BLOOD, BLOOD COMPONENTS AND BLOOD PRODUCTS

1. BLOOD, BLOOD COMPONENTS ANDBLOOD PRODUCTS

2. Most blood collected from donors is processed as follows: ■ Blood components, such as red cell and platelet concentrates, fresh frozen plasma (FFP) and cryoprecipitate, are prepared from a single donation of blood by simple separation methods such as centrifugation and are transfused without further processing. ■ Blood products, such as coagulation factor concentrates, albumin and immunoglobulin solutions, are prepared by complex processes using the plasma from many donors as the starting material (UK donor plasma is not used, see above)

3. In most circumstances it is preferable to transfuse only the blood component or product required by the patient (‘component therapy’) rather than use whole blood. This is the most effective way of using donor blood, which is a scarce resource, and reduces the risk of complications from transfusion of unnecessary components of the blood.

4. Red cell concentrates Virtually all the plasma is removed and is replaced by about 100 mL of an optimal additive solution, such as SAG-M, which contains sodium chloride, adenine, glucose and mannitol. The mean volume is about 330 mL. The PCV is about 0.57 L/L, but the viscosity is low as there are no plasma proteins in the additive solution, and this allows fast administration if necessary. All blood components (red cell and platelet concentrates, and plasma) are leucocyte-depleted in the UK by filtration within 48 hours of collection of the donor blood.

5. Washed red cell concentrates These are preparations of red cells suspended in saline, produced by cell separators to remove all but traces of plasma proteins. They are used in patients who have had severe recurrent urticarial or anaphylactic reactions

6. Platelet concentrates These are prepared either from whole blood by centrifugation or by plateletpheresis of single donors using cell separators. They may be stored for up to 5 days at 22°C with agitation. They are used to treat bleeding in patients with severe thrombocytopenia, and prophylactically to prevent bleeding in patients with bone marrow failure

7. Granulocyte concentrates These are prepared from single donors using cell separators and are used for patients with severe neutropenia with definite evidence of bacterial infection. The numbers of granulocytes are increased by treating donors with G-CSF and steroids.

8. Fresh frozen plasma FFP is prepared by freezing the plasma from 1 unit of blood at 30°C within 6 hours of donation. The volume is approximately 200 mL. FFP contains all the coagulation factors present in fresh plasma and is used mostly for replacementof coagulation factors in acquired coagulation factor deficiencies. It may be further treated by a pathogen-inactivation process, e.g. methylene blue or solvent detergent, to minimize the risk of disease transmission.

9. Cryoprecipitate This is obtained by allowing the frozen plasma from a single donation to thaw at 4–8°C and removing the supernatant. The volume is about 20 mL and it is stored at −30°C. It contains factor VIII:C, von Willebrand factor (VWF) and fibrinogen, and may be useful in DIC and other conditions where the fibrinogen level is very low. It is no longer used for the treatment of haemophilia A and von Willebrand disease because of the greater risk of virus transmission compared with virus-inactivated coagulation factor concentrates.

10. Factor VIII and IX concentrates These are freeze-dried preparations of specific coagulation factors prepared from large pools of plasma. They are used for treating patients with haemophilia and von Willebrand disease, where recombinant coagulation factor concentrates are unavailable. Recombinant coagulation factor concentrates, where they are available, are the treatment of choice for patients with inherited coagulation factor deficiencies

11. Albumin There are two preparations: ■ Human albumin solution 4.5%, previously called plasma protein fraction (PPF), contains 45 g/L albumin and 160 mmol/L sodium. It is available in 50, 100, 250 and 500 mL bottles. ■ Human albumin solution 20%, previously called ‘saltpoor’ albumin, contains approximately 200 g/L albumin and 130 mmol/L sodium and is available in 50 and 100 mL bottles.

