Dr PM Durand · June 2023
Platelets: clinical interpretation of abnormal counts in general practice
In general practice, one is sometimes faced with the (often unexpected) finding of a high or low platelet count. The finding may be accompanied by comments on platelet morphology. A decision needs to be made about what, if anything, needs to be done about the abnormal platelet count. In this newsletter, I outline a very brief approach to the differential diagnosis (DD) of abnormal platelet counts and some clinical ‘pointer’ questions I have used to refine the (DD). But first, a note about their structure and function.
PART 1: Platelet biogenesis, structure and function
Platelet biogenesis
Myelopoiesis comprises three broad cell lineages. These are erythropoiesis (red cells), granulopoiesis (neutrophils, eosinophils, and basophils) and megakaryopoiesis, which ultimately gives rise to the platelets observed in the peripheral blood.
Megakaryocytes are produced (primarily) in bone marrow tissue by differentiating from haemopoietic myeloid stem cells and under the control of thrombopoietin. As megakaryocytes mature, they migrate to the perivascular environment where the interactions with endothelial cells lead to pseudopodial extensions and fragments of proplatelets. Although the turbulence and shearing of blood flow cause some platelet formation, most of the megakaryocyte fragments embolize to the lungs where they interact with the pulmonary vasculature. Pulmonary tissue and vasculature are intimately associated with platelet shedding. Occasionally whole megakaryocytes may migrate to the lungs, but there is also mounting evidence that the megakaryocytes observed in lung interstitium may in fact originate from stem cells in the lungs themselves. Dissecting the origins of megakaryocytes and platelets is an ongoing research project.
Platelet structure and function
Platelets are small (1-3μm diameter), anucleate, and deceptively simple in appearance (Fig. 1). In addition to mitochondria, alpha-granules and dense granules are abundant intracellularly. Alpha granules contain high molecular weight molecules like factor V, von Willebrands Factor (vWF), and fibrinogen, while dense granules harbour low molecular weight compounds like serotonin, ADP, and calcium ions. Platelet bilayer membranes are convoluted with canalicular systems penetrating the cellular interior. Embedded in the membrane are two distinct functional receptor types: agonist and adhesion molecules. Agonist receptors recognize ligands like collagen or thrombin, while the adhesion receptors bind adhesins like vWF and fibrinogen.
Platelets play an active role in a range of physiological responses. In addition to haemostasis and thrombosis, platelets are involved in immunity and wound healing. I will focus on the coagulation functions only.
Platelets circulate close to the vessel walls and following a vascular insult or dysfunction in haemostasis cells adhere to the vessel wall matrix via receptors like GP1b/V/IX binding to vWF and GPVI binding to collagen. This binding activates platelets (they are also activated by thrombin, ADP, etc., which are released by granules or are present in the circulation) and they flatten out leading to a core thrombus, which rapidly incorporates additional platelets from the circulation.
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PART 2: Abnormalities in platelet count
In humans, 150–450 billion platelets are found in one litre of whole blood. Platelet lifespan is 8–10 days, which means a daily turnover of 100 billion platelets to maintain a count within the ‘normal’ reference range of 2 SDs from the mean. Normal for some patients (2.5% above and 2.5% below) may be slightly outside the standard reference range.
Abnormalities is platelet count are common in general practice, however, it should be borne in mind that normal platelet counts can be associated with abnormal platelet function. Abnormalities in function (with a normal platelet count) may be inherited or acquired and are not discussed below. If there is a suspicion of platelet functional abnormality, patients should be referred to a specialist haematologist.
Causes of thrombocytopaenia
Platelets are sometimes activated in the process of drawing blood into EDTA coated vacutainers because EDTA affects the GPIIb/IIIa receptor conformation facilitating its binding to fibrinogen. Small clots, fibrin strands and platelet clumps on the peripheral smear alert the haematologist that this has occurred, which means that the platelet count is artificially low. This cause of pseudo-thrombocytopaenia is first excluded before interpreting the cause of the low count.
As with all low cell counts, the causes of true thrombocytopaenia are divided into those with a central or peripheral origin. Central causes are due to either (A) ineffective megakaryopoiesis, or (B) displacement of the bone marrow and can be summarised, briefly, thus:
(A) Ineffective megakaryopoiesis
1. Medications: antibiotics, psychiatric medications, anti-inflammatories, antivirals, chemotherapy, quinine, etc.
2. Nutritional deficiencies: especially Vit B12 and folate.
3. Infections: herpesviruses, enteroviruses, retroviruses, hepatoviruses are commonly associated with decreased platelet production.
4. Autoimmune conditions: especially SLE and RA.
5. Poisons: alcohol, benzene, pesticides, heavy metals, etc.
6. Inherited / genetic conditions: More than 40 genetic conditions lead to thrombocytopaenia. Occasionally a diagnosis can be made based on the platelet morphology and associated features in other myeloid cells. Many conditions are asymptomatic, but in some cases a genetic diagnosis may be helpful for therapeutic and prognostic reasons. The more common inherited thrombocytopaenias are Bernard-Soulier, Glanzmann’s, von Willebrand’s type IIb, Gray platelet syndrome,
(B) Marrow displacement
1. Marrow fibrosis: Myelofibrosis and chemical insults that lead to fibrosis of the marrow architecture.
2. Neoplastic infiltrates: many solid tumours metastasize to the bone marrow.
3. Haemopoietic malignancies: malignancies of myeloid and lymphoid origins infiltrate and displace bone marrow. Acute promyelocytic leukaemia is not only a cause of marrow displacement, but is also associated with DIC, which is one of the peripheral causes of a low platelet count. Acute megakaryocytic leukaemia also results in a low platelet count because megakaryocytes fail to mature normally.
