Induction and monitoring of active delayed type hypersensitivity (DTH) in rats.
Delayed type hypersensitivity (DTH) is an inflammatory reaction mediated by CCR7- effector memory T lymphocytes that infiltrate the site of injection of an antigen against which the immune system has been primed. The inflammatory reaction is characterized by redness and swelling of the site of antigenic challenge. It is a convenient model to determine the in vivo efficacy of immunosuppressants. Cutaneous DTH can be induced either by adoptive transfer of antigen-specific T lymphocytes or by active immunization with an antigen, and subsequent intradermal challenge with the antigen to induce the inflammatory reaction in a given skin area. DTH responses can be induced to various antigens, for example ovalbumin, tuberculin, tetanus toxoid, or keyhole limpet hemocyanin (KLH).Here we demonstrate how to induce an active DTH reaction in Lewis rats. We will first prepare a water-in-oil emulsion of KLH, our antigen of interest, in complete Freund's adjuvant and inject this emulsion subcutaneously to rats. This will prime the immune system to develop memory T cells directed to KLH. Seven days later we will challenge the rats intradermally on the back with KLH on one side and with ovalbumin, an irrelevant antigen, on the other side. The inflammatory reaction will be visible 16-72 hours later and the red and swollen area will be measured as an indication of DTH severity.
Beeton
Christine
C
Department of Physiology and Biophysics, University of California, Irvine, USA. cbeeton@uci.edu
Chandy
K George
KG
D000276:Adjuvants, Immunologic / Q000008:administration & dosage; D000818:Animals; D003872:Dermatitis / Q000276:immunology / Q000473:pathology; D004655:Emulsions / Q000008:administration & dosage; D006433:Hemocyanins / Q000008:administration & dosage / Q000276:immunology*; D006968:Hypersensitivity, Delayed / Q000276:immunology* / Q000473:pathology*; D007156:Immunologic Memory; D007271:Injections, Intradermal; D007279:Injections, Subcutaneous; D051381:Rats; D011917:Rats, Inbred Lew; D012720:Severity of Illness Index; D013601:T-Lymphocytes / Q000276:immunology; D013997:Time Factors
D000276:Adjuvants, Immunologic; D004655:Emulsions; D006433:Hemocyanins; C032808:keyhole-limpet hemocyanin