12. Human albumin solutions are generally considered to be inappropriate fluids for acute volume replacement or for the treatment of shock because they are no more effective in these situations than synthetic colloid solutions such as polygelatins (Gelofusine) or hydroxyethyl starch (Haemaccel). However, albumin solutions are indicated for treatment of acute severe hypoalbuminaemia and as the replacement fluid for plasma exchange. The 20% albumin solution is particularly useful for patients with nephrotic syndrome or liver disease who are fluid overloaded and resistant to diuretics. Albumin solutions should not be used to treat patients with malnutrition or chronic renal or liver disease with low serum albumin.

13. Normal immunoglobulin This is prepared from normal plasma. It is used in patients with hypogammaglobulinaemia, to prevent infections, and in patients with, for example, immune thrombocytopenic purpura

14. Specific immunoglobulins These are obtained from donors with high titres of antibodies. Many preparations are available, such as anti-D, antihepatitis B and anti-varicella zoster

15. THE WHITE CELL The five types of leucocytes found in peripheral blood are neutrophils, eosinophils and basophils (which are all called granulocytes) and lymphocytes and monocytes

16. NEUTROPHILS The earliest morphologically identifiable precursors of neutrophils in the bone marrow are myeloblasts, which are large cells constituting up to 3.5% of the nucleated cells in the marrow. The nucleus is large and contains 2–5 nucleoli. The cytoplasm is scanty and contains no granules. Promyelocytes are similar to myeloblasts but have some primary cytoplasmic granules, containing enzymes such as myeloperoxidase. Myelocytes are smaller cells without nucleoli but with more abundant cytoplasm and both primary and secondary granules. Indentation of the nucleus marks the change from myelocyte to metamyelocyte. The mature neutrophil is a smaller cell with a nucleus with 2–5 lobes, with predominantly secondary granules in the cytoplasm which contain lysozyme, collagenase and lactoferrin.

17. Peripheral blood neutrophils are equally distributed into a circulating pool and a marginating pool lying along the endothelium of blood vessels. In contrast to the prolonged maturation time of about 10 days for neutrophils in the bone marrow, their half-life in the peripheral blood is extremely short, only 6–8 hours. In response to stimuli (e.g. infection, corticosteroid therapy), neutrophils are released into the circulating pool from both the marginating pool and the marrow. Immature white cells are released from the marrow when a rapid response (within hours) occurs in acute infection (described as a ‘shift to the left’ on a blood film).

18. Function The prime function of neutrophils is to ingest and kill bacteria, fungi and damaged cells. Neutrophils are attracted to sites of infection or inflammation by chemotaxins. Recognition of foreign or dead material is aided by coating of particles with immunoglobulin and complement (opsonization) as neutrophils have Fc and C3b receptors. The material is ingested into vacuoles where it is subjected to enzymic destruction, which is either oxygen-dependent with the generation of hydrogen peroxide (myeloperoxidase) or oxygenindependentoxygenindependent (lysosomal enzymes and lactoferrin). Leucocyte alkaline phosphatase (LAP) is an enzyme found in leucocytes. It is raised when there is a neutrophilia due to an acute illness. It is also raised in polycythaemia and myelofibrosis and reduced in CM

19. Neutrophil leucocytosis A rise in the number of circulating neutrophils to > 10 × 109/L occurs in bacterial infections or as a result of tissue damage. This may also be seen in pregnancy, during exercise and after corticosteroid administration. With any tissue necrosis there is a release of various soluble factors, causing a leucocytosis. Interleukin 1 is also released in tissue necrosis and causes a pyrexia. The pyrexia and leucocytosis accompanying a myocardial infarction are a good example of this and may be wrongly attributed to infection. A leukaemoid reaction (an overproduction of white cells, with many immature cells) may occur in severe infections, tuberculosis, malignant infiltration of the bone marrow and occasionally after haemorrhage or haemolysis. In leucoerythroblastic anaemia, nucleated red cells and white cell precursors are found in the peripheral blood. Causes include marrow infiltration with metastatic carcinoma, myelofibrosis, osteopetrosis, myeloma, lymphoma, and occasionally severe haemolytic or megaloblastic anaemia

