and dissolved in pbs containing 0 1 sodium azide 100 mg fecal pellet per 1 ml pbs sodium azide these samples were vortexed centrifuged and the supernatants were collected for analysis these mucosal and serum samples were accumulated at weekly intervals and analyzed for antigen e g ova specific iga igm igg ige igg1 igg2a igg2b and igg3 antibody titers mice were sacrificed on day 21 for analysis of ova specific antibody forming cells and t cell proliferative and cytokine profile responses submandibular and cervical lymph nodes sm cln mesenteric lymph nodes mln peyer 39 s patches pp vaginal ileal lymph nodes iln and spleen sp suspensions were made by passage of tissue through wire mesh after the excision of pps the small intestine was isolated to determine the ig secreting cells in the intestinal tract which directly relates to protection against intestinal pathogens the intestinal tissue was then gently cleaned minced and treated with 1 mm edta in pbs at 37 u00b0 c with agitation for 15 to 30 minutes next these tissues were treated with collagenase in rpmi media for approximately 1 hour finally lamina propria lymphocytes lpl were isolated using a percoll pharmacia uppsala sweden gradient the lower respiratory tract lung.