Hock immunization: A humane alternative to mouse footpad injections

Footpad injection is a commonly used immunization method in mice. Being relatively easy to do with well-characterized lymphatic drainage, it has become a very useful immunization protocol to study local immune responses in draining lymph nodes. However, its disadvantages include use of only hind feet as a routine site of immunization since mice use their fore feet for food handling, and exacerbation of inflammation and swelling at the injection site leading to unrelieved pain and distress since feet are weight-bearing structures. With increasingly stringent Institutional guidelines for animal manipulations, there is increasing need for more humane protocols. A novel immunization protocol involving injection into the hock, the lateral tarsal region just above the ankle, a non-weight bearing structure draining to the same lymph node as the footpad, retains the advantages of footpad immunization without its drawbacks. This study, comparing immune responses between footpad and hock immunization in six different inbred mouse strains to two different protein antigens and a heat-killed bacterium, shows that hock immunization is a better alternative to footpad immunization, inducing comparable immune responses and being considerably more humane.