20. Neutrophil leucocytosis Tissue necrosis, e.g. myocardial infarction, trauma Inflammation, e.g. gout, rheumatoid arthritis Drugs, e.g. corticosteroids, lithium Haematological: Myeloproliferative disease Leukaemoid reaction Leucoerythroblastic anaemia Physiological, e.g. pregnancy, exercise Malignant disease, e.g. bronchial, breast, gastric Metabolic, e.g. renal failure, acidosis Congenital, e.g. leucocyte adhesion deficiency, hereditary neutrophilia

21. Causes of neutropenia Acquired Viral infection Severe bacterial infection, e.g. typhoid Felty’s syndrome Immune neutropenia – autoimmune, autoimmune neonatal neutropenia Pancytopenia from any cause, including drug-induced marrow aplasia Pure white cell aplasia Inherited Ethnic (neutropenia is common in black races) Kostmann’s syndrome (severe infantile agranulocytosis) due to mutation in elastane 2 (ELA 2) gene Cyclical (genetic mutation in ELA2 gene with neutropenia every 2–3 weeks) Others, e.g. Schwachman–Diamond syndrome, dyskeratosis congenita, Chédiak–Higashi syndrome

22. HAEMATOLOGICALMALIGNANCIES The leukaemias, the lymphomas and multiple myeloma are an interrelated spectrum of malignancies of the myeloid and lymphoid systems. They are uncommon but not rare, the lymphomas alone being the fifth commonest cancer in the UK. The aetiology of these diseases for the most part is unknown, although viruses, irradiation, cytotoxic poisons and immune suppression have been implicated in a small proportion of casesThe pathogenesis involves at least one or usually more molecular abnormalities, and non-random chromosomal abnormalities have been detected in several leukaemias and lymphomas. Classification has become increasingly complex, with the universally applied WHO scheme demanding morphological, cytogenetic and sometimes molecular criteria to be fulfilled. Treatment options are multiple

23. THE LEUKAEMIAS These are relatively uncommon diseases with an incidence of about 10 per 100 000 per year; which can occur at any age. They are classified as being acute (short natural history) or chronic (long natural history), and of myeloid or lymphoid origin. More than half of the leukaemias present acutely (ALL, AML) with the remainder being chronic types (CLL, CML). The type of leukaemia varies with age; acute lymphoblastic leukaemia (ALL) is mainly seen in childhood and chronic lymphocytic leukaemia (CLL) is a disease of the elderly. The myelodysplastic syndromes are considered pre-leukaemic . Leukaemia can be diagnosed by examination of a stained slide of peripheral blood and bone marrow, but immune phenotyping, cytogenetics and molecular genetics are essential for complete subclassification and prognostication.

24. General classification The characteristics of leukaemic cells can be assessed by light microscopy, expression of cytosolic enzymes and expression of surface antigens. These will reflect the lineage and degree of maturity of the leukaemic clone. Thus, leukaemia can be divided on the basis of the speed of evolution of the disease into acute or chronic; each of these is then further subdivided into myeloid or lymphoid, according to the cell type involved. ■ acute myeloid leukaemia (AML) ■ acute lymphoblastic leukaemia (ALL) ■ chronic myeloid leukaemia (CML) ■ chronic lymphocytic leukaemia (CLL

25. Aetiology In the majority of patients this is unknown but several factors have been associated: ■ Radiation. This can induce genetic damage to haemopoietic precursors and ALL, AML and CML have been seen in increased incidences in survivors of Hiroshima and Nagasaki and in patients treated with ionizing radiation. ■ Chemical and drugs. Exposure to benzene used in industry may lead to marrow damage. AML occurs after treatment with alkylating agents (e.g. melphalan) and topoisomerase II inhibitors (e.g. etoposide). ■ Genetic. Leukaemia risk is highly elevated in a number of germline conditions that result in genetic instability or bone marrow failure. These include Fanconi anaemia, ataxia telangiectasia and Li–Fraumeni syndrome. The risk is elevated some 30 times in people with trisomy21. There is a high degree of concordance among monozygotic twins. ■ Viruses. Leukaemias are associated with human T cell lymphotropic virus type 1 (HTLV-1), which is found particularly in Japan and the Caribbean

