Materials and methods for the development of an antigen-specific immune non-responsiveness state
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example 1
[0027]Popliteal Lymph Node Assay (PLNA):
[0028]PLNA is a quantitative in vivo local graft vs host (GvH) allogeneic reaction that measures T cell activation. A mouse of a first strain (Strain A) is injected in the foot pad with lymphocytes from a mouse of a second stain (strain B). The injected lymphocytes from Stain B will cause a local GvH reaction in the popliteal lymph node of the first mouse (Stain A) causing swelling of the node due to the activation and proliferation of allogeneic T cells. The increase in mass of the popliteal lymph nodes can be measured by weighing the isolated node.
[0029]PLNA takes 1 week where at day 0, 1.5×106 lymph node lymphocytes from strain B mice are injected to the foot pad of strain A mice. Mice are sacrificed on day 7 and popliteal lymph nodes are removed and weighed. Typically, two to three-fold increase in the weight of popliteal lymph node can be observed in this GvH reaction. The immunosuppressive effect of compounds of intere...
example 2
Assessment of Anergy Induction by Compounds Using PLNA
[0030]In order to test the induction of anergy by the compounds, strain A mice can be re-challenged by strain B lymphocytes on day 7. Popliteal lymph nodes are harvested on day 14 and weighed. In the absence of anergy, popliteal lymph node remains swollen due to the re-challenge. If anergy is established by the compound in strain A mice against strain B lymphocytes, popliteal lymph node swelling in strain A mice will be suppressed upon strain B lymphocyte re-challenge, however, popliteal lymph node swelling should be observed if the strain A mice anergic to strain B lymphocytes are re-challenged by lymphocytes from strain C (FIG. 1A).
[0031]Experimental Design.
[0032]Out bred Swiss Webster mice were used as recipient. Lymphocytes from FVB mice were used for PLNA. For assessment of anergy induction by the compounds, Swiss mice were re-challenged using lymphocytes from FVB and SJL mice (FIG. 1B).
[0033]Blank control received PBS injec...
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