Reactivations of cytomegalovirus, human herpes virus 6, and Epstein-Barr virus differ with respect to risk factors and clinical outcome after hematopoietic stem cell transplantation.

One hundred two recipients of hematopoietic stem cell transplants (HSCTs) 45, from siblings and 57 from matched unrelated donors, were followed for cytomegalovirus (CMV), human herpes virus (HHV) 6, and Epstein-Barr Virus (EBV) reactivation by quantitative polymerase chain reaction in the context of immunologic reconstitution and posttransplantation complications. CMV, EBV, and HHV6 DNA copies (>100 copies/10(5) cells) were detected in 34%, 27%, and 26% of patients, respectively. The presence of 100 copies of EBV or CMV was associated with posttransplant complications: 29/66 versus 6/36 (P<.01) or 24/66 versus 4/36 (P=.01). CMV reactivation was more frequent among patients with acute graft-versus-host disease grade≥I: 17/35 versus 18/67 (P<.05). Older patient age of adults>16 year (2/16 versus 33/86; P<.05) and, to a lesser extent, CMV IgG positivity before HSCT (34/84 versus 1/10; P=.08) or an HLA-mismatched graft (9/16 versus 26/86; P=.08) constituted risk factors for CMV reactivation, which resulted in a higher rate of bacterial pneumonia (7/11 versus 28/91; P=.04). EBV reactivation risk was associated with donor EBV IgG seropositivity (28/84 versus 0/10; P=.03) and donor female gender (18/47 versus 10/55; P=.03). In contrast to EBV and CMV, EBV reactivation itself was associated with encephalitis (5/8 versus 23/94; P=.013), which was also seen as a trend among HHV6 reactivations (8/8 versus 46/94; P=.08). Multivariate analysis demonstrated that these factors play independent roles in the reactivation of the investigated herpes viruses.

Interleukin-17-producing cells increase among CD4+ lymphocytes before overt manifestation of acute graft-versus-host disease.

Interleukin-17A is a hallmark of a subset of CD4+ lymphocytes called T(H)17. Allogeneic hematopoietic stem cell transplantation (HSCT) induces an immune response that facilitates graft acceptance, but if clinically apparent as acute graft-versus-host disease (aGvHD), it may adversely affect transplantation outcomes. TH17 cells are involved in the inflammatory processes associated with several diseases, including inflammatory bowel disease (IBD) as a prototype. In this study we investigated the presence of IL-17-producing cells among peripheral blood mononuclear cells (PBMC) of patients after HSCT. The 48 patients of median age 45 years (range, 1.0-64 years), experienced hematologic malignancies (n=45) or nonmalignant disorders (n=3), treated with matched unrelated (n=24) or sibling (n=24) transplants. We examined IL-17-producing cells in alloreactive reactions after HSCT. PBMC were stimulated with BD Leukocyte Activation Cocktail (Ionomycin, Brefeldin A, and phorbol myristic acetate (PMA)) in the presence of BD GolgiStop. After stimulation the cells were labeled with anti-CD4 and intracellular anti-IL-17A monoclonal antibodies. IL-17+ cell proportions were analyzed in the CD4+ lymphocyte gate. We observed that patients at the time of hematologic reconstitution had higher proportions of IL-17-producing cells than healthy control subjects (0.73±0.13 vs 0.19±0.06%; P=.019). Fourteen patients displayed the first symptoms of aGvHD at the time of hematologic reconstitution, when they showed lower proportions of IL-17+ cells among CD4+ lymphocytes than their counterparts lacking aGvHD at a similar time after transplantation (0.29±0.09 vs 0.73±0.13%; P=.024). Eight patients developed aGvHD after hematologic reconstitution (median, 34 days). All of these patients displayed lower proportions of IL-17-producing CD4+ cells on the day of aGvHD compared with their initial measurements preceding this complication (0.34±0.14 vs 1.07±0.37%; P=.01).

