CHARACTERIZATION OF HUMAN GRANULOCYTIC EHRLICHIOSIS IN EXPERIMENTAL INFECTED HL-60 CELLS WITH ANAPLASMA PHAGOCYTOPHILUM USING NESTED PCR AND A. PHAGOCYTOPHILUM MAJOR SURFACE PROTEIN-2 MONOCLONAL ANTIBODY

The study was carried out to characterize the human granulocytic ehrlichiosis in experimental infected HL-60 cells with Anaplasma phagocytophilum using nested PCR and A. phagocytophilum major surface protein-2 monoclonal antibody. The nested PCR revealed only one band of 926 base pair DNA from A. phagocytophilum infected HL-60 cells. The western blot revealed several bands with a dominant 44-kDa. There were intense band of 100- and 160-kDa bands. The 44-kDa component was at least 10 times more abundant than the 100- and 160-kDa bands. In conclusion, the nested PCR would be a valuable tool for the characterization of human granulocytic ehrlichiosis.

Human Granulocytic Ehrlichiosis Complicating Early Pregnancy

Background. The goal of this case is to review the zoonotic infection, human granulocytic ehrlichiosis, presenting with pyrexia.Case. A 22-year-old multigravid female presented to the emergency department with a painful skin rash, high fever, and severe myalgias. The patient underwent a diagnostic evaluation for zoonotic infections due to her geographical and seasonal risk factors. Treatment of human granulocytic ehrlichiosis was successful though the patient spontaneously aborted presumably due to the severity of the acute illness.Conclusion. Treatment of human granulocytic ehrlichiosis in pregnancy presents unique challenges. Management of pyrexia during pregnancy is limited to external cooling in the setting of thrombocytopenia and elevated aminotransferases. Extensive counseling regarding teratogenic potential of medications allows the patient to weigh the pros and cons of treatment.

Human ehrlichiosis

Background. Human ehrlichiosis is a newly recognized disease. It is a tick-borne disease caused by several bacterial species of the genhus Erlichia. These are small gram-negative pleomorphic cocci, that are obligatory intracellular bacteria. Tick Ixodes is the principle vector in Europe, and Amblyomma americanum in the United States. Bacterial organisms replicate in a tick, and are transmited from infected cells in a vector to the blood cells of animals or humans. Human ehrlichiosis is a name for a group of diseases caused by different species of Ehrlichia. One of them is the disease named human monocytic ehrlichiosis, caused by Ehrlichia chaffeensis, and the other is a human granulocytic ehrlichiosis caused by Anaplasma phagocytophilia. Case report. We reported a 23-year-old patient admitted for the clinical treatment with the symptoms of high febrility (above 40 ?C), headache, vomiting, general weakness and exhaustion, but without data on a tick bite. The patient was treated with trimetoprim-sulfamethoxazole for a week when Ehrlichia chaffeensis was confirmed by the immunofluoroscence test, and the therapy contimed with doxacyclin. Conclusion. Human ehrlichiosis is also present in our country, so this disease should be considered everyday, especially in infectology practice.

Neutrophil NADPH Oxidase Is Reduced at the Anaplasma phagocytophilum Phagosome

ABSTRACT The intracellular organism Anaplasma phagocytophilum causes human granulocytic ehrlichiosis and specifically infects and multiplies in neutrophilic granulocytes. Previous reports have suggested that, for its survival, this bacterium suppresses the neutrophil respiratory burst. To investigate the mechanism of survival, we first assessed the kinetics of A. phagocytophilum entry into neutrophils by using double-labeling confocal microscopy. At 30, 60, 120, and 240 min of incubation, 25, 50, 55, and 70% of neutrophils contained bacteria, respectively. The neutrophil respiratory burst in the presence of A. phagocytophilum was assessed by a kinetic cytochrome c assay and by measurement of oxygen consumption. Neutrophils in the presence of A. phagocytophilum did not produce a significant respiratory burst, but A. phagocytophilum did not inhibit the neutrophil respiratory burst when phorbol myristate acetate was added. Immunoelectron microscopy of neutrophils infected with A. phagocytophilum or Escherichia coli revealed that NADPH oxidase subunits gp91 phox and p22 phox were significantly reduced at the A. phagocytophilum phagosome after 1 and 4 h of incubation. In neutrophils incubated simultaneously with A. phagocytophilum and E. coli for 30, 60, and 90 min, gp91 phox was present on 20, 14, and 10% of the A. phagocytophilum phagosomes, whereas p22 phox was present in 11, 5, and 4% of the phagosomes, respectively. Similarly, on E. coli phagosomes, gp91 phox was present in 62, 64, and 65%, whereas p22 phox was detected in 54, 48, and 48%. We conclude that A. phagocytophilum does not suppress a global respiratory burst and that, under identical conditions in the same cells, A. phagocytophilum, but not E. coli, significantly reduces gp91 phox and p22 phox from its phagosome membrane.