Blocking Assay, Produced, Enzyme-linked Immunosorbent Assay, Software