Should pre-transfusion screening RBC panels contain Wr(a+) cells?

Sometimes commercial RBC sets for the screening of irregular antibodies contain Wr(a+) cells. The aim of this study was to define the usefulness of employing RBC sets for the screening of irregular antibodies containing Wr(a+) cells in pre-transfusion tests. Anti-Wr(a) is a relatively common naturally occurring antibody in candidates to blood transfusion, although the risk of receiving a non-compatible unit is low. We have studied both the incidence of Wr(a) antibodies and the effects of having a Wr(a+) cell in the screening test on routine work in an unselected population of 787 patients requiring RBC transfusion and in 151 new blood donors. Irregular antibodies were found in 64 sera, 58 of which were specific for Wr(a) , 46 (5·8%) and 12 (7·9%) in patients and donors, respectively. The positive tested sera contained specific IgM in 16 cases, IgM + IgG in 13 cases and IgG in 27 cases. Anti-Wr(a) can usually be detected during cross-match procedures; therefore, the presence of Wr(a+) cells in pre-transfusion screening of blood recipients is not justified and it causes an undue increase in cost and time to unit release. Moreover, because of the rare association between anti-Wr(a) and haemolytic transfusion reaction, the use of Wr(a+) RBC-containing sets is also questionable in the countries that do not perform pre-transfusion cross-match tests.

Specific microRNAs are downregulated in human thyroid anaplastic carcinomas.

Thyroid carcinomas comprise a broad spectrum of tumors with different clinical behaviors. On the one side, there are occult papillary carcinomas (PTC), slow growing and clinically silent, and on the other side, rapidly growing anaplastic carcinomas (ATC), which are among the most lethal human neoplasms. We have analysed the microRNA (miR) profile of ATC in comparison to the normal thyroid using a microarray (miRNACHIP microarray). By this approach, we found an aberrant miR expression profile that clearly differentiates ATC from normal thyroid tissues and from PTC analysed in previous studies. In particular, a significant decrease in miR-30d, miR-125b, miR-26a and miR-30a-5p was detected in ATC in comparison to normal thyroid tissue. These results were further confirmed by northern blots, quantitative reverse transcription-PCR analyses and in situ hybridization. The overexpression of these four miRs in two human ATC-derived cell lines suggests a critical role of miR-125b and miR-26a downregulation in thyroid carcinogenesis, since a cell growth inhibition was achieved. Conversely, no effect on cell growth was observed after the overexpression of miR-30d and miR-30a-5p in the same cells. In conclusion, these data indicate a miR signature associated with ATC and suggest the miR deregulation as an important event in thyroid cell transformation.

Thyroid targeting of the N-ras(Gln61Lys) oncogene in transgenic mice results in follicular tumors that progress to poorly differentiated carcinomas.

Ras oncogenes are frequently mutated in thyroid carcinomas. To verify the role played by N-ras in thyroid carcinogenesis, we generated transgenic mice in which a human N-ras(Gln61Lys) oncogene (Tg-N-ras) was expressed in the thyroid follicular cells. Tg-N-ras mice developed thyroid follicular neoplasms; 11% developed follicular adenomas and approximately 40% developed invasive follicular carcinomas, in some cases with a mixed papillary/follicular morphology. About 25% of the Tg-N-ras carcinomas displayed large, poorly differentiated areas, featuring vascular invasion and forming lung, bone or liver distant metastases. N-ras(Gln61Lys) expression in cultured PC Cl 3 thyrocytes induced thyroid-stimulating hormone-independent proliferation and genomic instability with micronuclei formation and centrosome amplification. These findings support the notion that mutated ras oncogenes could be able to drive the formation of thyroid tumors that can progress to poorly differentiated, metastatic carcinomas.

Lymphocyte T subsets and natural killer cells in Italian and Philippino blood donors.

BACKGROUND AND OBJECTIVES: The characterization of lymphocyte subsets in blood donors has been utilized to determine the normal ranges that can be related to race. A study was performed in blood donors from two racial groups - Caucasian (Italians) and Asian (Philippinos) - to define respective T-lymphocyte subsets and levels of cytokines. MATERIALS AND METHODS: Ninety-two blood donors (46 Italians and 46 Philippinos) were enrolled. Blood count and immunophenotyping of lymphocytes by flow cytometry were carried out, and cytokine production was tested in six blood donors of each group. RESULTS: Philippino blood donors showed a significantly higher mean value of leucocytes (P = 0.01) and lymphocytes (P < 0.001) than Italians. The mean absolute count of lymphocyte subsets CD3- CD16+ CD56+ and CD3+ CD8+ were both significantly higher in Philippino than in Italian subjects, respectively, P < 0.01 and P < 0.0001. Philippinos showed a statistically significant higher frequency of lymphocytes producing interferon-gamma (IFN-gamma) compared to Italians (P = 0.02). CONCLUSIONS: T-lymphocyte subsets in Italian and Philippino blood donors seem to be correlated to ethnic background. The higher levels of CD3+ CD8+ T cells, natural killer (NK) cells and IFN-gamma-producing cells found in Philippinos suggest leucoreduction in Asian blood donors.