Association with the presence of naive T cells in chronic myeloid leukemia patients after allogeneic human stem cell transplantation and the lower incidence of chronic graft-versus host disease and relapse.

INTRODUCTION: Allotransplantation in chronic myeloid leukemia (CML) patients offers long-lasting remissions, which largely depend on immunologic surveillance of alloreactivity. Alloreactivity in CML patients has a durable potential. However a large proportion of relapsing patients, who have to undergo donor lymphocyte treatment is still abundant. METHODS: We studied a group of 31 CML patients post allogeneic transplantation for their level of T-cell receptor excision circles (TREC) and proportion of naive and memory/effector T cells in the peripheral blood (PB). TREC numbers were determined by quantitative PCR (qPCR) and T-cell subsets CD4(+)CD27(+)CD45RO(-), CD4(+)CD27(-)CD24RO(+), CD4(+)CCR7(+), and CD4(+)CCR7(-) by flow cytometry. Patients were analyzed for posttransplant chimerism, type of bcr-abl transcripts, and number of TREC in association with the presence of chronic graft-versus-host disease (cGVHD) and relapse. CML patients with TREC+ in PB had a higher proportion of CD4(+)CD27(+)CD45RO(-) cells (3.54 vs 2.45%; P = .105) and CD4(+)CCR7(+) cells (4.85 vs 2.67%; P = .007), and a lower proportion of CD4(+)CD27(-)CD45RO(+) cells (5.55 vs 9.09%; P = .037). The incidence of cGvHD was reduced among TREC+ CML patients (3/14 vs 11/17; P = .006). RESULTS: The 5 out of 31 CML patients who relapsed were characterized by the presence of b2a2, b3/a2 or both type of transcripts, a lack of TREC in the blood, and a lower proportion of naïve and effector/memory T cells. No association was observed between any of HLA specificities, type of bcr-abl transcripts and incidence of relapse. CONCLUSION: The presence of TREC is affected by chronic GvHD; TREC negativity may constitute a risk of mixed chimerism and relapse.

Outcome and prognostic factors in advanced Hodgkin's disease treated with high-dose chemotherapy and autologous stem cell transplantation: a study of 341 patients.

BACKGROUND: The reported probability of survival of patients with Hodgkin's disease (HD) following high-dose chemotherapy with autologous stem cell transplantation (HDC/ASCT) is 35-65% at 5 years. The Polish Lymphoma Research Group investigated retrospectively prognostic factors for overall survival (OS) and event-free survival (EFS), and the risk of secondary malignancies in a large series of patients who underwent HDC/ASCT. PATIENTS AND METHODS: The data of 341 consecutive patients treated in 10 centers from 1990 to 2002 were collected and analyzed. RESULTS: The actuarial 5-year OS and EFS were 64% [95% confidence interval (CI) 57% to 71%] and 45% (95% CI 39% to 51%), respectively. In the multivariate model, unfavorable prognostic factors for EFS were less than partial response at the time of ASCT [relative risk (RR), 2.92 (95% CI 1.68-5.08); P<0.001] and three or more previous chemotherapy lines (RR, 2.16; 95% CI 1.42-3.30; P<0.001). These two factors were also associated with unfavorable OS (RR, 3.32; 95% CI 1.90-5.79; P<0.001 and RR, 2.34, 95% CI 1.51-3.64; P<0.001). Five-year cumulative risk of secondary malignancy was 8.4% (95% CI 2% to 13%) and the only identified risk factor was splenectomy (P=0.02). CONCLUSIONS: HDC/ASCT should be considered early in the course of disease for patients with a response after standard therapy.

Application of microsatellite markers for monitoring of reconstitution pattern in patients after bone marrow transplantation/peripheral blood progenitor cell transplantation.

