Vascular endothelial cells are important regulators of thrombosis and hemostasis.

They actively influence not only the plasma coagulation system but platelet function as well. Under normal flow conditions, the endothelial cells do not act as a thrombogenic surface; thus, platelets do not adhere to them. However, under certain conditions they may become procoagulant and cause increased adherence of platelets, promoting increased thrombosis.1 For instance, when stimulated with endotoxin, interleukin-1, or tumor necrosis factor, endothelial cells synthesize tissue factor and other procoagulant proteins, which may cause thrombosis in patients with chronic inflammatory diseases. A wide variety of adhesion molecules are expressed on stimulated endothelial cells, including two selectins, P-selectin and E-selectin, that are involved in interactions with leukocytes and platelets. P-selectin is stored in the Weibel-Palade bodies in the endothelial cells as well as in the _-granules of the platelets. Expression of E-selectin is induced when the cells are stimulated with cytokines.2 During plateletendothelial interaction in inflamed mesenteric venules, both P-selectin and E-selectins are upregulated.3 Several investigators have reported platelet adhesion to thrombin-treated endothelial cells.47 Previous studies have shown the adherence of platelets to injured,8,9 or virally transformed10 endothelial cells.

Decreased platelet function and number may contribute to the severity of disease in some viral infections. One such disease is dengue, which is caused by a flavivirus transmitted by the Aedes aegyptimosquito. Infection with this virus leads to illness with various degrees of severity. One predominant feature of dengue is thrombocytopenia. Dengue virus appears to suppress marrow production of platelets: this may contribute to bleeding early in infection.11,12 However, just as platelet counts reach their nadir, marrow production of platelets resumes. Interaction of platelets with the endothelium could contribute to this thrombocytopenia. Most thrombocytopenia appears to result from peripheral use and platelet destruction. We have previously shown that the coagulopathic abnormalities are well compensated and hemorrhage in dengue is most likely due to activation of platelets.13 Therefore, vascular alteration may be the principal factor involved in the association of thrombocytopenia and hemorrhage with disease severity.

Dengue virus infection causes dengue fever,

Dengue Hemorrhagic Fever (DHF) and

Dengue Shock Syndrome (DSS). Thrombocytope nia is common in dengue fever and is always found in DHF/DSS. The pathogenesis of thrombocytopen ia is poorly understood. To further understand the relationship between antidengue virus antibody and anti-platelet antibody, we generated monoclonal anti-dengue virus antibodies from the dengue virus infected mice that developed transient thrombocytopen ia post dengue infection. The analysis of a panel of monoclonal antiNS-1 antibodies reveals three different patterns of platelet binding: strong, intermediate, or dull. Their isotypes are different, some are IgM while others are IgG1.

Most of antiplatelet antibodies are cross-reactive with NS-1 of dengue virus and can be competitively inhibited by recombinant NS1 protein, suggesting a molecular mimicry between dengue virus NS-1 protein and platelet. A clone, 13-F4-G5, preferentially bound activated platelets, can recognize two or three proteins around 150 kD on platelets. The binding to platelet would lyse the platelet in the presence of complement or enhance the ADP-induced platelet aggregation. Furthermore, some of these monoclonal antibodies would also react with the cellular antigens of BHK. Based on the data, we conclude that dengue virus infection

induces auto anti-platelet antibodies which thereafter may involve in the manifestation of thrombocytopen ia. A molecular mimicry between NS-1 and platelet is demonstrated. Dengue Fever (DF) is an acute infectious disease caused by dengue virus which has four serotypes. It is characterized by biphasic fever, headache, pain in various parts of the body, rash, lymphadenopat hy and leukopenia. In most cases, the disease of dengue fever is self-limited. However, there is risk to progress into Dengue Hemorrhagic Fever (DHF) or Dengue Shock Syndrome (DSS) especially when cross infection of

different serotypes occurs. DHF is a severe febrile disease characterized by abnormalities of hemostasis and increased vascular permeability, which in some instances results in DSS. DSS is a form of hypovolemic shock that is associated clinically with hemoconcentrat ion and frequently leads to death if appropriate care is not given[13]. Thrombocytope nia is common in dengue fever and is always found in DHF/DSS. Its pathogenesis is poorly understood. La Russa and Innis reported denguevirusinduced bone marrow suppression that depressed platelet synthesis[4]. Wang et al., found that dengue-2

virus can bind to human platelets in the presence of virus-specific antibody[5]. We also reported the presence of IgM anti-platelet auto-antibody in the sera of dengue patients and its titer is higher in DHF/DSS patients than in DF patients[6]. To further understand the relationship between antidengue virus antibody and anti-platelet antibody, a murine model of dengue virus infection was setup. Transient thrombocytopen ia developed at 10-13 days after primary or secondary infection and was associated with the generation of anti-platelet antibody[7]. A panel of monoclonal antibodies was generated from these

dengue virusinfected mice. In this study, it was reported that anti-dengue virus antibodies, especially antiNS-1 ones, could crossreact with platelet. The molecular mimicry between dengue virus and selfantigens was discussed.

appeared to be not related to 2001 Wiley-Liss, Inc. DHF/DSS development. J. Med. Virol. 63:143-149, 2001.

Abstract Dengue virus infection causes a wide range of diseases from dengue fever to life-threatening dengue hemorrhagic fever and dengue shock syndrome (DHF/DSS). The mechanisms involved in DHF/DSS pathogenesis remain unclear. Patient sera collected from an outbreak in southern Taiwan from November 1998 to January 1999 were studied. The presence of antibodies which cross-reacted with platelets could be detected in patient sera, and the isotype of these autoantibodies was IgM. The anti-platelet IgM levels were higher in DHF/DSS than in dengue fever patient sera in disease acute phase. These autoantibodies were still detectable in convalescent stage (1-3 weeks after acute phase) and even eight to nine months after illness. The platelet binding activity was not observed in other virus-infected patient sera tested. Further investigation showed that dengue patient sera caused platelet lysis in the presence of complement. The platelet cytotoxicity induced by DHF/DSS patient sera was higher than that by dengue fever sera. Dengue patient sera also inhibited platelet aggregation which, however,