Protocol detail

Murine Immunization Model for Antigen-Specific Responses

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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), C57BL/6-Tg(TcraTcrb)1100Mjb/J (OT-I), and B6.Cg-Tg(TcraTcrb)425Cbn/J (OT-II) mice were purchased from The Jackson Laboratory (Bar Harbor, ME). Thymectomized animals underwent adult thymectomy at the Jackson Laboratory (Bar Harbor, ME). E1/E3 deleted Ad5-SIINFEKL-Luc, Ad5-OVA, and Ad5HVR48(1–7)-Gag have been previously described (35 (link)–38 (link)). Mice were immunized intramuscularly with 10⁹ viral particles of the indicated vector in the quadriceps using 100 µl divided equally between the two legs. NYVAC-Env has been previously described (39 (link)) and was used at 10⁶ pfu per mouse. All animal experiments were performed in accordance with the Institutional Animal Care and Use Committee guidelines of Beth Israel Deaconess Medical Center.

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Provine N.M., Larocca R.A., Aid M., Penaloza-MacMaster P., Badamchi-Zadeh A., Borducchi E.N., Yates K.B., Abbink P., Kirilova M., Ng’ang’a D., Bramson J., Haining W.N, & Barouch D.H. (2016). Immediate Dysfunction of Vaccine-Elicited CD8+ T Cells Primed in the Absence of CD4+ T Cells. The Journal of Immunology Author Choice, 197(5), 1809-1822.

Publication 2016

B6.129S2-H2dlAb1-Ea/J C57BL/6-Tg(TcraTcrb)1100Mjb/J (OT B6.Cg-Tg(TcraTcrb)425Cbn/J

Adult Animals Institutional animal care and use committee Legs Mouse Quadriceps Thymectomy Vector Viral particles

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Variable analysis

independent variables

Mouse strains: C57BL/6, B6.SJL-ptprc^a (CD45.1+), B6.129S2-Cd4^tm1Mak/J (CD4 KO), B6.129S2-H2^dlAb1-Ea/J (MHC II KO), C57BL/6-Tg(TcraTcrb)1100Mjb/J (OT-I), and B6.Cg-Tg(TcraTcrb)425Cbn/J (OT-II)
Thymectomy: Thymectomized animals underwent adult thymectomy
Viral vectors: E1/E3 deleted Ad5-SIINFEKL-Luc, Ad5-OVA, Ad5HVR48(1–7)-Gag, and NYVAC-Env

dependent variables

Outcome or response measured from the mice immunized with the viral vectors

control variables

Mouse age: Six- to ten-week-old
Immunization route: Intramuscular in the quadriceps using 100 µl divided equally between the two legs
Viral particle or pfu doses: 10^9 viral particles for Ad5 vectors and 10^6 pfu for NYVAC-Env

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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