Expression of CD33 is a predictive factor for effect of gemtuzumab ozogamicin at different doses in adult acute myeloid leukaemia.

It remains unclear in adult acute myeloid leukaemia (AML) whether leukaemic expression of CD33, the target antigen for gemtuzumab ozogamicin (GO), adds prognostic information on GO effectiveness at different doses. CD33 expression quantified in 1583 patients recruited to UK-NCRI-AML17 (younger adults) and UK-NCRI-AML16 (older adults) trials was correlated with clinical outcomes and benefit from GO including a dose randomisation. CD33 expression associated with genetic subgroups, including lower levels in both adverse karyotype and core-binding factor (CBF)-AML, but was not independently prognostic. When comparing GO versus no GO (n=393, CBF-AMLs excluded) by stratified subgroup-adjusted analysis, patients with lowest quartile (Q1) %CD33-positivity had no benefit from GO (relapse risk, HR 2.41 (1.27-4.56), P=0.009 for trend; overall survival, HR 1.52 (0.92-2.52)). However, from the dose randomisation (NCRI-AML17, n=464, CBF-AMLs included), 6 mg/m2 GO only had a relapse benefit without increased early mortality in CD33-low (Q1) patients (relapse risk HR 0.64 (0.36-1.12) versus 1.70 (0.99-2.92) for CD33-high, P=0.007 for trend). Thus CD33 expression is a predictive factor for GO effect in adult AML; although GO does not appear to benefit the non-CBF AML patients with lowest CD33 expression a higher GO dose may be more effective for CD33-low but not CD33-high younger adults.

Laboratory clues to immunodeficiency; missed chances for early diagnosis?

Primary immunodeficiency is seen in an estimated one in 1200 people, and secondary immunodeficiency is increasingly common, particularly with the use of immunosuppresion, cancer therapies and the newer biological therapies such as rituximab. Delays in the diagnosis of immunodeficiency predictably lead to preventable organ damage. Examples of abnormal pathology tests that suggest immunodeficiency from all laboratory specialities are given, where vigilant interpretation of abnormal results may prompt earlier diagnosis. If immunodeficiency is suspected, suggested directed testing could include measuring immunoglobulins, a lymphocyte count and T-cell and B-cell subsets.

Azacitidine fails to eradicate leukemic stem/progenitor cell populations in patients with acute myeloid leukemia and myelodysplasia.

Epigenetic therapies demonstrate significant clinical activity in acute myeloid leukemia (AML) and myelodysplasia (MDS) and constitute an important new class of therapeutic agents. However hematological responses are not durable and disease relapse appears inevitable. Experimentally, leukemic stem/progenitor cells (LSC) propagate disease in animal models of AML and it has been postulated that their relative chemo-resistance contributes to disease relapse. We serially measured LSC numbers in patients with high-risk AML and MDS treated with 5'-azacitidine and sodium valproate (VAL-AZA). Fifteen out of seventy-nine patients achieved a complete remission (CR) or complete remission with incomplete blood count recovery (CRi) with VAL-AZA therapy. There was no significant reduction in the size of the LSC-containing population in non-responders. While the LSC-containing population was substantially reduced in all patients achieving a CR/CRi it was never eradicated and expansion of this population antedated morphological relapse. Similar studies were performed in seven patients with newly diagnosed AML treated with induction chemotherapy. Eradication of the LSC-containing population was observed in three patients all of whom achieved a durable CR in contrast to patients with resistant disease where LSC persistence was observed. LSC quantitation provides a novel biomarker of disease response and relapse in patients with AML treated with epigenetic therapies. New drugs that target this cellular population in vivo are required.

Pitfalls in the performance and interpretation of clinical immunology tests.

A broad overview, with examples, of the potential pitfalls encountered in the clinical immunology laboratory is presented. Illustrative examples and case scenarios are provided from autoimmunity, immunochemistry and cellular immunology, looking at both technical and interpretative pitfalls.

Early emergence of PNH-like T cells after allogeneic stem cell transplants utilising CAMPATH-1H for T cell depletion.

CAMPATH-1H (C-1H) is widely used in vivo and / or in vitro for T cell depletion in hematopoietic SCT. This humanised monoclonal antibody is specific for CD52, a marker coexpressed on the majority of human lymphocytes with CD48 and other glycosylphosphatidyl-inositol (GPI) anchored proteins. We detected CD52 / CD48 dual expression on >99% of CD3(+) lymphocytes from normal individuals and all 15 post-SCT patients whose transplants did not utilise C-1H. By contrast, 23 / 26 patients with transplants involving C-1H (in vivo, in vitro or both) exhibited populations lacking CD52 expression that accounted for 49.7% (4.2-86.2%) of the CD3+ lymphocytes (median and range) in samples evaluated at a median of 2 months post-SCT. Most CD52- cells also lacked CD48 expression. These GPI- T cells were of either donor or mixed donor / recipient origin. They were predominant in the early months after SCT at times of profound lymphopenia and inversely correlated with the recovery of the absolute lymphocyte count (r= - 0.663, P<0.0001). The presence of CD52- cells has been correlated previously with clinical outcome after CAMPATH therapy for both malignant and nonmalignant diseases.

