Mean Absolute Cell Count Research Articles

Altered immune reconstitution of B and T cells precedes the onset of clinical symptoms of chronic graft-versus-host disease and is influenced by the type of onset.

We analyzed lymphocyte subpopulations and cytokines 3months after allogeneic hematopoietic stem cell transplantation aiming to identify predictive cellular and serum markers for chronic graft-versus-host disease (cGVHD). Samples of 49 patients (pts) (no cGVHD (n=14), subsequent quiescent onset (n=16), de novo onset of cGVHD (n=19)) were analyzed in the absence of active GVHD by flow cytometry and enzyme-linked immunosorbent assay. All mean absolute cell counts are presented as cells per microliter; relative cell counts are presented as percentage of lymphocytes. Pts with subsequent de novo cGVHD had significantly higher relative and absolute counts of CD4+ T cells including higher absolute counts of CD4+ memory T cells (22.36%; 206.55/μl; 136/μl, respectively) compared to pts with subsequent quiescent onset of cGVHD (12.41%; 83.42/μl; 54.3/μl) and pts without cGVHD (10.55%) with regard to relative counts of CD4+ T cells. Similarly, significantly more relative and absolute regulatory T cell numbers (CD4+FOXP3+) were detected in pts with de novo onset of cGVHD (3.08% and 24.63/μl) compared to those in pts without (1.25% and 9.06/μl) or with quiescent onset of cGVHD (1.15% and 6.91/μl). Finally, relative B cell counts, including naïve and memory B cells, were also significantly decreased in pts developing quiescent cGVHD (0.85, 0.73, 0.12% resp.) when compared to pts with de novo onset (5.61, 5.24, 0.38%). The results demonstrate that alterations in immune reconstitution are already present before onset of clinical symptoms and differ between de novo and quiescent onset of disease.

Adipose Derived Stem Cells Characterization from Human Lipoaspirate: A Comparative Study

Background Numerous studies have provided data on the efficacy of ADSCs, supporting their use in current and future clinical applications. This is the first study to our knowledge, which aims at comparing the cell viability and the absolute number of mesenchymal stem cells and ADSCs from three different approaches of preparing adipose tissue for autologous transplantation. Patients & MethodsAdipose tissue was taken from the hip/thigh region of 8 female donors undergoing liposuction. From every patient, there was sent three different fat samples: lipoaspirated fat decanted (A), lipoaspirated fat prepared by normal saline washing (B) and stromal enriched lipograft (C). Multi-parameter flow cytometry to determine the absolute number and viability of ADSCs was performed. ResultsThe mean absolute cell counts per gram of adipose tissue were 8.33x10⁶ in samples A and 5.97x10⁶ in sample C. In B samples the mean absolute cell counts per gram of adipose tissue were 2.13x10⁶. The presence of ADSCs specific markers in all the C samples showed high expression (> 95%) in the positive markers and low expression (< 2%) in the negative markers and are essential to validate the purity of adipose stem cells in a sample. ConclusionThe results obtained from the analysis of eight different donors of lipoaspirate indicate that the highest absolute number of viable adipose derived stem cells is found in the Stromal Enriched Lipograft (sample C). Their purity was confirmed by the high expression (> 95%) in the positive markers and low expression (< 2%) in the negative markers.

Adipocytokines and CD34+ Progenitor Cells in Alzheimer's Disease

BackgroundAlzheimer's disease (AD) and atherosclerosis share common vascular risk factors such as arterial hypertension and hypercholesterolemia. Adipocytokines and CD34+ progenitor cells are associated with the progression and prognosis of atherosclerotic diseases. Their role in AD is not adequately elucidated.Methods and FindingsIn the present study, we measured in 41 patients with early AD and 37 age- and weight-matched healthy controls blood concentrations of adiponectin and leptin by enzyme linked immunoabsorbent assay and of CD34+ progenitor cells using flow cytometry. We found significantly lower plasma levels of leptin in AD patients compared with the controls, whereas plasma levels of adiponectin did not show any significant differences (AD vs. control (mean±SD): leptin:8.9±5.6 ng/mL vs.16.3±15.5 ng/mL;P = 0.038; adiponectin:18.5±18.1 µg/mL vs.16.7±8.9 µg/mL;P = 0.641). In contrast, circulating CD34+ cells were significantly upregulated in AD patients (mean absolute cell count±SD:253±51 vs. 203±37; P = 0.02) and showed an inverse correlation with plasma levels of leptin (r = −0.248; P = 0.037).In logistic regression analysis, decreased leptin concentration (P = 0.021) and increased number of CD34+ cells (P = 0.036) were both significantly associated with the presence of AD. According to multifactorial analysis of covariance, leptin serum levels were a significant independent predictor for the number of CD34+ cells (P = 0.002).ConclusionsOur findings suggest that low plasma levels of leptin and increased numbers of CD34+ progenitor cells are both associated with AD. In addition, the results of our study provide first evidence that increased leptin plasma levels are associated with a reduced number of CD34+ progenitor cells in AD patients. These findings point towards a combined involvement of leptin and CD34+ progenitor cells in the pathogenesis of AD. Thus, plasma levels of leptin and circulating CD34+ progenitor cells could represent an important molecular link between atherosclerotic diseases and AD. Further studies should clarify the pathophysiological role of both adipocytokines and progenitor cells in AD and possible diagnostic and therapeutic applications.

The effect of filtration on absolute and differential cell counts in fluid obtained at bronchoalveolar lavage

We have studied the effects of filtering mucus from bronchoalveolar lavage (BAL) fluid on its cellular contents. We examined BAL fluid from 18 patients without excessive endobronchial mucus, and from three patients with bronchiectasis. In the non-bronchiectatic subjects there was a fall of 18% in mean absolute cell count from 2.66 +/- SD 1.9 x 10(9) l-1 to 2.18 +/- 1.4 x 10(9) l-1 (P less than 0.05) as a result of filtration. This was due to a selective loss of macrophages and there were no significant changes in the absolute numbers of either lymphocytes or neutrophils. The differential macrophage count fell from a mean of 82 +/- 14% to 76 +/- 16%, while the mean differential lymphocyte count rose from 14 +/- 7% to 19 +/- 9% and the mean differential neutrophil count increased slightly from 4 +/- 4% to 5 +/- 4%. In the three patients with bronchiectasis the absolute cell counts were grossly raised, due almost entirely to an excess of neutrophils. In these patients, filtration reduced the absolute macrophage count by 61% and the absolute neutrophil count by 27%. Overall, change in absolute cell count correlated significantly with mucus volume (r = 0.76, P less than 0.05). Removal of mucus by filtration affects absolute cell count and may have a selective effect on cell numbers. If filtration of BAL fluid is unavoidable, its effect on cell counts should be clearly established if data from different centres are to be meaningfully compared.