B6.129s2 cd40lgtm1imx j

Manufactured by Jackson ImmunoResearch

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The B6.129S2-Cd40lgtm1Imx/J is a laboratory mouse strain. It is genetically modified to express a targeted mutation in the CD40 ligand (CD40L) gene.

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10 protocols using b6.129s2 cd40lgtm1imx j

Genetically Engineered Mouse Models

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All animal studies were performed according to the investigator’s protocol approved by the Institutional Animal Care and Use Committees of New York University School of Medicine. The Dnase1l3^−/− mouse model on C57BL/6 (B6) background [Dnase1l3^LacZ/LacZ] was described previously (20 (link)). Mice carrying a targeted allele of Dnase1 (Dnase1^{tm1.1(KOMP)Vlcg}) on B6 background were obtained from the Knockout Mouse Project and crossed with WT B6 and Dnase1l3^−/− mice to obtain Dnase1^−/− and Dnase1^−/−/Dnase1l3^−/− mice, respectively. Mice carrying a targeted allele of Cd40lg [B6.129S2-Cd40lg^tm1Imx/J] on B6 background were obtained from the The Jackson Laboratory and crossed to obtain Dnase1l3^−/−/Cd40lg^−/− mice. WT control mice of B6 background were bred in the same animal facility or purchased from Taconic, Inc. and maintained in the same facility. WT mice of BALB/c background for the pregnancy studies were obtained from Taconic, Inc. Pregnant dams were killed and exsanguinated for plasma collection, and pregnancy terms were estimated by embryo morphology.

Serpas L., Chan R.W., Jiang P., Ni M., Sun K., Rashidfarrokhi A., Soni C., Sisirak V., Lee W.S., Cheng S.H., Peng W., Chan K.C., Chiu R.W., Reizis B, & Lo Y.M. (2018). Dnase1l3 deletion causes aberrations in length and end-motif frequencies in plasma DNA. Proceedings of the National Academy of Sciences of the United States of America, 116(2), 641-649.

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Transgenic Mouse Models for Immunological Synapse

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AD10 TCR transgenic mice on a B10.BR background, specific for pigeon cytochrome c 88–104 and reactive against moth cytochrome c 88–103, were generated by S. Hedrick (University of California at San Diego, La Jolla, CA) and acquired from P. Marrack (National Jewish Center, Denver, CO). B10.A (Taconic), B10.A-Rag2tm1Fwa H2-T18a Tg (5CC7 TCR transgenic, Taconic), B6.129P2-Cd40tm1Kik/J (CD40 knock-out on B6 background, The Jackson Laboratory) and B6.129S2-Cd40lgtm1Imx/J (CD40L knock-out on a B6 background, The Jackson Laboratory) were purchased. B10.A CD40 knock-out mice were generated by breeding B10.A mice to CD40KO and selecting homozygous H-2^k CD40KO breeders from the F2 generation and subsequent generations. CD40L KO male mice were generated by breeding 5CC7 male mice to CD40LKO females. T cells from AD10 TCR transgenic mice were used to generate the Th2 and anti-IL-4 Th2 immunological synapse structures shown in Fig 1, while all other experimental results were generated using 5CC7 TCR transgenic mice. Mice were housed in specific-pathogen free conditions at Oregon Health and Science University according to institutional standards, and the research protocols, IS00003102 and IP00000656, were approved by the Oregon Health & Science University Institutional Animal Care and Use Committee.

Gardell J.L, & Parker D.C. (2017). Despite disorganized synapse structure, Th2 cells maintain directional delivery of CD40L to antigen-presenting B cells. PLoS ONE, 12(10), e0186573.

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Transgenic Mice for Cytochrome c Studies

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B10.A and B10.A-Rag2tm1Fwa H2-T18a (5CC7 TCR transgenic mice on a B10A.RAG KO background), specific for a pigeon cytochrome c peptide on I-E^k and reactive against moth cytochrome c, were purchased from Taconic (Germantown, NY) [51 (link)]. B6.Cg-Tg(TcraTcrb)425Cbn/J (OT-II TCR transgenic mice), specific for chicken ovalbumin residues 323-339 on I-A^b, B6;129S-Fcgr2btm1Ttk/J (FcR KO mice), B10.BR-H2k2 H2-T18a/SgSnJ, and B6.129P2-Cd40tm1Kik/J (CD40 KO mice on a B6 background) [6 (link)] were purchased from The Jackson Laboratory (Bar Harbor, ME). B6.129P2-Cd40tm1Kik/J were bred to B10.A mice, and heterozygous mice from this breeding were crossed and selected to produce I-E^k homozygous CD40 KO mice. 5CC7 TCR transgenic male mice were crossed to female B6.129S2-Cd40lgtm1Imx/J [52 (link)] (CD40L KO mice on a B6 background, purchased from Jackson) to produce 5CC7 CD40L KO male mice. Mice were housed in specific-pathogen free conditions at Oregon Health and Science University according to institutional standards.

Gardell J.L, & Parker D.C. (2016). Antigen-specific transfer of CD40L to antigen-presenting B cells during T cell help. European journal of immunology, 47(1), 41-50.

