Sammansättningen av leukocyter i perifert blod varierar med olika sjukdomstillstånd och analys av cellerna är avgörande vid diagnostisering och uppföljning av olika leukemier.  Manuell mikroskopering och utvärdering av leukocyterna är tidskrävande och inte alltid självklar eftersom det krävs personal med hög kompetens och erfarenhet för bedömning av cellerna. En tillförlitlig förklassificering av den manuella differentialräknaren CellaVision DM1200 är därför av stort intresse för att underlätta kvantifiering och klassificering av cellerna. För att se om instrumentets förklassificering påverkades av infärgningen undersöktes färglösningens inverkan genom ändringar i fosfatbuffertens pH. Blodutstryk som härstammade från 10 patientprov färgades in med buffertar med pH 6.0, 6.8 och 7.5. Studien visade på en god positiv korrelation i samtliga pH-justerade buffertar, men med bäst resultat för pH 6.0 och 6.8. CellaVision DM1200:s förklassificering av eosinofila granulocyter påverkades mest av pH ändringar, vilket kan förklaras av deras starkt acidofila granula. / In peripheral blood the composition of leukocytes vary due to diseases and therefore analysis of the cells is critical in diagnosing and monitoring various leukemias. Manual microscopy and evaluation of the leukocytes is time consuming and not always clear, and it requires educated staff with long experience to assess the cells. A reliable automatic pre-classification of the cells with the system CellaVision DM12000 is therefor of great interest to facilitate quantification and classification of leukocytes. To evaluate if the instrument’s pre-classification were affected by staining, changes of the current staining solution used in clinical assessment where made by adjustment of the phosphate buffer pH. Examined blood smears that were descendent from 10 different samples were stained in buffers with pH 6.0, 6.8 and 7.5. This study showed a high positive correlation in all of the pH-adjusted buffers with the best results in pH 6.0 and 6.8. CellaVision DM1200’s pre-classification of eosinophilic granulocytes were most affected by changes in pH, which can be explained by their strongly acidophilic granules.

<p>Hodgkin lymphoma (HL) is characterised histologically by a minority of malignant Hodgkin and Reed-Sternberg (HRS) cells surrounded by benign cells, and clinically by a relatively good prognosis. The treatment, however, leads to a risk of serious side effects. Knowledge about the biology of the disease, particularly the interaction between the HRS cells and the surrounding cells, is essential in order to improve diagnosis and treatment. </p><p>HL patients with abundant eosinophils in the tumours have a poor prognosis, therefore the eosinophil derived protein eosinophil cationic protein (ECP) was studied. Serum-ECP (S-ECP) was elevated in most HL patients. It correlated to number of tumour eosinophils, nodular sclerosis (NS) histology, and the negative prognostic factors high erythrocyte sedimentation rate (ESR) and blood leukocyte count (WBC). A polymorphism in the ECP gene (434(G>C)) was identified and the 434GG genotype correlated to NS histology and high ESR.</p><p>The poor prognosis in patients with abundant eosinophils in the tumours has been proposed to depend on HRS cell stimulation by the eosinophils via a CD30 ligand (CD30L)-CD30 interaction. However, CD30L mRNA and protein were detected in mast cells and the predominant CD30L expressing cell in HL is the mast cell. Mast cells were shown to stimulate HRS cell lines via CD30L-CD30 interaction. The number of mast cells in HL tumours correlated to worse relapse-free survival, NS histology, high WBC, and low blood haemoglobin. </p><p>Survival in patients with early and intermediate stage HL, diagnosed between 1985 and 1992, was generally favourable and comparatively limited treatment was sufficient to produce acceptable results for most stages. The majority of relapses could be salvaged. Patients treated with a short course of chemotherapy and radiotherapy had an excellent outcome.</p><p>In conclusion prognosis is favourable in early and intermediate stages and there are possibilities for further improvements based on the fact that mast cells and eosinophils affect the biology and prognosis of HL.</p>

Hodgkin lymphoma (HL) is characterised histologically by a minority of malignant Hodgkin and Reed-Sternberg (HRS) cells surrounded by benign cells, and clinically by a relatively good prognosis. The treatment, however, leads to a risk of serious side effects. Knowledge about the biology of the disease, particularly the interaction between the HRS cells and the surrounding cells, is essential in order to improve diagnosis and treatment. HL patients with abundant eosinophils in the tumours have a poor prognosis, therefore the eosinophil derived protein eosinophil cationic protein (ECP) was studied. Serum-ECP (S-ECP) was elevated in most HL patients. It correlated to number of tumour eosinophils, nodular sclerosis (NS) histology, and the negative prognostic factors high erythrocyte sedimentation rate (ESR) and blood leukocyte count (WBC). A polymorphism in the ECP gene (434(G&gt;C)) was identified and the 434GG genotype correlated to NS histology and high ESR. The poor prognosis in patients with abundant eosinophils in the tumours has been proposed to depend on HRS cell stimulation by the eosinophils via a CD30 ligand (CD30L)-CD30 interaction. However, CD30L mRNA and protein were detected in mast cells and the predominant CD30L expressing cell in HL is the mast cell. Mast cells were shown to stimulate HRS cell lines via CD30L-CD30 interaction. The number of mast cells in HL tumours correlated to worse relapse-free survival, NS histology, high WBC, and low blood haemoglobin. Survival in patients with early and intermediate stage HL, diagnosed between 1985 and 1992, was generally favourable and comparatively limited treatment was sufficient to produce acceptable results for most stages. The majority of relapses could be salvaged. Patients treated with a short course of chemotherapy and radiotherapy had an excellent outcome. In conclusion prognosis is favourable in early and intermediate stages and there are possibilities for further improvements based on the fact that mast cells and eosinophils affect the biology and prognosis of HL.