High peripheral blood progenitor cell counts enable autologous backup before stem cell transplantation for malignant infantile osteopetrosis.

Autosomal recessive osteopetrosis (OP) is a rare, lethal disorder in which osteoclasts are absent or nonfunctional, resulting in a bone marrow cavity insufficient to support hematopoiesis. Because osteoclasts are derived from hematopoietic precursors, allogeneic hematopoietic cell transplantation can cure the bony manifestations of the disorder. However, high rates of graft failure have been observed in this population. It is not possible to harvest bone marrow from these patients for reinfusion should graft failure be observed. We report that 8 of 10 patients with OP had high numbers of circulating CD34(+) cells (3% +/- 0.9%). This increased proportion of peripheral CD34(+) cells made it possible to harvest 2 x 10(6) CD34(+) cells per kilogram with a total volume of blood ranging from 8.3 to 83.7 mL (1.3-11.6 mL/kg). In addition, colony-forming assays documented significantly more colony-forming unit-granulocyte-macrophage and burst-forming unit-erythroid in the blood of osteopetrotic patients compared with controls; the numbers of colony-forming units approximated those found in control marrow. We conclude that OP patients with high levels of circulating CD34(+) are candidates for peripheral blood autologous harvest by limited exchange transfusion. These cells are then available for reinfusion should graft failure be observed in patients for whom retransplantation is impractical.

Rapid diagnosis and characterization of acute promyelocytic leukaemia in routine laboratory practice.

The rapid diagnosis of the t(15;17)(q22;q21) promyelocytic leukaemia is important in the early introduction of targeted therapy with all-trans retinoic acid plus chemotherapy. It has been noted that these are usually myeloperoxidase (MPO)-positive and HLA-DR-negative with homogenous expression of CD33 and heterogeneous expression of CD13. We evaluated the use of immunophenotyping, morphology and cytogenetics in our own practice. Cascade testing, using cytoplasmic MPO expression in a high percentage of blast cells in bone marrow as the primary screen and PML (promyelocytic leukaemia protein) expression as the secondary confirmatory test, allowed rapid identification of the cases with t(15;17). This approach allows early instigation of appropriate therapy.

A multicentre evaluation of the diagnostic efficiency of serological investigations for C1 inhibitor deficiency.

AIM: To determine the diagnostic efficiency of assays routinely used in the investigation of hereditary angio-oedema. METHODS: Over a four year period, 1144 samples were received for analysis from 907 patients suspected of C1 inhibitor deficiency. Analyses were performed for C4 and C1 inhibitor (functional and immunochemical). Notes were reviewed retrospectively on patients with low serological indicators to determine diagnosis. RESULTS: These are the first data to indicate the sensitivity, specificity, and predictive values of the assays most frequently used to screen for C1 inhibitor deficiency. A combination of low C4 and low C1 inhibitor function has 98% specificity for C1 inhibitor deficiency in this population and a 96% negative predictive value, and is thus a very effective screen. All patients with untreated C1 inhibitor deficiency had a low C4 value. CONCLUSIONS: All patients considered for a diagnosis of C1 inhibitor deficiency should have serum examined to measure both C4 and functional C1 inhibitor. If either is normal at presentation this essentially excludes a diagnosis of C1 inhibitor deficiency. These tests can be performed sequentially. If C4 is normal it is not necessary to proceed to C1 inhibitor analysis. If C1 inhibitor function and C4 are both low then a repeat sample should be obtained to confirm the findings.

Type 1 diabetes mellitus masking primary antibody deficiency.

A patient with a history of recurrent cutaneous and pulmonary infections, nephrotic syndrome, and an established diagnosis of type 1 diabetes was found to have unsuspected and unrecognised primary immunodeficiency. On review of the case, previous investigations pointed to the correct diagnosis over 10 years earlier. This combination of diagnoses has not previously been reported. The patient is now well on replacement intravenous immunoglobulin therapy, urinary loss of IgG having been specifically excluded before treatment. This case highlights how antibody deficiency can easily be missed despite an obvious infection history unless results are interpreted carefully and in context.

Spectrum of AIDS-associated malignant disorders.

BACKGROUND: To clarify which types of cancer result from AIDS, we compared the cancer experiences of people with AIDS with those of the general population by matching population-based cancer and AIDS registries in the USA and Puerto Rico. METHODS: We used a probabilistic matching algorithm to compare names, birth dates, and, where available, social-security numbers of 98,336 people with AIDS and 1,125,098 people with cancer aged less than 70 years. We defined AIDS-related cancers as those with both significantly raised incidence post-AIDS and increasing prevalence from 5 years pre-AIDS to 2 years post-AIDS. FINDINGS: Among people with AIDS, we found 7028 cases of Kaposi's sarcoma (KS), 1793 of non-Hodgkin lymphoma (NHL), and 712 other cases of histologically defined cancer. Incidence rates among people with AIDS were increased 310-fold for KS, 113-fold for NHL, and 1.9-fold (95% CI 1.5-2.3) for other cancers. Of 38 malignant disorders other than KS and NHL, only angiosarcoma (36.7-fold), Hodgkin's disease (7.6-fold), multiple myeloma (4.5-fold), brain cancer (3.5-fold), and seminoma (2.9-fold) were raised and increasing significantly (p<0.02) from the pre-AIDS to the post-AIDS period. INTERPRETATION: Interpretation is complicated by screening and shared risk factors, such as sexual behaviour and cigarette smoking. However, our data indicate that AIDS leads to a significantly increased risk of Hodgkin's disease, multiple myeloma, brain cancer, and seminoma. Immunological failure to control herpes or other viral infections may contribute to these malignant diseases.