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Murine Immune Response to Viral Vectors

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Six- to ten-week-old C57BL/6, B6.SJL-ptprc^a (CD45.1⁺), B6.129S2-Cd4^tm1Mak/J (CD4 KO), B6.129S2-H2^dlAb1-Ea/J (MHC II KO), B6.129S2-Cd40lg^tm1Imx/J (CD40L KO), and B6.129P2-Cd40^tm1Kik/J (CD40 KO) animals were purchased from The Jackson Laboratory (Bar Harbor, ME). Mice were immunized with the previously described E1/E3 deleted Ad26, or Ad5HVR48(1 –7 (link)) vectors expressing SIV Gag or SIV Env from the strain SIV_mac239, or LCMV GP (11 (link), 34 (link)–36 (link)). Mice were immunized i.m. in the quadriceps with 10⁹ viral particles of each vector in a volume of 100μl divided equally between the two legs. For co-administration of SIV Gag and SIV Env expressing vectors the final injection volume of 100μl was held constant. Mice were challenged with 1.75×10⁵ to 2.5×10⁵ cfu of recombinant Listeria monocytogenes expressing the LCMV epitope GP33-41 (Lm-GP33) by i.v. injection (a kind gift of Dr. Hao Shen)(37 (link)). Precise dose was back calculated following each experiment by plating on BHI-Agar plates, as previously described (38 ). All animal experiments were in accordance with Beth Israel Deaconess Medical Center Institutional Animal Care and Use Committee guidelines.

Provine N.M., Larocca R.A., Penaloza-MacMaster P., Borducchi E.N., McNally A., Parenteau L.R., Kaufman D.R, & Barouch D.H. (2014). Longitudinal Requirement for CD4+ T Cell Help for Adenovirus Vector–Elicited CD8+ T Cell Responses. The Journal of Immunology Author Choice, 192(11), 5214-5225.

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Humanized Mouse Models for Research

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C57Bl/6 Ly45.1, C57Bl/6 Ly45.2 mice were purchased from Charles River Laboratory. Cd40lg^-/- (B6.129S2-Cd40lgtm1Imx/J), humanized NSG or NSGW41 mice were purchased from The Jackson Laboratory and maintained in specific-pathogen-free (SPF) conditions. The procedures involving animals were designed and performed with the approval of the Animal Care and Use Committee of the San Raffaele Hospital (IACUC #818 for Cd40lg^-/-, #876 for NSGW41 and NSG) and communicated to the Italian Ministry of Health and local authorities according to Italian law.
Eight- to twelve- weeks old female mice were used for the experiments.

Omer-Javed A., Pedrazzani G., Albano L., Ghaus S., Latroche C., Manzi M., Ferrari S., Fiumara M., Jacob A., Vavassori V., Nonis A., Canarutto D, & Naldini L. (2022). Mobilization-based chemotherapy-free engraftment of gene-edited human hematopoietic stem cells. Cell, 185(13), 2248-2264.e21.

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Genetically Modified Mice for Immunotherapy

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All knockout and transgenic mice used in this study were female on a C57BL/6 background. C57BL/6J mice, Pmel-1 T-cell receptor (TCR)-transgenic mice (B6.Cg Thy1^a-Tg(TcraTcrb)8Rest/J), Batf3^−/− mice (B6.129S(C)-Batf3^tm1Kmm/J), CD40^−/− mice (B6.129S2-Cd40lg^tm1Imx/J), and CD80/CD86^−/− mice (B6.129S4-Cd80^tm1ShrCd86^tm2Shr/J) were purchased from the Jackson Laboratory. CD70^−/− mice have been previously described (36 (link), 37 (link)). CD27^−/− mice were originally generated by Dr. Jannie Borst (Netherlands Cancer Institute) and were backcrossed in the Cao lab with C57BL/6J mice from the Jackson Laboratory. These mice were bred in-house (Roswell Park Comprehensive Cancer Center). CD27^−/− mice were backcrossed onto Pmel-1 mice to generate CD27^−/− Pmel-1 mice. All mice were 7 to 10 weeks old at the beginning of each experiment and were maintained under specific pathogen-free conditions at the Roswell Park animal facility according to approved institutional guidelines.

Oba T., Hoki T., Yamauchi T., Keler T., Marsh H.C., Cao X, & Ito F. (2020). A critical role of CD40 and CD70 signaling in cDC1s in expansion and antitumor efficacy of adoptively transferred tumor-specific T cells. Journal of immunology (Baltimore, Md. : 1950), 205(7), 1867-1877.

