Comparison of lymphocyte immune phenotypes in bronchoalveolar lavage of non-smoking patients with sarcoidosis and other interstitial lung diseases.

BACKGROUND: Bronchoalveolar lavage (BAL) as complementary method is still used as ancillary tool in diagnosis of interstitial lung diseases. Tobacco smoking has been described to affect the BAL lavage cellular profile. To our knowledge, only few reports have so far investigated CD3+CD4+ and CD3+CD8+ lymphocyte subsets in non-smoking sarcoidosis patients additionally stratified according to CXR stage, and compared them to other non-smoking patients with interstitial lung diseases (ILDs). METHODS: We compared lymphocytes immune phenotypes, subsets, with CD3+, CD3+CD4+ and CD3+CD8+ cell markers, in the non-smoking subjects (n=297) including the patients with pulmonary sarcoidosis (S), idiopathic pulmonary fibrosis (IPF) (n=22), hypersensitivity pneumonitis (HP) (n=15), other interstitial idiopathic pneumonias (OIIPs) (n=39). According to prognosis, the patients with S were divided into four groups: 18 patients with Löfgren's syndrome (LS) in chest X-ray (CXR) ≤1 stage, 64 patients without LS in CXR ≤1 stage, 113 patients in CXR 2 stage and 26 patients with advanced CXR ≥3 stage. RESULTS: After the use of false discovery rate (FDR) correction, relative numbers (%) of CD3+, CD3+CD4+, CD3+CD8+ and CD3+CD4/CD3+CD8 ratio showed the most significant differences between the non-smokers with S (both with/without LS) and the non-smokers with other ILDs (IPF, OIIPs, HP). These lymphocytes subsets were further altered in the non-smokers with CXR stage 2 compared to the non-smokers with other ILDs (IPF, OIIPs, HP). We did not observe any differences in these lymphocyte subsets and CD3+CD4+/CD3+CD8+ ratio between the non-smokers with advanced sarcoidosis stage (CXR ≥3) and the non-smokers with IPF. CONCLUSIONS: Our data on the non-smokers confirmed the presence of the typical BAL cellular profile in sarcoidosis. The BAL cellular profile was helpful namely for differentiation of less advanced sarcoidosis. Its definite diagnostic utility should be the subject of further clinical studies with large numbers of the well characterized patients taking into consideration other clinical factors influencing BAL cellular profile, such as smoking or treatment.

Fetomaternal hemorrhage in normal vaginal delivery and in delivery by cesarean section.

BACKGROUND: The objective was to determine the incidence and volume of fetomaternal hemorrhage (FMH) in normal vaginal delivery and in delivery by cesarean section. Determination of these variables would enable optimalization of guidelines for D alloimmunization prophylaxis. STUDY DESIGN AND METHODS: In a prospective cohort study, a total of 3457 examinations were performed, 2413 after normal vaginal delivery and 1044 after cesarean delivery. FMH was assessed by flow cytometry. (FMH is fetal red blood cell [RBC] volume; fetal blood volume is double [expected fetal hematocrit is 50%].) RESULTS: The fetal RBC volume diagnosed in maternal circulation after delivery ranged from insignificant FMH of not more than 0.1 mL to excessive FMH of 65.9 mL (median, 0.7; mean, 0.78; SD, 1.48). FMH of more than 2.5 mL (immunoglobulin [Ig] G anti-D insufficient dose 50 µg) was observed in 1.4% (49/3457) and excessive volumes of FMH of more than 5 mL (insufficient dose, 100 µg) in 0.29% (10/3457). Delivery by cesarean section presented a higher risk of incidence of FMH of more than 2.5 mL (odds ratio, 2.2; p = 0.004) when compared with normal vaginal delivery. It did not, however, present a significant risk factor for the incidence of excessive volumes of FMH of more than 5 mL. CONCLUSION: During normal vaginal delivery as well as during delivery by cesarean section, FMH of less than 5 mL occurs in the great majority of cases, and thus for the prevention of D alloimmunization, an IgG anti-D dose of 100 µg should be sufficient. Contrarily, only rarely does greater FMH occur and delivery by cesarean section does not present a risk factor.

Combined measurement of plasma cell proliferative and apoptotic index in multiple myeloma defines patients with good and poor prognosis.