4. Granulomatous conditions: TB and sarcoidosis are the commonest causes of granulomata in the bone marrow, but, of course, there are others, e.g., leprosy, brucellosis, histoplasmosis etc.
5. Marrow necrosis: necrosis is relatively rare but may occur in a wide range of conditions including malignant infiltrates, medications and chemotherapy, poisons etc. Sickle cell disease is a common cause of necrosis in parts of Africa.
Peripheral causes of thrombocytopaenia result from the (1) consumption, (2) destruction, or (3) sequestration of platelets.
(1) Platelet consumption is due to the formation of clot. This may be normal, of course, but pathologically low numbers of platelets are typically due to the consumption of cells in thromboembolic conditions or in thrombotic microangiopathies (formation of microclots). Activation of the coagulation system and the production of microclots are potentially a very dangerous situation and can be associated with a range of conditions. The most common, and potentially most serious are TTP (thrombotic thrombocytopaenic purpura), DIC, HELLP syndrome, preeclampsia, and HUS (haemolytic ureamic syndrome). The clinical condition, patient history, and associated pathology results (U and E, FBC, LFTs, d-dimers, LDH, haptoglobin, peripheral smear, etc.) are essential for the differential diagnosis.
(2) The immune destruction of platelets is most commonly associated with viruses, although many medications may also bind to the surface of platelets and stimulate an immune response (heparin-induced thrombocytopaenia is well known, but other drugs may cause a similar picture). In South Africa, the common viruses associated with ITP (immune thrombocytopaenia purpura) are HIV, hepatitis, herpes viruses, and respiratory viruses. However, may others have been documented in IPT, e.g., SARS-CoV2.
(3) Platelet pooling (sequestration) typically occurs in the spleen. Any condition, therefore, that causes an enlarged spleen is often associated with a low platelet count. And the larger the spleen, the more platelets are sequestered. Massive splenomegaly occurs with MPDs (myeloproliferative conditions) and malaria with thrombocytopaenia a common complication. Of course, in malaria, platelet consumption (e.g., DIC) is also a concern.
Causes of thrombocytosis
Platelets are acute phase reactants. Conditions, therefore, that cause an inflammatory response or that activate the coagulation cascade stimulate megakaryopoiesis. Common conditions are infections, autoimmune diseases, iron deficiency, malignancies, trauma, etc. Such instances of thrombocytosis are secondary and are labelled as such. Primary causes refer to the haemopoietic clonal conditions.
As indicated above, megakaryopoiesis is one of the lineages in the myelopoiesis, the others being erythropoiesis and granulopoiesis. Clonal disorders of any components of the myeloid line may therefore spill over into megakaryopoiesis and cause dysplastic megakaryopoiesis and (sometimes) an elevated platelet count. Essential thrombocytopaenia is one of the myeloproliferaive conditions that primarily affects megakaryopoiesis, but occasionally one notices a high platelet count (or more commonly a normal platelet count with abnormal morphology) in other MPDs.
Because myeloproliferative conditions affect all the myeloid lineages, an elevated basophil count is often an associated feature. This is used as a useful clinical marker of potential MPDs. Of course, basophilia may also be reactive (allergies, medications, autoimmune conditions, etc.), but in the presence of splenomegaly or the suspicion of a clonal condition an elevated basophil count is a helpful marker.
PART 3: Clinical pointers / suggestions
Abnormalities in platelet counts are a common and often bewildering finding in general practice. If there is no obvious clinical cause, the question usually faced is: what, if anything, should be done about it? Below, I have listed some key questions to ask oneself that may guide one? These are only ‘pointer’ questions, but are often useful.
In the absence of any helpful further information from pathology results or from the questions below, it is suggested that cases are discussed with a pathologist. Further investigations and management can then be advised.
Some key ‘pointer’ questions to ask oneself on cases of thrombocytopaenia:
1. Is the low platelet count real? In other words, there is no platelet clumping reported.
2. Is there evidence of clotting (consumption of cells)?
3. Is there an enlarged spleen?
4. Has the patient had an infection? If so, consider immune-mediated destruction.
5. Are there petechiae, purpura?
6. Are the LFTs, Hb, WCC, U and E abnormal?
7. Is the patient taking a medication that could cause immune-destruction of platelets?
8. Is there possibly an inherited cause?
Some key ‘pointer’ questions to ask oneself on cases of thrombocytosis:
1. Is this reactive or a primary clonal problem?
2. Is the spleen enlarged?
3. Is there a basophilia?
4. Is the red cell count and granulocyte count abnormal?
5. Are the other pathology results abnormal? Iron studies, LFTs, U and E, etc.
Figure 1: Platelet structure with key agonist and adhesion receptors.