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Depletion of CX3CR1+ CD8+ T Cells

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Female C57BL/6 mice were purchased from the Jackson Laboratory. CD40^−/− mice (B6.129S2-Cd40lg^tm1Imx/J), CD80/86^−/− mice (B6.129S4-Cd80^tm1ShrCd86^tm2Shr/J), and Batf3^−/− mice (B6.129S(C)-Batf3^tm1Kmm/J), CD2-Cre mice (C57BL/6-Tg(CD2-cre)1Lov/J), and CX3CR1-DTR mice (B6N.129P2-Cx3cr1^tm3(DTR)Litt/J) on C57BL/6 background were purchased from the Jackson Laboratory, and bred in-house (Roswell Park Comprehensive Cancer Center). CD70^−/− mice on C57BL/6 background have been previously described (16 (link)). For inducible Cd2-cre/Cx3cr1^+/DTR mice, we crossed CD2-Cre mice with CX3CR1-DTR mice to generate Cd2-cre/Cx3cr1^+/DTR mice, allowing induction of DTR in CX3CR1⁺ CD8⁺ T cells. For depletion of CX3CR1⁺ CD8⁺ T cells, diphtheria toxin (Sigma) (250 ng/dose/mouse) was administered intraperitoneally (i.p.) every day starting from 1 day before neoantigen vaccination. All mice were 7 to 12 weeks old at the beginning of each experiment, maintained under specific pathogen-free, and housed in the Laboratory Animal Resources facility. All animal studies were conducted in accordance with and approved by the Institutional Animal Care and Use Committee (IACUC) at the Roswell Park Comprehensive Cancer Center.

Yamauchi T., Hoki T., Oba T., Kajihara R., Attwood K., Cao X, & Ito F. (2021). CD40 and CD80/86 signaling in cDC1s mediate effective neoantigen vaccination and generation of antigen-specific CX3CR1+ CD8+ T cells. Cancer immunology, immunotherapy : CII, 71(1), 137-151.

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Murine Models for Immunological Studies

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6–8 weeks old female C57BL/6J (strain code: 000664), BALB/cJ (strain code: 000651), Rag1 Knock-out (B6.129S7-Rag1^tm1Mom/J), CD4- Knockout (B6.129S2-Cd4 ^tm1Mak/J) and CD40 ligand knockout (B6.129S2-Cd40lg ^tm1Imx/J) were purchased from Jackson Laboratories Inc. Immunodeficient athymic Nude mice (Athymic NCr-nu/nu), SCID mice (NCI SCID/Ncr, BALB/c background) and immuno-competent SKH-1 elite mice (Crl:SKH1-Hr^hr) were obtained from Charles River, Inc. IFNAR knockout mice were a kind gift from Dr. G. Cheng (University of California, Los Angeles, CA) All animal studies were carried out in accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health and with the prior approval of the Animal Care and Use Committee of Johns Hopkins University (MO12M223).

Wang J.W., Jiang R., Peng S., Chang Y.N., Hung C.F, & Roden R.B. (2015). Immunologic Control of Mus musculus Papillomavirus Type 1. PLoS Pathogens, 11(10), e1005243.

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Ethical Evaluation of CD40L Mice

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CD40L^-/- mice (Jackson Laboratory, B6.129S2-Cd40lg^tm1Imx/J, Stock no. 002770)[69 (link)] were obtained from the Animal Facility at National Centre of Cell Science Pune, India. All experimental procedures and animal care and use were strictly regulated and reviewed in accordance with good animal ethics approved by the Institutional Animal Care and Use Committee at the Indian Institute of Science Education and Research Kolkata (AUP no. IISERK/ IAEC/AP/2019/45). Experiments were performed following the guidelines of the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), India.

Saadi F., Chakravarty D., Kumar S., Kamble M., Saha B., Shindler K.S, & Das Sarma J. (2021). CD40L protects against mouse hepatitis virus-induced neuroinflammatory demyelination. PLoS Pathogens, 17(12), e1010059.

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Genetic Modulation of CD8+ T Cells

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Female C57BL/6 mice were purchased from the Jackson Laboratory. CD40 -/-mice (B6.129S2-Cd40lg tm1Imx /J), CD80/86 -/-mice (B6.129S4-Cd80 tm1Shr Cd86 tm2Shr /J), and Batf3 -/- mice (B6.129S(C)-Batf3 tm1Kmm /J), CD2-Cre mice (C57BL/6-Tg(CD2-cre)1Lov/J), and CX3CR1-DTR mice (B6N.129P2-Cx3cr1 tm3(DTR)Litt /J) on C57BL/6 background were purchased from the Jackson Laboratory, and bred in-house (Roswell Park Comprehensive Cancer Center). CD70 -/- mice on C57BL/6 background have been previously described (14) (link). For inducible Cd2cre/Cx3cr1 +/DTR mice, we crossed CD2-Cre mice with CX3CR1-DTR mice to generate Cd2cre/Cx3cr1 +/DTR mice, allowing induction of DTR in CX3CR1 + CD8 + T cells. For depletion of CX3CR1 + CD8 + T cells, diphtheria toxin (Sigma) (250 ng/dose/mouse) was administered intraperitoneally (i.p.) every day starting from 1 day before neoantigen vaccination. All mice were 7 to 12 weeks old at the beginning of each experiment, and were maintained under specific pathogen-free conditions at the Roswell Park animal facility according to approved institutional guidelines.

Yamauchi T., Hoki T., Oba T., Attwood K., Cao X., & Ito F. (2020). CD40 and CD80/86 signaling in cDC1s mediate effective neoantigen vaccination and generation of antigen-specific CX3CR1⁺ CD8⁺ T cells in mice.

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