C57bl 6

For MSC/splenocyte co-culture assays, 10,000 MSC were seeded per well into 48-well plates. Freshly isolated splenocytes (1 × 10⁶ cells/well) from female C57BL/6 (Jackson Lab) mice were labeled with 10 μM CellTraceTM Violet (CTV) (Invitrogen, Warrington, UK, C34571) according to the manufacturer’s instructions, stimulated with ConA (1 µg/mL) (Sigma, C5275) for T-cell activation and co-cultured with MSC in a ratio (1:10, MSC: splenocytes) in complete RPMI medium (Gibco, 11875-093) with 10% FBS (Gibco, 10437028), 100 U/mL penicillin and 100 μg/mL streptomycin (Gibco, DW101203-031-1B) at 37 °C and 5% CO₂. After 5 days of co-culture, cells were washed and evaluated using flow cytometry. To assess T-cell proliferation, we used CTV. Each peak of the histograms corresponds to one cell division for CD3+ lymphocytes. The proliferation index was calculated as follows: the number of cells divided by the number of progenitors, as described by Roederer et al. [38 (link)]. Furthermore, cell survival was evaluated with SYTOX™ Green dye (Invitrogen, S7020). Flow cytometry was performed in a CytoFLEX (Beckman Coulter, Brea, CA, USA) and analyzed using the FlowJo 10.6 software.

Animals were housed under specific pathogen-free conditions as per the national animal testing regulations. All the murine experiments performed in this study were approved by the Institutional Animal Care and Use Committee (IACUC) of the University of Texas MD Anderson Cancer Center. Six-to-eight-week-old female C57BL/6 or BALB/c mice were purchased from Charles River Laboratory. Pmel-1 TCR transgenic mice on a C57BL/6 background (The Jackson Laboratory, Bar Harbor, ME) were crossed with CD90.1 congenic mice to yield pmel-1^+/+ × CD90.1^+/+ mice (hereafter referred to as pmel-1 mice)^{19 (link)}. All murine cell lines were obtained from ATCC and were maintained in complete medium, including RPMI 1640 with 10% FBS, 100 µg/ml streptomycin, and 100 µg/ml penicillin (Life Technologies, Carlsbad, CA).
Eight tumor models were evaluated in four different mouse strains: C57BL/6 strain for LLC (lung), Pan02 (pancreatic), and B16.F10 (melanoma); C3H strain for MBT-2 (bladder); BALB/c strain for CT26 (colon), H22 (liver), and EMT6 (breast); FVB strain for BR5FVB (ovarian). The latter cell line contains a BRCA and akt mutation similar to the genetics of many human serous ovarian tumors^{45 (link)}.

Male C57BL/6 (Taconic, Germantown, NY), B6-129S4-C3<tm1Crr>/j (C3^-/-), B6-129P2-Fcgr3^tm1Sjv/J (CD16^-/-), B6-Cg-Lgals3^tm1/poi/J (Gal3^-/-), B6-prf1^tm1sdz/1 (Prf1^-/-), B6-129P2-Tcrb^tm1momTcrd^tm1mom/J (TCR^-/-), B6-Tnfrs1a^tm1/mx/J (TNF R1^-/-), B6.129S7-Ifng^tm1ts/J (INFγ^-/-) mice were purchased from Jackson Laboratory (Bar Harbor, ME). All mice were housed under barrier conditions for at least 1 week after arrival at the university and were used at 7–9 week of age. Breeding pairs of B6-MyD88^-/- and B6-TRIF^-/- mice were a gift from Dr. Fitzgerald, University of Massachusetts School of Medicine (Worcester MA), were bred in the animal facilities at UMass Amherst and were used at 7 to 9 weeks of age. Mice were infected by intraperitoneal (i.p.) injection of 5000 exponentially growing pleomorphic Trypanosoma brucei Antat 1.1 or sham infected by i.p., injection of Dulbecco’s Phosphate Buffered Saline (PBS; GIBCO,Life Technologies); these parasites were derived from EATRO 1125 stock [88 (link)] and grown from cryopreserved parasites in immunocompromised (600r from a 127 Cesium source) C57BL/6 donors prior to injection. Parasitemia was assessed in tail blood by dilution in DPBS and counting using a hemocytometer.

Female C57BL/6 (6–8 weeks old) mice were used in this study and purchased from Animal Resources Centre (Perth, Western Australia). TC-1 cells (murine C57BL/6 lung epithelial cells transformed with HPV-16 E6/E7 and ras oncogenes).29 (link) TC-1 cells were cultured and maintained at 37 °C/5% CO₂ in RPMI 1640 medium (Gibco) supplemented with 10% heat inactivated fetal bovine serum (Gibco) and 1% nonessential amino acid (Sigma-Aldrich). The animal experiments were approved by the University of Queensland Animal Ethics committee (DI/034/11/NHMRC) and (UQDI/327/13/NHMRC) in accordance with National Health and Medical research Council (NHMRC) of Australia guidelines.