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Fifty-one patients were referred in one year (1992) for investigation of immediate type anaphylactic/anaphylactoid reactions during anaesthesia. Skin prick tests were made with 23 anaesthetic and associated agents in the concentrations used clinically. Definite or probable causes were identified by immediate type wealing reactions, supported by the clinical history in 36 of the 46 in whom a diagnosis of anaphylaxis was made. These comprised mainly the neuromuscular relaxants, chiefly suxamethonium (18); atracurium (6); gallamine (2); one each alcuronium; pancuronium; vecuronium and tubocurarine, as well as alfentanil (1); Gelofusine (2); cefuroxime (1) and latex (2). The materials for performing the skin prick test are readily available and it can be very helpful in making important aetiological diagnoses.

Alkaline phosphatase-labelled F(ab')2 polyvalent anti-human immunoglobulin stained a mean of 12% (s.d. 4.2) of lymphocytes in the whole peripheral blood of 15 normal individuals. However, when the sensitivity of detection of the bound anti-immunoglobulin reagent was enhanced by adding complexes of alkaline phosphatase with F(ab')2 anti-alkaline phosphatase, a mean of 22.1% (s.d. 7.8) of lymphocytes were positive. The mean number of T lymphocytes demonstrated in the same blood samples using a monoclonal anti-T lymphocyte antibody (OKT3) was 78% (s.d. 4.1) and was not increased by immunoenzyme enhancement. In five individuals the blood was washed at 37 degrees C to remove passively adsorbed IgG and was then studied using the enhanced method together with monoclonal anti-kappa and anti-lambda antibodies. The mean +/- s.d. number of kappa-positive lymphocytes was 15.5 +/- 4.6% and lambda-positive was 7.9 +/- 1.1%. The sum of these was the same as the number of cells stained either with anti-kappa and anti-lambda together or with the conventional polyvalent anti-immunoglobulin, confirming that the enhancement procedure was detecting integral membrane immunoglobulin and not passively adsorbed IgG. Application to the same blood sample of both the anti-T cell antibody and the enhancement procedure with polyvalent anti-immunoglobulin stained 99-100% of lymphocytes. The present observations confirm that there are two populations among the B-lymphocytes, the B-major cells with readily demonstrable surface immunoglobulin and the B-minor cells on which surface immunoglobulin is demonstrable only by very sensitive techniques (Haegert & Coombs, 1979). The B-major and B-minor cells together account for all the non-T lymphocytes and there are virtually no so-called 'null' cells in normal peripheral blood. These findings have significant implications for the use of surface membrane immunoglobulin as a marker in the typing of normal and abnormal lymphocyte populations.

The whole blood method for enumeration of T and B lymphocyte populations using alkaline phosphatase-labelled reagents (Druguel and Pepys, 1977) has been extended to the quantitation of lymphocytes bearing receptors for Fc(gamma) and for C3b. This technique, by avoiding the variable loss of cells which is unavoidably associated with separation of mononuclear cells from blood, permits accurate measurement of the proportions and absolute numbers of circulating lymphocytes bearing these receptors. Fc(gamma) receptors were detected using complexes of alkaline phosphatase (AP) and eluted rabbit IgG anti-AP antibody. Receptors for C3b were detected using isolated human C3b followed by conjugates of AP with F(ab)2 of sheep anti-human C3 antibody. The mean +/- S.D. of Fc(gamma) receptor-positive lymphocytes among 16 normal adults was 11.8 +/- 2.5%, 248 +/- 113/microliter and among 17 normal adults the values for C3b receptor-positive lymphocytes was 9.4 +/- 2.0%, 208 +/- 101/microliter.

Lymphocyte populations identified by surface markers were enumerated in whole peripheral blood of 3 patients with Goodpasture's syndrome (mediated by anti-glomerular basement membrane antibody) whilst on treatment with cyclosporin A (CSA). The whole blood method, using alkaline phosphatase-labelled reagents, permits precise enumeration of the relative and absolute numbers of lymphocyte subsets in the circulation. Using conventional rabbit and monoclonal mouse anti-human T-cell antibodies, a small decrease in the percentage of T-lymphocytes was noted following the start of CSA treatment. The absolute T-cell count fell, but only to just below normal. There was no marked or consistent effect on the percentages or absolute numbers of B-cells, C3b-receptor or Fc(gamma)-receptor bearing lymphocytes. These results are compatible with a mechanism of immunosuppression by CSA in which there is selective inhibition of a relatively minor subset of T-lymphocytes.

The proportions and absolute numbers of different lymphocyte populations were determined using alkaline phosphatase-labelled reagents in the whole peripheral blood of 22 patients with Crohn's disease. Monoclonal mouse anti-T cell antibody (OKT3) was used to identify T cells, polyvalent F(ab')2 anti-human immunoglobulin for B cells, C3b for C3b-receptor bearing cells, and soluble IgG antibody-antigen complexes for Fc(gamma)-receptor bearing cells. Endogenous myeloperoxidase served to distinguish monocytes. Application of this methodology to whole blood avoids the inevitable loss of cells which accompanies separation of mononuclear cells from blood and therefore permits precise enumeration of lymphocyte populations in the circulation. No significant difference from healthy adult controls was observed in any of the lymphocyte subsets tested.

Cells from 13 cases of T-prolymphocytic leukaemia (T-PLL) were studied with a battery of immunological techniques in order to define their membrane phenotype. All cases were E-rosette positive and were negative with OKT6, anti-HLA-DR, anti-Ig and M-rosettes; in 3, 20-30% of the cells had receptors for C3b. 7 cases had predominantly a 'helper/inducer' T-subset phenotype, (OKT4+, OKT8-) and 4 had a 'suppressor/cytotoxic' phenotype (OKT8+, OKT4-). Cells in 2 cases coexpressed OKT4 and OKT8 in 48% and 95% of prolymphocytes and in another, both OKT4 and OKT8 were negative. Terminal transferase (TdT) was negative by IF in all the cases, but a low positive level was detected biochemically in one. Although T-PLL appears to be heterogenous in respect of membrane phenotype, the observation of unexpected features in 8 of the cases raises the possibility that it may originate in a cell of intermediate maturation between late thymocytes and mature T-lymphocytes. These features plus the clinical manifestation of the disease - typical morphology, splenomegaly, lymphadenopathy, skin lesions, high WBC and aggressive clinical course - help define T-PLL as a distinct clinicopathological entity.