Abstract
BackgroundThrombocytopenia is commonly observed in severe leptospirosis. However, previous studies on coagulation alterations during leptospirosis resulted in inconsistent conclusions. Some findings showed that the prominent levels of thrombocytopenia observed in severe leptospirosis did not reflect the occurrence of disseminated intravascular coagulation (DIC) syndrome, while the others reached the conclusion that the hemorrhages observed in leptospirosis were due to DIC. The aim of this study is to elucidate whether DIC is an important feature of leptospirosis.MethodsThe leptospirosis model of guinea pig was established by intraperitoneal inoculation of Leptospira interrogans strain Lai. Hematoxylin and eosin (HE) staining, electron microscopy and immunohistochemistry staining were used to detect the pathologic changes. Platelet thrombus or fibrin thrombus was detected by HE, Martius Scarlet Blue (MSB) staining and electron microscopy. Hemostatic molecular markers such as 11-dehydrogenate thromboxane B2 (11-DH-TXB2), thrombomodulin (TM), thrombin-antithrombin III complex (TAT), D-Dimer and fibrin (ogen) degradation products (FDPs) in the plasma were examined by quantitative enzyme-linked immunosorbent assay (ELISA) to evaluate the hematological coagulative alterations in leptospirosis models.ResultsPulmonary hemorrhage appeared in the model guinea pig 24 hours after leptospires intraperitoneal inoculation, progressing to a peak at 96 hours after the infection. Leptospires were detected 24 hours post-inoculation in the liver, 48 hours in the lung and 72 hours in the kidney by immunohistochemistry staining. Spiral form of the bacteria was initially observed in the liver, lung and kidney suggestive of intact leptospires, granular form of leptospires was seen as the severity increased. Platelet aggregation in hepatic sinusoid as well as phagocytosis of erythrocytes and platelets by Kupffer cells were both observed. Neither platelet thrombus nor fibrin thrombus was found in the liver, lung or kidney via morphological observation. Thrombocytopenia was observed in all infected guinea pigs of our experimental leptospirosis study. Analysis of hematologic molecular markers showed that 11-DH-TXB2 and TM in the plasma were elevated significantly. TAT that reflects the thrombin activation had a trend of decline after infection. Although D-dimer and FDPs increased statistically, the increasing may not bear clinical significance.ConclusionPathologic and hematological studies for experimental leptospirosis of guinea pig indicated that the thrombocytopenia found in guinea pigs did not correlate with the occurrence of DIC. The platelet aggregation and Kupffer cells phagocytosis might be the potential causes of thrombocytopenia in severe leptospirosis.
Highlights
Thrombocytopenia is commonly observed in severe leptospirosis
Microscopic studies Hematoxylin and eosin (HE), Martius Scarlet Blue (MSB) staining and ultrastructural studies showed that there were conspicuous pathogenic changes in the liver, lung and kidney as described below, neither platelet thrombus nor fibrin thrombus was observed in these organs along with the course of leptospirosis
The platelet activation and injury of endothelial cells in leptospirosis were revealed, the thrombocytopenia found in experimental leptospirosis did not correlate with the occurrence of disseminated intravascular coagulation (DIC)
Summary
Thrombocytopenia is commonly observed in severe leptospirosis. previous studies on coagulation alterations during leptospirosis resulted in inconsistent conclusions. Some findings showed that the prominent levels of thrombocytopenia observed in severe leptospirosis did not reflect the occurrence of disseminated intravascular coagulation (DIC) syndrome, while the others reached the conclusion that the hemorrhages observed in leptospirosis were due to DIC. The aim of this study is to elucidate whether DIC is an important feature of leptospirosis. Leptospirosis is characterized by a broad spectrum of clinical manifestations, ranging from subclinical infection to Weil's syndrome, a severe and potentially fatal disease characterized by hemorrhage, acute renal failure and jaundice[3]. Some findings showed that thrombocytopenia occurs in up to 50% of Weil's disease patients, the prominent levels of thrombocytopenia seen in severe leptospirosis often do not reflect the occurrence of DIC syndrome [8,9,10,11,12]. The purpose of this study was to elucidate whether DIC is an important feature of experimental leptospirosis in guinea pig by pathologic and hematological studies
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