Enzyme histochemistry and immunohistochemistry on biopsy specimens of pathologic human bone marrow

A chronic lymphoproliferative disorder with distinctive features: a distinct variant of hairy-cell leukaemia.

Hairy-cell leukemia with T-cell features

The immunologic phenotyping of bone marrow biopsies and aspirates: frozen section techniques

T lymphocytes positively identified by alphanaphthyl acetate esterase (ANAE) (70%) were localized in perivascular “cuffs” in rheumatoid synovia. ANAE-negative lymphocytes (B lymphocytes) were primarily within the reactive lymphoid centers, whereas intra-cellular Ig-expressing plasma cells were outside the centers. Lysozyme or diffuse ANAE expressing mono-nuclear phagocytes (5–15%) were diffusely dispersed, but were seldom found in the lymphoid centers. There were few granulocytes (10%). On elution, plasma cells and lymphocytes were lost. Most granulocytes and mononuclear phagocytes in the eluate were immunoglobulin-positive. The in situ architecture is compatible with active immunologic inflammation and T-dependent immunglobulin synthesis.

Characterization of the immunocompetent cells of rheumatoid synovium from tissue sections and eluates

A technique has been devised which optimally demonstrates non-specific esterase activity in human blood lymphocytes. The reaction is carried out on smears fixed with formalin vapour, using alpha-naphthol butyrate as substrate at pH 8 and at a low concentration for a short incubation period. The pattern of esterase activity revealed by this method provides a discriminating marker for mature T-lymphocytes, which show dense, localised, dot-like positivity, and a probable marker for 'null' cells in the form of scattered granular positivity. B cells appear to be negative. Monocytes show a clearly different pattern of granular positivity.

Discrimination of B, T and null lymphocytes by esterase cytochemistry.

A distinctive pattern of alpha-naphthyl butyrate esterase positivity was observed in all of a series of 14 patients with HCL. This pattern was not observed in a range of other haematological malignancies including nine cases of CLL, two cases of Sezary's syndrome, six cases of null cell ALL, two cases of Schilling-type monocytic leukaemia and one case of lymphosarcoma cell leukaemia. The potential diagnostic value of this simple cytochemical test is discussed in relation to existing methods.

K. E. Higgy

Papers

Discrimination of B, T and null lymphocytes by esterase cytochemistry.

Identification of the hairy cells of leukaemic reticuloendotheliosis by an esterase method.

Hairy-cell leukaemia: A B-cell neoplasm with a severe deficiency of circulating normal B lymphocytes