| Summary: | Seven novel monoclonal antibodies reacting specifically against Anisakis larvae （type I, L3） were generated by fusing mouse myeloma NS-1 and spleen cells from Balb/c mice immunized with solubilized antigen of Anisakis larvae. All of these monoclonal antibodies （An1-An7） failed to show any reactivity against Ascaris suum, Dirofilaria immitis, Toxocara cati, Trichinella spiralis, Echinococcus multilocularis, and human tissues, except for a weak reactivity of An6 against intestine of Ascaris suum. These were reactive not only to solubilized antigens on ELISA but also to frozen sections of the larvae; Anl reacted with muscle （M） and the Renette cells （R）; An2 M, R and intestine （I） and pseudocoel （P） weakly; An3 M, P and R weakly; An4 M, R, P and I weakly; An5 M and P; An6 M, I and R weakly and An7 R and I. These monoclonal antibodies indicated a high specificity of reaction to Anisakis larvae because they did not react to Terranova larvae （type A） which is a worm of anisakid species. An1, An2 and An6 revealed a distinct reactivity against excretory-secretory antigen （ES antigen） using ELISA method. An2 and An6 recognized the antigen of 90kd and 68kd of molecular weight, respectively, as determined in ELISA with the antigens fractionated by gel filtration. Anl, An2, and An4 immunoprecipitated with the antigens of 32-36kd, 40 and 42-46kd, and 130kd of molecular weight, respectively, in Western blotting analysis. All of the antigens were apparently different from the hemoglobin of Anisakis larvae; this can be said because of the different reactivities in immunostaining and immunochemical studies. When An2 was applied for seroimmunodiagnosis of patients with anisakiasis, it was shown that this monoclonal antibody may be useful because of the improved specificity using ELISA analysis. Collectively, these monoclonal antibodies have potentially important values in investigating the taxonomical, pathological, and serodiagnostic features of Anisakis larvae. departmental bulletin paper
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