A Simple Splenic Reticuloendothelial Function Test:Counting Erythrocytes With Argyrophilic Inclusions

Abstract

The presently accepted methods for evaluation of splenic reticuloendothelial (RE) function include ""Tc sulfur colloid spleen scan, antibodycoated autologous erythrocyte clearance, and pocked erythrocyte count. All methods involve special equipment and/or risk and inconvenience to patients. A simple method of assessing splenic RE function was developed by counting erythrocytes with argyrophilic inclusions using a simple silver stain and an ordinary microscope.To test the validity of this method, blood samples were collected from patients suspected of having hyposplenia or asplenia, including patients with history of splenectomy, sickle cell disease or trait, and newborns. Blood samples were also collected from normal adults and from patients without hyposplenia or asplenia as controls. The samples were tested Reticuloendothelial (RE)) function is one of the many functions of the spleen, the importance of which is highlighted by the substantial risk of severe bacterial, systemic infection in asplenic individuals. Removal of the spleen for trauma increases the risk of death from overwhelming sepsis.1 Hyposplenia and asplenia occur in sickle cell disease and are associated with varying degrees of risk of bacterial sepsis.2 Hyposplenia also occurs in patients with bone marrow transplants, 3 alcoholic liver disease, 4 inflammatory bowel disease, 5 and in elderly people. 6 The tests often employed to evaluate splenic function include 99m Tc sulfur colloid spleen scan, antibody-coated autologous erythrocyte clearance, and pocked erythrocyte count. time consuming, involving patient inconvenience, or requiring special equipment.During our study of the argyrophilic nucleolar organizer region, we noticed that many erythrocytes from patients with histories of splenectomy or sickle cell disease contained argyrophilic inclusions. This prompted us to evaluate this feature as an indicator of splenic RE function. Toward this end, we compared the argyrophilic inclusion positive erythrocyte (AE) count with the pocked erythrocyte (PE) count for sensitivity, specificity, and correlation. We also studied the interobserver and intraobserver agreement for AE count. MATERIALS AND METHODSThirty-five EDTA anticoagulated blood samples were collected from patients with possible hyposplenia or asplenia. These included newborns up to 3 months of age (13 cases) and patients with histories of sickle cell disease (9 cases), sickle cell trait (2 cases), splenectomy (9 cases), chronic granulocytic leukemia (1 case), and autoimmune hemolytic anemia (1 case). Some of the cases first attracted our attention because of the presence of red cell inclusion bodies such as Howell Jolly (HJ) bodies and/or Pappenheimer (PH) bodies in their Wright-stained peripheral blood smears. The last two cases were selected 548 Downloaded from https://academic.oup.

The spectrum of splenic complications in patients with sickle cell disease in Africa: a systematic review

“…A simple method has been described, which is based on counting red cells containing argyrophilic inclusions using a light microscope. This method has a good correlation with the pitted red cells count method and a good inter‐ and intra‐observer reliability 100 . This method may be suitable to assess splenic function in patients with SCD in Africa, where resources for spleen scintigraphy and interference phase‐contrast microscopy are absent.…”

Assessment of splenic function

“…However, measuring the percentage of Howell–Jolly bodies via flow cytometry is a potentially more reliable parameter as large amounts of erythrocytes can be screened [19]. The percentage of argyrophilic inclusion-positive erythrocytes is a parameter that is simple and seems reliable [27]. Measuring the percentages of both Howell–Jolly bodies by flow cytometry as well as argyrophilic inclusion-positive erythrocytes does not require special equipment.…”

“…This method uses a silver stain and in comparison with a normal Wrights stain, the argyrophilic inclusions are shown to be Howell–Jolly bodies, Pappenheimer bodies and other inclusions visible in patients with a decreased or absent splenic function [27]. …”

“…The argyrophilic inclusion-positive erythrocyte count has sensitivity of 88.9% and specificity of 97.1% for splenic dysfunction when using the percentage of pitted erythrocytes as a gold standard [27]. …”

The Complete Spleen

“…A new technique has been described, which utilizes silver staining of erythrocytes (Tham et al, 1996). This shows up erythrocytic inclusions, such as Howell-Jolly bodies, Pappenheimer bodies, and intracellular organelles awaiting disposal.…”