We present data on application of microsatellite markers for detection of complete vs. mixed chimerism in individuals after bone marrow transplantation (BMT) and peripheral blood progenitor cell transplantation (PBPCT). Genotypes of the peripheral blood or bone marrow cells of three donor/recipient pairs were studied at several microsatellite loci. In two cases we identified complete chimerism in recipients' peripheral blood and bone marrow after transplantation. These two patients were alive with no symptoms of disease relapse and/or transplant rejection long time after BMT or PBPCT. In the third patient we observed mixed chimerism with the prevalence of donor's genotype while she was alive and well and the prevalence of recipient's genotype when she relapsed and required interferonotherapy. Results of our molecular studies correlated with the patients' clinical outcome. We show that molecular detection of mixed chimerism may be used for prediction of relapse in patients after BMT or PBPCT.

[Allogeneic bone marrow transplantation in myeloid leukemia: chemical conditioning, clinical course and results]. / Allogeniczny przeszczep szpiku w bialaczkach szpikowych: uwarunkowanie chemiczne przeszczepu, przebieg kliniczny, wyniki.

10 patients between the ages of 5 and 40 yrs with myeloid leukemia (4 acute, 6 chronic) in early (5 cases) or intermediate stage of the disease were given Cyclophosphamide and Busulfan (6 cases) or Cyclophosphamide, Busulfan and VP-16 (4 cases with CML) and bone marrow transplants from HLA-matched donors (in 9 cases from siblings and in one case from HLA phenotypically matched father). There was one transplant related death and 3 relapses in CML cases. In cases which relapsed GvHD was not observed. Altogether acute GvH and chronic GvHD was seen in 2 and 4 cases, respectively. All grafted cases with AML survive in continuous remission lasting more than 2 years (median 30.5 month).

Aggressive chemotherapy with autologous bone marrow transplantation in small cell lung carcinoma.

16 patients with an advanced stage SCLC were treated with the use of aggressive chemotherapy with bone marrow transplantation. With the gaining of experience the doses have been increased from standard-dose induction and 7 g/m2 of CTX in intensification to more aggressive induction (160 mg/kg of bw of CTX and 1.6 g/m2 VP-16 in two courses in 28 days interval) and intensification (CTX 7 g/m2 and VP-16 from 1.5 to 2.0 g/m2. Most recently, we used the following intensification which, in addition to the high dose CTX (6 g/m2), consisted of VP-16 0.9 g/m2 and BCNU 0.5 g/m2. The procedure proved to be safe. Hematological recovery emerged in all patients at a very similar time after autografting, irrespective to the late intensification regime. All cases, except one, received, after the hematological recovery, prophylactic cranial and at the primary tumor site irradiation as well as 2 to 4 courses of standard dose maintenance chemotherapy. The response rate was higher in the group receiving more aggressive induction and intensification. Long-term survival was seen only in patient which received more aggressive induction and intensification. Median survival of all cases was 13 months including 3 cases which are disease-free 24, 21 and 14 months after the beginning of the treatment.

Variants of SLE--a statistical approach for discrimination of a group of SLE cases into different subgroups sharing symptomatology. A pilot study.

Symptoms of two groups of Systemic lupus erythematosus (SLE) (Group 1 constituted 65 patients examined from 1975 to 1981; Group 2 constituted 104 patients examined from 1980 to 1988), which were diagnosed according to the American Rheumatism Association (ARA) preliminary criteria, were statistically analyzed with the use of a package of statistical programs which included computation of a matrix of correlation and cluster analysis. In both groups similar frequency and associations of SLE symptoms were seen. Hence, there was a positive correlation between kidney involvement and hematological abnormalities with nDNA Abs what was a hallmark of a severe SLE. In contrast, the symptoms of severe SLE disease was rarely seen in cases with Raynaud's phenomenon and discoid lesions. Our cluster analysis further distinguished groups on the basis of renal involvement, skin symptomatology, and polyserositis. These results also were similar in both groups of patients. That gave further credence to our results and continued to support the concept of SLE variants being distinguished on the basis of the clinical picture.