The Institutional Animal Care and Use Committee (Iacuc) s3

INSTITUTIONAL ANIMAL CARE AND USE COMMITTEE (IACUC)

IACUC/IIACUCLARC STANDARD PROCEDURE

IN VIVO LYMPHOCYTE HOMING IN MICE

Description of procedure:

Donor mice will be euthanized and lymphoid organs will be harvested to prepare leukocytes for injection into the recipients of the same genetic background. These leukocytes may be labeled with a non-toxic fluorescent dye for tracking purposes. Recipient mice will be kept warm to facilitate vasodilation and restrained briefly in a mouse restrainer. At time 0, antibody or other test compound in sterile saline or phosphate buffered saline may be injected i.v. into the tail vein. After 1 hr, the mouse is again briefly restrained in a mouse restrainer, and 3 x 10e6 leukocytes from the donor mice in sterile saline or phosphate buffered saline are injected i.v. (tail vein). The mouse is returned to the cage Endpoints for each experiment are varied, the timeline will be specified in the individual protocol.

Mice will also be evaluated for general appearance, ability to move normally around cage and reach food and water, ability to eat and drink, grooming habits, skin color, lethargy, aggression (animals will be housed individually if aggressive behavior is shown). Mice will be monitored for 15 min following each i.v. injection and the following day (in the case of the 18 hr experiments).

The mouse will be euthanized immediately if the mouse displays any evidence of respiratory distress or problems with normal movement. All mice will be euthanized at termination of the experiment.

Literature search words required:

Literature search was performed for refinement of this Standard Procedure on April 9, 2007

Key Words Search Site Years Covered Lymphocyte homing in mice Pubmed 1972- present

Agents:

This procedure requires fluorescent dye, test compound, saline. All agents administered to animals should be listed in the "Agents" section of RIO.

Adverse Effects:

Procedure, Agent or Potential Adverse Management Phenotype Effects Tail vein injections Reaction to agent, Euthanasia lethargy, labored breathing Monitoring Parameters:

Monitoring Parameters Frequency PI/Lab will Document General appearance/activity For 15 minutes after each No IV injection and then the next day for 18 hr experiments Describe the conditions, complications, and criteria (e.g. uncontrolled infection, loss of more than 15% body weight, etc.) that would lead to removal of an animal from the study, and describe how this will be accomplished (e.g. stopping treatment, euthanasia). Labored breathing, euthanasia. For all investigators housing animals with tumor formation, skin lesions, neurological deficits, or that are in Category E, list the expected characteristics/clinical presentations and endpoints of the animal model and the criteria for euthanasia. Note: The IACUC also requires such lists to be posted in the respective animal rooms and monitored by the IACUC compliance staff and LARC, to assure PI adherence to the endpoints listed. N/A.