Clonal Expansions of CD8(+) T Cells with IL-10 Secreting Capacity Occur during Chronic Mycobacterium tuberculosis Infection

Cyktor, Joshua C.; Carruthers, Bridget; Beamer, Gillian L.; Turner, Joanne

Specific pathogen-free, age/sex-matched CBA/J wild-type (National Cancer Institute, NIH, Frederick, MD), C57BL/6 wild-type (Jackson Laboratories, Bar Harbor, Maine), or CBA/J IL-10 2/2 mice were maintained in ventilated cages inside a biosafety level 3 (BSL3) facility and provided with sterile food and water ad libitum. To generate CBA/J IL-10 2/2 mice, CBA/J mice (Jackson laboratories, Bar Harbor, Maine) were crossed with C57BL/6 IL-10 2/2 mice (Jackson) for eight generations. At each cross progeny mice were ear-punched and DNA was screened for the presence of a neomycin cassette at the il10 gene locus. IL-10 +/ 2 mice were selected for further breeding. At the eighth generation, heterozygotes were crossed and IL-10-deficient homozygote CBA/J mice were selected. A homozygous breeder colony of CBA/J IL-10 2/2 mice was maintained thereafter. All protocols were approved by The Ohio State University's Institutional Laboratory Animal Care and Use Committee.