C c3 tlr4lps d j

T-cell/B-cell-deficient SCID (B6; 129S7-Rag1^tm1mom/J), TNF-deficient (B6;129S-Tnf^tm1Gkl/J), TNF receptor 1 (TNFR1)-deficient (B6;129S-Tnfrsf1a^tm1Imx Il1r1^tm1Imx/J), TNFR2-deficient (B6.129S2-Tnfrsf1b^tm1Mwm/J); CD14-deficient (B6.129S-Cd14^tm1Frm/J) and TLR4-deficient (C.C3-Tlr4^Lps-d/J) female mice were purchased from The Jackson Laboratory (Bar Harbor, ME). Mice were housed at the University of Pittsburgh Cancer Institute’s Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC) internationally accredited animal facility. The animal studies were performed in accordance with protocols approved by the University of Pittsburgh Institutional Animal Care and Use Committee.

BALB/c, C57BL/6, C.C3-Tlr4^Lps-d/J (Tlr4^-d), C.129-Il4^tm1Lky/J (4get), Tg(Cd4-cre)1Cwi/BfluJ (Cd4-Cre), B6.129P2-Tcrb^tm1Mom/J (Tcrb^−/−), B6Cr.129S4-Tnfsf4^tm1Sug/Pgn (Tnfsf4^−/−), B6;129S1-Il1rap^tm1Roml/J (Il1rap^−/−), and B6.129S7-Il1r1^tm1Imx/J (Il1r^−/−) mice were obtained from The Jackson Laboratory (Bar Harbor, ME, USA). Il1rap^−/− mice were subsequently backcrossed onto the BALB/c background for 10 generations. Il1rl1^−/− (ST2^−/−) mice and Crlf2^−/− (Tslpr^−/−) mice (both BALB/c background) were kindly provided by Dr. Andrew McKenzie (MRC Laboratory of Molecular Biology, Cambridge, UK) and Dr. Steven F. Ziegler (Benaroya Research Institute, Seattle, WA, USA), respectively, and were bred under specific pathogen-free (SPF) conditions at the Mayo Clinic (Rochester, MN, USA)^{21 (link), 22 (link)}. Bcl6^fl/fl mice were previously described^{23 (link)}. Animals used in this study were female and ranged from 6–12 weeks of age. All protocols and procedures for handling of the mice were reviewed and approved by the Mayo Institutional Animal Care and Use Committee, Mayo Clinic.

The Institutional Animal Care and Use Committee at the University of Florida approved all mice-related procedures and experiments (IACUC, protocol #201304539). Eight-week old male B6.129-Tlr2^tm1Kir/J and C.C3-Tlr4^Lps-d/J mice were obtained from the Jackson Laboratories (Bar Harbor, ME). Upon arrival, TLR2 and 4 mice were allowed to adapt to the new environment for at least 1 week prior to initiation of any experiment (Rivera et al., 2013 (link)). Mice were housed under specific-pathogen-free facility in University of Florida vivarium. Twenty-four mice were randomly assigned to the infection group and twenty-four to the control group for both TLR2^−/− and TLR4^−/− groups. The infected group orally inoculated by lavage with 10⁹ total cells per mL in RTF-4% CMC as previously described (Rivera et al., 2013 (link), Velsko et al., 2015 (link)), while sham-infected mice were mock-infected with RTF-4% CMC only. At twenty-four weeks of infection, mice were euthanized and sera, jaws and internal organs were collected.