We assessed plasma cell proliferation (by propidium iodide index, PC-PI) and apoptosis (by annexin-V index PC-AI) for the estimation of overall survival (OS) in multiple myeloma (MM) patients. 181 patients with newly diagnosed MM were assessed, treated using conventional chemotherapy. The values best discriminating patients with poor prognosis were PC-PI>3.0% and PC-AI<4.75%, and for good prognosis PC-PI≤3.0% and PC-AI≥4.75%. The median OS was 8 months vs 40 months, p=0.0002. Our results suggest that combined measurement of plasma cell proliferation and apoptosis creates a unique strong prognostic factor based on growth characteristics of the tumor clone.

Monitoring of plasma cell proliferative and apoptotic indices in the course of multiple myeloma.

In a group of 310 patients with multiple myeloma (MM) we assessed the proliferative (PC-PI) and apoptotic indices (PC-AI). Patients were divided according to the disease state, i.e. at the time of diagnosis, in relapse, and in MM remission. We assessed the behavior of both indices with respect to therapy response and activity of the disease. In patients who reached remission, there was a significant decrease of PC-PI together with an increase of PC-AI in comparison with initial measurements. In non-responders, there was a reverse trend with increasing PC-PI and decreasing PC-AI. The values of PC-PI and PC-AI in residual myeloma population were similar regardless of treatment, i.e. in patients treated using conventional chemotherapy and after high-dosed chemotherapy with autologous stem cell transplant. Patients in long-lasting remission phase maintained stable low values of PC-PI with high PC-AI without a significant change, whereas in the group of progressing/relapsing patients, there was a significant increase of PC-PI together with a decrease of PC-AI. Our results suggest that longitudinal measurement of proliferative and apoptotic indices in MM plasmocytes helps to estimate the behavior of the tumor population and may thus become a convenient auxiliary parameter for prognosis and a targeted, individualized treatment approach.

Prognostic significance of apoptotic index in multiple myeloma patients treated by conventional therapy and novel agents, thalidomide and bortezomib.

OBJECTIVE: To assess the outcome of the measurement of apoptotic index in myeloma patients treated by conventional chemotherapy and novel drugs with biological mechanism of action, thalidomide and bortezomib. PATIENTS AND METHODS: In a cohort of 189 patients with newly diagnosed multiple myeloma from November 1997 through February 2008, we assessed the prognostic significance of plasma cell apoptotic index (PC-AI) using annexin-V. The whole group was subsequently divided according to treatment approach (conventional chemotherapy only vs. inclusion of novel drugs, thalidomide and bortezomib), and curves of overall survival were constructed. RESULTS: In the whole group (n = 189), low levels of PC-AI <4.5% significantly separated patients with unfavorable prognosis (median OS 16 vs. 38 months, P = 0.004). In patients treated with conventional chemotherapy only (n = 139) the results were similar (median OS 10 vs. 25 months, P = 0.02), and the apoptotic index maintained its significance even within the group of 50 patients treated also with novel drugs (median OS 30 vs. 54 months, P = 0.027). PC-AI was found to be independent on both Durie-Salmon staging system and the International Prognostic Index. CONCLUSION: Presented results suggest the use of apoptotic index by flow cytometry measurement as a fast and accessible method for prognostic stratification of myeloma patients in routine practice.

The levels of sCD30 and of sCD40L in a group of patients with systemic lupus erythematodes and their diagnostic value.

CD30/CD30L and CD40/CD40L are molecules from the tumor necrosis factor (TNF) superfamily. They have a major effect on communications between the B and T cells, which leads to control of maturation, proliferation, and apoptosis of those cells. The aim of this study was to compare the levels of a soluble form of CD30 (sCD30) and a soluble ligand CD40 (sCD40L) in patients with systemic lupus erythematosus (SLE) (n=65) and healthy controls (sCD30 n=20, sCD40L n=10) with other parameters of SLE activity. Patients were divided into subgroups according to presence or absence of lupus nephritis (LN; 33 with LN, 32 without LN). The serum levels of selected parameters were assessed also in the subgroups with low active disease characterized by European Lupus Activity Measure (ECLAM) at most 3(n=29) and active disease with ECLAM more than 3 (n=36). The serum levels of sCD30 were 66.0+/-40.2 UI/ml in the whole group. The mean serum levels were 60.0+/-45.2 UI/ml in the subgroups with LN, 67.1+/-38.9 UI/ml in the subgroup without LN, 80.2+/-51.9 UI/ml in the subgroup with active disease, 55.4+/-24.1 UI/ml in the subgroup with low active disease, and finally, 40.1+/-19.2 U/ml in the controls. Significant differences were found between the SLE patients and controls (p=0.0001) and between the active and nonactive groups (p=0.002). A correlation was found between levels of CD30 and ECLAM (r=0.25, p

Percentage of eosinophils with surface CD44 and levels of eosinophil cationic protein in the peripheral blood in children with bronchial asthma and their changes after montelukast therapy.

