Two local isolates of Eimeria tenella gametocytes against coccidiosis were immunologically evaluated in chickens. Cell mediated immune response was detected by modified splenic cell migration inhibition assay (MSCMIA) and data were expressed in terms of per cent migration index. No significant difference in per cent migration index was detected for the chickens immunized either with Vaccine-I (local isolate-I) or with Vaccine-II (local isolate-II); however per cent migration index was comparatively lower in chickens immunized with Vaccine-II as compared to Vaccine-I; indicating a somewhat higher cell mediated immune (CMI) response. Humoral immune response was monitored by ELISA in vaccinated and control chickens. Significantly elevated (P<0.05) antibody titer (IgG) in chickens immunized with Vaccine-II as compared to Vaccine-I was detected. Significantly higher protection (67%) in chickens immunized with Vaccine-II followed by vaccine-I (49%) was recorded. Further, oocyst count was significantly lower (P<0.05) in chickens immunized with Vaccine-II as compared to those immunized with Vaccine-I. It was concluded that vaccinal strain (Vaccine-II) contained additional protein of high molecular weight (49.23 kDa) in its gametocytes provided cross protection and can be used to prepare commercial vaccine against coccidiosis in poultry