Autoimmune hemolytic anemia in chronic lymphocytic leukemia: clinical, therapeutic, and prognostic features.

Fifty-two cases of autoimmune hemolytic anemia (AHA) were observed within a series of 1203 patients (4.3%) with chronic lymphocytic leukemia (CLL) followed at a single institution. Nineteen were observed at the time of CLL diagnosis and 33 during the clinical follow-up. Ninety percent of the patients with CLL/AHA showed active CLL and 25% had been treated previously. The antierythrocyte autoantibody (AeAb) was an IgG in 87% of cases and an IgM in 13%. A lymphocyte count more than 60 x 10(9)/L (P <.00001), age above 65 years (P <.01), and male gender (P <.01) emerged as independent parameters that correlated significantly with an increased rate of AHA at CLL diagnosis. Patients previously treated with chlorambucil (CB) plus prednisone (PDN) and with fludarabine plus PDN showed a similar rate of AHA (1.8% and 2.5%, respectively). After steroid therapy associated with CB in case of active CLL, 70% of patients achieved the complete disappearance of the AeAb. The actuarial AHA relapse-free survival probability was 54% at 5 years and the median survival probability after AHA was 41 months. Infections represented the main cause of morbidity and mortality. IgG AHA and the occurrence of AHA at the same time of CLL diagnosis emerged as independent factors significantly correlated with a better survival probability of AHA/CLL patients. Taken together, this study indicates that in CLL, AHA is a rare event with no independent effect on survival for which steroids, associated with CB if required, and a careful management of infections may successfully control the 2 conditions. Cooperative studies are needed to better define the optimal steroid schedule and the therapeutic role of other immunosuppressive agents and splenectomy. (Blood. 2000;95:2786-2792)

P-VABEC: a prospective study of a new weekly chemotherapy regimen for elderly aggressive non-Hodgkin's lymphoma.

PURPOSE: To evaluate, in a prospective trial, a new combination chemotherapy specifically designed for elderly patients. PATIENTS AND METHODS: From October 1988 to December 1990, 60 previously untreated patients older than 60 years of age with aggressive non-Hodgkin's lymphoma (NHL) were treated at our institution with a new weekly alternating six-drug chemotherapy regimen, P-VABEC. The schedule consisted of doxorubicin, etoposide, and cyclophosphamide alternated weekly with vincristine and bleomycin. Oral prednisone was administered daily during the entire treatment period. Twenty-six of 60 patients were treated for a total of eight courses and 34 of 60 for 12 courses. RESULTS: A total of 45 patients (75%) achieved a complete response (CR), 10 (17%) a partial response (PR), and five (8%) no response. So far, 20 of 45 CR patients have relapsed, four of 10 PR patients have progressed, and three patients have died while in CR. Twenty-eight patients are still alive and responding (22 CRs, six PRs) after a median follow-up of 25 months. The projected overall survival (OS), disease-free survival (DFS), and event-free survival (EFS) rates at 2 years were 64%, 57%, and 55%, respectively. The outcome of patients treated with eight courses was similar to that of those who received 12 courses of P-VABEC in terms of CR rate and actuarial curves of OS, DFS, and EFS. Hematologic toxicity was mild in all patients. CONCLUSION: The P-VABEC regimen is active, well tolerated, and one of the briefest first-line chemotherapy regimens so far reported in the treatment of elderly patients with aggressive NHL. However, prospective randomized trials are needed to establish the real advantage of this regimen compared with other standard chemotherapy regimens.

A metastatic breast carcinoma presenting as autoimmune hemolytic anemia.

The authors describe the case of a 75-year-old female who was hospitalized for anemia of unknown origin. Physical examination revealed a swelling in the right mammary region, where a mastectomy scar was present from surgery for a breast carcinoma. On admission, laboratory tests disclosed anemia (Hb, 8.5 g/dl), with a reticulocyte count of 65,000/mm3 and slightly increased bilirubin. Immunohematologic study revealed the presence of a red cell autoantibody with anti-D specificity in the serum and in the eluate from the patient's erythrocytes. A biopsy of the swelling was performed and histologic examination showed the presence of metastatic cells of breast carcinoma. The patient was given chemotherapy and radiotherapy. At this writing the anemia was absent, the immunohematologic study was negative, the swelling was greatly reduced, and no other metastatic lesions of breast carcinoma were present.

Prognostic value of autoantibodies against erythrocytes and platelets in chronic lymphocytic leukemia (CLL).

