Evaluation of antibodies reactive with porcine lymphocytes and lymphoma cells in formalin-fixed, paraffin-embedded, antigen-retrieved tissue sections

Tadashi Tanimoto From the Chuo Meat Inspection Center, Kochi Prefectural Government, 38-1 Ebinomaru, Kochi City 780 (Tanimoto), and Department of Pathology II, Kochi Medical School, Kohasu, Oko-cho, Nankoku City 783 (Ohtsuki), Japan.

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 DVM, PhD
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Yuji Ohtsuki From the Chuo Meat Inspection Center, Kochi Prefectural Government, 38-1 Ebinomaru, Kochi City 780 (Tanimoto), and Department of Pathology II, Kochi Medical School, Kohasu, Oko-cho, Nankoku City 783 (Ohtsuki), Japan.

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 MD, PhD

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Abstract

Objectives

To determine whether antibodies raised against human or porcine lymphocytes are reactive with porcine lymphocytes and lymphoma cells in formalin-fixed, paraffin-embedded, antigen-retrieved tissue sections, and to determine the conditions suitable for antigen retrieval.

Design

We evaluated reactivities of 27 anti-lymphocyte antibodies with porcine tissue sections, including those of lymphomas, treated with 11 antigen-retrieval methods.

Animals

Swine.

Procedure

We used 19 anti-human and 8 anti-porcine lymphocyte antibodies. For antigen retrieval, we tested 11 methods: heating with 6 soaking solutions in a microwave oven, heating with 2 commercially available soaking solutions in a water bath, and enzyme digestion with 3 proteases. After these treatments, sections were immunostained along with untreated sections.

Results

Of 27 anti-lymphocyte antibodies tested, CDw75 (clone LN-1), CD79α (mb-1, clone HM57), CD79β(B29, clone B29/123), HLA-DR (clone TAL. 1B5), and polyclonal CD3 were strongly reactive with porcine lymphoid tissues, including lymphomas, when sections were deparaffinized and heated in 0.1M Tns-HCI buffer (pH 8.0) in a microwave oven or with antigen-retrieval solution in a water bath. All patterns of positive staining were essentially the same as those previously described in human beings.

Conclusion

These 5 antibodies, with the antigen retrieval sequences, are helpful in identifying, distinguishing, and characterizing lymphocyte subsets in archival sections of normal and pathologic porcine lymphoid tissues including lymphomas. In addition, our findings indicated that antigen retrieval by microwave heating of the sections can appreciably expand the range of antibodies useful in paraffin immunohistochemistry, including those that otherwise work only on sections from frozen tissues or from an antigen-derived species. (Am J Vet Res 1996;57:853–859)

Abstract

Objectives

To determine whether antibodies raised against human or porcine lymphocytes are reactive with porcine lymphocytes and lymphoma cells in formalin-fixed, paraffin-embedded, antigen-retrieved tissue sections, and to determine the conditions suitable for antigen retrieval.

Design

We evaluated reactivities of 27 anti-lymphocyte antibodies with porcine tissue sections, including those of lymphomas, treated with 11 antigen-retrieval methods.

Animals

Swine.

Procedure

We used 19 anti-human and 8 anti-porcine lymphocyte antibodies. For antigen retrieval, we tested 11 methods: heating with 6 soaking solutions in a microwave oven, heating with 2 commercially available soaking solutions in a water bath, and enzyme digestion with 3 proteases. After these treatments, sections were immunostained along with untreated sections.

Results

Of 27 anti-lymphocyte antibodies tested, CDw75 (clone LN-1), CD79α (mb-1, clone HM57), CD79β(B29, clone B29/123), HLA-DR (clone TAL. 1B5), and polyclonal CD3 were strongly reactive with porcine lymphoid tissues, including lymphomas, when sections were deparaffinized and heated in 0.1M Tns-HCI buffer (pH 8.0) in a microwave oven or with antigen-retrieval solution in a water bath. All patterns of positive staining were essentially the same as those previously described in human beings.

Conclusion

These 5 antibodies, with the antigen retrieval sequences, are helpful in identifying, distinguishing, and characterizing lymphocyte subsets in archival sections of normal and pathologic porcine lymphoid tissues including lymphomas. In addition, our findings indicated that antigen retrieval by microwave heating of the sections can appreciably expand the range of antibodies useful in paraffin immunohistochemistry, including those that otherwise work only on sections from frozen tissues or from an antigen-derived species. (Am J Vet Res 1996;57:853–859)

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