Boston University (BU) is committed to observing Federal policies and regulations and AAALAC International standards for the humane care and use of animals. This policy describes adjuvants. Adherence to this policy is mandatory unless a specific exception has been approved by the IACUC.<\/p>\n
This policy is applicable to all persons responsible for conducting research, teaching, training, breeding, and related activities, hereinafter referred to collectively as \u201cactivities\u201d, involving vertebrate animals at or under the auspices of Boston University.<\/p>\n
The use of adjuvants in animal research studies of basic and applied immunology to induce or enhance an immune response requires careful consideration. The requirement for relatively nonspecific inflammation to elicit robust immunity obliges the investigator to balance the cost of potential, local, and\/or systemic pain and\/or distress of the research animal due to the inflammation with the presumed scientific benefit to be gained from the experiment. The validity and applicability of the scientific knowledge gained must be tempered with acknowledgment that the use of potent inflammatory agents, particularly Complete Freund\u2019s Adjuvant (CFA), can result in side effects. Whenever possible, alternatives to CFA must be used.1, 2 However, use of CFA remains scientifically justified in many systems, especially in the induction of autoimmune disease models for which no comparable alternatives exist.<\/p>\n
The use of adjuvants must be described and scientifically justified in the IACUC protocol, to include volume, site and frequency of administration. Pre-emptive analgesia at the discretion of the Principal Investigator (PI) should be described in the IACUC protocol and administered when adjuvants are being given. Analgesia is required for CFA administration unless scientifically justified, in which case animals will be placed in Category E. Some injection locations (e.g. footpad injection) may also require placement in Category E and special husbandry care.<\/p>\n
Intervals between immunizations depend upon the research goal and the animal\u2019s response. It may be necessary to administer frequent injections if the research goal is to create an animal model for autoimmune disease. In contrast, if the goal is to produce antibodies to harvest for other projects, the PI is advised to monitor the antibody titer regularly (recommended every two weeks), and adjust the immunization schedule accordingly. Finally, the immunization schedule and animal response may also be dependent upon the size and antigenicity of the antigen and on the adjuvant used. As a general rule, administration of CFA followed by boosters of Incomplete Freund\u2019s Adjuvant (IFA) for antibody production are separated by 4\u20136 weeks. In summary, injections should be separated in time as much as is commensurate with the goal of immunization.<\/p>\n
There are many commercially available immunologic adjuvants. Selection of specific adjuvants or types depends on whether they are to be used for research and antibody production or in vaccine development. Adjuvants for vaccine use only need to produce protective antibodies and good systemic memory while those for antiserum production need to rapidly induce high titer and high avidity antibodies. No single adjuvant is ideal for all purposes and all have advantages and disadvantages. Adjuvant use is generally accompanied by undesirable side effects of varying severity and duration. Research on new adjuvants focuses on substances which have minimal toxicity while retaining maximum immunostimulation. Investigators should always be aware of potential pain and distress associated with adjuvant use in laboratory animals.<\/p>\n
It is beyond the scope of this policy to describe all adjuvants within this policy. CFA should be the last resort and others tried first. Below are some that are currently used in animal studies by investigators at Boston University.<\/p>\n
NM PorB (Neisseria meningitidis membrane protein) or porin Ag complex:
\nOn days 1, 14, and 28, administer 0.05\u20130.2ml SC in the flank of mice.<\/p>\n
At six\u2013ten weeks old, mice are immunized subcutaneously with 10\u2013200ug of the Ag protein emulsified in 200ul Ribi.
\nFour weeks later, mice are boosted with the same Ag protein mixture emulsified in IFA.
\nMice are euthanized three\u2013four days after the second immunization.<\/p>\n
Administer 200ul of a 1:1 emulsion of TiterMax and the Ag IP in mice.
\nRepeat once.<\/p>\n
Mice are immunized three times at two-week intervals with the Ag in 100ul of IFA. Two weeks after the last immunization, mice are euthanized.<\/p>\n