26. ACUTE LEUKAEMIAS The acute leukaemias increase in incidence with advancing age. Acute myeloid (myeloblastic, myelogenous) leukaemia (AML) has a median age at presentation of 65 years and may arise ‘de novo’ or against a background of myelodysplasia, either of unknown aetiology or related to cytotoxic chemotherapy. Acute lymphoid (lymphoblastic) leukaemia (ALL) has a substantially lower median age at presentation and in addition is the commonest malignancy in childhood

27. Clinical features The majority of patients with acute leukaemia, regardless of subtype, present with symptoms reflecting inadequate haematopoiesis secondary to leukaemic cells infiltration of the bone marrow: ■ anaemia – shortness of breath on effort, excessive tiredness, weakness ■ leucopenia – recurrent infections ■ thrombocytopenia – bleeding and bruising (particularly acute promyelocytic leukaemia) ■ marrow infiltration – bone pain. Examination may be unremarkable, but features include: ■ pallor ■ fever (due to infection, not the disease itself) ■ petechiae, purpura, bruises, fundal haemorrhage (particularly acute promyelocytic leukaemia) ■ lymphadenopathy, hepatosplenomegaly (more notable in lymphoblastic leukaemia) ■ violaceous skin lesions (acute myelomonocytic ■ testicular enlargement (acute lymphoblastic leukaemia) ■ cranial nerve palsies occasionally found (acute lymphoblastic leukaemia leukaemia)

28. WHO classification of acute leukaemia Acute myeloid leukaemia AML with recurrent genetic abnormalities AML with t(8;21)(q22;q22), (AML1/ETO) AML with abnormal bone marrow eosinophils and inv(16)(p13;q22) or t(16;16)(p13;q22), (CBFβ/MYH11) Acute promyelocytic leukaemia with t(15;17)(q22;q12), PML/ RAR-alpha and variants AML with 11q23 (MLL) abnormalities AML with multilineage dysplasia Following MDS or MDS/MDP Without antecedent MDS or MDS/MDP, but with dysplasia in at least 50% of cells in two or more myeloid lineages AML and myelodysplastic syndromes, therapy related Alkylating agent/radiation-related type Topoisomerase II inhibitor-related type Other AML, not otherwise categorized* AML, minimally differentiated AML without maturation AML with maturation Acute myelomonocytic leukaemia Acute monoblastic/acute monocytic leukaemia Acute erythroid leukaemia (erythroid/myeloid and pure erythroleukaemia variants) Acute megakaryoblastic leukaemia Acute basophilic leukaemia Acute panmyelosis with myelofibrosis Myeloid sarcoma

29. Acute lymphoid leukaemia Precursor B cell acute lymphoblastic leukaemia t(9;22)(q34;q11); BCR/ABL fusion gene t(4;11)(q21;q23); MLL-AF4 fusion gene t(1;19)(q23;p13.3); E2A/PBX1 fusion gene t(12;21)(p13;q22); TEL/AMLI Precursor T cell acute lymphoblastic leukaemia Burkitt-cell leukaemia

30. Investigations Confirmation of diagnosis ■ Blood count. Hb low, WBC raised usually (sometimes low), platelets low. ■ Blood film. Blast cells almost invariably seen,. , lineage identified morphologically, confirmed with immunophenotyping. ■ Bone marrow aspirate. Increased cellularity, reduced erythropoiesis, reduced megakaryocytes, sometimes trilineage dysplasia. Replacement by blast cells > 20%