Antigen expression in canine tissues, recognized by a monoclonal antibody generated against canine melanoma cells

J. L. Oliver III From the Department of Pathobiology, College of Veterinary Medicine, Auburn University, AL 36849-5519 (Oliver, Wolfe).

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 DVM, PhD
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L. G. Wolfe From the Department of Pathobiology, College of Veterinary Medicine, Auburn University, AL 36849-5519 (Oliver, Wolfe).

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Summary

A murine hybridoma monoclonal antibody (mab), IBF9, was generated by fusing myeloma cells (P3X63Ag8.653) with spleen cells from a BALB/c mouse immunized with the canine melanoma cell line CML-10c7. Initial screening of hybridoma antibodies was performed by use of an indirect immunoperoxidase assay on formalin-fixed CML- 10c7 cells. The isotype of mab IBF9 was IgGl as determined by radial gel immunodiffusion. The antibody was tested for reactivity against a panel of formalin-fixed, paraffin-embedded normal and neoplastic canine tissues, using immunoperoxidase staining. Immunostaining was observed in melanomas (24 of 38), a few carcinomas, basal cell tumors, and cutaneous lymphosarcomas. Immunostaining was not observed in fibrosarcomas, hemangiosarcomas, hemangiopericytomas, or histiocytomas. Staining of normal adult canine tissues was limited to a few epithelial tissues and a small percentage of lymphocytes. Fetal tissues were not reactive with mab IBF9. There were statistically significant differences in frequency of reactivity among melanomas with regard to oral vs nonoral, malignant vs benign, and mitotic indices ≥ 1 vs mitotic indices < 1. Differences were not significant when tumors were compared for degree of pigmentation or histologic type.

On the basis of these findings, we suggest that mab IBF9 may be of assistance in diagnosis of nonpigmented melanomas and in assessing the malignant potential of melanomas.

Summary

A murine hybridoma monoclonal antibody (mab), IBF9, was generated by fusing myeloma cells (P3X63Ag8.653) with spleen cells from a BALB/c mouse immunized with the canine melanoma cell line CML-10c7. Initial screening of hybridoma antibodies was performed by use of an indirect immunoperoxidase assay on formalin-fixed CML- 10c7 cells. The isotype of mab IBF9 was IgGl as determined by radial gel immunodiffusion. The antibody was tested for reactivity against a panel of formalin-fixed, paraffin-embedded normal and neoplastic canine tissues, using immunoperoxidase staining. Immunostaining was observed in melanomas (24 of 38), a few carcinomas, basal cell tumors, and cutaneous lymphosarcomas. Immunostaining was not observed in fibrosarcomas, hemangiosarcomas, hemangiopericytomas, or histiocytomas. Staining of normal adult canine tissues was limited to a few epithelial tissues and a small percentage of lymphocytes. Fetal tissues were not reactive with mab IBF9. There were statistically significant differences in frequency of reactivity among melanomas with regard to oral vs nonoral, malignant vs benign, and mitotic indices ≥ 1 vs mitotic indices < 1. Differences were not significant when tumors were compared for degree of pigmentation or histologic type.

On the basis of these findings, we suggest that mab IBF9 may be of assistance in diagnosis of nonpigmented melanomas and in assessing the malignant potential of melanomas.

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