NOVEL IMMUNIZATION PROCEDURES USED TO DEVELOP MONOCLONAL-ANTIBODIES THAT BIND TO SPECIFIC STRUCTURES IN MELOIDOGYNE SPP

Homogenates of the anterior regions of Meloidogyne incognita adult females injected into the spleen of Balb/c mice stimulated the production of nine monoclonal antibodies (MAbs) that bound to secretory granules formed in the nematode dorsal esophageal gland. Five of these nine MAbs were female-specific and bound to secretory granules in M. incognita, M. javanica, and M. arenaria, but not to those in M. hapla or Heterodera glycines. None of these nine MAbs recognized antigens in the dorsal gland of second-stage juveniles of the Meloidogyne spp. Two MAbs that bound to secretory granules within the subventral glands of females of M. incognita, M. javanica, and M. arenaria, but not to those in M. hapla or H. glycines, were obtained from an intrasplenic nitrocellulose implant carrying stylet secretions from M. incognita females. No MAbs to nematode esophageal glands were produced by a subcutaneous implant of nematode stylet secretions on nitrocellulose or through intraperitoneal injection using cyclophosphamide to favor the immune response to homogenates of the anterior region of M. incognita females. Antibodies to nematode nuclei, amphids, esophago-intestinal cells, muscle, and other structures also were produced. All antibody binding was confirmed by indirect immunofluorescence microscopy of nematode specimens. Intrasplenic immunizations required the least time, used small quantities of immunogen, and produced the most desirable MAbs.