Bronchial asthma (BA) is chronic inflammation of the respiratory tract with a role played by a variety of cells, particularly mast cells, eosinophils (Eo), and T lymphocytes. The serum levels of Eo cationic protein (S-ECP) reflect the severity of bronchial inflammation and the level of bronchial hyperreactivity in asthma patients. One of the most important adhesion molecules is CD44. We examined S-ECP, the percentage of Eo with surface CD44 expression (EoCD44), and Eo count in the peripheral blood of newly diagnosed pediatric atopic patients with intermittent and persistent mild BA according to the Global Iniative for Asthma 2002, in a proportion of patients 3 months after initiation of montelukast therapy. Ninety-seven children with BA had their medical history taken, and S-ECP, with the percentage of EoCD44 determined by direct fluorescence from whole blood using flow cytometry with a Coulter EPICS XL cytometer, and Eo count, total serum immunoglobulin E levels (S-IgE) were determined. Therapy with montelukast (5 mg daily) was started in 23 children. Three months after the first collection, a second S-ECP level and EoCD44 count determinations were made. An inverse correlation between S-ECP and EoCD44 (-0.602; p < 0.0001) was found in the 97 children with BA. In the 23 children receiving montelukast we documented inverse correlation of fluctuation on S-ECP and EoCD44 after 3 months. These results were not significant. An inverse correlation between S-ECP and percent of EoCD44 was established in the 97 children with asthma before therapy initiation. The lower percentage of EoCD44 in peripheral blood in asthmatic children is due to Eo inflammation activity and attests to massive Eo invasion into the airways. Determination of the percentage proportion of EoCD44 is another potential indirect marker of the multiple features of Eo inflammation.

The relationship between proliferation and apoptosis in patients with monoclonal gammopathy of undetermined significance or multiple myeloma.

Multiple myeloma (MM) is a clonal neoplastic lymphoproliferative disease affecting terminally differentiated B cells i.e. plasma cells characterized by slow proliferation activity and different resistance to apoptosis with latent accumulation of myeloma cells in the bone marrow. This process is induced by failure of normal tissue homeostatic mechanisms. We compared plasma cell proliferation and apoptic indices in various phases of MM and in monoclonal gammophaty of untetermined significance.

Evaluation of plasma cell propidium-iodide and annexin-V indices: their relation to prognosis in multiple myeloma.

In a group of 117 patients with multiple myeloma (MM) examined at the time of diagnosis, i.e. excluding previous chemotherapy, we analysed the levels of propidium-iodide (proliferative) - PC-PI/CD(138) and annexin-V (apoptotic) - PC-AI/CD(138) indices of myeloma plasmocytes using the method of flow-cytometry to determine their relationship to prognosis. It was revealed that patients with high values of PC-PI/CD(138) had substantially worse overall survival than those with low values. Patients with a level of propidium-iodide index > or = 2,8 % exprimed a median survival of 13 months only in comparison with 42 months in patients with levels < 2,8 % (p = 0,0005). In the PC-AI/CD(138) index a reverse trend was registered. Patients with PC-AI/CD(138) > or = 4,0 % had long overall survival (median was not assessable at the time of evaluation), whereas patients with low apoptosis values < 4,0 % had median overall survival 16 months only (p = 0,01). Based on the sequentional graphic analysis of the curves of overall survival was found that the optimal discrimination level sequestering patients with good and poor prognosis was, in the case of PC-PI/CD(138) value 2,8 %, whereas in the case of PC-AI/CD(138) value 4,0 %. Among patients with good prognosis, there were no statistically significant differences in overall survival according to different levels of proliferative and apoptotic index. We conclude that evaluation of the propidium-iodide and annexin-V index using flow-cytometry is a quick, useful, and easily accessible method for the evaluation of plasma cell kinetics and thus prognosis of the disease, multiple myeloma.