Antiglobulin test (AT) and Dixon tests were performed in 100 patients with CLL. Thirty-five of them had Rai stages 0 or 1, 19 stage 2, 13 stage 3, and 33 stage 4. Twelve patients showed red blood cells autoantibodies (RBCAb) positivity; positivity at Dixon test (direct, indirect, or both) was observed in 74%. The presence of autoantibodies against erythrocytes and platelets did not influence survival curves, but anemia and thrombocytopenia are considered risk factors, independently of the presence of an autoimmune disorder. Nine RBCAb positive patients with positive Dixon test had the worst survival curves, 5 of these were anemic and 1 thrombocytopenic and anemic.

Clinical and pathological restaging in aggressive non-Hodgkin's lymphomas.

Residual tumor masses are sometimes found in non Hodgkin's lymphoma (NHL) patients after chemo-radiotherapy treatment. Radiologic findings do not differentiate lymphoma from fibrosis necrotic tissue. Surgical and histology reevaluation is the most reliable method of evaluation the viability of tumor residual masses. Sixty-three consecutive high-grade NHL were treated with F-MACHOP regimen. After 3 courses of F-MACHOP the response was evaluated clinically: twenty two (36%) achieved a clinical complete remission, 32 a clinical partial remission, 7 were non responders and two were not evaluable. An early pathological response evaluation was performed in 23 clinical partial responders. Fifteen (65%) patients had no evidence of NHL after pathological restaging and were considered as pathologic complete response and completed treatment with 3 additional courses F-MACHOP. Eight (35%), histologically positive, were defined as pathologic partial response and were crossed to salvage regimens. The 1 year actuarial event-free survival differed significantly according to clinical response after 3 courses of F-MACHOP and the outcome of clinical partial responders was highly dependent on the result of pathological restaging (87% for negative and 23% for positive). Our experience suggests that a significant proportion of clinical partial responses are histologically confirmed at pathological restaging when this procedure is performed early during treatment.

Combination of Cytosine-Arabinoside (ARA-C), Cyclophosphamide and Prednisone in the Treatment of B-Chronic Lymphocytic Leukemia in Advanced Stages and Progressive Disease.

Recently the major advances in B-chronic lymphocytic leukemia (B-CLL) have been in defining biological characteristics and prognostic criteria. However it remains to be established which is the best therapeutic approach following the first line treatment, particularly when the patients are completely unresponsive to the standard treatment using Chlorambucil (CHL) and Prednisone (PDN) and the disease is progressive. We report the results of a combination regimen using Cytosine-Arabinoside (ARA-C), Cyclophosphamide (CTX) and PDN in 19 B-CLL patients with advanced disease, resistant to CHL + PDN. The treatment schedules were as follows: Schedule A) ARA-C 60mg/sqm from day 1 to 4 s.c., CTX 75 mg/sqm from day 1 to 4 i.v., PDN 40 mg/sqm from day 1 to 4 p.o.: courses were repeated every 4 weeks for 6 months; Schedule B) ARA-C 100 mg/sqm from day 1 to 7 s.c., CTX 100 mg/sqm from day 8 to 14 i.v., PDN 40 mg/sqm from day 1 to 21 p.o.: courses were repeated every 4 weeks for 6 months. Fourteen pts were treated with schedule A and 5 pts with schedule B. We observed 9 partial remissions (PR), 5 cases with no response and 5 patients with progression. The median duration of PR is 20 months and two patients remain in PR after 18 and 60 months, respectively. The combination of ARA-C, CTX plus PDN is highly effective in advanced stages and in pretreated B-CLL but we need larger randomized studies to draw more definitive conclusions.

Immunological definition of acute promyelocytic leukemia (FAB M3): a study of 39 cases.

Acute promyelocytic leukemia (FAB-M3) is a distinct entity among acute non-lymphoid leukemias (ANLL) with peculiar morphological, biological, clinical and prognostic features. An atypical form of M3 (M3v) could be confused with other FAB ANLL and therefore the diagnosis of this variant requires ultrastructural analysis and/or cytogenetic study and/or selective gene rearrangement studies. The immunological phenotype of blast cells in 39 APL patients was studied at diagnosis. The diagnosis of M3 FAB type was ascertained in 32 and the diagnosis of M3v in 7 cases. Using a large series of monoclonal antibodies (mAb), the APL blast cells were B and T cell antigens-negative, HLA-DR constantly negative, CD13- and/or CD33-positive, CD9-positive. Among ANLL this phenotype seems to be closely related to APL both in M3 type and M3v subtype. Because the diagnosis of APL (M3 or M3v) is important in order to establish the specific therapeutic approach, the discriminant capacity of the immunological typing to identify M3 and mainly M3v (hypogranular) could be determinant for a "quick" diagnosis.

A second example of hemolysis due to IgA autoantibody with anti-e specificity.

A case of warm autoimmune hemolytic anemia due to IgA antibody is described. The patient had clinical and hematologic signs of hyperhemolysis, but all specific tests were negative. The direct antiglobulin test was positive only when it was performed with anti-IgA monospecific antiserum. The autoantibody eluted from the patient's red cells showed anti-e specificity. The sensitivity of broad-spectrum antiglobulin serum and the possible hemolytic mechanisms of IgA-coated red cells are discussed.