Seasonal variation in intake of carotenoids and vegetables and fruits among white men in New Jersey.

In a population-based case-control study of lung cancer among New Jersey men, usual adult consumption of many vegetables and fruits was included in the interview to assess the protective potential of carotenoids. With data from 900 controls the percentage of New Jersey white men who eat specific vegetables and fruits primarily in certain seasons, the relative importance of in-season and out-of-season consumption, and the median length of season were determined. Although first asking whether a food item was consumed all-year-round or primarily in certain seasons and then asking for the appropriate frequency of consumption facilitated the interview, obtaining out-of-season frequency of consumption and length of season was not necessary. Substituting 0 for reported out-of-season frequencies and 3 mo for reported season lengths reduced slightly the observed associations between diet and lung cancer risk but did not modify the overall pattern noted. Carotenoid intake in winter-fall was estimated to be about two-thirds that in summer-spring.

Carotenoid intake, vegetables, and the risk of lung cancer among white men in New Jersey.

A population-based incident case-control study of lung cancer in white males was conducted during 1980-1981 in six high-risk areas in New Jersey. Interviews were completed for 763 cases and 900 controls. To assess whether dietary intake of carotenoids, preformed retinol, or total vitamin A influences the risk of lung cancer, the authors asked the respondents about the usual frequency of consumption, approximately four years earlier, of 44 food items which provide 83% of the vitamin A in the US diet and about the use of vitamin supplements. The men in the lowest quartile of carotenoid intake had a relative risk of 1.3 compared with those in the highest quartile after adjusting for smoking. No increase in risk was associated with low consumption of retinol or total vitamin A. Intake of vegetables, dark green vegetables, and dark yellow-orange vegetables showed stronger associations than did the carotenoid index; the smoking-adjusted risks of those in the lowest quartiles of consumption of these food groups reached relative risks of 1.4-1.5 compared with the risks of those in the highest quartiles. The protective effect of vegetables was limited to current and recent cigarette smokers; the smoking-adjusted relative risks for low consumers reached 1.7, 1.8, and 2.2 compared with the risks for high consumers for vegetables, dark green vegetables, and dark yellow-orange vegetables, respectively. The reduction in risk with vegetable intake was most apparent for squamous cell carcinomas, but it extended to adenocarcinomas and most other cell types when only current and recent smokers were analyzed. This protection among current and recent smokers is consistent with the model that vegetable intake prevents a late-stage event of carcinogenesis. Consumption of dark yellow-orange vegetables was consistently more predictive of reduced risk than consumption of any other food group or the total carotenoid index, possibly because of the high content of beta-carotene relative to other carotenoids in this particular food group.

Dietary carotene and vitamin A and risk of lung cancer among white men in New Jersey.

A population-based incident case-control study of lung cancer in white males was conducted during 1980-81 in six high-risk areas of New Jersey. Interviews were completed with 763 cases and 900 controls or with their next of kin. In order to assess whether dietary intake of carotene, preformed retinol, or total vitamin A modified the risk of lung cancer, subjects were asked about their usual frequency of consumption, several years earlier, of 44 food items, which provides 83% of the vitamin A in the American diet, and about their use of vitamin supplements. The men in the lowest quartile of carotene intake had 1.3 the risk (P-value for trend = .05) of those in the highest quartile after adjustment was made for smoking duration and intensity and education. No association was seen for retinol (P-value for trend = .11) or total vitamin A (P-value for trend = .30). The inverse association between carotene intake and lung cancer was most compelling for squamous cell carcinoma, with the smoking-and education-adjusted risk of those in the lowest quartile reaching 1.4 (P-value for trend = .03) the risk of those men in the highest quartile. Risk of lung adenocarcinoma was not related to carotene intake. The reduction in risk of squamous cell lung cancer with increasing carotene intake was noted in pipe and cigar smokers and cigarette smokers of different intensities. Among nonsmokers adenocarcinoma predominated. The inverse association between carotene and risk of squamous cell lung cancer was not especially strong or graded in response; but it was consistent and could be noted in each stratum when the subjects were divided by education, age, or mode of interview (direct vs. next of kin). The results of the other 4 case-control and 3 cohort studies that have looked at diet and risk of lung cancer are not consistent, and the question whether dietary carotene or total vitamin A reduces the risk of lung cancer is not yet resolved.